The frequency of epitope specific naïve CD4+ T cells in humans has not been extensively examined. In this study, a systematic approach was used to examine the frequency of CD4+ T cells that recognize the Protective Antigen of Bacillus anthracis in both Anthrax Vaccine Adsorbed vaccinees and non-vaccinees with HLA-DRB1*01:01 haplotypes. Three epitopes were identified that had distinct degrees of immunodominance in subjects that had received the vaccine. Average naïve precursor frequencies of T cells specific for these different epitopes in the human repertoire ranged from 0.2 to 10 per million naïve CD4+ T cells, which is comparable to precursor frequencies observed in the murine repertoire. Frequencies of protective Antigen-specific T cells were two orders of magnitude higher in immunized subjects than in nonvaccinees. The frequencies of epitope specific memory CD4+ T cells in vaccinees were directly correlated with the frequencies of precursors in the naïve repertoire. At the level of TCR usage, at least one preferred Vβ in the naïve repertoire was present in the memory repertoire. These findings implicate naïve frequencies as a crucial factor in shaping the epitope specificity of memory CD4+ T cell responses.

The results of our in vitro study demonstrate the capacity of dexamethasone to mitigate the deleterious impact of endometriotic PF on embryo development. If confirmed in vivo, dexamethasone may prove a useful adjunct for the treatment of endometriosis-associated infertility.

STUDY QUESTION Are there phase-specific changes in the early secretory (ES) phase human tubal lavage proteome that can inform and potentially optimize IVF culture media? SUMMARY ANSWER The human tubal lavage proteome during the ES phase relative to the menstrual phase reveals substantial differential protein abundance in pathways such as glycolysis, redox homeostasis and activation of 14-3-3 zeta-mediated signaling. WHAT IS KNOWN ALREADY The Fallopian tube is uniquely suited to the development of the preimplantation embryo as it transits the tube during the ES phase of the menstrual cycle. Euploid cleavage-stage embryo arrest may reflect incomplete recapitulation of in-vivo conditions by current media formulations. STUDY DESIGN, SIZE, DURATION Proteome-wide analysis of distal tubal lavage specimens collected from 26 healthy women undergoing open microtubal anastomosis surgery from January 2013 to January 2018 was performed. Specimens were grouped by menstrual cycle phase in order to analyze phase-specific differences in protein abundance. For the murine embryo assay, single-cell embryos (N = 482) were collected from superovulated wild type C57BL/6 female mice and cultured in microdrops over 5 days for the assessment of blastocyst development. PARTICIPANTS/MATERIALS, SETTING, METHODS Human tubal lavage specimens were processed for label-free mass spectrometry. Reported menstrual cycle day was confirmed by measuring serum hormones. Key protein targets in the ES phase were validated via immunoblot. The ES phase-specific increase in 14-3-3 zeta protein was confirmed via ELISA of conditioned media obtained from primary human Fallopian tube epithelial cell culture. A murine embryo assay was performed to investigate the impact of graduated concentrations of 14-3-3 zeta on the blastocyst development rate. MAIN RESULTS AND THE ROLE OF CHANCE Comparison of the ES and menstrual phase human tubal lavage proteomes revealed 74 differentially expressed proteins with enrichment of pathways and biological processes involved in the regulation of carbohydrate metabolism, oxidative stress and cell survival. The adapter-regulator protein 14-3-3 zeta was among the most significantly increased in the ES phase. Supplementation of embryo culture media with 14-3-3 zeta at concentrations tested did not significantly improve the murine blastocyst development. LIMITATIONS, REASONS FOR CAUTION Although select associations were recapitulated in the conditioned media from sex steroid exposed primary human tubal epithelial cells, cell culture represents an in-vitro approximation. Changes to embryo culture media, such as protein supplementation, must undergo rigorous preclinical safety testing prior to adoption for human use. WIDER IMPLICATIONS OF THE FINDINGS This study represents the first description of the human Fallopian tube lavage proteome across the menstrual cycle, revealing a unique proteomic signature during the ES phase. Although supplementation of culture media with 14-3-3 zeta at appropriate concentrations showed no significant impact on the murine blastocyst development rate, other biologically plausible candidate proteins for individual or high throughput testing strategies are identified. STUDY FUNDING/COMPETING INTEREST(S) This work was funded in part by an Army Medical Department Advanced Medical Technology Initiative grant from the United States Army Medical Research and Materiel Command’s Telemedicine and Advanced Technology Research Center. There are no competing interests. TRIAL REGISTRATION NUMBER N/A

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Objectives 1) Examine the effectiveness of gene-based nutrition counseling on health-related behaviors in service members as measured by body weight, body mass index, blood glucose, lipids, 25-hydroxyvitamin D, % body fat, waist circumference, and blood pressure; 2) Evaluate the feasibility of a digital application to accurately capture diet, activity, and sleep behaviors; 3) Describe military-unique characteristics in demographics, diet, and lifestyle for Northwest Army and Southwest Air Force cohorts Methods In this prospective RCT, a baseline genomic profile built from 70 genes/80 variants following amplicon sequencing on a MiSeq platform is augmented by serum biomarkers specific to diet-related chronic disease risk (metabolic syndrome and vitamin D deficiency) for each subject. Treatment group receives gene-based nutrition counseling; control group receives evidence-based nutrition pamphlets. A digital app is used for real-time health data capture, validated by in-person RD interviews. After the initial 6 weeks, both groups use study resources independently and return for final body composition, BP, and serum biomarkers at 12 weeks. Results Preliminary findings: Northwest cohort has enrolled 26 of 75 active duty soldiers. Sample has slightly more males (54%), mean age 29.6 years, 64% married, 77% Caucasian, non-Hispanic, and 60% report some college. About 50% lost an average of 15 lbs to join the Army. Mean anthropometrics for females: BMI 27.4, 30.3% body fat, waist circ 32.2 in; for males: BMI 30.7, 28% body fat, waist circ 40 in. High variant presence noted for obesity and hypertension. Signals of abnormal biomarkers are insignificant although 60% of sample has baseline serum vitamin D less than 30 ng/mL. Conclusions This initiative has begun to advance the precision nutrition agenda in the military setting where advanced practice RNs and RDs will soon use genomic risk data to guide diet and lifestyle counseling to support personal and military readiness and optimize mission performance. Funding Sources TriService Nursing Research Program.