concanavalin-a and Mycobacterium-avium-intracellulare-Infection

Mycobacterium avium complex-induced immunosuppressive macrophages (MAC-MPhis) exhibit suppressor activity against concanavalin A-induced T cell mitogenesis (T cell Con A mitogenesis). We examined the profiles of the MAC-MPhi-mediated suppression of lipopolysaccharide-induced B cell mitogenesis (B cell LPS mitogenesis) and found the following. First, although N(G)-monomethyl-L-arginine and carboxy-PTIO effectively blocked the MAC-MPhi's suppressor activity against T cell Con A mitogenesis, MAC-MPhi's action against B cell LPS mitogenesis was only weakly affected by these NO-reducing agents. Second, B cell LPS mitogenesis was remarkably more susceptible to MAC-MPhi-derived reactive oxygen intermediates than T cell Con A mitogenesis. Third, B cell LPS mitogenesis was less susceptible to the inhibitory effects of the other MAC-MPhi-derived suppressor mediators, including free fatty acids, TGF-beta and prostaglandin E(2), than T cell Con A mitogenesis. Fourth, MAC-MPhi's suppressor activity was strongly dependent on B7-1 like molecule-mediated cell contact with target cells only in the case of T cell Con A mitogenesis. Therefore, there are significant differences in the modes of suppressor action of MAC-MPhis against T cell and B cell mitogenesis.

Topics: Animals; B-Lymphocytes; Cell Communication; Cells, Cultured; Concanavalin A; Enzyme Inhibitors; Immune Tolerance; Immunity, Cellular; Lymphocyte Activation; Macrophages; Male; Mice; Mice, Inbred BALB C; Mycobacterium avium Complex; Mycobacterium avium-intracellulare Infection; Nitric Oxide Synthase; omega-N-Methylarginine; Reactive Oxygen Species; T-Lymphocytes

The suppressor activity of splenic macrophages induced by Mycobacterium intracellulare infection (MI-M phi s) against T cell concanavalin A (Con A) mitogenesis is mediated by MI-M phi's mediators, such as reactive nitrogen intermediates (RNIs), phosphatidylserine, free fatty acids, prostaglandin E(2) and to a minor extent TGF-beta. Here, we have compared the roles of RNIs and TGF-beta in the expression of MI-M phi's suppressor activity against Con A mitogenesis and anti-CD3 monoclonal antibody (mAb)- and anti-CD28 mAb-induced mitogenesis (TCR signal-induced mitogenesis) of the target T cells, and have found the following. First, N(G)-monomethyl-L-arginine (NMMA) inhibited MI-M phi's suppressor activity against TCR signal-induced mitogenesis as well as Con A mitogenesis. Second, anti-TGF-beta mAb weakly restored the MI-M phi-mediated suppression only in the case of Con A mitogenesis, under limited conditions, such as very low cell densities of MI-M phi s. Third, the blocking effects of NMMA plus anti-TGF-beta mAb were somewhat more prominent in the case of Con A mitogenesis than in the case of TCR signal-induced mitogenesis. Fourth, Con A- or TCR signal-stimulated MI-M phi s secreted significant amounts of the latent TGF-beta but not the active one. These findings indicate that RNIs, but not TGF-beta, play important roles in the MI-M phi-mediated suppression of TCR signal-induced mitogenesis, as well as Con A mitogenesis, of the target T cells.

Topics: Animals; Antibodies, Monoclonal; Antigen Presentation; CD28 Antigens; CD3 Complex; Concanavalin A; Dose-Response Relationship, Drug; Enzyme Inhibitors; Humans; Immune Tolerance; Immunosuppressive Agents; Lymphocyte Activation; Macrophage Activation; Macrophages; Male; Mice; Mice, Inbred BALB C; Mycobacterium avium Complex; Mycobacterium avium-intracellulare Infection; Nitrogen; omega-N-Methylarginine; Reactive Nitrogen Species; Receptors, Antigen, T-Cell; Signal Transduction; Spleen; T-Lymphocytes; Transforming Growth Factor beta

We found previously that immunosuppressive macrophages (Mphis) induced by Mycobacterium intracellulare infection (MI-Mphis) transmitted their suppressor signals to target T cells through cell contact with target T cells. In this study, we examined what kinds of Mphi surface molecules are required for such cell-to-cell interaction. First, it was found that a B7-1-like molecule (B7-1LM) recognizable with one of three test clones of anti-B7-1 monoclonal antibodies (mAbs) was required for expression of the Mphi suppressor activity. Neither anti-B7-2, anti-ICAM-1, nor anti-VCAM-1 mAb blocked the Mphi suppressor activity. Second, MI-Mphis increased the expression of B7-1LM in parallel with the acquisition of the suppressor activity. Moreover, MI-Mphis bound with target T cells in a B7-1LM-dependent fashion. Third, mAb blocking of CTLA-4 on target T cells did not reduce the suppressor activity of MI-Mphis, suggesting the role of a putative molecule on target T cells other than CTLA-4 as the receptor for B7-1LM of MI-Mphis. Fourth, concanavalin A (Con A) stimulation of MI-Mphis was needed for effective cell contact with target T cells and subsequent expression of the suppressor activity of MI-Mphis. Fifth, the Con A-induced increase in the suppressor activity of MI-Mphis was inhibited by KN-62 but not by herbimycin A, H-7, nor H-88, indicating that Con A-induced up-regulation of MI-Mphi function is mediated by calmodulin-dependent protein kinase II or ATP/P2Z receptors, but independent of protein tyrosine kinase, protein kinase C, and protein kinase A. These findings indicate that a B7/CTLA-4-independent mechanism is needed for the transmission of the suppressor signals from MI-Mphis to target T cells.

Topics: Animals; Antigens, CD; Antigens, Differentiation; B7-1 Antigen; Blotting, Western; Cell Communication; Cells, Cultured; Concanavalin A; CTLA-4 Antigen; Immune Tolerance; Lymphocyte Activation; Macrophage Activation; Macrophages; Male; Mice; Mice, Inbred BALB C; Mycobacterium avium-intracellulare Infection; T-Lymphocytes

Previously, we found that phospholipids and reactive nitrogen intermediates (RNI) collaborated in expression of the T cell mitogenesis-inhibitory activity of immunosuppressive macrophages induced by Mycobacterium avium-intracellulare complex (MAIC) infection. In this study, we examined the roles of free fatty acids (FFA) and prostaglandins (PG) as effectors of MAIC-induced macrophages, and moreover, their collaborating effects with RNI. First, treatment of MAIC-induced macrophages with quinacrine (phospholipase A2 (PLA2) inhibitor), dexamethasone (inhibitor of PLA2 and PG synthesis) or indomethacin (PG synthesis inhibitor) attenuated their suppressor activity against concanavalin A (Con A)-induced mitogenesis of splenocytes (SPC), indicating important roles of FFA liberated from membrane phospholipids and PG, as effectors. Second, oleic acid, PGE2, RNI generated from NOR 4 (a new nitric oxide (NO) donor), and phosphatidylserine (PS) exhibited suppressor activity against SPC mitogenesis without showing significant cytotoxicity, in an irreversible manner. Third, the suppressor activities of RNI and PGE2 were potentiated by combined use with oleic acid in a synergistic manner. Fourth, a dual-chamber experiment in which target SPC were separated from MAIC-induced macrophages by a Millipore filter revealed a requirement for cell-to-cell contact for expression of the suppressor function of MAIC-induced macrophages. These findings indicate that RNI, FFA, PG, and phospholipids (presumably PS) and their collaboration play central roles in expression of the T cell mitogenesis-inhibitory function of MAIC-induced suppressor macrophages.

Topics: Animals; Cell Communication; Concanavalin A; Fatty Acids, Nonesterified; Female; Humans; Immune Tolerance; Macrophages; Mice; Mice, Inbred BALB C; Mitogens; Mycobacterium avium Complex; Mycobacterium avium-intracellulare Infection; Phospholipids; Prostaglandins; Spleen; T-Lymphocytes

The profile of generation and characteristics of immunosuppressive macrophages (M phi s), which suppress the ConA-mitogenic response of spleen cells (SPCs), in host CBA/JN mice during the course of Mycobacterium avium complex (MAC) and M. tuberculosis (MT) infections were investigated. In both infections, a marked reduction in ConA mitogenic response of splenic T cells was seen around 2 weeks after infection, and this was accompanied by generation of potent immunosuppressive M phi s in the SPCs of infected mice. The suppressive activity was much stronger in MT-infected mice than in MAC-infected ones. In both infections, the large part of the suppressive M phi s exhibited suppressor activity that depended on the arachidonic acid cascade, particularly mediated by prostaglandins (PGs), and the remainder showed the suppressor action independent from PGs. The unique finding of this study is that the generation of IL-2 reactive T cell populations in SPCs in response to ConA signal was markedly inhibited by the MAC- and MT-induced immunosuppressive M phi s, whereas the suppressive M phi s failed to reduce the IL-2-producing ability of splenic T cells. In any case, the present results indicate a close similarity in immunosuppressive M phi s induced by MAC and MT infections.

Topics: Animals; Cells, Cultured; Concanavalin A; Histocompatibility Antigens Class II; Immunosuppression Therapy; Indomethacin; Interleukin-2; Macrophage Activation; Macrophages; Mice; Mycobacterium avium-intracellulare Infection; Receptors, Interleukin-2; T-Lymphocytes; Tuberculosis