ovalbumin and Nephritis

Regulatory T cells (Treg) are important for maintaining immune homeostasis. Adoptive transfer of Tregs is protective in renal disease models in both immunocompetent and immunodeficient mice. However the involvement of TCR recognition of renal antigens remains to be clarified. To address this question, we made use of Tregs from the DO11.10 mouse (a TCR transgenic (Tg) mouse), that recognise the non-murine antigen Ovalbumin (OVA) and therefore are not activated by renal antigens. DO11.10 Tregs were assessed functionally in vitro and demonstrated equivalent suppression to WT BALB/c Tregs. Adriamycin Nephropathy (AN) was induced in mice which had been transfused with CD4+CD25+Tregs isolated from DO11.10 or BALB/c mice. To eliminate the memory/activation state as a cause of differences in activity, the protective capacity of DO11.10 Tregs pre-activated with OVA in vivo was assessed. Transfer of WT BALB/c Tregs significantly attenuated the development of AN with less glomerulosclerosis, tubular atrophy and macrophage infiltration as compared to AN mice without Treg transfer. However, mice receiving either naïve or pre-activated DO11.10 Tregs were not protected from AN. The lack of protection by DO11.10 Tregs was not due to failure to traffic to the affected kidney. These results suggest that antigen recognition in the kidney is important for Treg protection against injury.

Topics: Animals; Antigens; Disease Models, Animal; Flow Cytometry; Immunohistochemistry; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mice, Transgenic; Nephritis; Ovalbumin; Real-Time Polymerase Chain Reaction; Receptors, Antigen, T-Cell; T-Lymphocytes, Regulatory

Autoantibody-mediated diseases like myasthenia gravis, autoimmune hemolytic anemia and systemic lupus erythematosus represent a therapeutic challenge. In particular, long-lived plasma cells producing autoantibodies resist current therapeutic and experimental approaches. Recently, we showed that the sensitivity of myeloma cells toward proteasome inhibitors directly correlates with their immunoglobulin synthesis rates. Therefore, we hypothesized that normal plasma cells are also hypersensitive to proteasome inhibition owing to their extremely high amount of protein biosynthesis. Here we show that the proteasome inhibitor bortezomib, which is approved for the treatment of multiple myeloma, eliminates both short- and long-lived plasma cells by activation of the terminal unfolded protein response. Treatment with bortezomib depleted plasma cells producing antibodies to double-stranded DNA, eliminated autoantibody production, ameliorated glomerulonephritis and prolonged survival of two mouse strains with lupus-like disease, NZB/W F1 and MRL/lpr mice. Hence, the elimination of autoreactive plasma cells by proteasome inhibitors might represent a new treatment strategy for antibody-mediated diseases.

Topics: Animals; Boronic Acids; Bortezomib; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Inbred MRL lpr; Mice, Inbred NZB; Models, Immunological; Nephritis; Ovalbumin; Plasma Cells; Protease Inhibitors; Proteasome Inhibitors; Pyrazines; Time Factors

Treatment with cyclosporin A (CsA) improves proteinuria and reduces renal cellular infiltration in chronic serum sickness (CSS). We examined if these effects were associated with a reduced renal expression of CD54 and its ligands, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and MHC class II molecules. We studied two groups of rats in which CSS was induced by daily injections of ovalbumin (OVA): a group treated with CsA (OVA.CsA group, n = 11) and a group that received no treatment (OVA.CSS group, n = 11). An additional group of five rats (control group) received only phosphate buffer. Immunostaining techniques were used to follow CSS and to study the expression of CD54, CD18, CD11b/c, IFN-gamma, TNF-alpha and MHC class molecules. Proteinuria (mg/24 h) was reduced from 248.2 +/- 73.1 (OVA.CCS group) to 14.5 +/- 13.1 with CsA treatment (P < 0.0001). The renal expression of CD54 and its ligands (CD18 and CD11b/c) was reduced by 50% to 75%. Correspondingly, there was a 60% to 85% reduction in the number of infiltrating leucocytes. The number of cells expressing TNF-alpha, IFN-gamma and MHC II molecules was also reduced. CsA reduces expression of CD54 and its ligands. This effect is associated with a reduction of cellular infiltration, IFN-gamma, TNF-alpha-producing cells and with MHC II expression in the kidney. These findings suggest that expression of adhesion molecules plays a critical role in CSS and underline the importance of cellular immunity in this experimental model.

Topics: Animals; Cell Adhesion Molecules; Chemotaxis, Leukocyte; Chronic Disease; Creatinine; Cyclosporine; Disease Models, Animal; Gene Expression Regulation; Histocompatibility Antigens; Immune Complex Diseases; Immunization; Immunosuppressive Agents; Intercellular Adhesion Molecule-1; Interferon-gamma; Kidney; Kidney Glomerulus; Male; Microscopy, Fluorescence; Nephritis; Ovalbumin; Proteinuria; Rats; Rats, Sprague-Dawley; Serum Sickness; T-Lymphocytes; Tumor Necrosis Factor-alpha

This investigation was designed to clarify the potential role of basic antigens in immune complex nephritis. The GBM is negatively charged and binds cationic substances. We studied 1) whether chemically cationised proteins can bind to the GBM, act as planted antigen, and induce 'in situ' immune complex formation or 2) if a pre-formed cationic antigen antibody complex can localise in the GBM. Results obtained with 5 chemically cationised proteins support the first possibility.

Topics: Animals; Antigen-Antibody Complex; Basement Membrane; Cations; Ferritins; Humans; Immune Complex Diseases; Immunoglobulin G; Immunoglobulin M; Kidney Glomerulus; Nephritis; Ovalbumin; Proteins; Rats; Serum Albumin

Topics: Animals; Antibodies, Antinuclear; Antigen-Antibody Complex; Electrophoresis; Female; Fluorescent Antibody Technique; Glomerulonephritis; Graft vs Host Disease; Immune Sera; Immunization; Immunoglobulin G; Kidney Glomerulus; Lupus Erythematosus, Systemic; Male; Mice; Microscopy, Electron; Nephritis; Ovalbumin; Rabbits

Topics: Antigen-Antibody Complex; Chromatography; Chromatography, Gel; Complement System Proteins; Edetic Acid; Glomerulonephritis; Humans; Immune Sera; Immunodiffusion; Lupus Erythematosus, Systemic; Molecular Weight; Nephritis; Ovalbumin; Radioimmunoassay

Young adult mice (30 to 43 days old) of autoimmune NZB and BIW F(1) strains failed to develop immunologic tolerance when treated with ultracentrifuged bovine gamma globulin and then challenged 12 days later. By contrast, weanling NZB and B/ W F(1) mice (18 to 25 days) as well as weanling C3H mice (16 to 19 days) became tolerant and had no serum antibody 12 days after challenge. The C3H mice remained unresponsive, but NZB and B! W F(1) mice produced antibody between days 27 and 41. The rapid loss of previously established tolerance to foreign antigens could have its parallel in the loss of tolerance to autoantigens with subsequent development of lupus nephritis and Coombs' positive hemolytic anemia in these animals.

Topics: Anemia, Hemolytic, Autoimmune; Animals; Animals, Newborn; Antibody Formation; Autoantibodies; Cattle; gamma-Globulins; Immune Tolerance; Lupus Erythematosus, Systemic; Mice; Nephritis; Ovalbumin

Topics: Animals; Antigen-Antibody Reactions; Complement System Proteins; gamma-Globulins; Glomerulonephritis; Kidney; Metabolism; Nephritis; Ovalbumin; Pepsin A; Proteinuria; Rabbits; Rats; Research; Tissue Extracts