Effects on the local immunity in the testis by exposure to di-(2-ethylhexyl) phthalate (DEHP) in mice.

Exposure to di-(2-ethylhexyl) phthalate (DEHP) has been reported to induce spermatogenic disturbance through oxidant stress and affect the immune system as an adjuvant. However, the effect of DEHP on the testicular immune microenvironment has not yet been investigated. In the present study, we examined the testicular immune microenvironment after exposure to doses of DEHP, previously identified as no-observed-adverse-effect levels. Adult male mice were administered food containing 0%, 0.01% or 0.1% DEHP and then testes were analyzed. The results showed that a slight but significant spermatogenic disturbance appeared in the 0.1% DEHP group but not in the 0.01% DEHP group at 8 weeks. It was also demonstrated that lymphocytes and F4/80- and MHC class II- positive cells were significantly increased with the elevation of IL-10 and IFN-γ mRNA expressions in the testes of not only the 0.1% DEHP group but also the 0.01% DEHP group at 8 weeks. Histochemical analyses involving horseradish peroxidase (HRP) as a tracer showed that a little blood-borne HRP had infiltrated into the lumen of a few seminiferous tubules beyond the blood-testis-barrier in both the 0.1% and 0.01% DEHP groups at 8 weeks. This indicates that a dose of DEHP that has little effects on spermatogenesis can change the testicular immune microenvironment with functional damage of the blood-testis barrier.

Postinflammation stage of autoimmune orchitis induced by immunization with syngeneic testicular germ cells alone in mice.

We previously established an immunological infertility model, experimental autoimmune orchitis (EAO), which can be induced by two subcutaneous injections of viable syngeneic testicular germ cells on days 0 and 14 in mice without using any adjuvant. In this EAO model, CD4+ T-cell-dependent lymphocytic infiltration and immune deposits were found with spermatogenic disturbance on day 120. However, the late stage of EAO (= postactive inflammation stage on day 365) has not yet been investigated. Therefore, we investigated the histopathological characteristics of the late stage. The results revealed that the lymphocytic infiltration finally resolved; however, the seminiferous epithelium persistently showed maturation arrest and the Sertoli cell-only feature. In the seminiferous tubules showing maturation arrest, both proliferation and apoptosis of germ cells had occurred simultaneously. It was also noted that there were deposits of immunoglobulin G and the third component of complement on the thickened basement membrane of seminiferous tubules in the late stage of EAO. These results indicate that histopathology after active inflammation in EAO comprises persistent damage to the seminiferous epithelium and may resemble the histopathology of "idiopathic disturbance of spermatogenesis" in man.

Intratesticular expression of mRNAs of both interferon γ and tumor necrosis factor α is significantly increased in experimental autoimmune orchitis in mice.

Experimental autoimmune orchitis (EAO) is one of the models of immunological male infertility. Murine EAO is CD4+T cell-dependent and classically induced by immunization with a testicular homogenate and adjuvants. We previously established that immunization with viable syngeneic testicular germ cells (TGC) can also induce murine EAO with no use of any adjuvant. Analyses of this EAO model have already revealed that cultured spleen cells of immunized mice secreted interferon (IFN)-γ and that treatment of the immunized mice with anti-IFN-γ monoclonal antibodies significantly suppressed the EAO. It is known that both IFN-γ and tumor necrosis factor (TNF)-α are representative cytokines of Th1 cells and exhibit local toxicity toward the seminiferous epithelium in vivo. However, changes in these two cytokines in EAO-affected testes have not yet been investigated. Therefore, in the present study, we investigated the expression of intratesticular IFN-γ and TNF- α mRNAs in TGC-induced EAO using real-time RT-PCR. The results demonstrated that the intratesticular mRNAs for both IFN-γ and TNF-α significantly increased, while other cytokines such as IL-1α, IL-1ß, IL-6 and TGF-ß did not show dramatic changes in the immunized mice. These results suggest that secretion of significant amounts of IFN-γ and TNF-α in situ contributes to the spermatogenic disturbance in EAO.

Histological development of human testicular cords from 70 to 90 days of gestation.

The development of the testicular cord structure was investigated in 4 human fetuses between 70 and 90 days of gestation, in which the testicular cords are differentiating into the seminiferous tubules. Histological examinations were performed using stains with haematoxylin-eosin (HE), Masson's trichrome (MT), periodic acid schiff (PAS), anti-proliferating cell nuclear antigen (PCNA) monoclonal antibodies, and TUNEL methods. It was found that the testicular cords structures were indefinitely observed in HE-stained sections of four fetuses. However, the basement membranes of the testicular cord were clearly stained with MT, showing the tubular structure. Furthermore, cells in the testicular cords were positive with PAS, but the interstitial tissues outside the testicular cords were negative. PCNA-positive cells were detected not only inside but also outside the testicular cords, however, TUNEL positive cells are not detected throughout all testicular tissues.

Caput epididymitis but not orchitis was induced by vasectomy in a murine model of experimental autoimmune orchitis.

Immunization of mice with viable syngeneic testicular germ cells (TGC) alone can induce autoimmune responses against autoantigens of both round and elongating spermatids, resulting in the development of experimental autoimmune orchitis (EAO). Histological lesions in this EAO model without an adjuvant are characterized by lymphocytic infiltration into the testes, spermatogenic disturbance, and a complete lack of epididymitis. In this study, we investigated the effects of vasectomy (Vx) on TGC-induced EAO expecting that Vx augments the severity of testicular inflammation in A/J mice. The results showed that mice receiving Vx alone exhibited no significant inflammatory cell response in either the testes or epididymides, and mice receiving shamVx+TGC immunization had EAO with no epididymitis. In sharp contrast, no EAO was found in the testes of any mice receiving Vx+TGC immunization. Instead, caput epididymitis involving CD4+T cells, CD8+T cells, B cells, and macrophages were induced in them with striking elevation of the tissue levels of both IL6 and IL10 mRNA. Furthermore, serum autoantibodies induced by shamVx+TGC immunization were reactive with both round (immature) and elongating (mature) spermatids; however, those induced by Vx+TGC immunization were specific to acrosomes of mature spermatids and spermatozoa. These unexpected results indicate that Vx may induce the mode by which autoreactive lymphocytes gain access to TGC autoantigens in the epididymides, leading to autoimmune responses against the autoantigens of mature rather than immature spermatids.

Developmental ontogeny of autoantigens associated with localized autoimmunity in murine testis and epididymis.

Experimental autoimmune orchitis (EAO) with experimental autoimmune epididymitis (EAE) can be induced in mice by immunization with testicular antigens emulsified in adjuvants. On immunization with syngeneic testicular germ cells (TGC) alone, EAO with no EAE is induced in mice. Recently, we found that EAE with no EAO can be induced in vasectomized mice by immunization with TGC. In the present study, we investigated the appearance of autoantigens relevant to EAO and EAE by reacting each immune serum sample with testes and epididymides extracts from normal mice of various ages by immunoblotting. The results showed that the antisera obtained from mice with EAO lesions specifically defined testicular antigens with molecular weights of 15 kDa, 40 kDa, 75 kDa and >200 kDa from 4 weeks of age, but the antisera obtained from mice with EAE strongly defined both testicular and epididymal antigens of 25 kDa from 5 and 8 weeks of ages, respectively. These results suggest that vasectomy changes the target autoantigens in TGC-induced autoimmunity.

Right gonadal arteries passing dorsally to the inferior vena cava: embryological hypotheses.

The right gonadal artery (RGA) usually arises from the anterior wall of the abdominal aorta below the level of the renal arteries and veins, passes ventrally to the inferior vena cava (IVC), and then runs obliquely downward to reach the pelvic cavity. In this study, we observed 59 Japanese cadavers and found that in eight of them (13.6%), the RGA's passed dorsally to the IVC. Together with the various courses of RGAs reported in the literature, we here divided the courses into four subtypes, due to the relationship to the left renal vein and the IVC, of superior, middle and lower types (L1 = ventrally, L2 = dorsal of IVC). Superior type was seen in 3.6%. Lower types were fount to in the most cases (L1: 83.5%, L2: 13.6%). The middle type was not realized. The classification of this variations make it easy to describe the RGA's passing below the level of the left renal vein. RGA can pass either ventrally or dorsally to the IVC. RGAs passing at or above the level of the left renal vein run dorsally but not ventrally to the IVC. The variations in the origin, course and branching of RGAs are attributed to development of the IVC.