Humoral immune response to membrane components of Chlamydia trachomatis and expression of human 60 kDa heat shock protein in follicular fluid of in-vitro fertilization patients

Abstract

Recent evidence suggests that Chlamydia trachomatis can persist in the female upper genital tract in an unculturable state. Since unsuspected C. trachomatis infection has been associated with adverse in-vitro fertilization (IVF) outcome we sought to detect further evidence of C. trachomatis in the genital tracts of women undergoing IVF. The prevalence and distribution of antibodies to the major structural proteins of C. trachomatis in paired follicular fluid and sera of women undergoing IVF were examined. Sera and follicular fluid samples from 149 women were assayed for immunoglobulin (Ig)G and IgA antibodies to two C. trachomatis antigens, the major outer membrane protein (MOMP) and a recombinant lipopolysaccharide (rLPS) fragment. Additionally, the expression of human 60 kDa heat shock protein (hsp 60) in follicular fluid was determined. All cervical and follicular fluid samples were negative for C. trachomatis by polymerase chain reaction, ligase chain reaction and DNA probe. Sera from 60% of the subjects were positive for antichlamydial rLPS IgG; 36% were positive for anti-MOMP IgG. Similarly, rLPS-directed and MOMP-directed IgA were detected in sera of 34 and 14% of the subjects respectively. IgG antibodies to MOMP and rLPS were detected in 42 and 41% of the follicular fluid examined respectively. Anti-MOMP IgA was identified in 8.7% of the follicular fluid while 27.5% were positive for anti-rLPS IgA. Human hsp 60 expression was documented in 11.6% of the follicular fluid tested. IgA antibodies to both MOMP (P = 0.03) and rLPS (P = 0.02) in follicular fluid were associated with a failure to become pregnant after embryo transfer. IgG antibodies in sera and follicular fluid and IgA antibodies in sera were unrelated to IVF outcome. Similarly only anti-MOMP IgA (P = 0.02) and anti-rLPS IgA (P = 0.04) in follicular fluid were correlated with human hsp 60 expression in follicular fluid. The unique association between IgA antibodies to two chlamydial antigens in follicular fluid and both hsp 60 expression and IVF failure provides further support for the possibility that a persistent upper genital tract chlamydial infection contributes to IVF failure in some women.

Immunopathogenic consequences of Chlamydia trachomatis 60 kDa heat shock protein expression in the female reproductive tract

“…In human disease, elevated antibody responses to the chlamydial hsp60 have been strongly correlated with the presence of pelvic inflammatory disease, ectopic pregnancy, scarring trachoma, and tubal infertility (Neuer et al 1997;Peeling et al 1998;Tiitinen et al 2006;Jakus et al 2008;Mascellino et al 2008;Ondondo et al 2009). Similarly, cell-mediated immune reactivity to the chlamydial hsp60 is also demonstrable in women with salpingitis or tubal infertility (Witkin et al 1993;Witkin et al 1994a, b;Kinnunen et al 2000;Tiitinen et al 2006).…”

Human granulosa–luteal cells initiate an innate immune response to pathogen-associated molecules

“…Retrospective analysis of patients undergoing fertility treatment suggest that increased endometrial LPS reduces IVF success (Kamiyama et al 2004). More recently, it has been demonstrated that bacteria can be isolated from the follicular fluid of women undergoing infertility treatment, with the extent of bacterial colonization of the follicle being correlated with the success of fertility treatment (Neuer et al 1997, Pelzer et al 2013.…”

Hyaluronan in follicular fluid and embryo implantation following in vitro fertilization and embryo transfer

“…There was no difference in the level of estradiol among all investigated groups. Transvaginal oocyte retrieval and follicular fluid collection was initiated 34-36 h following ovulation induction, as reported previously [11]. After oocyte retrieval, the cumulus-oocyte complexes were collected in Gamete medium (Vitrolife, Goteberg, Sweden) and incubated at 37 o C with 5% CO 2 .…”