Multifocal damage of the testis induced in rats by passive transfer of antibodies prepared against non-collagenous fraction of basement membrane.

Multifocal damage of the testis was induced in 70% of the rats injected with an antiserum against a non-collagenous fraction (D-STBM) obtained from a preparation enriched in basement membranes of seminiferous tubules. The damaged areas were characterized by perivascular and peritubular cell infiltrates, changes in the walls of small vessels and seminiferous tubules, and sloughing of the germinal epithelium. By electron microscopy, the most frequent changes observed in basement membrane of the seminiferous tubules were folding, focal thickening, and delamination. By immunofluorescence, discontinuous linear deposits of rabbit IgG were observed along the walls of the seminiferous tubules. In the same localization, faint immunofluorescence showing the presence of rat IgG was also detected. The same pattern was obtained when rabbit and rat IgG eluted from the testes of these rats were layered on sections of normal rat testis. Moreover, by immunoelectron microscopy, discontinuous deposits of rabbit IgG were detected along the basement membranes of the seminiferous tubules. Neither C3 deposits nor changes in the serum CH 50 were observed. By leucocyte migration inhibition reaction (LMIR) a cellular immune response to basement membrane antigens was detected. In the control group, 12% of the rats injected with normal rabbit serum presented mild interstitial cell infiltrates and occasional sloughing of the germinal epithelium. Neither deposits of rat IgG or rabbit IgG nor a cellular immune response were observed.

[Experimental autoimmune orchitis]. / Orquitis autoinmune experimental.

Experimental autoimmune orchitis (EAO) has been extensively studied in spite of which its pathogenic mechanisms are still poorly understood. It has been mostly induced in guinea pigs using spermatozoa or a testicular homogenate plus adjuvants. Initially, the aim of our work was to establish if non-spermatic antigens, such as extracellular components of the walls of seminiferous tubules, were able to induce an autoimmune orchitis. For this purpose, we obtained from rat testes: a) a preparation rich in basement membranes of seminiferous tubules (STBM) and b) a soluble fraction of STBM, non-related to collagen (D-STBM), presenting common antigenic determinants with laminin, the main non-collagen glycoprotein of basement membranes. Fifty per cent of rats immunized with STBM, D-STBM or a murine laminin, developed a multifocal and moderate damage of the testes characterized by mild interstitial cell infiltrates, alterations of the basement membranes of seminiferous tubules and Sertoli cells, sloughing of the germinal epithelium and tubular atrophy. Circulating antibodies and a specific cellular immune response were also detected. Moreover, rats passively injected with an heterologous anti-D-STBM serum developed a similar testicular lesion and showed Ig deposits on the basement membranes of seminiferous tubules. In relation to the pathogenic mechanisms of EAO, we studied the variations of T and B cell populations, at the immunization draining lymph nodes, during the development of orchitis. A severe EAO was induced in Wistar rats by immunization with an homologous testes homogenate plus adjuvants.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation and immunological reactivity of soluble fractions from rat seminiferous tubule basement membrane.

A preparation rich in basement membranes isolated from rat testes (STBM) was exposed to pepsin, collagenase, trypsin, and pronase to obtain soluble fractions. The immunological reactivity of these fractions was studied by gel immunodiffusion or by passive hemagglutination tests against an anti-STBM serum. All fractions reacted with the antiserum, but the highest titer was detected when the antiserum was reacted with a fraction that contained only traces of hydroxyproline (fraction 1), whereas low titers were obtained with collagen or collagen fragments isolated from STBM. Antibodies in the anti-STBM serum were mainly directed to the glycoproteins of STBM not related to collagen. Fraction 1, obtained by subsequent collagenase and trypsin digestion of STBM and purification by Sephadex G-200, was a high molecular weight glycoprotein that was free of half-cystine and methionine, had only traces of hydroxyproline, and contained 7.2% neutral sugars, 0.26% sialic acid, and 8.7 residues of glucosamine per 1000 residues of amino acids.

Experimental orchitis induced in rats by passive transfer of an antiserum to seminiferous tubule basement membrane.

A multifocal damage of the testis was obtained when rats were injected intravenously or under the tunica albuginea of the testis with a rabbit antiseminiferous tubule basement membrane serum. The damage was characterized by foci of perivascular and peritubular infiltrates of mononuclear round cells, infolding, thickening, and rupture of the seminiferous tubular wall and different degrees of injury of the germinal epithelium such as, cell disorganization, cell sloughing, and atrophy. Delamination and thickening of seminiferous tubule basement membrane and vacuolization of the Sertoli cell cytoplasm was often observed by electron microscopy. A linear deposit of rabbit gamma-globulin was detected by immunohistochemical techniques along the basement membranes of the seminiferous tubules and vessels. Testicular damage was not detected in rats injected with normal rabbit serum, used as control. In the kidneys of rats injected intravenously with the immune serum, a deposit of rabbit gamma-globulin was detected along glomerular basement membrane. Focal areas of mononuclear cell infiltrates, hypercellularity of glomeruli and thickening of glomerular capillary walls and Bowman's capsule were also observed.

Partial chemical and immunological characterization of pepsin-solubilized collagen from the tunica albuginea of rat testis.

A pepsin-solubilized collagen was isolated from the tunica albuginea of rat testis. Intrinsic viscosity, amino acid and carbohydrate composition were similar to data reported for other soluble collagens. An antiserlm to the pepsin-solubilized collagen was obtained and by indirect immunofluorescence an antigen-antibody reaction was observed in the collagen fibers of the tunica albuginea and in the intertubular collagen fibers of rat testis. By serological and immunofluorescence techniques, cross-reactions with other collagens indicates a lack of organ specificity. A strong cross-reaction between this antiserum and basement membranes from different organs of the rat was also found.

Orquitis autoinmune experimental / Experimental autoimmune orchitis

A pesar de los numerosos trabajos realizados en el modelo de orquitis autoinmune experimental (OAE) aún existen varias incógnitas acerca de su patogenia. Clásicamente, la OAE se indujo en cobayos utilizando como antígenos, espermatozoides u homogenado testicular. Nuestro primer interés fue determinar si antígenos no espermáticos como los componentes extracelulares de la pared del túbulo seminífero, utilizados como inmunógenos, eran capaces de inducir un cuadro autoinmune testicular. Obtuvimos, a partir del testículo de la rataÑ a) una preparación rica en membranas basales del túbulos seminífero (STBM) y b) a partir del STBM, una fracción no relacionada al componente colágeno (D-STBM) que demostró poseer determinantes antigénicos comunes con la laminina, principal glicoproteína no colágena de las membranas basales. El 50% de las ratas inmunizadas con STBM, D-STBM o laminina, desarrolaron una lesión testicular moderada, multifocal, caracterizada por leves infiltrados intersticiales, alteraciones de las membranas basales del túbulo seminífero y de la célula de Sertoli, descamación del epitelio germinal y atrofia tubular. También se detectaron anticuerpos circulantes y una respuesta de inmunidad celular específica. A su vez, ratas inyectadas con un suero heterólogo anti-D-STBM desarrollaron una lesión similar. Por otra parte, se indujo una orquitis severa inmunizando ratas Wistar con un hmogenado testicular homólogo y adyuvantes y se estudiaron las poblaciones linfáticas de los ganglios...