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A role for autoimmune regulator (AIRE) in female fertility
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Symposium Abstracts / Journal of Reproductive Immunology 86 (2010) 11–34
O6 A role for autoimmune regulator (AIRE) in female fertility S. Jasti a , B.K. Petroff b , M.G. Petroff a,∗ a
Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas, USA b Department of Internal Medicine, University of Kansas Medical Center, Kansas City, Kansas, USA The discrimination between self and foreign antigens by developing T cells is a hallmark function of the immune system, and is established in the thymus during development of central tolerance. Along with ubiquitous antigens, many antigens otherwise restricted to peripheral tissues are expressed by medullary thymic epithelial cells under the control of the transcriptional regulator AIRE (AutoImmune REgulator). Functional deletion of AIRE in mice and humans causes a breakdown of self-tolerance and organspecific autoimmune disease. Mice lacking AIRE have been reported to have defects in fertility, but whether this gene plays a role in immune tolerance to female reproductive tissues is not understood. We used AIRE-deficient mice on the Balb/c genetic background to evaluate a possible role of central tolerance in female reproduction. Mice lacking AIRE exhibited delayed puberty in comparison to wild-type controls. Only 37.5% of AIRE-deficient mice (n = 8) gave litters, whereas 100% of wild-type females (n = 6) delivered litters. Litter size in AIRE-deficient mice was also reduced in comparison to controls (4.6 pups vs. 6.8 pups, respectively). Following birth of the first litter, none of the AIRE-deficient mice delivered a second litter, whereas all control females delivered 2nd litters. To assess whether AIRE influenced ovarian follicular reserves, virgin females were sacrificed at 4, 8, 12, 16 and 20 weeks of age, and follicles were counted using histochemical techniques. While there was no clear relationship between follicular reserves and age, approximately 60% of mice exhibited ovarian inflammatory infiltrates; of these, 45% showed complete follicular loss (n = 18). In conclusion, deletion of AIRE in mice severely impaired female fertility. This may be caused, at least in part, by a loss in ovarian follicular reserves and/or associated inflammation. Supported by KUMC Research Institute Lied Pilot Grant and NIH R21HD062879. doi:10.1016/j.jri.2010.06.053 O7 Rat testicular macrophages constitutively produce interleukin-10 and have an ‘alternatively activated’ response to inflammatory stimulation in vitro W.R. Winnall a,∗ , J.A. Muir a , P. Hertzog b , M.P. Hedger a
a
Centre for Reproduction and Development, Monash Institute of Medical Research, Clayton, VIC, Australia b Centre for Innate Immunity and Infectious Diseases, Monash Institute of Medical Research, Clayton, VIC, Australia The testis is a site of immune privilege, providing protection for the developing germ cells from innate and
adaptive immune responses which can reduce fertility. Testicular macrophages (TMs) may play a role in the maintenance of testicular immune privilege, but the mechanisms by which they might do this are incompletely understood. Macrophages are classified into two general phenotypes based on their responses to stimulation; “classically activated” (M1) macrophages, which undergo strong inflammatory responses, and “alternatively activated” (M2) macrophages, which have anti-inflammatory and tissue repair properties. In order to characterise their phenotype, we have isolated rat TMs and cultured them for 2–16 h, comparing them to bone marrow-derived macrophages (BMMs) by measuring inflammatory regulator production using real-time PCR and ELISA. M1 responses were stimulated with the bacterial product lipopolysaccharide (LPS) and interferon-␥ (IFN␥). M2 responses were stimulated using the anti-inflammatory cytokine interleukin-4 (IL-4). TMs constitutively expressed mRNA for anti-inflammatory IL-10 and produced IL-10 protein, whereas BMMs produced no detectable IL-10. IL-10 mRNA was highest in TMs stimulated with LPS and IFN␥, at both 6 and 16 h. TMs had a greatly diminished response to LPS/IFN␥ at 2 h, in terms of pro-inflammatory TNF␣ and IL-1. TMs had little IL-10, TNF␣ or IL-1 response to IL-4 at 2 h, but produced the suppressor of cytokine signaling, SOCS1, suggesting a novel role for this factor in the testis. Interestingly, after 16 h of TM IL-4 treatment, there was a downregulation of IL-1 mRNA, but no significant change in IL-10 levels. TMs therefore exhibited an ‘alternatively activated’ phenotype, involving IL-10, consistent with a role in supporting testicular immune privilege. M2 macrophages contribute to fibrosis in other tissues, implicating a possible involvement of these macrophages in testicular fibrosis, which is a major cause of testicular damage in conditions such as vasectomy, cryptorchidism and infertility. doi:10.1016/j.jri.2010.06.054 O8 Microbial colonisation of follicular fluid: alterations in cytokine expression and adverse assisted reproductive technology outcomes E.S. Pelzer a,∗ , K. Cunningham a , J.J. Allan c , K. Mengersen b , J.M. Allan c , T. Launchbury c , K. Beagley a , C.L. Knox a a
Institute of Health and Biomedical Innovation, Brisbane, QLD, Australia b Mathematical Sciences, Queensland University of Technology, Brisbane, QLD, Australia c Wesley Monash IVF, Brisbane, QLD, Australia Previous studies have measured cytokine expression within follicular fluid collected at the time of transvaginal oocyte retrieval and compared the profiles with the aetiology of infertility and/or successful or unsuccessful assisted reproductive technology (ART) outcomes. Seventy-one paired follicular fluid and vaginal swab specimens collected from ART patients were cultured to detect microorganisms and then were tested for the presence of cytokines by multiplex fluorescence bead assays. Spec-
Symposium Abstracts / Journal of Reproductive Immunology 86 (2010) 11–34
O6 A role for autoimmune regulator (AIRE) in female fertility S. Jasti a , B.K. Petroff b , M.G. Petroff a,∗ a
Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas, USA b Department of Internal Medicine, University of Kansas Medical Center, Kansas City, Kansas, USA The discrimination between self and foreign antigens by developing T cells is a hallmark function of the immune system, and is established in the thymus during development of central tolerance. Along with ubiquitous antigens, many antigens otherwise restricted to peripheral tissues are expressed by medullary thymic epithelial cells under the control of the transcriptional regulator AIRE (AutoImmune REgulator). Functional deletion of AIRE in mice and humans causes a breakdown of self-tolerance and organspecific autoimmune disease. Mice lacking AIRE have been reported to have defects in fertility, but whether this gene plays a role in immune tolerance to female reproductive tissues is not understood. We used AIRE-deficient mice on the Balb/c genetic background to evaluate a possible role of central tolerance in female reproduction. Mice lacking AIRE exhibited delayed puberty in comparison to wild-type controls. Only 37.5% of AIRE-deficient mice (n = 8) gave litters, whereas 100% of wild-type females (n = 6) delivered litters. Litter size in AIRE-deficient mice was also reduced in comparison to controls (4.6 pups vs. 6.8 pups, respectively). Following birth of the first litter, none of the AIRE-deficient mice delivered a second litter, whereas all control females delivered 2nd litters. To assess whether AIRE influenced ovarian follicular reserves, virgin females were sacrificed at 4, 8, 12, 16 and 20 weeks of age, and follicles were counted using histochemical techniques. While there was no clear relationship between follicular reserves and age, approximately 60% of mice exhibited ovarian inflammatory infiltrates; of these, 45% showed complete follicular loss (n = 18). In conclusion, deletion of AIRE in mice severely impaired female fertility. This may be caused, at least in part, by a loss in ovarian follicular reserves and/or associated inflammation. Supported by KUMC Research Institute Lied Pilot Grant and NIH R21HD062879. doi:10.1016/j.jri.2010.06.053 O7 Rat testicular macrophages constitutively produce interleukin-10 and have an ‘alternatively activated’ response to inflammatory stimulation in vitro W.R. Winnall a,∗ , J.A. Muir a , P. Hertzog b , M.P. Hedger a
a
Centre for Reproduction and Development, Monash Institute of Medical Research, Clayton, VIC, Australia b Centre for Innate Immunity and Infectious Diseases, Monash Institute of Medical Research, Clayton, VIC, Australia The testis is a site of immune privilege, providing protection for the developing germ cells from innate and
adaptive immune responses which can reduce fertility. Testicular macrophages (TMs) may play a role in the maintenance of testicular immune privilege, but the mechanisms by which they might do this are incompletely understood. Macrophages are classified into two general phenotypes based on their responses to stimulation; “classically activated” (M1) macrophages, which undergo strong inflammatory responses, and “alternatively activated” (M2) macrophages, which have anti-inflammatory and tissue repair properties. In order to characterise their phenotype, we have isolated rat TMs and cultured them for 2–16 h, comparing them to bone marrow-derived macrophages (BMMs) by measuring inflammatory regulator production using real-time PCR and ELISA. M1 responses were stimulated with the bacterial product lipopolysaccharide (LPS) and interferon-␥ (IFN␥). M2 responses were stimulated using the anti-inflammatory cytokine interleukin-4 (IL-4). TMs constitutively expressed mRNA for anti-inflammatory IL-10 and produced IL-10 protein, whereas BMMs produced no detectable IL-10. IL-10 mRNA was highest in TMs stimulated with LPS and IFN␥, at both 6 and 16 h. TMs had a greatly diminished response to LPS/IFN␥ at 2 h, in terms of pro-inflammatory TNF␣ and IL-1. TMs had little IL-10, TNF␣ or IL-1 response to IL-4 at 2 h, but produced the suppressor of cytokine signaling, SOCS1, suggesting a novel role for this factor in the testis. Interestingly, after 16 h of TM IL-4 treatment, there was a downregulation of IL-1 mRNA, but no significant change in IL-10 levels. TMs therefore exhibited an ‘alternatively activated’ phenotype, involving IL-10, consistent with a role in supporting testicular immune privilege. M2 macrophages contribute to fibrosis in other tissues, implicating a possible involvement of these macrophages in testicular fibrosis, which is a major cause of testicular damage in conditions such as vasectomy, cryptorchidism and infertility. doi:10.1016/j.jri.2010.06.054 O8 Microbial colonisation of follicular fluid: alterations in cytokine expression and adverse assisted reproductive technology outcomes E.S. Pelzer a,∗ , K. Cunningham a , J.J. Allan c , K. Mengersen b , J.M. Allan c , T. Launchbury c , K. Beagley a , C.L. Knox a a
Institute of Health and Biomedical Innovation, Brisbane, QLD, Australia b Mathematical Sciences, Queensland University of Technology, Brisbane, QLD, Australia c Wesley Monash IVF, Brisbane, QLD, Australia Previous studies have measured cytokine expression within follicular fluid collected at the time of transvaginal oocyte retrieval and compared the profiles with the aetiology of infertility and/or successful or unsuccessful assisted reproductive technology (ART) outcomes. Seventy-one paired follicular fluid and vaginal swab specimens collected from ART patients were cultured to detect microorganisms and then were tested for the presence of cytokines by multiplex fluorescence bead assays. Spec-