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INTRODUCTION
The HIV-1 envelope (Env) trimer is protected by multiple mechanisms of humoral evasion, including conformational masking, glycan shielding, and sequence variation.Stabilization of the Env trimer through the incorporation of a disulfide (SOS) between residues 501 of gp120 and 605 of gp41 and the substitution of an isole...
PMC10491024
RESULTS
PMC10491024
Sorting of VRC 018 B cells from donor N751 reveal that only a small fraction of elicited antibodies recognize the glycan-dense surface of the Env trimer
In the Vaccine Research Center (VRC) 018 clinical trial (To quantify the number of cells binding to the glycan-dense Env-trimer surface relative to those bound the glycan-free base, we performed cell cytometry on immunoglobulin G-positive (IgGFrom two plates of single-sorted B cells, 96 wells each, specific for the gly...
PMC10491024
Vaccine-elicited human antibodies N751-2C06.01 and N751-2C09.01 neutralize HIV strain BG505
VIRUS
We further characterized the purified antibodies for neutralization activity and found that the antibodies N751-2C06.01 and N751-2C09.01 (named for donor-lineage.clone, hereafter referred to as 2C06 and 2C09) neutralized both BG505 and a version of BG505 virus with glycan at residue 611 missing (BG505.N611Q), found to ...
PMC10491024
Cryo-EM structure of N751-2C06.01 in complex with BG505 DS-SOSIP reveals the antibody to bind at the fusion-peptide site of vulnerability
To define recognition details, we determined the cryo-EM structure of 2C06 in complex with BG505 DS-SOSIP. We obtained a 3D-reconstruction map at 2.95 Å from 645,442 particles utilizing C1 symmetry (Antibody 2C06 bound at the fusion-peptide site of vulnerability (The fusion peptide was flanked by CDRs H1 and H2 on one ...
PMC10491024
Cryo-EM structure of N751-2C09.01 in complex with BG505 DS-SOSIP reveals epitope similarity to N751-2C06.01
To understand the differences and similarities between antibodies 2C06 and 2C09, we determined the cryo-EM structure of 2C09 in complex with BG505 DS-SOSIP. 2C09 binding to the envelope trimer did not induce noticeable asymmetry, and we obtained a 3D-reconstruction map at 2.81 Å with C3 symmetry from 445,733 particles ...
PMC10491024
Antibodies 2C06 and 2C09 form a reproducible antibody class
Antibodies 2C06 and 2C09 originated from similar heavy-chain variable genes, IGHV3-64D and IGHV3-64, respectively (The similarity in origin genes and in Env-trimer recognition suggested that 2C06 and 2C09 might be members of a reproducible antibody class. Indeed, most residues involved in trimer binding were identical ...
PMC10491024
Antibody 2C06 partially accommodates, but antibody 2C09 clashes with, glycan241
MUTANT
We next investigated the parameters governing breadth of recognition by antibodies 2C06 and 2C09. As described in the aforementioned structural analysis, both 2C06 and 2C09 epitopes involved S241, a rare glycan hole in BG505 among HIV isolates; specifically, in both 2C06 and 2C09, heavy-chain residues D61 and R64 were ...
PMC10491024
High-entropy epitope residues contribute to the strain specificity of 2C06 and 2C09
MUTANTS
While recognition of the 241 glycan hole explains much of the strain specificity of 2C06 and 2C09, there were seven strains other than BG505 in our 208-isolate panel missing glycan241, and we next investigated whether HIV sequence diversity of the epitope residues contributed to the strain-specific neutralization of th...
PMC10491024
DISCUSSION
As noted above, the HIV Env trimer evades antibody-mediated neutralization by conformational masking, glycan shielding, and sequence variation. Env trimers stabilized in the prefusion-closed conformation, however, induce recognition of the neutralization-susceptible shape of Env, and their use in human vaccine trials s...
PMC10491024
Limitations of the study
somatic hypermutations
This study describes antibodies isolated from a single donor at a single time point. The quality of isolated antibodies is also limited by the source of the study, the VRC 018 clinical trial, which was designed for the primary goal of testing the safety of BG505 DS-SOSIP adjuvanted by alum as a vaccine candidate. As a ...
PMC10491024
STAR★METHODS
PMC10491024
RESOURCE AVAILABILITY
PMC10491024
Lead contact
Further information and requests for resources and reagents should be directed to and will be fulfilled by Peter D. Kwong (
PMC10491024
Materials availability
Plasmids generated in this study are available upon request.
PMC10491024
Data and code availability
Cryo-EM maps have been deposited to the EMDB with accession codes EMD-29725 and EMD-29731, and fitted coordinates have been deposited to PDB with accession codes 8G4M and 8G4T.This paper does not report original code.Any additional information required to reanalyze the data reported in this paper is available from the ...
PMC10491024
EXPERIMENTAL MODEL AND SUBJECT DETAILS
PMC10491024
Serum samples
INFECTIOUS DISEASES, ALLERGY
The samples were collected under the Vaccine Research Center’s (VRC), National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health protocol VRC 018 (
PMC10491024
Cell lines
HEK293F, Expi293F and FreeStyle 293-F cells were purchased from Thermo Fisher Scientific. The cells were used directly from the commercial sources following manufacturer suggestions as described in detail below.
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METHOD DETAILS
PMC10491024
Preparation of Env trimers and antigen-specific probes
BG505 DS-SOSIP and ConC trimers were expressed in stable CHO cell lines and purified by non-affinity chromatography.
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B cell sorting
Cryopreserved PBMC from VRC 018 clinical trial (
PMC10491024
Rapid assembly, transfection, and production of immunoglobulins (RATP-Ig)
The sorted B cells were subjected to RATP-Ig following the procedures as described previously.
PMC10491024
AlphaLISA screening of RATP-Ig supernatants
RATP-Ig supernatants from glycan-base BG505-positive B cells were diluted into AlphaLISA buffer (PBS +0.05% Tween 20 + 0.5 mg/mL BSA), and 5 μL of each were transferred to an OptiPlate-384 assay plate (white opaque, PerkinElmer, Waltham, MA). To each well was added 10 μL of biotinylated Env trimer probe at 10 nM final ...
PMC10491024
Antibody preparation
DNA encoding antibody heavy- and light-chain variable regions were synthesized and cloned into the pVRC8400 vectors.
PMC10491024
Env-pseudovirus neutralization assays
single-round infection
Monoclonal antibody neutralization was assessed based on the single-round infection assay of TZM-bl cells with HIV-1 Env-pseudoviruses as described previously.
PMC10491024
Enzyme-linked immunosorbent assay (ELISA)
AVGIGA
Anti-fusion peptide ELISA was performed by coating 96-well plates (Costar High Binding Half-Area; Corning, Kennebunk, ME) overnight at 4°C with 50 μL/well of 2 μg/mL FP8v1-rTTHC (AVGIGAVF), FP7v1-rTTHC (AVGIGAV), or FP6v1-rTTHC (AVGIGA) in PBS. Between each subsequent step, plates were washed five times with PBS-T (PBS...
PMC10491024
IgG binding affinity measured by Carterra
A medium density HC30M sensor chip was preconditioned with 1-min pulses of each of three solutions: 50 mM NaOH, 1 M NaCl, and 10 mM Glycine pH 2.0. After preconditioning, the chip was activated with a 10-min injection of a freshly prepared 1:1:1 (v/v/v) mixture of 0.4 M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hy...
PMC10491024
Bio-layer interferometry (BLI)
AHC
An Octet RED instrument (FortéBio) instrument was used to measure recognition of isolated antibodies to HIV-1 BG505 DS-SOSIP, glycan-base BG505, ConC and glycan-base BG505-N241 trimers. Isolated antibodies (50 μg/mL) were immobilized for 300 s on AHC biosensor tips and equilibrated for 60 s in Octet Kinetics Buffer (Sa...
PMC10491024
Surface plasmon resonance measurements of Fab binding affinity
Binding affinities and kinetics of 2C06 and 2C09 antibodies were assessed by surface plasmon resonance (SPR) on a Biacore S-200 (GE Healthcare) at 25°C in HBS-P+ buffer (10 mM HEPES, pH 7.4, 150 mM NaCl and 0.05% surfactant P-20). Glycan-reactive 2G12 IgG was immobilized onto a CM5 chip by amine coupling to 8000–10000 ...
PMC10491024
Cryo-EM sample preparation and data collection
Samples for cryo-EM grid preparation of the 2C06-BG505 DS-SOSIP complex were produced by first mixing 15 μL of purified BG505 SOSIP at 4 mg/mL with 55 μL of 2C06 Fab at 1 mg/mL; n-Dodecyl β-D-maltoside (DDM) was added to have a final concentration of 0.005% (w/v) to prevent preferred orientation and aggregation during ...
PMC10491024
Model building and refinement
For structural determination, a model of the antibody Fab was generated using AlphaFold2 python notebook (
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Antibody recombination and sequence signature analyses
EVENT
IgBlast was used to annotate antibody germline gene and to analyze V(D)J recombination event.The precursor frequency was calculated using OLGA software.
PMC10491024
QUANTIFICATION AND STATISTICAL ANALYSIS
Cryo-EM data were processed and analyzed using CryoSparc. Cryo-EM structural statistics were analyzed with Phenix and Molprobity. Statistical details of experiments are described in
PMC10491024
Supplementary Material
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ACKNOWLEDGMENTS
AIDS, Rosales-Zavala, Cancer
INFECTIOUS DISEASES, ALLERGY, AIDS, FRANK, PLUMMER, CANCER
We thank Jonathan Stuckey for assistance with figures, members of the Virology Laboratory and Vector Core, Vaccine Research Center, for discussions and comments on the manuscript, and David Ho and members of the Aaron Diamond AIDS Research Center for helpful comments. We thank Preeti Apte, Anita Arthur, Allison Beck, N...
PMC10491024
REFERENCES
PMC10491024
Sorting of VRC 018 B cells from donor N751 identifies antibodies 2C06 and 2C09 that neutralize BG505
Langmuir
(A) VRC 018 clinical regimen and a flowchart for B cell sorting and antibody screening.(B) Single B cell sorting with glycan-base BG505 trimer probes reveals only a small fraction of memory B cells from VRC 018 clinical trial to be directed to the glycan-dense surface of the Env trimer versus its glycan-free base.(C) A...
PMC10491024
Cryo-EM structure of N751-2C06.01 in complex with BG505 DS-SOSIP reveals antibody recognition at the fusion-peptide site of vulnerability
(A) Cartoon representation of 2C06 in complex with BG505 DS-SOSIP. 2C06 bound at the fusion-peptide site and interacted with fusion peptide (FP), shown in red. Antibody heavy chain (H) is shown in blue, light chain (L) in pale cyan, gp41 in dark gray, and gp120 in light gray.(B) Density map for fusion peptide N-termina...
PMC10491024
Cryo-EM structure of N751-2C09.01 in complex with BG505 DS-SOSIP reveals epitope similarity to N751-2C06.01
(A) Cartoon representation of 2C09 Fab in complex with BG505 DS-SOSIP. 2C09 bound at the fusion-peptide site and interacted with fusion peptide extensively. Heavy chain (H) is shown in orange, light chain (L) in yellow-orange, fusion peptide (FP) in red, the rest of gp41 in darker gray, and gp120 in lighter gray.(B) De...
PMC10491024
Antibodies 2C06 and 2C09 form a reproducible class
(A and B) Analysis of junction regions and sequence alignment of 2C06 and 2C09. Germline gene nucleotide and amino acid residues are shown in black. The somatic hypermutations are colored red. Nucleotides removed by exonuclease trimming are crossed out. Germline amino acid differences between IGHV3-64 and IGHV3-64D are...
PMC10491024
Antibody 2C06 partially accommodates, but antibody 2C09 clashes with, glycan241
PyMOL
MUTANT
(A) BLI binding analysis of 2C06 and 2C09 with Env trimers with or without glycan241 in comparison with other fusion-peptide-directed antibodies. Both BG505 DS-SOSIP and glycan-base BG505 have serine at 241. Glycan-base BG505-N241 is an S241N mutant of glycan-base BG505 containing (B) Structures of VRC34.01 (PDB: 6nc3)...
PMC10491024
Recognition requirements for high-entropy residues contribute to strain specificity of 2C06 and 2C09
(A) While neutralization breadth generally correlates with epitope entropy, antibodies directed to the fusion-peptide sites of vulnerability have similar epitope entropy. Neutralization breadth represents the percentage of the 208-strain panel neutralized with IC(B) Analyses of sequence entropy and contact surface area...
PMC10491024
Highlights
HIV-1 neutralizing antibodies 2C06 and 2C09 from VRC 018 clinical trial are isolatedCryo-EM structures of 2C06 and 2C09 in complex with HIV envelope trimer are determined2C06 and 2C09 binding requires a glycan hole at 241 and BG505-specific residuesAntibodies 2C06 and 2C09 form a reproducible class, recognizing fusion ...
PMC10491024
Background:
toxicity, nonbacterial breast inflammation
PATHOGENESIS, PLASMA CELL MASTITIS, PCM
These authors contributed equally to this work.Plasma cell mastitis (PCM) is a nonbacterial breast inflammation with severe and intense clinical manifestation, yet treatment methods for PCM are still rather limited. Although the mechanism of PCM remains unclear, mounting evidence suggests that the dysregulation of immu...
PMC10063228
Methods:
PCM
We have developed a knowledge graph architecture toward immunotherapy and systematic immunity that consists of herbal drug–target interactions with a novel scoring system to select drug combinations based on target-hitting rates and phenotype relativeness. To this end, we employed this knowledge graph to identify an he...
PMC10063228
Results:
RECURRENCE, PCM
Our clinical data suggests that the herbal drug combination could significantly reduce the serum level of various inflammatory cytokines, downregulate serum IgA and IgG level, reduce the recurrence rate, and reverse the clinical symptoms of PCM patients with improvements in general health status.
PMC10063228
Conclusions:
RECURRENCE, PCM
In summary, we reported that an herbal drug combination identified by knowledge graph can alleviate the clinical symptoms of PCM patients. We demonstrated that the herbal drug combination holds great promise as an effective remedy for PCM, acting through the regulation of immunoinflammatory pathways and improvement of ...
PMC10063228
Funding:
C. Liu’s lab was supported by grants from the Public Health Science and Technology Project of Shenyang (grant: 22-321-32-18); Y. Yang’s laboratory was supported by the National Natural Science Foundation of China (grant: 81874301), the Fundamental Research Funds for Central University (grant: DUT22YG122), and the Key R...
PMC10063228
Research organism
PMC10063228
Introduction
TCM
DISEASES, PLASMA CELL MASTITIS, PCM
Plasma cell mastitis (PCM) represents a serious inflammatory condition of breast that occurs in young and middle-aged females at nonpregnant and nonlactating period (Traditional Chinese Medicine (TCM) has a rather long history for the prevention and treatment of complex diseases in eastern Asia (Knowledge graph has eme...
PMC10063228
Methods
PMC10063228
Construction of knowledge graph toward immunotherapy
We employed data mining techniques to collect and compile 240 targets of immunotherapy and systematic immunity from the PubMed database. Next, we collected and compiled 345 herbal drug entities officially released by the National Health Commission of China and the National Administration of Traditional Chinese Medicine...
PMC10063228
Design of the clinical trial
PCM
In brief, 160 female patients diagnosed with PCM at the Shengjing Hospital Affiliated to China Medical University were recruited in the clinical trial between January 2021 and February 2022. The patients were divided 1:1 into experimental group (EG) and control group (CG). It is noteworthy that, in order to demonstrate...
PMC10063228
Clinical trial protocol
The clinical trial for the herbal drug combination was registered at
PMC10063228
Measurement of serum inflammatory cytokines by ELSIA assay
STERILE
Venous blood of the CG and EG was collected in a sterile non-anticoagulant test tube before and after treatment. The immune transmission turbidimetry was used according to the procedure of CRP kit, and automatic biochemical analyzer was used to detect the level of CRP. The levels of serum cytokines were measured by ELI...
PMC10063228
Measurement of serum immunoglobulin level
STERILE, PCM
The venous blood of PCM patients in the two groups was collected in a sterile non-anticoagulant tube before and after treatment. The serum IgG and IgA were measured by rate scattering turbidimetry using Array 360 System automatic-specific protein analyzer (Beckman Company, USA).
PMC10063228
Assessment of clinical symptoms of PCM patients
PATHOLOGY, PCM
The clinical symptoms were evaluated by an attending physician with board certification in pathology. The patients were scored before and after treatment according to the standard rating scale for PCM (
PMC10063228
Statistics
All data were evaluated as mean ± SEM. Statistical analysis of the quantitative multiple group comparisons was performed using ANOVA, followed by Tukeyʼs test, whereas pairwise comparisons were performed using the
PMC10063228
Results
In a previous study, we collected and compiled 240 targets for immunotherapy and systematic immunity from literature data (
PMC10063228
Snapshot of the medical knowledge graph toward immunotherapy.
The knowledge graph was constructed using Neo4j and Python tools (
PMC10063228
CONSORT flowchart diagram of the clinical trial for the herbal drug combination.
Next, we extracted the subgraph for the herbal drug combination mentioned above and created a network diagram for the drug combination using Cytoscape tools (
PMC10063228
The score statistics of 10 rounds of random herbal drug combinations (1000 random combination from each round) for the treatment of plasma cell mastitis.
It is noteworthy that the scoring results of the 10 rounds are presented as normal distributions. The herbal drug combination identified for the clinical trial was among the top 20 choices in all the 10 rounds and is marked with red dots.
PMC10063228
Pathway analysis for the potential cellular targets of the herbal drug combination via Reactome Knowledgebase.
The statistically significant pathways are highlighted and displayed in yellow color (p<0.05).Efficacy was assessed every two cycles, and the results were summarized after 6 mo of treatment. The baseline characteristics are shown in
PMC10063228
The change of percentage for numerous immunological cytokines, including IL-2, IL-4, IL-6, IL-10, IL-17, CRP, IFN-γ, IL-1β, and TNF-α, from the control group and the treatment group (with Traditional Chinese Medicine [TCM] treatment).
TNF-α
Notably, a few key cytokines such as IL-2, IL-4, IFN-γ, IL-1β, and TNF-α were significantly downregulated in the treatment group compared to the control group.
PMC10063228
Patients’ raw data of serum cytokines from the control group (without Traditional Chinese Medicine [TCM] treatment) and the treatment group (with TCM treatment) in the clinical trial (
PMC10063228
The change of percentage for IgA level and IgG level from the control group and the treatment group (with Traditional Chinese Medicine [TCM] treatment).
PMC10063228
Patients’ raw data of serum IgA and IgG level from the control group (without Traditional Chinese Medicine [TCM] treatment) and the treatment group (with TCM treatment) in the clinical trial (
PMC10063228
The change of scores from items, including symptom, pain, and living quality.
pain
The scores of symptoms, pain, and living quality from the treatment group were significantly improved compared to the control group.
PMC10063228
Patients’ raw data of symptom scores from the control group (without Traditional Chinese Medicine [TCM] treatment) and the treatment group (with TCM treatment) in the clinical trial (
PMC10063228
Patients’ raw data of pain scores from the control group (without Traditional Chinese Medicine [TCM] treatment) and the treatment group (with TCM treatment) in the clinical trial (
PMC10063228
The comparison of score of life quality between different groups.
Control group (CG) baseline, experimental group (EG) baseline, CG treated with methylprednisolone, and EG treated with herbal drug combinations.
PMC10063228
Comparison of operation rate, recurrence rate, and incidence of adverse reactions between the two groups (EG: experimental group; CG: control group).
PMC10063228
The comparison of whole breast for six representative plasma cell mastitis (PCM) patients in the experimental group (EG) before treatment and after treatment.
(
PMC10063228
Clinical symptom scores between the two groups in the clinical trial (EG: experimental group; CG: control group; The clinical symptom rating scale for plasma cell mastitis (PCM) is displayed in
PMC10063228
Discussion
pain
RECURRENCE, BREAST CANCER, ADVERSE EVENTS, PCM, PATHOGENESIS
With the increasing amount of biomedical data, the traditional drug discovery campaign has been revolutionized with the aid of artificial intelligence techniques to accelerate the process and reduce the cost (Although the pathogenesis of PCM remains largely unclear, there have been numerous reports implicating that the...
PMC10063228
Conclusion
human disorders
RECURRENCE, PCM
In summary, we report the identification and clinical assessment of an herbal drug combination toward PCM. We demonstrated that the herbal drug combination holds great promise as an effective remedy for PCM, acting through the regulation of multiple cellular targets and immunoinflammatory pathways, which leads to the i...
PMC10063228
Ethical statement
The protocol was approved by the Institutional Review Board (IRB) of the China Medical University (approval number: 2021PS024T). This study was registered with
PMC10063228
Acknowledgements
Cancer
CANCER
Y Yang’s laboratory was supported by the National Natural Science Foundation of China (grant: 81874301), the Fundamental Research Funds for Central University (grant: DUT22YG122), and the Key Research project of ‘be Recruited and be in Command’ in Liaoning Province (2021JH1/10400050); C Liu’s lab was supported by grant...
PMC10063228
Additional information
PMC10063228
Competing interests
Senior editor, No competing interests declared.No competing interests declared.is an employee of China Resources Sanjiu Medical & Pharmaceutical.is an employee of Shanghai BeautMed Corporation.Reviewing editor,
PMC10063228
Author contributions
Conceptualization, Resources, Supervision, Funding acquisition, Project administration.Data curation, Supervision, Investigation, Project administration.Validation, Visualization, Project administration.Data curation, Software, Visualization, Methodology.Software, Methodology.Validation, Project administration.Resource...
PMC10063228
Ethics
Clinical trial registration ClinicalTrials.gov: NCT05530226.Human subjects: The protocol was approved by the Institutional Review Board (IRB) of the China Medical University (approval number: 2021PS024T). This study was registered with
PMC10063228
Additional files
PMC10063228
Baseline charateristics and rating scale for the clinical trial.
(
PMC10063228
Clinical protocol for the clinical trial study.
PMC10063228
Some detailed information for the six patients displayed in
PMC10063228
Python source code for the scoring system of knowledge graph to assess and select appropriate drug combinations.
PMC10063228
Data availability
Figure 1–3 are computational and therefore no data have been generated. In addition, Figure 4—source data 1, Figure 5—source data 1, Figure 6—source data 1, Figure 6—source data 2 and Figure 6—source data 3 contain the numerical data used to generate the figures.
PMC10063228
References
’, pain, human disorders, Astragalus’, Fructus forsythiae’, breast inflammation, human disease, Danshen’, TCM
PLASMA CELL MASTITIS, BREAST CANCER, DISEASE COURSE, DISEASE, MINOR, SYNDROME, SYNDROME, BREAST INFLAMMATION, PCM, PATHOGENESIS, PLASMA CELL MASTITIS
This study presents a valuable identification of an herbal drug combination for treating plasma cell mastitis (PCM), a breast inflammation with severe and intense clinical symptoms. The data were collected and analyzed using a solid approach in a clinical trial of 160 patients (NCT05530226). They can be used to underst...
PMC10063228
1. Introduction
sore throat, fever, inflammation, URTIs, infection, scratchy
SORE THROAT, UPPER RESPIRATORY TRACT INFECTIONS, INFLAMMATION, FLU SYMPTOMS, INFECTION, COMMON COLD, COLD, INFLUENZA
Introduction: Upper respiratory tract infections (URTIs) are caused by bacteria or viruses, with the most common causes being the common cold and influenza. The high occurrence of URTI means therapies that are effective with minimal side effects are in constant demand. Palmitoylethanolamide (PEA) is a signaling lipid p...
PMC10609976
2. Methods
cancer, HIV, or the chronic, sore throat, allergic, mood disorders, fever, malignancy, URTIs, cognitive damage, cough, scratchy
SORE THROAT, NEUROLOGICAL DISORDERS, ADVERSE EVENTS, CHRONIC ASTHMA, EVENT, RECRUITMENT, MULTIPLE SCLEROSIS, EVENTS, INFLAMMATORY RESPONSE
This was a double-blind, randomised, placebo-controlled trial conducted over 12 weeks that utilised an active group (Levagen+) and a placebo group (maltodextrin). Potential participants were provided with the participant information sheet, and following initial screening via a telehealth consultation, acceptable partic...
PMC10609976
3. Results
cough, diarrhoea, hoarseness, scratchy throat
ADVERSE EVENTS
Four hundred and twenty-six participants enrolled in the study, with 398 participants completing full trial requirements. There were 19 withdrawals in the active group and 9 in the placebo group. The Levagen+ group reported four adverse events (diarrhoea, A total of 87 participants experienced at least one URTI during ...
PMC10609976
4. Discussion
headaches, fever, infections, pain, sore throats, infection, scratchy throats
DISEASE, EVENT, INFECTIONS, LONG COVID, INFECTION, COMMON COLD, EVENTS, FLU
The aim of this study was to assess the effectiveness of PEA (Levagen+) on URTI incidence, duration, and symptom severity in otherwise healthy adults over a 12-week period. Both groups were equally matched, with no between-group differences in participant demographics (A study by Masek and colleagues (1974) supplemente...
PMC10609976
Author Contributions
A.R.: Conceptualization, methodology, software, formal analysis, resources, data curation, writing—review and editing, visualization, supervision, project administration, and funding acquisition. R.S.: Investigation, writing—original draft, and writing—review and editing. D.B.: Conceptualization, methodology, data cura...
PMC10609976
Institutional Review Board Statement
The study was conducted in accordance with the Declaration of Helsinki, and approved by the Institutional Review Board Bellberry Limited, Committee H on 6 July 2020: approval number 2020-04-340.
PMC10609976
Informed Consent Statement
Informed consent was obtained from all subjects involved in the study.
PMC10609976
Data Availability Statement
The datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request.
PMC10609976
Conflicts of Interest
The authors declare no conflict of interest.
PMC10609976
References
EVENT
SF-8 outcome measures for each question (Q1 to Q8). There were no significant differences for any question at any collection time point.SF-8 scoring based on participants’ responses to each question.Summary of participants’ baseline demographics.Values are represented as mean ± SD.Trial event outcome measures.Symptom o...
PMC10609976