Input: Therefore, we describe a microtitre assay system for glucocorticoid receptors which is a whole-cell competitive binding radioassay using [3H]-dexamethasone as radioligand.

glucocorticoid receptors


Input: This modification of a previously described protocol simplifies and reduces laboratory work and allows assay reproducibility to be controlled more reliably.


Input: Thus enabled to perform the test on multiple blood samples in parallel, we investigated cardiac infarction patients over a 12-day period to test if glucocorticoid receptor binding is altered in this'stressful' disease.

blood samples

glucocorticoid receptor


Input: On the first day of the disease, glucocorticoid receptor capacity was significantly decreased without alteration of the receptor-ligand affinity, whereas on days 4 and 12 the number of receptor sites was normal again.

glucocorticoid receptor


Input: This result fits well into the general observation of stress-induced down-regulation of immune responses.


Input: SRC-related proto-oncogenes and transcription factors in primary human T cells: modulation by cyclosporin A and FK506.

SRC-related proto-oncogenes

transcription factors

human T cells


Input: Activation of T lymphocytes induces transcription of genes encoding for lymphokines.

T lymphocytes

genes

lymphokines


Input: Interleukin-2 (IL-2) gene expression is controlled transcriptionally by the cooperative activity of specific trans-activating factors that bind to the IL-2 enhancer.

trans-activating factors

(IL-2) gene

IL-2 enhancer


Input: Cyclosporin A (CsA) and FK506 inhibit the production of IL-2 in T lymphocytes at the level of gene transcription.

T lymphocytes


Input: A member of the src gene family, the lymphocyte-specific protein tyrosine kinase, p56lck, has been implicated in IL-2 production.

src gene family

lymphocyte-specific protein tyrosine kinase


Input: CsA was found not to inhibit lck gene expression, nor the activity of the lck gene product.

lck

lck gene product


Input: However, CsA and FK506 inhibit the appearance of DNA binding activity of factors that bind to the NF-AT and AP-1 sites in the IL-2 enhancer.

NF-AT

AP-1 sites

IL-2 enhancer


Input: Since the induction of NF-AT and AP-1 is induced by the same stimuli that stimulate IL-2 production, these results indicate that the immunosuppressant action of CsA and FK506 is exerted at the level of these trans-activating factors.

NF-AT

trans-activating factors


Input: The AP-1 site at -150 bp, but not the NF-kappa B site, is likely to represent the major target of protein kinase C in the interleukin 2 promoter.

AP-1 site

-150 bp

NF-kappa B site

promoter

protein kinase C

interleukin 2


Input: Stimulation of T cells with antigen results in activation of several kinases, including protein kinase C (PKC ), that may mediate the later induction of activation-related genes.

T cells

kinases

protein kinase C

PKC


Input: We have examined the potential role of PKC in induction of the interleukin 2 (IL-2) gene in T cells stimulated through the T cell receptor/CD3 complex.

PKC

T cell receptor/CD3 complex

interleukin 2 (IL-2) gene

T cells


Input: We have previously shown that prolonged treatment of the untransformed T cell clone Ar-5 with phorbol esters results in downmodulation of the alpha and beta isozymes of PKC, and abrogates induction of IL-2 mRNA and protein.

T cell clone Ar-5

PKC

protein

IL-2 mRNA


Input: Here we show that phorbol ester treatment also abolishes induction of chloramphenicol acetyltransferase activity in Ar-5 cells transfected with a plasmid containing the IL-2 promoter linked to this reporter gene.

chloramphenicol acetyltransferase

Ar-5 cells

plasmid

IL-2 promoter

reporter gene


Input: The IL-2 promoter contains binding sites for nuclear factors including NFAT-1, Oct, NF-kappa B, and AP-1, which are all potentially sensitive to activation of PKC.

IL-2 promoter

binding sites

nuclear factors

Oct

NF-kappa B

PKC


Input: We show that induction of a trimer of the NFAT and Oct sites is not sensitive to phorbol ester treatment, and that mutations in the NF-kappa B site have no effect on inducibility of the IL-2 promoter.

NF-kappa B

site

IL-2 promoter


Input: In contrast, mutations in the AP-1 site located at -150 bp almost completely abrogate induction of the IL-2 promoter, and appearance of an inducible nuclear factor binding to this site is sensitive to PKC depletion.

AP-1 site

-150 bp

IL-2 promoter

inducible nuclear factor

PKC


Input: Moreover, cotransfections with c-fos and c-jun expression plasmids markedly enhance induction of the IL-2 promoter in minimally stimulated T cells.

IL-2 promoter

minimally stimulated

T cells


Input: Our results indicate that the AP-1 site at -150 bp represents a major, if not the only, site of PKC responsiveness in the IL-2 promoter.

AP-1 site

-150 bp

IL-2 promoter

PKC


Input: [Effect of antihypertensive therapy with captopril on gluco- and mineralo corticoid receptors of peripheral blood lymphocytes in hypertensive patients of various age]

peripheral blood lymphocytes


Input: Binding of 3H-dexamethasone and 3H-aldosterone by peripheral lymphocyte receptors was investigated in healthy persons and hypertensive patients before and after 2-week captopril treatment.

peripheral lymphocyte receptors


Input: The number of glucocorticoid and mineralocorticoid binding sites was increased in hypertensives vs normotensives.

glucocorticoid and

mineralocorticoid binding sites


Input: The treatment with the ACE inhibitor captopril led to activation of hormone-receptor interactions.


Input: There was a more marked rise of the number of receptors in middle-aged (44-55 years) hypertensives vs elderly (61-80 years) subjects after captopril treatment.

receptors


Input: The use of interferon-gamma-treated U937 cells in chemiluminescence assays to detect red cell, platelet and granulocyte antibodies of potential clinical significance.

interferon-gamma-treated U937 cells


Input: The chemiluminescent (CL) response of interferon-gamma-treated U937 (IFN-U937) cells to sensitized target cells has been used to detect red cell, platelet and granulocyte antibodies.

interferon-gamma-treated U937 (

IFN-U937) cells

red cell

platelet

granulocyte antibodies


Input: A clone of U937 cells was selected which expressed Fc receptor I (Fc gamma RI) and which, after incubation with IFN-gamma for 72 h, was capable of generating high levels of lucigenin-enhanced CL.

U937 cells

Fc receptor I

Fc gamma RI

IFN-gamma


Input: The CL responses of IFN-U937 cells and peripheral blood human monocytes to sensitized red cells, platelets or granulocytes were then compared.

IFN-U937 cells

peripheral blood human monocytes

sensitized red cells

platelets

granulocytes


Input: Assays using monocytes or IFN-U937 cells were of comparable sensitivity for detection of antibodies against all three types of target cell.

monocytes

IFN-U937 cells

antibodies


Input: In addition, the use of IFN-U937 cells reduced interassay variation and simplified assay performance.

IFN-U937 cells


Input: The potential clinical usefulness of these CL assays was suggested by the ability of both monocytes and IFN-U937 cells to respond to red cells, platelets or granulocytes sensitized with sera from pregnant women whose babies had either haemolytic disease of the newborn (HDN ), alloimmune thrombocytopenia or alloimmune neutropenia respectively.

monocytes

red cells

platelets

granulocytes

IFN-U937 cells


Input: In addition, monocytes and IFN-U937 cells both responded to red cells sensitized with antibodies against a variety of specificities of assumed (although not documented) clinical significance for blood transfusion recipients.

monocytes

red cells

IFN-U937 cells

antibodies


Input: In contrast, monocytes and IFN-U937 cells responded only weakly to red cells sensitized with either anti-D in sera from mothers of babies unaffected by HDN, or with antisera containing high titre antibodies with specificities not normally associated with significantly reduced red cell survival.

monocytes

red cells

red cell

IFN-U937 cells

anti-D

high titre antibodies


Input: Ablation of transplanted HTLV-I Tax-transformed tumors in mice by antisense inhibition of NF-kappa B [published erratum appears in Science 1993 Mar 12;259(5101):1523]

NF-kappa B


Input: Mice transgenic for the human T cell leukemia virus (HTLV-I) Tax gene develop fibroblastic tumors that express NF-kappa B-inducible early genes.

human T cell leukemia virus (HTLV-I)

gene

NF-kappa B-inducible early genes

Tax


Input: In vitro inhibition of NF-kappa B expression by antisense oligodeoxynucleotides (ODNs) inhibited growth of these culture-adapted Tax-transformed fibroblasts as well as an HTLV-I -transformed human lymphocyte line.

NF-kappa B

culture-adapted Tax-transformed fibroblasts

HTLV-I -transformed human lymphocyte line


Input: In contrast, antisense inhibition of Tax itself had no apparent effect on cell growth.

Tax


Input: Mice treated with antisense to NF-kappa B ODNs showed rapid regression of transplanted fibrosarcomas.

NF-kappa B


Input: This suggests that NF-kappa B expression may be necessary for the maintenance of the malignant phenotype and provides a therapeutic approach for HTLV-I -associated disease.

NF-kappa B


Input: Membrane receptors for aldosterone: a novel pathway for mineralocorticoid action.

Membrane receptors


Input: Rapid nongenomic in vitro effects of aldosterone on intracellular electrolytes, cell volume, and Na(+)-H+ antiport have been found in human mononuclear leukocytes (HML ).

Na(+)-H+ antiport

human mononuclear leukocytes

HML


Input: Binding of 125I-labeled aldosterone to plasma membranes of HML shares important features with these functional data.

HML


Input: This includes a very low apparent dissociation constant (Kd) of 0.1 nM for both aldosterone and the effect on the Na(+)-H(+)-antiport, a high turnover rate, and the almost exclusive binding selectivity for aldosterone.

Na(+)-H(+)-antiport


Input: Dexamethasone, RU 26988, corticosterone, ouabain, amiloride, and 18-hydroxyprogesterone were inactive as ligands.


Input: Deoxycorticosterone acetate had an intermediate activity with an apparent Kd of 100 nM.


Input: These findings are the first to demonstrate membrane binding of aldosterone being compatible with major aspects of its nongenomic effects.


Input: Leukotriene B4 transcriptionally activates interleukin-6 expression involving NK-chi B and NF-IL6.

NK-chi B


Input: Leukotriene B4 (LTB4) is a notable participant in inflammation and chemotaxis.


Input: It is, however, still unclear whether LTB4 acts in this regard directly or indirectly by stimulating the release of chemotactic and inflammatory cytokines.

inflammatory cytokines


Input: Here we report that LTB4 induces synthesis of interleukin (IL)-6 by human blood monocytes through transcriptional activation of the IL-6 gene.

interleukin (IL)-6

human blood monocytes

gene


Input: We furthermore demonstrate that this process involves activation of the transcription factor NF-chi B and, to a lesser extent, of NF-IL6, while the activity of the transcription factor AP-1, shown to otherwise confer IL-6 inducibility, appeared to be unaffected by LTB4.

transcription factor

NF-chi B

transcription factor


Input: Involvement of NF-chi B and NF-IL6 in induction of IL-6 transcription by monocytes was demonstrated using deleted forms of the IL-6 promoter.

NF-chi B

promoter


Input: Activation of the IL-6 promoter by LTB4 was not only associated with accumulation of the respective transcripts but resulted in synthesis of functional IL-6 protein as well.

IL-6 promoter

IL-6 protein


Input: In addition, LTB4 mediated transactivation of a heterologous promoter construct containing the NF-chi B or the NF-IL6 enhancer, but not the AP-1 enhancer.

heterologous promoter construct

NF-IL6 enhancer

AP-1 enhancer

NF-chi B


Input: The signaling events mediating this effect appeared to involve the release of H2O2, since LTB4 failed to induce NF-chi B or NF-IL6 in the presence of the scavenger of H2O2, N-acetyl-L-cysteine.

NF-chi B


Input: Bcl-2: a repressor of lymphocyte death.

lymphocyte


Input: The genes and mechanisms that control programmed cell death are currently the subject of intense study.


Input: The bcl-2 gene, a repressor of lymphocyte death, is perhaps the best understood of the programmed cell death associated genes.

bcl-2 gene

lymphocyte


Input: Here, Stanley Korsmeyer provides a brief overview of bcl-2, concentrating on its roles in B- and T- cell development and in oncogenesis.


Input: Cytoplasmic domain heterogeneity and functions of IgG Fc receptors in B lymphocytes.

IgG Fc receptors

B lymphocytes


Input: B lymphocytes and macrophages express closely related immunoglobulin G (IgG) Fc receptors (Fc gamma RII) that differ only in the structures of their cytoplasmic domains.

B lymphocytes

immunoglobulin G (

IgG) Fc receptors

Fc gamma RII

cytoplasmic domains


Input: Because of cell type-specific alternative messenger RNA splicing, B-cell Fc gamma RII contains an insertion of 47 amino acids that participates in determining receptor function in these cells.

B-cell

Fc gamma RII


Input: Transfection of an Fc gamma RII -negative B-cell line with complementary DNA's encoding the two splice products and various receptor mutants indicated that the insertion was responsible for preventing both Fc gamma RII -mediated endocytosis and Fc gamma RII -mediated antigen presentation.

Fc gamma RII

Fc gamma RII

Fc gamma RII

-negative B-cell line


Input: The insertion was not required for Fc gamma RII to modulate surface immunoglobulin -triggered B-cell activation.

Fc gamma RII

surface immunoglobulin


Input: Instead, regulation of activation involved a region of the cytoplasmic domain common to both the lymphocyte and macrophage receptor isoforms.

cytoplasmic domain


Input: In contrast, the insertion did contribute to the formation of caps in response to receptor cross-linking, consistent with suggestions that the lymphocyte but not macrophage form of the receptor can associate with the detergent-insoluble cytoskeleton.


Input: [Age-related changes in glucocorticoid and mineralocorticoid receptors in lymphocytes of healthy persons and patients with hypertension]

lymphocytes


Input: It has been found that the number of glucocorticoid receptors in lymphocytes of the peripheral blood of healthy elderly subjects increases, while the number of mineralocorticoid receptors decreases.

glucocorticoid receptors

lymphocytes


Input: The mechanisms of hormone-receptor interactions in hypertension are activated: the number of glucocorticoid and mineralocorticoid binding sites grows in hypertensive patients.


Input: Still a more essential rise in the number of receptors is observed in mid-age hypertensive patients than in elderly ones.


Input: The mechanism of action of cyclosporin A and FK506.


Input: CsA and FK506 are powerful suppressors of the immune system, most notably of T cells.

T cells


Input: They act at a point in activation that lies between receptor ligation and the transcription of early genes.

early genes


Input: Here, Stuart Schreiber and Gerald Crabtree review recent findings that indicate CsA and FK506 operate as prodrugs: they bind endogenous intracellular receptors, the immunophilins, and the resulting complex targets the protein phosphatase, calcineurin, to exert the immunosuppressive effect.

endogenous intracellular receptors

immunophilins

protein phosphatase,

calcineurin


Input: Induction of monocytic differentiation and NF-kappa B -like activities by human immunodeficiency virus 1 infection of myelomonoblastic cells.

NF-kappa B

myelomonoblastic cells


Input: The effects of human immunodeficiency virus 1 (HIV-1) infection on cellular differentiation and NF-kappa B DNA binding activity have been investigated in a new model of myeloid differentiation.

NF-kappa B


Input: PLB-985 cells represent a bipotential myelomonoblastic cell population capable of either granulocytic or monocytic differentiation after induction with appropriate inducers.

PLB-985 cells

bipotential myelomonoblastic cell population


Input: By virtue of the presence of CD4 on the cell surface, PLB-985 cells were chronically infected with HIV-1 strain IIIB.

PLB-985 cells


Input: PLB-IIIB cells clearly possessed a more monocytic phenotype than the parental myeloblasts, as determined by differential staining, increased expression of the myeloid-specific surface markers, and transcription of the c-fms proto-oncogene.

PLB-IIIB cells

parental myeloblasts

myeloid-specific surface markers

c-fms proto-oncogene


Input: NF-kappa B binding activity was inducible by tumor necrosis factor and phorbol myristate acetate in PLB-985.

NF-kappa B

tumor necrosis factor


Input: However, in PLB-IIIB cells, constitutive expression of a novel NF-kappa B complex was detected, composed of proteins ranging between 70 and 110 kD.

PLB-IIIB cells

NF-kappa B complex

proteins


Input: These proteins interacted specifically with the symmetric NF-kappa B site from the interferon beta (IFN-beta) promoter.

proteins

symmetric NF-kappa B site

interferon beta (IFN-beta) promoter


Input: Mutations affecting the 5' guanine residues of the kappa B site were unable to compete for these NF-kappa B -related proteins.

5' guanine residues

kappa B site

NF-kappa B -related proteins


Input: Inducibility of endogenous IFN-beta and IFN-alpha RNA was also increased in PLB-IIIB cells.

PLB-IIIB cells


Input: These studies indicate that HIV-1 infection of myelomonoblastic cells may select for a more mature monocytic phenotype and that unique subunit associations of NF-kappa B DNA binding proteins may contribute to differential NF-kappa B -mediated gene expression.

myelomonoblastic cells

NF-kappa B DNA binding proteins

NF-kappa B


Input: Cortisol receptor resistance: the variability of its clinical presentation and response to treatment.


Input: Primary (partial) cortisol receptor resistance was previously reported in a total of 7 patients and 14 asymptomatic family members.

cortisol receptor


Input: Its occurrence is considered to be extremely rare.


Input: In the present study we report on 6 patients (2 males and 4 females) with the syndrome.


Input: The first male patient presented with mild hypertension.


Input: Hydrochlorothiazide therapy resulted in life-threatening hypokalemia.


Input: The second male patient had slight hypertension without hypokalemia.


Input: All four female patients presented between the age of 20-30 yr with acne, hirsutism, and irregular menstruations.


Input: Low dose dexamethasone therapy (1-1.5 mg/day) was of clinical benefit in these patients.


Input: All patients showed insufficient suppression of serum cortisol concentrations in the overnight 1-mg dexamethasone test.


Input: The diurnal rhythm of ACTH and cortisol was intact, albeit at an elevated level.


Input: There was a normal increase in ACTH, cortisol, and GH (except in one obese patient) in response to insulin-induced hypoglycemia, while cortisol production was elevated in three patients.


Input: Circulating adrenal androgen levels were increased in all patients.


Input: Glucocorticoid receptors were investigated in a whole cell dexamethasone binding assay in mononuclear leukocytes.

Glucocorticoid receptors

mononuclear leukocytes


Input: In the first male patient, the number of receptors was very low, while the affinity was lower than that in controls.


Input: A lowered affinity to dexamethasone was found in one female patient, while a lowered number of receptors was found in three patients.


Input: In the second male patient, no abnormalities were found.


Input: As a bioassay for glucocorticoid action we also measured dexamethasone suppressibility of mitogen-stimulated incorporation of [3H]thymidine in mononuclear leukocytes.

mononuclear leukocytes


Input: In the male patient with normal receptor status, dexamethasone suppressibility of [3H]thymidine incorporation was significantly lower than that in healthy controls with respect to both maximal suppression and IC50.


Input: Partial cortisol receptor resistance might be less rare than previously thought.


Input: In the six patients presented, at least three different forms can be recognized.


Input: Therapy with dexamethasone was successful in female patients with acne and hirsutism, as the secondary increase in the production of adrenal androgens was effectively controlled.


Input: The development of functionally responsive T cells.

T cells


Input: The work reviewed in this article separates T cell development into four phases.


Input: First is an expansion phase prior to TCR rearrangement, which appears to be correlated with programming of at least some response genes for inducibility.

TCR


Input: This phase can occur to some extent outside of the thymus.


Input: However, the profound T cell deficit of nude mice indicates that the thymus is by far the most potent site for inducing the expansion per se, even if other sites can induce some response acquisition.


Input: Second is a controlled phase of TCR gene rearrangement.

TCR gene


Input: The details of the regulatory mechanism that selects particular loci for rearrangement are still not known.

loci


Input: It seems that the rearrangement of the TCR gamma loci in the gamma delta lineage may not always take place at a developmental stage strictly equivalent to the rearrangement of TCR beta in the alpha beta lineage, and it is not clear just how early the two lineages diverge.

TCR gamma loci

TCR beta

gamma delta lineage

alpha beta lineage


Input: In the TCR alpha beta lineage, however, the final gene rearrangement events are accompanied by rapid proliferation and an interruption in cellular response gene inducibility.

TCR alpha beta lineage,


Input: The loss of conventional responsiveness is probably caused by alterations at the level of signaling, and may be a manifestation of the physiological state that is a precondition for selection.


Input: Third is the complex process of selection.


Input: Whereas peripheral T cells can undergo forms of positive selection (by antigen-driven clonal expansion) and negative selection (by abortive stimulation leading to anergy or death), neither is exactly the same phenomenon that occurs in the thymic cortex.

peripheral

T cells


Input: Negative selection in the cortex appears to be a suicidal inversion of antigen responsiveness: instead of turning on IL-2 expression, the activated cell destroys its own chromatin.

activated cell

chromatin


Input: The genes that need to be induced for this response are not yet identified, but it is unquestionably a form of activation.


Input: It is interesting that in humans and rats, cortical thymocytes undergoing negative selection can still induce IL-2R alpha expression and even be rescued in vitro, if exogenous IL-2 is provided.

cortical thymocytes

IL-2R alpha

exogenous IL-2


Input: Perhaps murine thymocytes are denied this form of rescue because they shut off IL-2R beta chain expression at an earlier stage or because they may be uncommonly Bcl-2 deficient (cf. Sentman et al., 1991; Strasser et al., 1991).

murine thymocytes

IL-2R beta chain


Input: Even so, medullary thymocytes remain at least partially susceptible to negative selection even as they continue to mature.

medullary thymocytes


Input: I kappa B/MAD-3 masks the nuclear localization signal of NF-kappa B p65 and requires the transactivation domain to inhibit NF-kappa B p65 DNA binding.

I kappa B/MAD-3

NF-kappa B p65

transactivation domain

NF-kappa B p65


Input: The active nuclear form of the NF-kappa B transcription factor complex is composed of two DNA binding subunits, NF-kappa B p65 and NF-kappa B p50, both of which share extensive N-terminal sequence homology with the v-rel oncogene product.

NF-kappa B transcription factor complex

DNA binding subunits

NF-kappa B p65

NF-kappa B p50

v-rel oncogene product


Input: The NF-kappa B p65 subunit provides the transactivation activity in this complex and serves as an intracellular receptor for a cytoplasmic inhibitor of NF-kappa B, termed I kappa B.

NF-kappa B p65 subunit

NF-kappa B


Input: In contrast, NF-kappa B p50 alone fails to stimulate kappa B-directed transcription, and based on prior in vitro studies, is not directly regulated by I kappa B.

NF-kappa B p50

I kappa B


Input: To investigate the molecular basis for the critical regulatory interaction between NF-kappa B and I kappa B/MAD-3, a series of human NF-kappa B p65 mutants was identified that functionally segregated DNA binding, I kappa B -mediated inhibition, and I kappa B -induced nuclear exclusion of this transcription factor.

NF-kappa B

I kappa B/MAD-3

human

NF-kappa B p65 mutants

I kappa B

I kappa B

transcription factor


Input: Results from in vivo expression studies performed with these NF-kappa B p65 mutants revealed the following: 1) I kappa B/MAD-3 completely inhibits NF-kappa B p65 -dependent transcriptional activation mediated through the human immunodeficiency virus type 1 kappa B enhancer in human T lymphocytes, 2) the binding of I kappa B/MAD-3 to NF-kappa B p65 is sufficient to retarget NF-kappa B p65 from the nucleus to the cytoplasm, 3) selective deletion of the functional nuclear localization signal present in the Rel homology domain of NF-kappa B p65 disrupts its ability to engage I kappa B/MAD-3, and 4) the unique C-terminus of NF-kappa B p65 attenuates its own nuclear localization and contains sequences that are required for I kappa B -mediated inhibition of NF-kappa B p65 DNA binding activity.

NF-kappa B p65 mutants

I kappa B/MAD-3

NF-kappa B p65

I kappa B/MAD-3

NF-kappa B p65

NF-kappa B p65

Rel homology domain

NF-kappa B p65

I kappa B/MAD-3

C-terminus

NF-kappa B p65

I kappa B

NF-kappa B p65

human immunodeficiency virus type 1 kappa B enhancer

human T lymphocytes


Input: Together, these findings suggest that the nuclear localization signal and transactivation domain of NF-kappa B p65 constitute a bipartite system that is critically involved in the inhibitory function of I kappa B/MAD-3.

transactivation domain

NF-kappa B p65

I kappa B/MAD-3


Input: Unexpectedly, our in vivo studies also demonstrate that I kappa B/MAD-3 binds directly to NF-kappa B p50.

I kappa B/MAD-3

NF-kappa B p50


Input: This interaction is functional as it leads to retargeting of NF-kappa B p50 from the nucleus to the cytoplasm.

NF-kappa B p50


Input: However, no loss of DNA binding activity is observed, presumably reflecting the unique C-terminal domain that is distinct from that present in NF-kappa B p65.

C-terminal domain

NF-kappa B p65


Input: Characterization of a new tissue-specific transcription factor binding to the simian virus 40 enhancer TC-II (NF-kappa B) element.

transcription factor

simian virus 40 enhancer TC-II (NF-kappa B) element


Input: We have biochemically and functionally characterized a new transcription factor, NP-TCII, which is present in nuclei from unstimulated T and B lymphocytes but is not found in nonhematopoietic cells.

transcription factor

NP-TCII

unstimulated T and

B lymphocytes

nonhematopoietic cells


Input: This factor has a DNA-binding specificity similar to that of NF-kappa B but is unrelated to this or other Rel proteins by functional and biochemical criteria.

NF-kappa B

Rel proteins


Input: It can also be distinguished from other previously described lymphocyte-specific DNA-binding proteins.

lymphocyte-specific DNA-binding proteins


Input: The candidate oncoprotein Bcl-3 is an antagonist of p50/NF-kappa B -mediated inhibition.

candidate oncoprotein Bcl-3

p50/NF-kappa B


Input: The candidate oncogene bcl-3 was discovered as a translocation into the immunoglobulin alpha-locus in some cases of B-cell chronic lymphocytic leukaemias.

candidate oncogene bcl-3

immunoglobulin alpha-locus


Input: The protein Bcl-3 contains seven so-called ankyrin repeats.

protein Bcl-3

ankyrin repeats


Input: Similar repeat motifs are found in a number of diverse regulatory proteins but the motifs of Bcl-3 are most closely related to those found in I kappa B proteins in which the ankyrin repeat domain is thought to be directly involved in inhibition of NF-kappa B activity.

repeat motifs

diverse regulatory proteins

I kappa B proteins

ankyrin repeat

NF-kappa B


Input: No biological function has yet been described for Bcl-3, but it was noted recently that Bcl-3 interferes with DNA-binding of the p50 subunit of NF-kappa B in vitro.

p50 subunit

NF-kappa B


Input: Here we demonstrate that Bcl-3 can aid kappa B site -dependent transcription in vivo by counteracting the inhibitory effects of p50/NF-kappa B homodimers.

p50/NF-kappa B homodimers

kappa B site


Input: Bcl-3 may therefore aid activation of select NF-kappa B -regulated genes, including those of the human immunodeficiency virus.

NF-kappa B

-regulated genes


Input: [Mechanism of action of steroid hormones. I. Estrogens]


Input: The steroid hormone are very versatile molecules: although they are related among them by their chemical structure, they have very diverse functions and including antagonic.


Input: Their action mechanism is not completely cleared.


Input: The estrogens participate in the regulation of practically all the reproductive and sexual events of the female, although the intracellular actions by which they take place are not well known and the proposed models do not adequately satisfy the questions.


Input: Currently it is accepted the existence of a cytoplasmic and/or nuclear receptor, without explaining satisfactorily how the hormones come to the nucleus.


Input: The endocrine events that are rapidly expressed (seconds) are due to a possible interaction with cellular membrane.


Input: The purpose of this review is to analyze and concilliate the reported data on the mechanism of action of estrogens.


Input: Reduced susceptibility to HIV-1 infection of ethyl-methanesulfonate-treated CEM subclones correlates with a blockade in their protein kinase C signaling pathway.

ethyl-methanesulfonate-treated CEM subclones

protein kinase C


Input: We have described the isolation of chemically induced CEM subclones that express CD4 receptors and bind soluble gp120, yet show a markedly reduced susceptibility to infection with HIV-1.

CEM subclones

CD4 receptors


Input: Two subclones were found to have an abnormal response to the protein kinase C (PKC) activator PMA.

protein kinase C

PKC


Input: PMA treatment induced CD3 and CD25 (IL-2R) receptors on the parental line and on other ethyl-methanesulfonate-derived subclones, but not on these two mutants.

ethyl-methanesulfonate-derived subclones


Input: Direct assays of PKC activity were conducted.

PKC


Input: Total cellular PKC enzymatic activity was found to be normal in these subclones.

PKC


Input: PMA -induced CD4 down-modulation occurred normally.


Input: In addition, activation of c-raf kinase was normal.

c-raf kinase


Input: Since HIV-1 long terminal repeat contains two functional nuclear factor kB (NF-kB) regulatory elements, we studied the ability of PMA to induce NF-kB binding activity by different assays.

HIV-1 long terminal repeat

nuclear factor kB (NF-kB) regulatory elements

NF-kB


Input: Chloramphenicol acetyl transferase (CAT) assays using the HIV-1 (-139)long terminal repeat-CAT construct showed no PMA induction of CAT activity in these subclones (unlike the parental line and other subclones).

Chloramphenicol acetyl transferase

CAT

CAT

HIV-1 (-139)long terminal repeat-CAT construct


Input: Okadaic acid, an inhibitor of phosphatases 1 and 2A, did not overcome the defect in these subclones.


Input: Gel retardation assays, using a 32P-probe containing the HIV-1 NF-kB probe and nuclear extracts from PMA -treated cells, showed significantly reduced induction of nuclear NF-kB binding proteins in these two subclones compared with wild type CEM and a control subclone.

NF-kB

nuclear NF-kB binding proteins

wild type

CEM

PMA -treated cells


Input: Deoxycholate treatment of cytoplasmic extracts from these subclones released much reduced NF-kB binding proteins from their cytoplasmic pools.

NF-kB binding proteins


Input: Thus, reduced levels of PKC -induced nuclear NF-kB activity in two T cell subclones did not affect their normal cell growth, but correlated with a pronounced reduction in their susceptibility to HIV-1 infection.

PKC

NF-kB


Input: Nuclear factor of activated T cells contains Fos and Jun.

Nuclear factor

Fos

Jun

activated T cells


Input: The nuclear factor NF-AT (ref. 1) is induced in T cells stimulated through the T-cell receptor/CD3 complex, and is required for interleukin-2 (IL-2) gene induction.

nuclear factor NF-AT

T-cell receptor/CD3 complex

T cells

(IL-2) gene


Input: Although NF-AT has not been cloned or purified, there is evidence that it is a major target for immunosuppression by cyclosporin A (CsA) and FK506 (refs 2-7).

NF-AT


Input: NF-AT induction may require two activation-dependent events: the CsA -sensitive translocation of a pre-existing component and the CsA -resistant synthesis of a nuclear component.

NF-AT

nuclear component


Input: Here we report that the newly synthesized nuclear component of NF-AT is the transcription factor AP-1.

nuclear component

NF-AT

transcription factor


Input: We show that the inducible nuclear form of NF-AT contains Fos and Jun proteins.

NF-AT

Fos

Jun


Input: Furthermore, we identify a pre-existing NF-AT -binding factor that is present in hypotonic extracts of unstimulated T cells.

pre-existing NF-AT -binding factor

unstimulated

T cells


Input: On the basis of binding, reconstitution and cotransfection experiments, we propose that activation of NF-AT occurs in at least two stages: a CsA -sensitive stage involving modification and/or translocation of the pre-existing NF-AT complex, and a CsA -insensitive stage involving the addition of newly synthesized Fos or Fos/Jun proteins to the pre-existing complex.

NF-AT

NF-AT complex

Fos

Fos/Jun proteins

pre-existing complex


Input: A lymphoid cell-specific nuclear factor containing c-Rel-like proteins preferentially interacts with interleukin-6 kappa B-related motifs whose activities are repressed in lymphoid cells.

lymphoid cell-specific nuclear factor

c-Rel-like proteins

interleukin-6 kappa B-related motifs

lymphoid cells


Input: The proto-oncoprotein c-Rel is a member of the nuclear factor kappa B transcription factor family, which includes the p50 and p65 subunits of nuclear factor kappa B.

proto-oncoprotein c-Rel

nuclear factor kappa B transcription factor family

p65 subunits

nuclear factor kappa B


Input: We show here that c-Rel binds to kappa B sites as homodimers as well as heterodimers with p50.

c-Rel

homodimers

heterodimers

kappa B sites


Input: These homodimers and heterodimers show distinct DNA-binding specificities and affinities for various kappa B motifs.

homodimers

heterodimers

kappa B motifs


Input: In particular, the c-Rel homodimer has a high affinity for interleukin-6 (IL-6) and beta interferon kappa B sites.

c-Rel homodimer

beta interferon

kappa B sites


Input: In spite of its association with p50 in vitro, however, we found a lymphoid cell-specific nuclear factor in vivo that contains c-Rel but not p50 epitopes; this factor, termed IL-6 kappa B binding factor II, appears to contain the c-Rel homodimer and preferentially recognizes several IL-6 kappa B-related kappa B motifs.

lymphoid cell-specific nuclear factor

c-Rel

p50 epitopes

IL-6 kappa B binding factor II

c-Rel homodimer

kappa B-related kappa B motifs


Input: Although it has been previously shown that the IL-6 kappa B motif functions as a potent IL-1/tumor necrosis factor-responsive element in nonlymphoid cells, its activity was found to be repressed in lymphoid cells such as a Jurkat T-cell line.

IL-6 kappa B motif

IL-1/tumor necrosis factor-responsive element

nonlymphoid cells

lymphoid cells

Jurkat T-cell line


Input: We also present evidence that IL-6 kappa B binding factor II functions as a repressor specific for IL-6 kappa B-related kappa B motifs in lymphoid cells.

IL-6 kappa B binding factor II

IL-6 kappa B-related

kappa B motifs

lymphoid cells


Input: Cortisol resistance in acquired immunodeficiency syndrome.


Input: This study concerns 9 iv drug abusers with acquired immunodeficiency syndrome (AIDS) who developed hypercortisolism without the clinical signs or metabolic consequences of hypercortisolism.


Input: All patients were characterized by an Addisonian picture (weakness, weight loss, hypotension, hyponatremia, and intense mucocutaneous melanosis ).


Input: An acquired form of peripheral resistance to glucocorticoids was suspected.


Input: We, therefore, examined glucocorticoid receptor characteristics on mononuclear leukocytes by measuring [3H]dexamethasone binding and the effect of dexamethasone on [3H]thymidine incorporation, which is one of the effects of glucocorticoid receptor activation.

glucocorticoid receptor

glucocorticoid receptor

mononuclear leukocytes


Input: Glucocorticoid receptor density was increased in AIDS patients with an Addisonian picture (group 1; 16.2 +/- 9.4 fmol/million cells) compared to values in 12 AIDS patients without an Addisonian picture (group 2; 6.05 +/- 2.6 fmol/million cells; P less than 0.01) and sex- and age-matched controls (3.15 +/- 2.3 fmol/million cells; P less than 0.01).

Glucocorticoid receptor


Input: The affinity of glucocorticoid receptors (Kd) was strikingly decreased (9.36 +/- 3.44 nM in group 1; 3.2 +/- 1.5 nM in group 2; 2.0 +/- 0.8 nM in controls; P less than 0.01).

glucocorticoid receptors


Input: [3H]Thymidine incorporation was decreased dose-dependently by dexamethasone in controls and patients; the effect was significantly blunted (P less than 0.05) in group 1 patients, which suggests that activation of glucocorticoid receptor is impaired as a result of the glucocorticoid receptor abnormality.

glucocorticoid receptor

glucocorticoid receptor


Input: In conclusion, AIDS patients with hypercortisolism and clinical features of peripheral resistance to glucocorticoids are characterized by abnormal glucocorticoid receptors on lymphocytes.

abnormal glucocorticoid receptors

lymphocytes


Input: Resistance to glucocorticoids implies a complex change in immune-endocrine function, which may be important in the course of immunodeficiency syndrome.


Input: A novel primer extension method to detect the number of CAG repeats in the androgen receptor gene in families with X-linked spinal and bulbar muscular atrophy.

androgen receptor gene


Input: X-linked spinal and bulbar muscular atrophy (SBMA), an adult-onset form of motor neuron disease, was recently reported to be caused by amplification of the CAG repeats in the androgen receptor gene.

androgen receptor gene


Input: We report here a simple and rapid strategy to detect the precise number of the CAGs.


Input: After the DNA fragment containing the CAG repeats is amplified by the polymerase chain reaction, a primer extension is carried out; the extension of the end-labelled reverse primer adjacent to 3' end of CAG repeats stops at the first T after CAG repeats with the incorporation of dideoxy ATP in the reaction mixture.

DNA fragment

CAG repeats

end-labelled reverse primer

3' end

CAG repeats

CAG repeats


Input: The resultant primer products are analysed by denaturing polyacrylamide gel electrophoresis and autoradiography.

primer products


Input: This method could be quite useful to detect not only CAG repeats in SBMA but also other polymorphic dinucleotide and trinucleotide repeats.

CAG repeats

polymorphic dinucleotide

trinucleotide repeats


Input: Cellular immune and cytokine pathways resulting in tissue factor expression and relevance to septic shock.

tissue factor


Input: Cells of monocyte lineage serve as effector cells in the cellular immune response.

effector cells


Input: In addition, they respond to LPS and cytokines with activation and expression of inflammatory effector gene products similar to those elicited by the antigen driven response.

inflammatory effector gene products


Input: The response to antigen proceeds at the T helper cell level through two independent forms of cellular collaboration, contact and lymphokine.

T helper cell

lymphokine


Input: We review the control of expression of the Tissue Factor (TF) gene and the function of the TF protein.

Tissue Factor

TF

TF protein

) gene


Input: The enhanced initiation of transcription of the TF gene appears to require engagement of a 56 bp LPS Response Element, an enhancer that is engaged by both AP-1 type heterodimeric complexes as well as NF kappa B like heterodimeric complexes.

TF gene

LPS Response Element

AP-1 type heterodimeric complexes

NF kappa B like heterodimeric complexes


Input: Dissociation of NF kappa B from Ig kappa B by cytokine and LPS stimulation, and possibly activated T cells, may represent a common pathway to induction of the TF and other inflammatory genes.

NF kappa B

Ig kappa B

TF

activated T cells


Input: Enhancement of expression of TF is observed upon adhesion of Mo to endothelial cells and extracellular matrix proteins, as well as upon engagement of leukocyte integrins.

TF

leukocyte integrins

endothelial cells


Input: The biological effects that follow from expression of TF by vascular cells have been resolved by analysis of function aided by the use of recombinant full length TF and truncated surface domain of TF.

TF

recombinant full length TF

truncated surface domain

TF


Input: The rules of assembly of the cognate ligands of TF, namely the zymogen plasma factors VII and the serine protease factor VIIa, with the soluble surface domain of TF in free solution, in the presence of phospholipid surfaces and cell surface and of the anchored TF molecule have been described.

TF

zymogen plasma factors VII

serine protease factor VIIa

soluble surface domain

TF

anchored TF molecule


Input: It is evident that assembly of the surface domain of TF with VIIa to form the binary TF.VIIa complex induces a significant increase in the Kcat of the catalytic domain of VIIa for small peptidyl substrates and more profoundly for protein substrate.

surface domain

TF

VIIa

TF.VIIa complex

catalytic domain

VIIa

protein substrate


Input: This provides substantial evidence for an allosteric effect on the catalytic cleft of VIIa that is imparted by binding to TF, its cognate catalytic cofactor.

VIIa

TF

cofactor


Input: It is also evident that the TF.VIIa complex is proteolytically active and can activate the zymogen plasma factor X to the serine protease Xa in free solution, inferring that extended substrate recognition by induced structural loci of the TF.VIIa complex are created from either or both proteins to constitute a new recognition structure.

TF.VIIa complex

zymogen plasma factor X

serine protease Xa

TF.VIIa complex

new recognition structure


Input: It is also evident that association of X with charged phospholipid surfaces enhances the proteolytic activation of this zymogen by increasing recognition and susceptibility of the sessile peptide bond deduced from the markedly decreased Km and increased Kcat.

X

zymogen


Input: Phorbol ester reduces constitutive nuclear NF kappa B and inhibits HIV-1 production in mature human monocytic cells.

constitutive nuclear

NF kappa B

mature human

monocytic cells


Input: NF kappa B is a potent mediator of specific gene expression in human monocytes and has been shown to play a role in transcription of the HIV-1 genome in promonocytic leukemias.

NF kappa B


Input: There is little information available on the response of NF kappa B to cytokines in normal human monocytes.

NF kappa B

cytokines

normal human monocytes


Input: We have used a 32P-labeled oligonucleotide derived from human immunodeficiency virus (HIV-1) long terminal repeat, which contains a tandem repeat of the NF kappa B binding sequence, as a probe in a gel retardation assay to study this transcription factor.

human immunodeficiency virus (HIV-1) long terminal repeat

tandem repeat

NF kappa B binding sequence

transcription factor


Input: Using this assay, we have detected NF kappa B in extracts of nuclei from normal human monocytes.

NF kappa B

normal human monocytes


Input: Treatment of normal monocytes with 12-0-tetradecanoyl phorbol-13-acetate (TPA) for 4-24 h caused the complete disappearance of NF kappa B from nuclear extracts of monocytes.

normal monocytes

NF kappa B


Input: A similar result was obtained with the mature monocytic leukemia cell line THP-1.

mature monocytic leukemia cell line


Input: The constitutive transcription factor SP1 was unaffected by addition of TPA.

constitutive transcription factor


Input: The disappearance of NF kappa B from the nucleus was concentration dependent between 10 and 50 ng/ml of phorbol ester.

NF kappa B


Input: In THP-1 cells, TPA also induced a new, faster-migrating NF kappa B species not induced in monocytes.

faster-migrating NF kappa B species

monocytes


Input: Protein kinase C inhibitor staurosporine, but not cyclic nucleotide-dependent protein kinase inhibitor HA-1004, also dramatically reduced constitutive levels of nuclear NF kappa B.

nuclear

NF kappa B


Input: Finally, TPA addition to monocytes infected with HIV-1 inhibited HIV-1 replication, as determined by reverse transcriptase assays, in a concentration-dependent manner.

monocytes

reverse transcriptase


Input: These results are in striking contrast to the increase in nuclear NF kappa B and HIV-1 replication induced by phorbol esters in promonocytic leukemia cells U937 and HL-60, and emphasize the importance of studying cytokine regulation of HIV-1 in normal monocytes.

NF kappa B

cytokine

promonocytic leukemia cells U937

normal monocytes


Input: A mechanism for the antiinflammatory effects of corticosteroids: the glucocorticoid receptor regulates leukocyte adhesion to endothelial cells and expression of endothelial-leukocyte adhesion molecule 1 and intercellular adhesion molecule 1.

glucocorticoid receptor

endothelial-leukocyte adhesion molecule 1

intercellular adhesion molecule 1

endothelial cells


Input: Corticosteroids are the preeminent antiinflammatory agents although the molecular mechanisms that impart their efficacy have not been defined.


Input: The endothelium plays a critical role in inflammation by directing circulating leukocytes into extravascular tissues by expressing adhesive molecules for leukocytes [e.g., endothelial-leukocyte adhesion molecule 1 (ELAM-1) and intercellular adhesion molecule 1 (ICAM-1 )].

circulating leukocytes

leukocytes

adhesive molecules

endothelial-leukocyte adhesion molecule 1

intercellular adhesion molecule 1


Input: We therefore determined whether corticosteroids suppress inflammation by inhibiting endothelial expression of adhesion molecules for neutrophils (polymorphonuclear leukocytes ).

adhesion molecules

neutrophils

polymorphonuclear

leukocytes


Input: Preincubation of endothelial cells with endotoxin [lipopolysaccharide (LPS ), 1 microgram/ml] led to a 4-fold increase in subsequent adherence of polymorphonuclear leukocytes (P < 0.0001, n = 10) to endothelial cells, an increase that was markedly attenuated when endothelial cells were treated with dexamethasone (IC50 < 1 nM, P < 0.0001, n = 6 or 7) during preincubation with LPS.

endothelial cells

polymorphonuclear leukocytes

endothelial cells

endothelial cells


Input: Moreover, the steroid receptor agonist cortisol (10 microM), but not its inactive metabolite tetrahydrocortisol (10 microM), diminished LPS -induced endothelial cell adhesiveness.

steroid receptor


Input: Further evidence that the action of dexamethasone was mediated through ligation of corticosteroid receptors [human glucocorticoid receptors (hGRs )] was provided by experiments utilizing the steroid antagonist RU-486.

corticosteroid receptors

human glucocorticoid receptors

hGRs


Input: RU-486 (10 microM), which prevents translocation of ligated hGR to the nucleus by inhibiting dissociation of hGR from heat shock protein 90, completely aborted the effect of dexamethasone on adhesiveness of endothelial cells (P < 0.0005, n = 3).

hGR

hGR

endothelial cells


Input: Treatment of endothelial cells with LPS (1 microgram/ml) stimulated transcription of ELAM-1, as shown by Northern blot analysis, and expression of membrane-associated ELAM-1 and ICAM-1, as shown by quantitative immunofluorescence (both P < 0.001, n = 9).

endothelial cells

membrane-associated


Input: Dexamethasone markedly inhibited LPS -stimulated accumulation of mRNA for ELAM-1 and expression of ELAM-1 and ICAM-1 (IC50 < 10 nM, both P < 0.001, n = 4-9); inhibition of expression by dexamethasone was reversed by RU-486 (both P < 0.005, n = 4-6).


Input: As in the adhesion studies, cortisol but not tetrahydrocortisol inhibited expression of ELAM-1 and ICAM-1 (both P < 0.005, n = 3 or 4).


Input: In contrast, sodium salicylate (1 mM) inhibited neither adhesion nor expression of these adhesion molecules.

adhesion molecules


Input: These studies suggest that antagonism by dexamethasone of endotoxin -induced inflammation is a specific instance of the general biological principle that the glucocorticoid receptor is a hormone-dependent regulator of transcription.

endotoxin

glucocorticoid receptor


Input: Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and adult testis.

transcription factor

BSAP

B lymphocytes


Input: BSAP has been identified previously as a transcription factor that is expressed at early, but not late, stages of B-cell differentiation.

BSAP

transcription factor


Input: Biochemical purification and cDNA cloning has now revealed that BSAP belongs to the family of paired domain proteins.

BSAP

paired domain proteins


Input: BSAP is encoded by the Pax-5 gene and has been highly conserved between human and mouse.

BSAP

Pax-5 gene


Input: An intact paired domain was shown to be both necessary and sufficient for DNA binding of BSAP.

BSAP


Input: Binding studies with several BSAP recognition sequences demonstrated that the sequence specificity of BSAP differs from that of the distantly related paired domain protein Pax-1.

BSAP

BSAP

paired domain protein Pax-1

recognition sequences


Input: During embryogenesis, the BSAP gene is transiently expressed in the mesencephalon and spinal cord with a spatial and temporal expression pattern that is distinct from that of other Pax genes in the developing central nervous system (CNS ).

BSAP gene

Pax genes


Input: Later, the expression of the BSAP gene shifts to the fetal liver where it correlates with the onset of B lymphopoiesis.

BSAP gene


Input: BSAP expression persists in B lymphocytes and is also seen in the testis of the adult mouse.

BSAP

B lymphocytes


Input: All of this evidence indicates that the transcription factor BSAP may not only play an important role in B-cell differentiation but also in neural development and spermatogenesis.

transcription factor

BSAP


Input: Cell cycle-dependent initiation and lineage-dependent abrogation of GATA-1 expression in pure differentiating hematopoietic progenitors.

pure differentiating hematopoietic progenitors


Input: The programmed activation/repression of transcription factors in early hematopoietic differentiation has not yet been explored.

transcription factors


Input: The DNA-binding protein GATA-1 is required for normal erythroid development and regulates erythroid-expressed genes in maturing erythroblasts.

DNA-binding protein

erythroid-expressed genes

maturing erythroblasts


Input: We analyzed GATA-1 expression in early human adult hematopoiesis by using an in vitro system in which "pure" early hematopoietic progenitors are induced to gradual and synchronized differentiation selectively along the erythroid or granulocyte-macrophage pathway by differential treatment with hematopoietic growth factors.

early hematopoietic progenitors


Input: The GATA-1 gene, though virtually silent in quiescent progenitors, is activated after entrance into the cell cycle upon stimulation with hematopoietic growth factors.

GATA-1 gene

quiescent progenitors


Input: Subsequently, increasing expression along the erythroid pathway contrasts with an abrupt downregulation in the granulocyte-macrophage lineage.

granulocyte-macrophage lineage


Input: These results suggest a microenvironment-directed, two-step model for GATA-1 expression in differentiating hematopoietic progenitors that involves (i) cycle-dependent initiation and (ii) lineage-dependent maintenance or suppression.

differentiating hematopoietic progenitors


Input: Hypothetically, on/off switches of lineage-restricted transactivators may underlie the binary fate decisions of hematopoietic progenitors.

lineage-restricted transactivators

hematopoietic progenitors


Input: Specific NF-kappa B subunits act in concert with Tat to stimulate human immunodeficiency virus type 1 transcription.

NF-kappa B subunits

Tat


Input: NF-kappa B is a protein complex which functions in concert with the tat-I gene product to stimulate human immunodeficiency virus (HIV) transcription.

NF-kappa B

protein complex

tat-I gene product


Input: To determine whether specific members of the NF-kappa B family contribute to this effect, we have examined the abilities of different NF-kappa B subunits to act with Tat-I to stimulate transcription of HIV in Jurkat T-leukemia cells.

NF-kappa B family

NF-kappa B subunits

Tat-I

Jurkat T-leukemia cells


Input: We have found that the p49(100) DNA binding subunit, together with p65, can act in concert with Tat-I to stimulate the expression of HIV -CAT plasmid.

p49(100) DNA binding subunit

Tat-I

HIV -CAT plasmid


Input: Little effect was observed with 50-kDa forms of p105 NF-kappa B or rel, in combination with p65 or full-length c-rel, which do not stimulate the HIV enhancer in these cells.

50-kDa forms

NF-kappa B

rel

full-length c-rel

HIV enhancer


Input: These findings suggest that the combination of p49(100) and p65 NF-kappa B can act in concert with the tat-I gene product to stimulate the synthesis of HIV RNA.

NF-kappa B

tat-I gene product

HIV RNA


Input: Stable expression of HB24, a diverged human homeobox gene, in T lymphocytes induces genes involved in T cell activation and growth.

diverged human homeobox gene


Input: A diverged homeobox gene, HB24, which is known to be induced following lymphocyte activation, was introduced into Jurkat T cells under the control of a constitutive promoter.

diverged homeobox gene

constitutive promoter

Jurkat T cells


Input: Stable transfectants of HB24 were established that expressed high levels of HB24 mRNA and possessed an altered phenotype suggestive of activated T cells.

HB24 mRNA

activated T cells


Input: A number of genes known to be induced following T cell activation and associated with cell growth were increased in the transfectants, including c-fos, c-myc, c-myb, HLA-DR, lck, NF-kappa B, interleukin-2 and interleukin-2 receptor alpha (IL-2R alpha ).

transfectants

c-fos

c-myc

c-myb

HLA-DR

lck

interleukin-2 receptor alpha

IL-2R alpha

NF-kappa B


Input: Analysis of IL-2R alpha expression by transient transfection of IL-2R alpha promoter constructs into the HB24 transfectants revealed constitutive expression (about 60% of phytohemagglutinin- and phorbol ester- activated Jurkat cells) that was dependent on the kappa B site in the IL-2R alpha promoter.

IL-2R alpha

IL-2R alpha promoter constructs

kappa B site

IL-2R alpha promoter

HB24 transfectants


Input: Furthermore, as a consequence of the increased HB24 mRNA levels, the Jurkat HB24 transfectants proliferated more rapidly than control cell lines.

HB24 mRNA

Jurkat HB24 transfectants

control cell lines


Input: Thus, stable expression of HB24 confers an activation phenotype on a human T cell line, implicating this gene as an important transcriptional factor during T cell activation and growth.

human T cell line

transcriptional factor


Input: Functional interaction between the two zinc finger domains of the v-erb A oncoprotein.

zinc finger domains

v-erb A oncoprotein


Input: The v-erb A oncogene of avian erythroblastosis virus is a mutated and virally transduced copy of a host cell gene encoding a thyroid hormone receptor.

v-erb A oncogene

host cell gene

thyroid hormone receptor


Input: The protein expressed by the v-erb A oncogene binds to DNA and acts as a dominant negative inhibitor of both the thyroid hormone receptor and the closely related retinoic acid receptor.

v-erb A oncogene

DNA

thyroid hormone receptor

retinoic acid receptor


Input: The v-erb A protein has sustained two amino acid alterations within its DNA-binding domain relative to that of c-erb A, one of which, at serine 61, is known to be important for v-erb A function in the neoplastic cell.

v-erb A protein

DNA-binding domain

c-erb A

v-erb A

neoplastic cell


Input: We report here that the second alteration, at threonine 78, also plays an important, although more indirect, role: alteration of the sequence at threonine 78 such that it resembles that of c-erb A can act as an intragenic suppressor and can partially restore function to a v-erb A protein rendered defective due to a mutation at position 61.

c-erb A

v-erb A protein

position 61


Input: Threonine 78 lies within the D-box of the v-erb A protein, a region thought to mediate receptor-receptor dimerizations, and is not in physical proximity to the serine at position 61.

D-box

v-erb A protein

position 61


Input: It therefore appears that an indirect interaction occurs between these two sites and that this interaction is crucial for v-erb A function.

v-erb A


Input: cis-acting sequences required for inducible interleukin-2 enhancer function bind a novel Ets-related protein, Elf-1.

cis-acting sequences

interleukin-2 enhancer

Ets-related protein


Input: The recent definition of a consensus DNA binding sequence for the Ets family of transcription factors has allowed the identification of potential Ets binding sites in the promoters and enhancers of many inducible T-cell genes.

DNA binding sequence

Ets binding sites

promoters

enhancers

inducible T-cell genes

Ets family

transcription factors


Input: In the studies described in this report, we have identified two potential Ets binding sites, EBS1 and EBS2, which are conserved in both the human and murine interleukin-2 enhancers.

Ets binding sites


Input: Within the human enhancer, these two sites are located within the previously defined DNase I footprints, NFAT-1 and NFIL-2B, respectively.

human enhancer

DNase I


Input: Electrophoretic mobility shift and methylation interference analyses demonstrated that EBS1 and EBS2 are essential for the formation of the NFAT-1 and NFIL-2B nuclear protein complexes.

NFAT-1 and NFIL-2B nuclear protein complexes


Input: Furthermore, in vitro mutagenesis experiments demonstrated that inducible interleukin-2 enhancer function requires the presence of either EBS1 or EBS2.

interleukin-2 enhancer


Input: Two well-characterized Ets family members, Ets-1 and Ets-2, are reciprocally expressed during T-cell activation.

Ets family members


Input: Surprisingly, however, neither of these proteins bound in vitro to EBS1 or EBS2.


Input: We therefore screened a T-cell cDNA library under low-stringency conditions with a probe from the DNA binding domain of Ets-1 and isolated a novel Ets family member, Elf-1.

T-cell cDNA

DNA binding domain

novel

Ets family member


Input: Elf-1 contains a DNA binding domain that is nearly identical to that of E74, the ecdysone-inducible Drosophila transcription factor required for metamorphosis (hence the name Elf-1, for E74-like factor 1 ).

DNA binding domain

ecdysone-inducible Drosophila transcription factor

E74-like factor 1


Input: Elf-1 bound specifically to both EBS1 and EBS2 in electrophoretic mobility shift assays.


Input: It also bound to the purine-rich CD3R element from the human immunodeficiency virus type 2 long terminal repeat, which is required for inducible virus expression in response to signalling through the T-cell receptor.

purine-rich CD3R element

human immunodeficiency virus type 2 long terminal repeat

T-cell receptor


Input: Taken together, these results demonstrate that multiple Ets family members with apparently distinct DNA binding specificities regulate differential gene expression in resting and activated T cells.

multiple Ets family members


Input: Binding of erythroid and non-erythroid nuclear proteins to the silencer of the human epsilon-globin-encoding gene.

silencer

human epsilon-globin-encoding gene


Input: To clarify the molecular mechanisms involved in the developmental control of hemoglobin-encoding genes we have been studying the expression of these genes in human cells in continuous culture.

hemoglobin-encoding genes

genes

human cells


Input: We have previously reported the presence of a transcriptional control element with the properties of a silencer extending from -392 to -177 bp relative to the cap site of the human epsilon-globin-encoding gene [Cao et al., Proc.Natl.Acad.Sci.USA 86 (1989) 5306-5309].

transcriptional control element

silencer

-392 to -177 bp

cap site

human epsilon-globin-encoding gene


Input: We also showed that this silencer has stronger inhibitory activity in HeLa cells, as compared to K562 human erythroleukemia cells.

silencer

HeLa cells

K562 human erythroleukemia cells


Input: Using deletion mutants and cis-cloned synthetic oligodeoxyribonucleotides in transient expression assays, nucleotide sequences responsible for this effect have now been further delimited to 44 bp located from -294 to -251 bp.

deletion mutants

-294 to -251 bp


Input: Gel electrophoresis mobility shift assays and DNaseI footprinting assays demonstrate that these negative regulatory sequences are recognized differently by proteins present in nuclear extracts obtained from HeLa and K562 cells.

DNaseI

proteins

HeLa

K562 cells


Input: Two binding proteins are detected in K562 nuclear extracts, while only one is found in extracts from HeLa cells.

HeLa cells


Input: Possible mechanisms by which these proteins may regulate transcription of the epsilon-globin-encoding gene in erythroid and non-erythroid cells are discussed.

epsilon-globin-encoding gene

erythroid and

non-erythroid cells


Input: Calcitriol: a hematolymphopoietrope? [editorial]


Input: A MEDLINE search of the English-language literature was conducted using the indexing terms' immunology, calcitriol and vitamin D' to identify studies indicating a role for calcitriol as a primary immunomodulator.


Input: Sixty-six papers published between January 1956 and June 1991 were identified.


Input: Forty-five of these reports are cited in this review.


Input: The data strongly suggest an endocrine, autocrine and/or paracrine role for calcitriol in immune regulation.


Input: No unifying hypothesis has yet emerged explaining this collection of data.


Input: This paper provides a brief review of immune properties currently attributed to calcitriol.


Input: Transcription of the hypersensitive site HS2 enhancer in erythroid cells.

hypersensitive site HS2 enhancer

erythroid cells


Input: In the human genome, the erythroid-specific hypersensitive site HS2 enhancer regulates the transcription of the downstream beta-like globin genes 10-50 kilobases away.

human genome

erythroid-specific hypersensitive site HS2 enhancer

downstream beta-like globin genes

10-50 kilobases


Input: The mechanism of HS2 enhancer function is not known.

HS2 enhancer


Input: The present study employs RNA protection assays to analyze the transcriptional status of the HS2 enhancer in transfected recombinant chloramphenicol acetyltransferase (CAT) plasmids.

HS2 enhancer

transfected recombinant

) plasmids

chloramphenicol acetyltransferase

CAT


Input: In erythroid K562 cells in which the HS2 enhancer is active, the HS2 sequence directs the synthesis of long enhancer transcripts that are initiated apparently from within the enhancer and elongated through the intervening DNA into the cis-linked CAT gene.

erythroid K562 cells

HS2 enhancer

HS2 sequence

enhancer

cis-linked CAT gene


Input: In nonerythroid HL-60 cells in which the HS2 enhancer is inactive, long enhancer transcripts are not detectable.

nonerythroid HL-60 cells

HS2 enhancer

long enhancer transcripts


Input: Splitting the HS2 enhancer between two tandem Ap1 sites abolishes the synthesis of a group of long enhancer transcripts and results in loss of enhancer function and transcriptional silencing of the cis-linked CAT gene.

HS2 enhancer

tandem Ap1 sites

cis-linked CAT gene

long enhancer transcripts


Input: In directing the synthesis of RNA through the intervening DNA and the gene by a tracking and transcription mechanism, the HS2 enhancer may (i) open up the chromatin structure of a gene domain and (ii) deliver enhancer binding proteins to the promoter sequence where they may stimulate the transcription of the gene at the cap site.

RNA

DNA

HS2 enhancer

chromatin structure

gene domain

cap site

enhancer binding proteins


Input: Characterization of a novel T lymphocyte protein which binds to a site related to steroid/thyroid hormone receptor response elements in the negative regulatory sequence of the human immunodeficiency virus long terminal repeat.

novel T lymphocyte protein

steroid/thyroid hormone receptor response elements

negative regulatory sequence

human immunodeficiency virus long terminal repeat


Input: We have previously identified a T lymphocyte protein which binds to a site within the LTR of the human immunodeficiency virus type 1 (HIV-1) and exerts an inhibitory effect on virus gene expression.

T lymphocyte protein

LTR


Input: The palindromic site (site B) recognized by this protein is related to the palindromic binding sites of members of the steroid/thyroid hormone receptor family.

palindromic site

site B

palindromic binding sites

steroid/thyroid hormone receptor family


Input: Here we characterize the T cell protein binding to this site as a 100 kD protein which is most abundant in T cells and which binds to site B as a 200 kD complex.

T cell protein

200 kD complex

T cells

site B


Input: This protein is distinct from other members of the steroid/thyroid hormone receptor family including the COUP protein which has a closely related DNA binding specificity.

steroid/thyroid hormone receptor family

COUP protein


Input: TAR -independent transactivation by Tat in cells derived from the CNS: a novel mechanism of HIV-1 gene regulation.

TAR

Tat


Input: The Tat protein of human immunodeficiency virus type 1 (HIV-1) is essential for productive infection and is a potential target for antiviral therapy.

Tat protein


Input: Tat, a potent activator of HIV-1 gene expression, serves to greatly increase the rate of transcription directed by the viral promoter.

Tat

viral promoter


Input: This induction, which seems to be an important component in the progression of acquired immune deficiency syndrome (AIDS ), may be due to increased transcriptional initiation, increased transcriptional elongation, or a combination of these processes.


Input: Much attention has been focused on the interaction of Tat with a specific RNA target termed TAR (transactivation responsive) which is present in the leader sequence of all HIV-1 mRNAs.

Tat

RNA target

TAR

transactivation responsive

HIV-1 mRNAs


Input: This interaction is believed to be an important component of the mechanism of transactivation.


Input: In this report we demonstrate that in certain CNS-derived cells Tat is capable of activating HIV-1 through a TAR -independent pathway.

CNS-derived cells

Tat

TAR


Input: A Tat -responsive element is found upstream within the viral promoter that in glial-derived cell lines allows transactivation in the absence of TAR.

Tat -responsive element

viral promoter

glial-derived cell lines

TAR


Input: Deletion mapping and hybrid promoter constructs demonstrate that the newly identified Tat -responsive element corresponds to a sequence within the viral long terminal repeat (LTR) previously identified as the HIV-1 enhancer, or NF-kappa B domain.

hybrid promoter constructs

Tat -responsive element

viral long terminal repeat

LTR

HIV-1 enhancer

NF-kappa B domain


Input: DNA band-shift analysis reveals NF-kappa B binding activity in glial cells that differs from that present in T lymphoid cells.

NF-kappa B

T lymphoid cells


Input: Further, we observe that TAR-deleted mutants of HIV-1 demonstrate normal late gene expression in glial cells as evidenced by syncytia formation and production of viral p24 antigen.

TAR-deleted mutants

glial cells

viral p24 antigen


Input: (ABSTRACT TRUNCATED AT 250 WORDS)


Input: Transcription factor AP-2 activates gene expression of HTLV-I.

Transcription factor AP-2


Input: The HTLV-I LTR contains three conserved regulatory elements known as 21 base pair repeats which are required for stimulation of gene expression by the transactivator protein tax.

HTLV-I LTR

regulatory elements

21 base pair repeats

transactivator protein

tax


Input: Mutagenesis indicates that the 21 bp repeats can be subdivided into three motifs, A, B and C, each of which influences the level of tax activation.

21 bp repeats

A

B

C

tax


Input: The A site in the 21 bp repeat has strong homology with previously described binding sites for the transcription factor AP-2.

A site

21 bp repeat

binding sites

transcription factor AP-2


Input: We demonstrated that AP-2 mRNA was present in T-lymphocytes and that cellular factors from both non-transformed and transformed T-lymphocytes specifically bound to the consensus motif for AP-2 in each 21 bp.

AP-2 mRNA

T-lymphocytes

T-lymphocytes

consensus motif


Input: To determine the role of AP-2 in the regulation of the HTLV-I LTR gene expression, we used an AP-2 cDNA in DNA binding and transient expression assays.

HTLV-I LTR

cDNA


Input: Gel retardation and methylation interference studies revealed that bacterially produced AP-2 bound specifically and with high affinity to all three 21 bp repeats, and that it required the core sequence AGGC for specific binding.

bacterially produced

21 bp repeats

core sequence


Input: Binding of AP-2 prevented the subsequent binding of members of the CREB/ATF family to an adjacent regulatory motif in the 21 bp repeat.

CREB/ATF family

adjacent regulatory motif

21 bp repeat


Input: Transfection of an AP-2 expression construct into T-lymphocytes activated gene expression from the HTLV-I LTR.

AP-2 expression construct

HTLV-I LTR

T-lymphocytes


Input: At least two 21 bp repeats were required for high levels of AP-2 activation and mutagenesis of the AP-2 consensus binding sequences in the 21 bp repeats eliminate this activation.

21 bp repeats

consensus binding sequences

21 bp repeats


Input: (ABSTRACT TRUNCATED AT 250 WORDS)


Input: Activation of the human immunodeficiency virus type 1 enhancer is not dependent on NFAT-1.

human immunodeficiency virus type 1 enhancer


Input: The function of a putative NFAT-1 site in the human immunodeficiency virus type 1 enhancer has been analyzed.

putative NFAT-1 site

human immunodeficiency virus type 1 enhancer


Input: Activation by the T-cell antigen receptor is minimal in Jurkat cells and is mediated by the kappa B sites.

T-cell antigen receptor

Jurkat cells

kappa B sites


Input: The putative NFAT-1 region is not required for the response to anti-CD3 or to mitogens in T-cell, B-cell, or monocyte/macrophage leukemia lines, nor is it a cis-acting negative regulatory element.

putative

region

cis-acting negative regulatory element

mitogens

T-cell

B-cell

monocyte/macrophage leukemia lines


Input: The B cell-specific nuclear factor OTF-2 positively regulates transcription of the human class II transplantation gene, DRA.

B cell-specific nuclear factor

human class II transplantation gene

DRA


Input: The promoter of the major histocompatibility class II gene DRA contains an octamer element (ATTTGCAT) that is required for efficient DRA expression in B cells.

major histocompatibility class II gene

DRA

octamer element

DRA

B cells


Input: Several DNA-binding proteins are known to bind this sequence.

DNA-binding proteins


Input: The best characterized are the B cell-specific OTF-2 and the ubiquitous OTF-1.

B cell-specific OTF-2


Input: This report directly demonstrates that OTF-2 but not OTF-1 regulates the DRA gene.

DRA gene


Input: In vitro transcription analysis using protein fractions enriched for the octamer-binding protein OTF-2 demonstrate a positive functional role for OTF-2 in DRA gene transcription.

DRA gene


Input: In contrast, OTF-1 -enriched protein fractions did not affect DRA gene transcription although it functionally enhanced the transcription of another gene.

OTF-1 -enriched protein fractions

DRA gene


Input: Recombinant OTF-2 protein produced by in vitro transcription /translation could also enhance DRA gene transcription in vitro.

Recombinant

OTF-2 protein

DRA gene


Input: In vivo transient transfection studies utilizing an OTF-2 expression vector resulted in similar findings: that OTF-2 protein enhanced DRA gene transcription, and that this effect requires an intact octamer element.

OTF-2 expression vector

DRA gene

octamer element

OTF-2 protein


Input: Together these results constitute the first direct evidence of a positive role for the lymphoid-specific octamer-binding factor in DRA gene transcription.

lymphoid-specific octamer-binding factor


Input: Interferon-gamma potentiates the antiviral activity and the expression of interferon-stimulated genes induced by interferon-alpha in U937 cells.

Interferon-gamma

interferon-stimulated genes

interferon-alpha in U937 cells


Input: Binding of type I interferon (IFN-alpha/beta) to specific receptors results in the rapid transcriptional activation, independent of protein synthesis, of IFN-alpha-stimulated genes (ISGs) in human fibroblasts and HeLa and Daudi cell lines.

type I interferon (IFN-alpha/beta) to specific receptors

IFN-alpha-stimulated genes

ISGs

human fibroblasts

HeLa

Daudi cell lines


Input: The binding of ISGF3 (IFN-stimulated gene factor 3) to the conserved IFN-stimulated response element (ISRE) results in transcriptional activation.

IFN-stimulated gene factor 3

IFN-stimulated response element

ISRE


Input: This factor is composed of a DNA-binding protein (ISGF3 gamma ), which normally is present in the cytoplasm, and other IFN-alpha-activated proteins which preexist as latent cytoplasmic precursors (ISGF3 alpha ).

DNA-binding protein

ISGF3 gamma

IFN-alpha-activated proteins

latent cytoplasmic precursors

ISGF3 alpha


Input: We have found that ISG expression in the monocytic U937 cell line differs from most cell lines previously examined.

monocytic U937 cell line

cell lines


Input: U937 cells express both type I and type II IFN receptors, but only IFN-alpha is capable of inducing antiviral protection in these cells.

U937 cells

type I and type II IFN receptors

IFN-alpha


Input: Pretreatment with IFN-gamma potentiates the IFN-alpha -induced protection, but IFN-gamma alone does not have any antiviral activity.

IFN-gamma

IFN-alpha

IFN-gamma


Input: ISG15 mRNA accumulation in U937 cells is not detectable before 6 h of IFN-alpha treatment, peaks at 24 h, and requires protein synthesis.

ISG15 mRNA

U937 cells

IFN-alpha


Input: Although IFN-gamma alone does not induce ISG expression, IFN-gamma pretreatment markedly increases and hastens ISG expression and transcriptional induction.

IFN-gamma

IFN-gamma


Input: Nuclear extracts assayed for the presence of ISRE binding factors by electrophoretic mobility shift assays show that ISGF3 is induced by IFN-alpha within 6 h from undetectable basal levels in untreated U937 cells.

ISRE

binding factors

IFN-alpha

U937 cells


Input: Activation of ISGF3 alpha, the latent component of ISGF3, occurs rapidly.

ISGF3 alpha


Input: However, the increase in ISGF3 activity ultimately correlates with the accumulation of ISGF3 gamma induced by IFN-alpha or IFN-gamma.

ISGF3 gamma

IFN-alpha

IFN-gamma


Input: (ABSTRACT TRUNCATED AT 250 WORDS)


Input: Single point estimation of glucocorticoid receptors in lymphocytes of normal subjects and of children under long term glucocorticoid treatment.

glucocorticoid receptors

lymphocytes


Input: A single point assay of glucocorticoid receptors (GR) in human lymphocytes based on the measurement of specific dexamethasone binding has been developed and compared with a common multi-point Scatchard analysis.

glucocorticoid receptors

GR

lymphocytes


Input: The assay conditions-concentration of the ligand 20 nmol/l, incubation time 2 h and the cell count 2-6 mil.


Input: cells/tube in the assay volume 0.25 ml were found to be optimal.


Input: An attempt was also undertaken to use a cell harvester for the separation of cells from unbound ligand.


Input: Though specifically bound dexamethasone measured by whole-cell assay and that using cell harvester correlated well, almost by one order lower values obtained with the latter method render it non-applicable for receptor quantitation.


Input: The results from 9 healthy volunteers (average GR concentration 7131 +/- 1256 sites/cell) correlated excellently with those obtained by the Scatchard analysis.

GR


Input: The single point assay has been also applied for determination of GH in 10 children treated with large doses of prednisone.


Input: The average values from healthy volunteers did not differ significantly from those found in these children, though much broader range was found in patients.


Input: Glucocorticoid receptor and inhibition of 3-O-methyl-D-glucose uptake by glucocorticoids in peripheral blood leukocytes from normal humans: correlation between receptor level and hormone effect in vitro.

Glucocorticoid receptor

peripheral blood leukocytes


Input: We have measured the glucocorticoid receptor concentration in mononuclear and polymorphonuclear leukocytes, both of which were isolated from peripheral blood from ten healthy male volunteers.

glucocorticoid receptor

mononuclear

polymorphonuclear leukocytes


Input: In parallel, the inhibitory effect of dexamethasone on 3-O-methyl-D-glucose uptake was assayed in the corresponding mononuclear leukocytes.

mononuclear leukocytes


Input: The glucocorticoid receptor levels in mononuclear leukocytes correlated with those in polymorphonuclear leukocytes, and there was a linear relationship between the cellular glucocorticoid receptor levels and glucocorticoid -mediated inhibition of the uptake of 3-O-methyl-D-glucose in mononuclear leukocytes.

glucocorticoid receptor

glucocorticoid receptor

mononuclear leukocytes

polymorphonuclear leukocytes

mononuclear leukocytes


Input: When mononuclear leukocytes were incubated in the presence of 8-bromo-cAMP, cellular glucocorticoid receptor levels increased and a more pronounced inhibitory effect of dexamethasone was observed on the transport of 3-O-methyl-D-glucose.

mononuclear leukocytes

glucocorticoid receptor


Input: We conclude that the cellular glucocorticoid receptor levels in peripheral blood leukocytes reflect in vitro responsiveness to glucocorticoids in mononuclear leukocytes from healthy males, and that the individual responsiveness may alter upon changes in the cellular levels of glucocorticoid receptor.

glucocorticoid receptor

glucocorticoid receptor

peripheral blood leukocytes

mononuclear leukocytes


Input: Transcription factor activation and functional stimulation of human monocytes.

Transcription factor

human monocytes


Input: Activation of expression of genes encoding transcription factors: c-fos and c-jun and formation of AP1 transcriptional complex in human monocytes was investigated.

genes

c-fos

c-jun

transcription factors

AP1 transcriptional complex

human monocytes


Input: It was found that lipopolysaccharide induced strongly both c-fos and c-jun expression as well as AP1 formation.

c-fos

c-jun


Input: Interferon gamma activated strongly c-fos and weakly c-jun and AP1.

Interferon gamma

c-fos

c-jun


Input: Tumor necrosis factor induced slightly c-fos and had almost no effect on c-jun and AP1.

Tumor necrosis factor

c-fos

c-jun


Input: The data suggest that differences in functional responses elicited in monocytes by all three factors may be dependent on different routes on nuclear signalling employed by the factors.

monocytes

factors

factors


Input: Regulation of interleukin-1 beta production by glucocorticoids in human monocytes: the mechanism of action depends on the activation signal.

interleukin-1 beta

human monocytes


Input: Glucocorticoids are known to downregulate interleukin-1 beta production in monocytic cells by two different mechanims: direct inhibition of the gene transcription and destabilization of the preformed interleukin-1 beta mRNA.

interleukin-1 beta

interleukin-1 beta

monocytic cells

mRNA


Input: Now we have examined the effect of the nature of the monocyte activating signal on these two inhibitory mechanims.


Input: When human monocytes were preincubated with dexamethasone for 1 hour and then stimulated either with bacterial lipopolysaccharide or phorbol myristate, it was found that dexamethasone inhibited the lipopolysaccharide-induced interleukin-1 beta protein production, but the phorbol myristate -induced production was increased 3-10 fold.

human monocytes

interleukin-1 beta


Input: This difference was also seen at the mRNA level.


Input: When dexamethasone was added to the cultures 3 hours after the stimulators, it clearly decreased the interleukin-1 beta mRNA levels regardless of the stimulator used (although the effect was clearly weaker on the PMA-induced mRNA ).

interleukin-1 beta mRNA

PMA-induced mRNA


Input: Thus these data suggest that the phorbol myristate-induced signal (prolonged protein kinase C activation ?) cannot be inhibited by prior incubation with dexamethasone and it also protects the induced mRNA for the degradative action of dexamethasone.

protein kinase C

induced mRNA


Input: Nuclear transcription factors that bind to elements of the IL-2 promoter.

Nuclear transcription factors

IL-2 promoter


Input: Induction requirements in primary human T cells.

primary human T cells


Input: Prior studies have identified several elements that contribute to the activity of the IL-2 promoter in the stimulated T cell line, Jurkat.

IL-2 promoter

T cell line

Jurkat


Input: The sites and their corresponding nuclear binding factors include: NF-kappa B, AP-1, AP-3, OCT-1, and NF-AT.

NF-kappa B

NF-AT


Input: The latter " nuclear factor for activated T cells " likely contributes to the tissue specificity of IL-2 gene expression.

nuclear factor for

activated T cells

gene


Input: Using electrophoretic mobility shift assays, we have studied these transcription factors in primary T cells from human blood to verify their presence in a physiologic setting and to identify the signals that stimulate factor activity.

transcription factors

primary T cells


Input: All factors are induced in the nuclei of T cells upon activation with mitogens but not with exogenous IL-2 growth factor.

T cells

mitogens

exogenous IL-2 growth factor


Input: However, the signaling requirements and sensitivity to protein synthesis inhibitors differ considerably.


Input: Only the activities for NF-AT and AP-1 sites require two signals for optimal induction, i.e., PMA plus either lectin or antibody to the CD3 or CD28 surface molecules.

lectin

antibody

CD28 surface molecules


Input: Other factors are induced by lectin, antibody, and/or PMA alone.

lectin

antibody


Input: After appropriate stimulation, both NF-AT and AP-1 are peculiarly sensitive to the protein synthesis inhibitor anisomycin.

NF-AT


Input: Our data correlate the activity of NF-AT and AP-1 in gel shift assays with the two signals requirements for IL-2 gene expression.

NF-AT

IL-2 gene


Input: NF-kappa B activity in T cells stably expressing the Tax protein of human T cell lymphotropic virus type I.

NF-kappa B

Tax protein

T cells


Input: The effect of constitutive Tax expression on the interaction of NF-kappa B with its recognition sequence and on NF-kappa B -dependent gene expression was examined in T lymphoid Jurkat cell lines (19D and 9J) stably transformed with a Tax expression vector.

Tax

NF-kappa B

NF-kappa B

recognition sequence

Tax expression vector

T lymphoid Jurkat cell lines


Input: Tax expressing T cell lines contained a constitutive level of NF-kappa B binding activity, detectable by mobility shift assay and uv cross-linking using a palindromic NF-kappa B probe homologous to the interferon beta PRDII site.

Tax

NF-kappa B

expressing T cell lines

palindromic NF-kappa B probe

interferon beta PRDII site


Input: In Jurkat and NC2.10 induction with phorbol esters resulted in the appearance of new DNA binding proteins of 85, 75, and 54 kDa, whereas in Tax expressing cells the 85-kDa protein and a 92-kDa DNA binding protein were constitutively induced.

DNA binding proteins

Tax

85-kDa protein

92-kDa DNA binding protein

expressing cells


Input: Expression of Tax protein in 19D and 9J resulted in transcription of the endogenous NF-kappa B -dependent granulocyte-macrophage colony stimulating factor gene and increased basal level expression of transfected NF-kappa B -regulated promoters.

Tax protein

NF-kappa B

endogenous NF-kappa B -dependent granulocyte-macrophage colony stimulating factor gene

-regulated promoters


Input: Nonetheless transcription of both the endogenous and the transfected gene was inducible by PMA treatment.


Input: Tax expression in Jurkat T cells may alter the stoichiometry of NF-kappa B DNA binding proteins and thus change the expression of NF-kappa B -regulated promoters.

Tax

NF-kappa B

DNA binding proteins

NF-kappa B

Jurkat T cells

-regulated promoters


Input: A nuclear factor NF-GM2 that interacts with a regulatory region of the GM-CSF gene essential for its induction in responses to T-cell activation: purification from human T-cell leukemia line Jurkat cells and similarity to NF-kappa B.

nuclear factor

NF-kappa B

regulatory region

GM-CSF gene

T-cell leukemia line Jurkat cells


Input: Activation of T cells by antigen, lectin, or a combination of phorbol-12-myristate acetate (PMA) and calcium ionophore (A23187) leads to the induction of genes for a set of lymphokines, including granulocyte-macrophage colony-stimulating factor (GM-CSF ).

T cells

lectin

lymphokines

granulocyte-macrophage colony-stimulating factor

GM-CSF


Input: We demonstrated in earlier studies that the upstream region of the mouse GM-CSF promoter at positions between -95 and -73 is essential for transcriptional activation in response to PMA/A23187.

upstream region

mouse

promoter

-95 and -73

GM-CSF


Input: This region contains two DNA-binding motifs, GM2 and GC-box.

DNA-binding motifs

GC-box


Input: The GM2 sequence (GGTAGTTCCC) is recognized by an inducible factor NF-GM2; the other (CCGCCC) by constitutive factors A1, A2, and B.

GM2 sequence

inducible factor NF-GM2

constitutive factors

B


Input: To elucidate the mechanism of GM-CSF gene activation, we have purified the inducible factor NF-GM2 from the nuclear extract of stimulated Jurkat cells on the basis of specific DNA-binding activity.

GM-CSF

inducible factor NF-GM2

Jurkat cells


Input: The purified NF-GM2 consists of 50 (p50) and 65 kDa (p65) polypeptides and has a binding activity specific for both the GM-CSF and immunoglobulin kappa (GGAAAGTCCC) enhancers.

GM-CSF

immunoglobulin kappa (GGAAAGTCCC) enhancers


Input: Electrophoretically purified p50 alone can form a protein-DNA complex, but in the mixture, p50 associates preferentially with p65 to form the NF-GM2 complex.

protein-DNA complex

NF-GM2 complex


Input: In addition, p65 gave per se, with low affinity, a protein-DNA complex that migrated more slowly than native NF-GM2 complex.

protein-DNA complex

NF-GM2 complex


Input: Furthermore, an antiserum against KBF1 (identical to 50 kDa NF-kappa B protein) reacted with the p50 of NF-GM2, indicating that the NF-GM2 polypeptide cannot be immunologically differentiated from the 50 kDa subunit of NF-kappa B.

50 kDa NF-kappa B protein

NF-GM2 polypeptide

50 kDa subunit

NF-kappa B


Input: The purified NF-GM2 activated in vitro transcription from the kappa B enhancer, while it failed to stimulate transcription from the GM-CSF promoter harboring the GM2 sequence.

kappa B enhancer

GM-CSF promoter

GM2 sequence


Input: This suggests that the activation mechanism of the GM-CSF gene through the GM2/GC-box sequence is different from that of genes carrying the kappa B enhancer alone.

GM-CSF gene

GM2/GC-box sequence

kappa B enhancer


Input: Characterization of an immediate-early gene induced in adherent monocytes that encodes I kappa B -like activity.

immediate-early gene

adherent monocytes

I kappa B


Input: We have cloned a group of cDNAs representing mRNAs that are rapidly induced following adherence of human monocytes.

cDNAs

mRNAs

human monocytes


Input: One of the induced transcripts (MAD-3) encodes a protein of 317 amino acids with one domain containing five tandem repeats of the cdc10/ankyrin motif, which is 60% similar (46% identical) to the ankyrin repeat region of the precursor of NF-kappa B /KBF1 p50.

five tandem repeats

cdc10/ankyrin motif

ankyrin repeat region

NF-kappa B

/KBF1 p50


Input: The C-terminus has a putative protein kinase C phosphorylation site.

C-terminus

protein kinase C phosphorylation site


Input: In vitro translated MAD-3 protein was found to specifically inhibit the DNA-binding activity of the p50/p65 NF-kappa B complex but not that of the p50/p50 KBF1 factor or of other DNA-binding proteins.

MAD-3 protein

p50/p65 NF-kappa B complex

p50/p50 KBF1 factor

DNA-binding proteins


Input: The MAD-3 cDNA encodes an I kappa B-like protein that is likely to be involved in regulation of transcriptional responses to NF-kappa B, including adhesion-dependent pathways of monocyte activation.

MAD-3 cDNA

I kappa B-like protein

NF-kappa B


Input: Cortivazol mediated induction of glucocorticoid receptor messenger ribonucleic acid in wild-type and dexamethasone -resistant human leukemic (CEM) cells.

glucocorticoid receptor

messenger ribonucleic acid

wild-type

dexamethasone -resistant human leukemic (CEM) cells


Input: Cortivazol is a phenylpyrazolo glucocorticoid of high potency and unusual structure.


Input: In both wild-type and highly dexamethasone (dex)-resistant clones of the human leukemic cell line CEM, exposure to cortivazol leads to cell death.

wild-type

dexamethasone (dex)-resistant clones

human leukemic cell line CEM


Input: It has been shown recently that in wild-type CEM cells but not in a dex-resistant, glucocorticoid receptor (GR)-defective clone ICR-27 TK-3, dex induces GR mRNA.

wild-type CEM cells

dex-resistant,

(GR)-defective clone

ICR-27 TK-3

glucocorticoid receptor

GR mRNA


Input: To test the hypothesis that cortivazol acts in dex -resistant cells by making use of the residual GR found there, wild-type and dex -resistant clones were treated with various concentrations of cortivazol and induction of GR mRNA was studied.

dex -resistant cells

dex -resistant clones

GR

GR mRNA


Input: Cortivazol significantly induced GR mRNA in the normal CEM-C7 as well as in two classes of dex -resistant clones, although the dex -resistant clones needed at least 10 times more cortivazol than the normal cells for significant GR mRNA induction.

GR mRNA

mRNA

normal CEM-C7

dex -resistant clones

dex -resistant clones

normal cells

GR


Input: Increased levels of GR mRNA were noticed as early as 3 h after treatment.

GR mRNA


Input: A general correlation between induction of GR mRNA and lysis of the normal and dex -resistant cells was found.

GR

mRNA

dex -resistant cells


Input: Positive induction of GR mRNA might be one of the earliest crucial steps in the lysis of normal and dex-resistant CEM cells, or might serve as a marker for the process.

GR mRNA


Input: However, the lysis pathway in the dex -resistant cells is defective in that dex -resistant clones needed significantly more cortivazol than the normal cells for lysis of the cells.

dex -resistant cells

dex -resistant clones

normal cells


Input: HIV enhancer activity perpetuated by NF-kappa B induction on infection of monocytes [see comments]

HIV enhancer

NF-kappa B

monocytes


Input: Permissiveness to replication of human immunodeficiency virus (HIV) differs in T lymphocytes and macrophages.

T lymphocytes

macrophages


Input: In T cells, HIV transcription is poorly detected in vivo.

T cells


Input: Cloned, normal T lymphocytes show very little, if any, basal activity of the HIV enhancer and low nuclear expression of NF-kappa B, a potent transcriptional activator of the HIV enhancer.

T lymphocytes

HIV enhancer

HIV enhancer

NF-kappa B

potent transcriptional activator


Input: In contrast, fixed tissue macrophages express detectable HIV proteins, indicating permanent virus transcription.

macrophages

HIV proteins


Input: One explanation for the perpetuation of virus infection in macrophages could be sustained nuclear NF-kappa B expression.

macrophages

NF-kappa B


Input: However, the U937 monocytic cell line, which is fully permissive to HIV replication, is known to express only low levels of nuclear NF-kappa B.

U937 monocytic cell line

nuclear NF-kappa B


Input: We show here that chronic HIV infection results in both induction of a nuclear factor with antigenic properties indistinguishable from those of NF-kappa B and permanently increased HIV enhancer activity.

nuclear factor

NF-kappa B

HIV enhancer


Input: This phenomenon, which is independent of tumour necrosis factor, is associated with HIV replication, and is thus likely to explain at least in part the perpetuation of HIV infection in monocytes.

tumour necrosis factor

monocytes


Input: Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NF-kappa B [published erratum appears in Science 1991 Oct 4;254(5028):11]

rel-related human cDNA

65-kD subunit

NF-kappa B


Input: A DNA probe that spanned a domain conserved among the proto-oncogene c-rel, the Drosophila morphogen dorsal, and the p50 DNA binding subunit of NF-kappa B was generated from Jurkat T cell complementary DNA with the polymerase chain reaction (PCR) and degenerate oligonucleotides.

DNA probe

proto-oncogene c-rel

Jurkat T cell complementary DNA

Drosophila morphogen dorsal

p50 DNA binding subunit

NF-kappa B


Input: This probe was used to identify a rel-related complementary DNA that hybridized to a 2.6-kilobase messenger RNA present in human T and B lymphocytes.

rel-related complementary DNA

2.6-kilobase messenger RNA


Input: In vitro transcription and translation of the complementary DNA resulted in the synthesis of a protein with an apparent molecular size of 65 kilodaltons (kD).

complementary DNA

65 kilodaltons


Input: The translated protein showed weak DNA binding with a specificity for the kappa B binding motif.

kappa B binding motif


Input: This protein-DNA complex comigrated with the complex obtained with the purified human p65 NF-kappa B subunit and binding was inhibited by I kappa B-alpha and -beta proteins.

protein-DNA complex

purified human p65 NF-kappa B subunit

I kappa B-alpha

-beta proteins


Input: In addition, the 65-kD protein associated with the p50 subunit of NF-kappa B and the kappa B probe to form a complex with the same electrophoretic mobility as the NF-kappa B -DNA complex.

65-kD protein

p50 subunit

NF-kappa B

NF-kappa B -DNA complex

kappa B probe


Input: Therefore the rel-related 65-kD protein may represent the p65 subunit of the active NF-kappa B transcription factor complex.

rel-related 65-kD protein

p65 subunit

active NF-kappa B transcription factor complex


Input: The human myelomonocytic cell line U-937 as a model for studying alterations in steroid-induced monokine gene expression: marked enhancement of lipopolysaccharide-stimulated interleukin-1 beta messenger RNA levels by 1,25-dihydroxyvitamin D3.

human myelomonocytic cell line U-937

lipopolysaccharide-stimulated interleukin-1 beta messenger RNA


Input: The active metabolite of vitamin D, 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3 ], is a potent regulator of human monocyte/macrophage function in vitro.


Input: To establish a model for 1,25-(OH)2D3 regulation of human monocyte monokine synthesis, three human cell lines (U-937, THP-1, and HL-60) were examined for: 1) the presence of functional 1,25-(OH)2D3 receptors; 2) the accumulation of interleukin-1 beta (IL-1 beta) mRNA and IL-1 beta protein in response to lipopolysaccharide (LPS ); and 3) the regulation of this response by 1,25-(OH)2D3.

human cell lines

interleukin-1 beta (

) mRNA

IL-1 beta

IL-1 beta protein


Input: All three cell lines expressed vitamin D receptor and had increased levels of IL-1 beta mRNA in response to LPS.

IL-1 beta mRNA


Input: Preincubation of cells with 1,25-(OH)2D3 augmented IL-1 beta mRNA levels only in U-937 and HL-60 cells.

IL-1 beta

mRNA


Input: From these data, and taking into consideration their state of differentiation and relative ease of culture, U-937 was chosen over HL-60 and THP-1 as the cell line we further characterized.


Input: In U-937 cells, optimum time and dose of pretreatment with 1,25-(OH)2D3 were determined to be 12-24 h at a receptor saturating concentration of 1,25-(OH)2D3 (10 nM).

U-937 cells


Input: Preincubation of cells with 1,25-(OH)2D3 had no effect on the time course of IL-1 beta mRNA appearance in response to LPS.

IL-1 beta

mRNA


Input: However, exposure of U-937 cells to 1,25-(OH)2D3 increased by 200% the level of IL-1 beta mRNA detected and decreased by three orders of magnitude the concentration of LPS required to achieve steady state mRNA levels equivalent to those observed in U-937 cells not preincubated with the hormone.2+o

U-937 cells

U-937 cells

IL-1 beta

mRNA


Input: Regulation of M-CSF expression by M-CSF: role of protein kinase C and transcription factor NF kappa B.

M-CSF

M-CSF

protein kinase C

transcription factor NF kappa B


Input: Macrophage-colony-stimulating factor (M-CSF ), also referred to as CSF-1, regulates the survival, growth, differentiation and functional activity of monocytes by binding to a single class of high-affinity cell surface receptors, known to be the product of the c-fms protooncogene.

Macrophage-colony-stimulating factor

M-CSF

high-affinity cell surface receptors

monocytes

c-fms protooncogene


Input: The detection of both M-CSF and c-fms expression by cells of the monocyte lineage has suggested that M-CSF may act by an autocrine mechanism.

monocyte lineage

M-CSF


Input: Interestingly, it has been shown that M-CSF can induce the expression of its own gene.

M-CSF