Rescue of autoimmune hepatitis by soluble MHC class II molecules in an altered concanavalin A-induced experimental model.

BACKGROUND: Soluble major histocompatibility complex class II (sMHCII) molecules have been described to maintain tolerance through the suppression of autoreactive T lymphocytes. In order to evaluate their ability to rescue autoimmune hepatitis (AIH) symptoms, the present work attempted to administer sMHCII molecules to an in vitro as well as in vivo concanavalin A (ConA)-induced AIH model. METHODS: The in vitro AIH model consisted of splenocyte stimulation with ConA in the presence or absence of serum-isolated sMHCII molecules. An in vivo ConA-modified model with or without sMHCII treatment was developed. The cytokine profile in culture supernatants and serum was tested by ELISA. Cell markers were evaluated by immunofluorescence, while cell proliferation by tritiated thymidine uptake. AIH symptoms were assessed by daily observations for the establishment of a disease severity scoring system and liver histology was evaluated using a biomolecular imager. RESULTS: The presence of sMHCII molecules in the ConA-stimulated cell cultures leads to a significant reduction of cell proliferation. The administration of sMHCII molecules to the ConA-treated animals showed a significant reduction in the levels of IL-2, IL-4, and IL-10, as well as a decrease in the number of spleen CD4+ and CD8+ cells. Upon development of a scoring system, it was shown that the sMHCII treatment was accompanied by a slower progression of the disease, while rescuing fibrotic liver morphology. CONCLUSION: The results presented in this study confirm the ability of sMHCII proteins to alleviate autoimmune hepatitis, possibly highlighting new therapeutic approaches for autoimmune diseases.

Soluble major histocompatibility complex molecules in immune regulation: highlighting class II antigens.

The involvement of major histocompatibility complex (MHC) antigens in the development and regulation of immune response has been well defined over the years, starting from maturation, antigenic peptide loading, migration to the cell membrane for recognition by the T-cell receptor and recycling for immune response cessation. During this intracellular trafficking, MHC antigens find a way to be excreted by the cells, because they can be found as soluble MHC class I (sMHC-I) and class II (sMHC-II) molecules in all body fluids. Although secretion mechanisms have not been sufficiently studied, sMHC molecules have been shown to display important immunoregulatory properties. Their levels in the serum have been shown to be altered in a variety of diseases, including viral infections, inflammation, autoimmunities and cancer, etc. while they seem to be involved in a number of physiological reactions, including maintenance of tolerance, reproduction, as well as mate choice vis-à-vis species evolution. The present review aims to present the thus far existing literature on sMHC molecules and point out the importance of these molecules in the maintenance of immune homeostasis.

Soluble MHC-II proteins promote suppressive activity in CD4+ T cells.

Soluble MHCII (sMHCII) molecules are present in body fluids of healthy individuals and are considered to be involved in the maintenance of self tolerance, and are also related to various diseases. Their concentration increases during in vivo antigen-specific tolerogenic stimulation and it was recently shown that exosome-mediated tolerance is MHCII dependent. At the cellular level, sMHCII proteins compete with membrane MHCII for T-cell receptor binding on CD4(+) T cells. Immunoaffinity purification techniques isolated sMHCII antigens from the serum of human serum albumin (HSA) -tolerant mice as a single highly glycosylated protein of ~ 60,000 molecular weight, specifically interacting with anti-class II antibodies in Western blotting and ELISA. Mass spectroscopy showed that these sMHCII proteins were loaded with the tolerogenic peptide as well as multiple self peptides. At the cellular level, sMHCII suppressed antigen-specific, and to a lesser degree antigen-non-specific, spleen cell proliferation and induced CD25 in naive T cells. In T cells activated by antigen-seeded macrophages, sMHCII decreased CD28 and increased CTLA-4 protein expression, while decreasing interleukin-2 and increasing interleukin-10 production. In this case, sMHCII proteins were shown to decrease ZAP-70 and LAT phosphorylation. The results presented here for the first time provide evidence for the role of sMHCII proteins in immune response suppression and maintenance of tolerance, revealing novel regulatory mechanisms for immune system manipulation.

Characterization of antigen-binding and MHC class II-bearing T cells with suppressive activity in response to tolerogenic stimulus.

Antigen specific non-responsiveness is generally developed through clonal deletion, anergy, and suppression. The term "suppression" is being considered as a functional immune deficit that can be adoptively transferred by regulatory/suppressor T cells. Following tolerance induction protocols the aim of the present study was to characterize the T cells involved in antigen-specific suppression. After defining the immunogenic and tolerogenic protocols in vitro and in vivo, it was shown that CD90(+) cells from tolerogenic mice were able to reduce specific antibody production when adaptively transferred to immunized mice. These cells were shown to highly express CD25 and Foxp3, co-localizing with CD4 and MHC class II antigens (MHCII), while a small percentage of these cells (8-14%) was binding free antigen. Further isolation of CD90(+)MHCII(+) and CD90(+)HSA(+) from mice having received the tolerogenic treatment and adaptive transfer to immunized mice showed that CD90(+)MHCII(+) cells were able to suppress antigen-specific response only when transferred along with the second antigenic challenge, while CD90(+)HSA(+) cells were suppressive only when given along with the first antigenic challenge. The suppressive effect of these two sub-populations could also be obtained in in vitro spleen cell proliferation assays in response to the HSA antigenic stimulus. Both in vitro and in vivo tolerogenic treatments were shown to correlate with soluble MHCII production in culture supernatants or serum respectively. Increase of MHCII in the serum could only be detected upon adaptive transfer of CD90(+)HSA(+) cells, whereas transfer of CD90(+)MHCII(+) cells resulted in increased levels of IL-10 and IFN-γ in the serum. These results defined at least two different levels of suppression, one during the onset which was antigen-specific and MHC restricted, and another non-specific at the end of an immune response.