concanavalin-a and Histoplasmosis

Splenic macrophages from Histoplasma capsulatum-infected mice express inducible nitric oxide synthase (iNOS), and the iNOS expression correlates with severity of the infection. We examined whether production of NO is responsible for apoptosis and the anti-lymphoproliferative response of splenocytes from mice infected with H. capsulatum. In situ terminal deoxynucleotidyl transferase nick end labeling revealed apoptotic nuclei in cryosections of spleen from infected but not normal mice. Splenocytes of infected mice were unresponsive to stimulation by either concanavalin A or heat-killed H. capsulatum yeast cells. Splenocyte responsiveness was restored by addition to the medium of NG-monomethyl-L-arginine, a known inhibitor of NO production. The proliferative response of splenocytes from infected mice was also restored by depletion of macrophages or by replacement with macrophages from normal mice. In addition, expression of iNOS returned to its basal level when the animals had recovered from infection. These results suggest that suppressor cell activity of macrophages is associated with production of NO, which also appears to be an effector molecule for apoptosis of cultured splenocytes from infected mice.

Topics: Animals; Apoptosis; Concanavalin A; Histoplasma; Histoplasmosis; Immune Tolerance; Macrophage Activation; Macrophages; Mice; Mice, Inbred C57BL; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Spleen

The incorporation of (3H) thymidine and the biosynthesis of interleukin-2(IL-2) were investigated in Concanavalin A (ConA) and histoplasmin stimulated lymphocytes from spleen of infected Balb/c mice with the yeast phases of Histoplasma capsulatum. The ability to incorporate (3H) thymidine of Con A stimulated lymphocytes in culture from spleen of Histoplasma capsulatum infected mice, as well as the IL-2 content present in the supernatants of that cultures, were depressed along the first three weeks of the experiments, but starting week five, normal values were restored or even discretly increased. Incorporation of (3H) thymidine in histoplasmin stimulated lymphocytes remained inhibited along the seven weeks the experiment lasted. Results showed that inoculation of H. capsulatum yeast in mice provoked a temporary immunosuppression on cell mediated immunity, that can be explained by means of the inability of T cells to produce enough IL-2 necessary for the proliferation of T cells in culture.

Topics: Animals; Cells, Cultured; Concanavalin A; Histoplasmin; Histoplasmosis; Immunity, Cellular; Interleukin-2; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Thymidine

Blastogenic responses of spleen cells to histoplasmin and ribosomal antigens and to the mitogens concanavalin A. phytohemagglutinin, and lipopolysaccharide were studied in normal and immunized mice (10(5) live yeast cells of Histoplasma capsulatum given by the subcutaneous route). Cells (10(6) per well) were cultured with and without antigens and mitogens in microtiter plates with RPMI 1640-5% heat-inactivated normal mouse serum for 72 h at 37 degrees C. Cells were harvested after a 16- to 18-h pulse with 1 microCi of [3H]thymidine (6.7 Ci/mol), and thymidine incorporation was measured by scintillation counting. The initial blastogenic response to concanavalin A (54 X 10(3) cpm) was suppressed (P less than 0.001) from 4 to 14 days post-immunization and returned to control levels on day 21. The response to phytohemagglutinin was suppressed up to 21 days. Lipopolysaccharide responses, however, were affected to a lesser degree. Blastogenic responses to histoplasmin and H. capsulatum ribosomes were similar on day 0 in normal and immune lymphocytes, but by day 4 cells from immunized mice were more responsive (P less than 0.01). The maximum response to H. capsulatum antigens was detected on day 42 and was 9- to 16-fold higher than in controls. These results demonstrate in vitro responses of primed lymphocytes on exposure to H. capsulatum antigens and suppressed responses to mitogens during early stages of the immune response.

Topics: Animals; Concanavalin A; Dose-Response Relationship, Immunologic; Histoplasma; Histoplasmin; Histoplasmosis; Immunization; Lipopolysaccharides; Lymphocyte Activation; Mice; Phytohemagglutinins; Ribosomes; Spleen; Time Factors

The effect of granulomatous infections upon the activity of a T lymphocyte subclass in human peripheral blood that can be induced by concanavalin A (Con A) to function in a suppressor mode was studied. Peripheral blood lymphocytes (PBL) from eleven patients with disseminated mycotic or mycobacterial infections or from controls were preincubated with and without Con A, washed and cultured with allogeneic PBL freshly drawn from healthy donors sensitive to histoplasmin. DNA synthesis was then measured in co-cultures stimulated by Con A, histoplasmin, or by the mixed lymphocyte culture (MLC) reaction alone. As compared with cells preincubated without Con A, the Con A-pretreated cells were significantly less effective in suppressing the responses of normal PBL to histoplasmin (P < 0.01), and in a one-way MLC reaction (P < 0.05). The Con A-induced suppressor activity of PBL from nine patients with localized granulomatous infections did not differ significantly from that exerted by PBL of normal controls in two of the three co-culture systems employed. These studies suggest that either dysfunction or a reduction of the Con A-inducible T-suppressor cell subpopulation in peripheral blood is frequent among patients was disseminated granulomatous infections.

Topics: Adolescent; Adult; Aged; Blastomycosis; Concanavalin A; Histoplasmin; Histoplasmosis; Humans; Hypersensitivity, Delayed; Leprosy; Leukocyte Count; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Middle Aged; Skin Tests; T-Lymphocytes, Regulatory; Tuberculosis