Search

Vitamin D3, via its active metabolite 1,25-dihydroxyvitamin D3, plays a critical part in male and female reproduction in the rat. 1,25-Dihydroxyvitamin D3 activity is mediated by an intracellular receptor (VDR). VDR distribution in reproductive tissue has not been studied using antibodies against the receptor. We developed a polyclonal antibody against the VDR and used it to examine VDR distribution in male and female rat reproductive tissues. In rat testes, VDR epitopes were observed in seminiferous tubules, specifically in spermatogonia, Sertoli cells and spermatocytes. Spermatozoa stained faintly. Epithelial cells of the epididymis, seminal vesicles and prostate also expressed VDR epitopes. In the female rat reproductive tract, immunostaining for VDR was seen in ovarian follicles, specifically in granulosa cells. Weaker VDR immunostaining was observed in follicular thecal cells and in the ovarian stroma and germinal epithelium. Corpus luteal cells stained intensely for VDR. Epithelium of fallopian tubes and the uterus also contained VDR epitopes. Both nuclear and cytoplasmic VDR immunostaining was observed in male and female rat reproductive tissues. We conclude that the VDR is widely distributed in male and female reproductive tissues and that it is likely to mediate actions of 1,25-dihydroxyvitamin D3 in the tissues.

Male human genital tract was treated for the immunohistochemical demonstration of secretory IgA in order to verify its own possible antimicrobial properties. An intensely positive immunoreaction for sIgA in the epithelial cells of prostate and urethral glands was observed; in the same organs the presence of immunoreactive cell clusters in the subepithelial layers was noted. Immunostaining in the epithelia of deferent duct, seminal vesicle, deferential ampulla, ejaculatory duct and bulbourethral glands was absent. The findings suggest that a local immune response sIgA-mediated is present in the lower male genital tract.

Male human genitourinary tract was treated for the immunohistochemical demonstration of secretory IgA (sIgA) in order to verify its own possible antimicrobial properties. An intense immunoreactivity for sIgA was observed in the apical cells of the urethral and vesical epithelia. The ureteric epithelium, at the luminal surface, showed discontinuous areas of less dense or completely absent reaction product. A less intense immunoreactivity in the pelvic apical epithelial cells was found; an intensely positive immunoreaction for sIgA in the epithelial cells of prostate and urethral glands was also observed. The results suggest that a local immune response sIgA-mediated is present in the lower male urogenital tract.

Human immunodeficiency virus (HIV) can be transmitted through infected seminal fluid or vaginal or rectal secretions during heterosexual or homosexual intercourse. To prevent mucosal transmission and spread to the regional lymph nodes, an effective vaccine may need to stimulate immune responses at the genitourinary mucosa. In this study, we have developed a mucosal model of genital immunization in male rhesus macaques, by topical urethral immunization with recombinant simian immunodeficiency virus p27gag, expressed as a hybrid Ty virus-like particle (Ty-VLP) and covalently linked to cholera toxin B subunit. This treatment was augmented by oral immunization with the same vaccine but with added killed cholera vibrios. Polymeric secretory immunoglobulin A (sIgA) and IgG antibodies to p27 were induced in urethral secretions, urine, and seminal fluid. This raises the possibility that the antibodies may function as a primary mucosal defense barrier against SIV (HIV) infection. The regional lymph nodes which constitute the genital-associated lymphoid tissue contained p27-specific CD4+ proliferative and helper T cells for antibody synthesis by B cells, which may function as a secondary immune barrier to infection. Blood and splenic lymphocytes also showed p27-sensitized CD4+ T cells and B cells in addition to serum IgG and IgA p27-specific antibodies; this constitutes a third level of immunity against dissemination of the virus. A comparison of genito-oral with recto-oral and intramuscular routes of immunization suggests that only genito-oral immunization elicits specific sIgA and IgG antibodies in the urine, urethra, and seminal fluid. Both genito-oral and recto-oral immunizations induced T-cell and B-cell immune responses in regional lymph nodes, with preferential IgA antibody synthesis. The mucosal route of immunization may prevent not only virus transmission through the genital mucosa but also dissemination and latency of the virus in the draining lymph nodes.

Immunoglobulin A (IgA) is present at mucosal sites of the body which are exposed to the external environment. In this study we evaluated the levels of IgA and its transport protein secretory component (SC) in organs of the male reproductive tract of both intact and castrate-hormone-treated rats. Our goals were to determine whether these proteins are present in the male reproductive tract and whether sex hormones can influence the amounts of IgA and SC in selected organs. We found that in intact animals, IgA was present in the prostate, epididymis, vas deferens and testis and that SC levels in the prostate were 22-fold greater than in these same organs. Dihydrotestosterone (DHT) and/or estradiol, when administered to castrate animals, dramatically increased the levels of prostatic SC. In contrast, the levels of IgA were only minimally affected. DHT administration also resulted in a significant increase in SC found in the seminal vesicles. These studies demonstrate that IgA and SC are present in the male reproductive tract of the rat. Further, they show that androgens and estrogens act at selected sites in the male reproductive tract to play an important role in maintaining SC levels and thereby suggest that these hormones influence the movement of IgA from tissues into secretions.

The establishment of a hybridoma (1C4) producing sperm immobilizing monoclonal antibody to human seminal plasma No. 7 antigen (HSP No. 7 Ag.) and the isolation of the pure antigen by immunoaffinity chromatography bound monoclonal antibodies have been reported previously. In the present investigation, HSP No. 7 Ag. has been termed 'Ferrisplan' and its distribution in male genital organs, spermatozoa and body fluids has been studied. The amount of Ferrisplan in the body fluids was determined by radioimmunoassay. Large amounts were detected in seminal plasma and milk, trace amounts in saliva, and none in the serum and urine. The concentration of Ferrisplan was highest in the seminal plasma of azoospermic patients and gradually decreased from oligospermia to normospermia. Using an immunofluorescent method with anti-Ferrisplan monoclonal antibody, strong staining was observed on the epithelial layers of human seminal vesicles, no staining on testes and bright staining on the post-nuclear cap and mid-piece segment of spermatozoa. These results indicate that Ferrisplan is excreted mainly from the seminal vesicle and adheres to the post-nuclear cap and mid-piece of the spermatozoa as a sperm-coating antigen.

A series of monoclonal antibodies that react with human lymphocyte subsets was used in an indirect immunoperoxidase technique to study representative blocks from normal human testis, epididymis, vas deferens, prostate and seminal vesicles. Biopsies of testis, epididymis and vas obtained during surgical procedures directed at the investigation and treatment of infertile males were also studied. In all normal tissues, apart from the peripheral testis where no lymphocytes were identified, T lymphocytes were the predominant cell type (Leu 4+). These lymphocytes were largely of the suppressor/cytotoxic phenotype (Leu 2a+) and were more abundant in between the epithelial cells in the rete testis, epididymis, vas deferens, seminal vesicles and prostatic acini. Cells of the helper/inducer phenotype (Leu 3a+) were identified mainly within the interstitium of the epididymis and the prostate. B-lymphocytes (Leu 12+) were few in number and were mainly in the stroma of the prostate. In each organ the ratio of the T-cell subsets was determined and changes in this ratio were observed in epididymal and vasal biopsies from some infertile males. Finally, in testis biopsies from infertile men, suppressor/cytotoxic T-cells were demonstrated between the germinal epithelium and the fibrous tunica of the seminiferous tubules and as focal aggregates in the interstitium.

Experimental autoimmune vesiculo-prostatitis (EAVP) was transferred within inbred Wistar rats by a relatively small number of spleen cells obtained 30 days after immunization of donors with MAG extract in CFA and injected in the internal jugular vein of recipients. Specific tissue alterations identical to those present in the glands of the donors were observed 7 days after the transfer of the cells. The main alteration present was infiltration of mononuclear cells in the interstitium accompanied by fibrosis in more severe cases and flattening of epithelium in the ventral and dorsal prostate, seminal vesicle, and occasionally in coagulating gland. Nine out of ten recipients developed sexual accessory glandular lesions of various degrees. No histological alterations were observed in the absence of cell-mediated immune response and extensive damage was only observed in rats expressing two or more positive assays. The separation of the transferred cells into enriched population of T and B cells has proved that T lymphocytes are required for the production of the tissue lesions. Depletion of T lymphocytes by nylon wool separation and anti-rat thymocytes serum and complement completely abrogate their capacity to transfer the glandular alterations. Furthermore, specific antibody precursor cells on their own seem to be incapable to promote the antibody synthesis and to initiate the glandular damage. We conclude that T lymphocytes are required for the adoptive transfer of specific tissue damage observed in autoimmune vesiculo-prostatitis as well as for the development of a cell-mediated immune response to MAG antigens. Whether the same or distinct T-cell subsets are involved in these two effects is discussed.

Single cryoinjuries to the accessory reproductive organs and to the gonads of normal adult male rabbits elicit antibody responses in 35.8 and 11.9 per cent of the cases, respectively. The antibodies display agglutinating and precipitating properties against extracts of the organ that underwent the freezing injury, but do not react in the presence of extracts of other organs of the rabbit. Repeated cryoinjuries raise both the percentage of reacting animals (up to 100 per cent after four successive cryostimulations, in some cases) and the titer of the humoral responses, which, in addition, endure longer in blood than do responses to single stimuli. These humoral phenomena are the expression of immunologic reactions mounted by the host animals against "self" components, regarded, however, as "nonself" by their surveillance apparatus.

Using the indirect immunofluorescent technique, the occurrence and distribution of the low molecular weight acid-stable acrosin inhibitors from boar seminal plasma (BSAI) in the boar genital tract was studied applying specific inhibitor-directed rabbit-immunoglobulins. The acrosin inhibitors are localized in the mucosa cells of the cauda epididymis, the vas deferens, the seminal vesicles, the urethra and distinct glandular units of the prostate. The wide distribution of the acrosin inhibitors within the boar genital tract indicates a protective function towards the destructive proteolytic potential of acrosin liberated e.g. during sperm aging from desintegrating spermatozoa.

An isolated soluble antigen involved in the autoimmune response against rat male accessory glands migrated as a double band when it was submitted to analytical disc polyacrylamide gel electrophoresis and as a homogeneous molecular species in SDS electrophoresis. The purified antigen had a molecular weight of approximately 78 K by SDS electrophoresis and 80 K by gel filtration chromatography. The antigen was identified as a protein and displays androgen dependency. The purified fraction was pathogenically active in microgram doses and induced humoral and delayed-type hypersensitivity responses. Besides, lesions were observed in the target organ. The cellular infiltration was located in prostate glands according to the localization of antigen by immunofluorescence findings.

The peroxidase-antiperoxidase (PAP) technique was used to identify cytoplasmic immunoglobulins in the accessory sex glands of 15 normal bulls and 13 bulls with inflammation of the ASG. Immunoglobulin containing cells (ICC) of the types IgA, IgM, total IgG, IgG1 and IgG2 were measured and their percentages expressed. In accessory sex glands from normal bulls, IgA containing cells were the most frequent in prostate and bulbourethral glands (86.7% and 86.1%, respectively of all ICC present) whereas in the ampulla, IgG containing cells comprised 78.6% of the ICC. IgG1 and IgG2 containing cells were present in all the accessory sex glands in approximately equal numbers. Frequencies of IgM containing cells in the ampulla, prostate and bulbourethral glands were 6.3%, 4.0% and 3.7%, respectively. Although all isotypes of ICC were present in the seminal vesicle, the very low number precluded accurate quantification. In inflamed ampulla, seminal vesicle, bulbourethral gland and colliculus seminalis, IgG containing cells were the most frequent ICC with values of 66.2%, 83.0%, 69.0% and 53.5%, respectively; IgA containing cells were the second in prevalence with values of 21.5%, 10.3% 19.3% and 40.5%, respectively. The contribution of ICC to the locally protective immunoglobulins in accessory sex gland secretions is discussed.