FA 5 Progress Report WV-INBRE - Joan C. Edwards School of ...

Program Director/Principal Investigator (Last, First, Middle): Rankin, Gary O 42during stress conditions of the model.Aim #2: To assess the effect of cold-stress on Chlamydia-induced infertility in the mouse modelThis aim was undertaken to examine the effect of stress on induction of infertility in a mouse model.Cold-stress was induced by immersing mice in cold water for 5 minutes daily for 24 days.Stressed and non-stressed groups of mice which received Depo-Provera on day 17 were infectedintravaginally with 107 IFU of C. trachomatis in a volume of 30 μl of phosphate-buffered saline(PBS) while under anesthesia. Infected mice were monitored along with cervico-vaginal swabbingat 3-day intervals during primary and secondary course of infections. After recovery fromChlamydia genital infection, all mice were mated with proven breeder male mice and monitored for19-21 days. Our results showed that exposure of mice to 24-day stress resulted in a greaterintensity of Chlamydia genital infection. The percentage of pregnant mice and the number of pupswas calculated. We noted that 10 out of 19 (52%) stressed mice were infertile compared to 2 of 19(10%) non-stress mice. Cold-water induced stress also resulted in a decreased mean number ofembryos (4 to 6) in stressed mice compared to mean number of embryos (7 to 9) in non-stressedmice. These results suggest that stress may increase complications of immunopathogenesisresulting from C. trachomatis genital infection and infertility. Further experimentation is underway todetermine the effect and mechanism of stress in modulation of fertility in mice. (Abstract submittedto the General Meeting of American Society for Microbiology, San Francisco, CA, June 2012).Aim #3: To determine the effect of cold-induced stress on the histopathological changes of thegenital tract during Chlamydia infection in the mouse modelComparative analysis of the immune responses and histopathologic analysis in primary infectionversus secondary Chlamydia genital infection in stressed and non-stressed mice was initiated atthe Pathology Laboratory Service in the Medical School of Marshall University. Preliminary data ofthe cervical regions of mice representing stressed and non-stressed mice showed no significantdifference of hispathology. However, gross examination revealed that stressed and infected micehad fluid-filled and heavily extended uterus unlike non-stressed infected mice. Evaluation of eachanatomical site of uterine horn and oviduct for acute inflammation, chronic (lymphocytes) andplasma cells and fibrosis for chronic inflammation was initiated before the relocation of thePathology Laboratory Service in the Medical School of Marshall University.Aim #4: To elucidate the mechanism(s) by which stress increases susceptibility to Chlamydiatrachomatis genital infection in the modelOur study showed that increased susceptibility to Chlamydia infection is accompanied by multipleimmunosuppressive effects may be due to the inhibition of recruitment of T cells into genital tract,suppression of proinflammatory cytokines, chemokines by stress hormones such as noradrenaline(norepinephrine). The primary focus of this aim is to investigate how the stress hormone,norepinephrine (noradrenaline) impacts immune-mediated protection against Chlamydia trachomatisinfection. We will test whether ADR antagonists; propranolol (non-selective), or atenolol (selective)blockade affects Chlamydia clearance during genital infection. We speculate that administration ofantagonists will reduce the Chlamydia burden in genital tract of stressed mice to levels similar tonon-stressed infected mice. An alternative approach, using a noradrenaline receptor knockoutmouse, is under consideration.PROTECTION AGAINST RESEARCH RISKSN 1. Will human subjects be involved next year?Y 2. Will vertebrate animals be used next year?If yes, provide the date of Institutional Animal Care and Use Committee (IACUC) approval and enclosewith transmittal.02/09/2011PHS 2590 (Rev. 06/09)Continuation Format Page