Kenneth S. K. Tung

Adoptive transfer of murine autoimmune orchitis to naive recipients with immune lymphocytes

A protocol was developed for reproducibly transferring experimental autoimmune orchitis (EAO) to naive recipient mice. Cell donors were (C57BL/6 x A/J)F1 mice immunized about 14 days earlier with mouse testicular homogenate with Freunds adjuvant and an extract of Bordetella pertussis. Lymphocytes from lymph nodes and spleens were equally capable of transferring disease. As few as 5 X 10(6) cells were able to transfer EAO, which began on Day 5-7 after transfer. Infiltrate of lymphocytes and macrophages in the region of the rete testis and straight tubules was the most reproducible early lesion, suggesting that this is the initial site of T cell-antigen interaction. It was not necessary to use both Mycobacteria and B. pertussis adjuvants in donor immunization to achieve transfer of EAO. Disease transfer was antigen specific since only cells from donors immunized with TH could transfer disease. In vitro stimulation of the cells with testicular antigens and/or concanavalin A was a prerequisite to successful transfer of EAO, which was dependent on the presence of L3T4+ T cells since depletion of these cells greatly diminished EAO in recipients and the lymphocyte proliferation response to testicular antigens. Disease did not depend on an antibody response by the recipients. The results imply that effector cells, once generated by immunization and fully activated or selected by in vitro stimulation, can home to specific locations in the testis, locate relevant autoantigens, and cause disease.

Experimental allergic orchitis in mice

Inbred strains of mice were studied for their susceptibility to the induction of experimental allergic orchitis after sensitization with mouse testicular homogenate in complete Freunds adjuvant accompanied by injections of extract from Bordetella pertussis. Susceptibility to autoimmune orchitis was found to be linked to the major histocompatibility complex in BALB/c and C57BL/10 mice and mapped to genes encoded within the H-2Ddregion. In five of six groups of bidirectional (susceptible × resistant) F1 hybrids, H-2Dd-linked susceptibility was inherited as a dominant autosomal trait. However, in (BALB/cByJ × DBA/2J)F1 and (DBA/2J × BALB/cByJ)F1 hybrids, dominant autosomal resistance to the induction of autoimmune orchitis was observed. Backcross analysis between the resistant F1 hybrid and the susceptible BALB/cByJ parent suggests that a single independently segregating DBA/2J locus is capable of negating H-2Dd-linked susceptibility, and controls resistance to the induction of autoimmune orchitis.

Murine Sertoli cells: major histocompatibility antigens and glycoconjugates

Sertoli cells, cultured from testes of 2-3-week-old Balb/c mice, contain tripartite nucleoli, exhibit phagocytic function, and have the typical morphologic appearance of Sertoli cells by light microscopy, transmission and scanning electron microscopy. Fluorescence-activated cell sorter analysis indicated the presence on mouse. Sertoli cells of H-2 but not Ia antigens. Alpha-D-mannose and N-acetyl-D-glucosamine determinants are detected on Sertoli cell surface by inhibition of lectin binding using appropriate sugars. Interaction of Sertoli cells with concanavalin A (Con A) or wheatgerm agglutinin (WGA) results in rapid patching of the labeled lectin, which become internalized as perinuclear vesicles. These changes are accompanied by rounding of the Sertoli cell, mimicking cellular changes known to occur when fat Sertoli cells are stimulated in vitro by FSH or cyclic AMP. Thus, Sertoli cells have surface alloantigens that permit them to serve as target to cytotoxic T lymphocytes, but not as antigen presenting cells.

Reduced T-lymphocyte subsets in systemic lupus erythematosus: Effects of immune complexes and lymphocytotoxic antibodies

Abstract Patients with active systemic lupus erythematosus (SLE) show significant reductions of T cells with receptors for the Fc portion of IgG, Tγ cells, which have been found to suppress in vitro B-cell responses to pokeweed mitogen. T cells with receptors for the Fc portion of IgM, Tμ cells, which show both helper and suppressor functions, are also reduced in SLE. Levels of both Tγ and Tμ cells in patients with inactive disease are intermediate between those of patients with active SLE and normal individuals. In a majority of individual patients, levels of Tγ cells were found to fluctuate with disease activity. SLE patients with reduced levels of Tγ cells showed high levels of circulating immune complexes, although a uniform inverse correlation was not detected. The pathophysiologic mechanisms in the observed decrease of T-cell subsets in SLE patients were studied by determination of the effect of aggregated IgG (an in vitro model of immune complexes) and lymphocytotoxic antibodies on T-cell subsets from normal individuals. Addition of aggregated IgG led to irreversible reduction of Tγ cells with no effect on Tμ cells. By contrast, both types of T-cell subsets were reversibly decreased by SLE sera with lymphocytotoxic antibody activity. Thus the observed decrease in T-cell subsets appears to be secondary to both immune complexes and lymphocytotoxic antibodies in SLE patients, and is reversible with remission of disease activity. Analysis of T-cell subpopulations in SLE appears to reflect other laboratory parameters associated with disease activity in these patients.

Experimental allergic orchitis in mice: III. Differential susceptibility and resistance among BALB/c sublines

Significant differences in susceptibility to the induction of experimental allergic orchitis (EAO) were found to exist among the various substrains of BALB/c mice. Substrains which were susceptible to disease induction include BALB/cKa, BALB/cByJ, BALB/cWtJ, BALB/cCmcCr, BALB/cCrgl, BALB/cHeA, BALB/cAnNCr, BALB/cPt, BALB/cWehiUnm, BALB/cOrnl and BALB/cArg whereas BALB/cJ and BALB/cSki BoyPt were resistant to EAO. Aspermatogenesis and autoimmune vasitis, two additional lesions characteristic of murine EAO, were found to correlate with susceptibility to autoimmune orchitis. Susceptibility and/or disease resistance did not segregate within any of the evolutionary branches of the BALB/c genealogic tree nor did it correlate with any of the previously described allelic differences distinguishing BALB/c substrains. Therefore, differential disease susceptibility may represent the manifestation of mutational and/or retroviral induced genotypic differences in the regulatory genes controlling testicular autoimmunity.

Experimental allergic orchitis in mice: IV. Preliminary characterization of the major murine testis specific aspermatogenic autoantigen(s).

Active experimental allergic orchitis (EAO), characterized by inflammation of the testes (autoimmune orchitis), aspermatogenesis, epididymitis and vasitis was induced in mice using a panel of tissue antigens as immunogens. Immunization with allogeneic murine tissue homogenates emulsified in complete Freunds adjuvant (CFA) accompanied by the injection of pertussigen revealed that only adult murine testicular and epididymal homogenates are capable of eliciting murine EAO. All other tissue antigens studied including prepubertal mouse and epididymal homogenates failed to elicit significant disease. Immunization with xenogeneic testicular antigens also failed to elicit significant disease indicating that the major murine aspermatogenic autoantigen(s) is also highly species specific. Sensitization with allogeneic mouse testicular homogenates (MTH) from different disease resistant strains was for the most part no less potent in inducing significant disease than was immunization with mouse testicular homogenates from disease susceptible strains. However, testicular homogenates from NZB/B1NJUnm and MRL/MpJ-/+Unm mice were significantly less potent at inducing autoimmune epididymitis as compared to other strains, indicating possible interstrain differences in the immunogenicity of the aspermatogenic autoantigen(s) relevant to eliciting epididymitis. Attempts at solubilization and purification of the major murine aspermatogenic autoantigen(s) utilizing techniques employed for the purification of aspermatogenic autoantigens such as AP3 from guinea pig (GP) testes were unsuccessful. Additional extraction procedures resulted in solubilization of the relevant autoantigen(s) only after reduction in the presence of 6 M guanidine hydrochloride. These data suggest that: (1) there may be a much more limited number of aspermatogenic autoantigens in murine testes as compared to GP testes; (2) the disease inducing determinant(s) may be expressed as either a sequential antigenic determinant(s) or as an antigenic determinant(s) in the carbohydrate portion of a glycoprotein or glycopeptide; and (3) the disease inducing autoantigen(s) may be present in situ in a highly insoluble form requiring active processing within the target organ in order to generate soluble antigen capable of being seen by immune reactants.

Lupus nephritis in a pediatric renal transplant recipient

A case of aggressive lupus nephritis in a pediatric renal transplant patient is described. She initially presented with end-stage glomerulonephritis for which an underlying etiology could not be determined. Ten months after cadaveric renal transplantation, systemic lupus erythematosus was diagnosed, when she developed diffuse proliferative glomerulonephritis in association with antinuclear antibody, anti-double-stranded DNA antibody and extrarenal manifestations of lupus. It is plausible that she developed recurrent rather thande novo lupus nephritis following transplantation. Reactivation of lupus nephritis in a renal transplant is unusual in adults, and is previously unreported in children.

A useful technique applicable to the immunofluorescence test

A simple techinque for carrying out the immunofluorescence procedure is described that eliminates the problem of drying of antiserum and reduces cost per test.

Transfer of susceptibility to experimental autoimmune orchitis from responder to non-responder substrains of BALB/c mice.

Substrains of BALB/c mice differ in their susceptibility to experimental autoimmune orchitis (EAO), with BALB/cJ representative of the non-responders and BALB/cBy representative of the responders. We examined whether the susceptibility of these two substrains could be altered by reciprocal adoptive transfer of lymphoid cells. The cells transferred were of three types, normal spleen cells, T cell-enriched spleen and lymph node cells from mice immunized with testis homogenate (TH) in complete Freunds adjuvant (CFA) and given an extract of Bordetella pertussis (BP) and the latter cells activated by in vitro culture with TH antigen for 48 h. Controls were given buffer alone. Cell or buffer recipients were immunized with TH + CFA + BP three weeks later and examined for testicular histopathology 25-28 days after immunization. The cultured, immune T-enriched cells were consistently effective in transferring susceptibility from BALB/cBy to BALB/cJ. In the reverse experiments, non-responsiveness could be transferred from BALB/cJs to BALB/cBys most effectively with immune, non-cultured T-enriched cells. Transfer of cultured, immune T-enriched cells from BALB/cJs to other BALB/cJs had no significant effect on susceptibility to EAO. The results suggest that susceptibility to EAO in BALB/c mice depends on the T cell responses in the mice and not on differences at the level of the testis.

Renal transplant obstruction by B cell lymphoproliferative disorder: case report and review of the literature

We report a case of renal transplant ureteral obstruction caused by a cyclosporine-associated B cell lymphoproliferative disorder. The interaction of cyclosporine with Epstein-Barr virus infections and the predisposition towards B cell lymphoproliferative disorders are reviewed. The characterization of these tumors by deoxyribonucleic acid hybridization studies and a review of the clinical experience treating these disorders also are presented.