A Central Role for αβ T Cells in the Pathogenesis of Murine Lupus (original) (raw)
Subsets of Transgenic T Cells That Recognize CD1 Induce or Prevent Murine Lupus: Role of Cytokines
Journal of Experimental Medicine, 1998
T cells with T cell receptor (TCR) transgenes that recognized CD1 on syngeneic B cells stimulated B cells to secrete immunoglobulins in vitro. The CD4 ϩ , CD8 ϩ , or CD4 Ϫ CD8 Ϫ T cells from the spleen of the TCR transgenic BALB/c donors induced lupus with anti-double stranded DNA antibodies, proteinuria, and immune complex glomerulonephritis in irradiated BALB/c nude mice reconstituted with nude bone marrow. Injection of purified CD4 Ϫ CD8 Ϫ T cells from the marrow of transgenic donors prevented the induction of lupus by the transgenic T cells. Transgenic T cells that induced lupus secreted large amounts of interferon (IFN)-␥ and little interleukin (IL)-4, and those that prevented lupus secreted large amounts of IL-4 and little IFN-␥ or IL-10.
Cellular Immunology, 2002
Lupus-prone (MRL Â C57BL/6) F 1 mice lacking cd T cells show more severe lupus than their T cell-intact counterparts, suggesting that cd T cells down-modulate murine lupus. To determine the mechanisms for this effect, we assessed the capacity of cd T cell lines derived from spleens of ab T cell-deficient MRL/Mp-Fas lpr (MRL/Fas lpr) mice to down-regulate anti-dsDNA production generated by CD4 þ ab T helper cell lines and activated B cells from wild-type MRL/Fas lpr mice. One line, GD12 (gd TCR þ , CD4 À CD8 À), had the capacity to reduce anti-dsDNA production in a contact-dependent manner. GD12 also killed activated MRL/Fas lpr (H-2 k) B cells, with less cytolysis of resting B cells than that generated by in comparison to cytokine-matched cd T cell lines. In addition, GD12 also killed activated B cells derived from C57BL/6-Fas lpr (H-2 b) or b 2-microglobulin (b 2 M)-deficient MRL/ Fas lpr mice, suggesting cytolysis was neither MHC-nor CD1-restricted. Killing by GD12 was inhibited by anti-TNFa and anti-TNF-R1, and partially blocked by anti-gd TCR Fab fragments, but not by anti-FasL, anti-TNF-R2 (p75) or concanamycin A. IL-10 produced by GD12 also partially inhibited ab Th1-dependent but not ab Th2-dependent autoantibody production. These findings prove that we have identtified a cd T cell line that suppresses autoantibody synthesis by ab T-B cell collaboration in vitro.
T cell autoimmunity in Ig transgenic mice
Journal of immunology (Baltimore, Md. : 1950), 1999
Autoantibodies directed at a diverse group of proteins of the U1/Sm ribonucleoprotein (snRNP) are characteristic of systemic lupus erythematosus and are found in the MRL murine model of this disease. This study examines the role of transgenic B lymphocytes in the regulation of autoreactive T cells to the snRNP autoantigen. Transgenic mice were developed bearing an Ig heavy chain gene specific for the D protein component of murine snRNP. B lymphocytes in these mice are neither deleted nor anergic and are of an immature (heat-stable Aghigh) phenotype. T lymphocytes from anti-snRNP transgenic mice were examined using a recombinant form of the D protein of the murine snRNP complex. Our results revealed that transgenic anti-snRNP B cell APCs stimulated CD4 T cells from wild-type C57BL/6 and MRL lpr/lpr mice, while nonspecific APCs failed to stimulate CD4 T cells. This study demonstrates that autoreactive T cells are not deleted from wild-type mice, although their activation is facilitate...
Interferon-α accelerates murine systemic lupus erythematosus in a T cell-dependent manner
Arthritis & Rheumatism, 2010
Objective-To investigate the mechanism for lupus acceleration by interferon alpha (IFNα) in NZB/W mice. Methods-NZB/W mice were treated with an adenovirus expressing IFNα. T cells were depleted in some mice with an anti-CD4 antibody. The production of anti-dsDNA antibodies was measured by ELISA and ELISpot assays. Germinal centers and antibody-secreting cells (ASCs) in spleens and IgG deposition and leukocyte infiltrates in kidneys were visualized by immunofluorescence staining. The phenotype of splenic cells was determined by flow cytometry. Finally, somatic hypermutation and gene usage in heavy chain variable regions of IgG2a and IgG3 were studied by single cell PCR. Results-IFNα accelerated lupus in NZB/W mice is associated with elevated serum levels of IgG2 and IgG3 anti-dsDNA antibodies, and accumulation of many IgG ASCs in the spleen, which do not develop into long-lived plasma cells. Furthermore, IgG2a and IgG3 antibodies in these mice are highly somatically mutated and use distinct repertoires of VH genes. The induction of SLE in these mice is associated with an increase in B cell TLR7 expression, increased serum levels of BAFF, IL-6 and TNFα, and induction of T cells expressing IL-21. Although IFNα drives a T-independent increase in serum levels of IgG, autoantibody induction and the development of nephritis are both completely dependent on CD4 T cell help. Conclusion-Our study shows that although IFNα activates both innate and adaptive immune responses in NZB/W mice, CD4 T cells are necessary for IFNα driven induction of anti-dsDNA antibodies and clinical SLE.
Immunity, 1999
reviewed by Miller and Flavell, 1994). Similar, if less tight, self-tolerance mechanisms are also considered to operate in B cells (reviewed by Goodnow et al., 1995). Systemic lupus erythematosus (SLE), a prototype of human systemic autoimmune diseases, is characterized Diseases by a wide variety of multiorgan injuries (reviewed by ‡ Department of Immunology and Cell Biology Kotzin, 1996; Vyse and Kotzin, 1998), among which the Faculty of Medicine hallmarks are proliferative glomerulonephritis and arthri-Kyoto University tis. For decades, a number of implications have been Yoshida-Konoe, Sakyo-ku made about the breakdown of self-tolerance in lupus Kyoto, 606-8501 diseases of both humans and mice (Roark et al., 1995; Japan reviewed by Kotzin, 1996). The exact cellular and molec- § Department of Pathology ular mechanisms leading to the characteristic disease Ehime University School of Medicine complex of lupus, however, remain unknown. Several Shigenobu-cho, Onsen-gun murine models that spontaneously develop lupus-like Ehime, 791-0295 diseases have been extensively studied for decades. Japan NZB, MRL/Mp, and BXSB mice develop lupus-like diseases of rather late onset with a similar yet distinct disease spectrum (reviewed by Theofilopoulos and Summary Dixon, 1985). NZBxNZW F1 (reviewed by Drake et al., 1995), MRL/Mp-lpr/lpr (reviewed by Cohen and Eisen-PD-1, a 55 kDa transmembrane protein containing an berg, 1991), and BXSB/Yaa (reviewed by Izui et al., 1995), immunoreceptor tyrosine-based inhibitory motif, is inwhich are generated by the introduction of additional duced in lymphocytes and monocytic cells following genetic factors into the above model mice, develop by activation. Aged C57BL/6(B6)-PD-1 Ϫ/ Ϫ congenic mice far more aggressive diseases of earlier onset. Recent spontaneously developed characteristic lupus-like progenome-wide linkage analysis of these lupus-prone liferative arthritis and glomerulonephritis with premice has revealed a number of loci linked with the indidominant IgG3 deposition, which were markedly acvidual disease manifestations, although none of these celerated by introduction of a Fas mutation (lpr). susceptible genes have been identified to date (re-Introduction of a PD-1 null mutation into the 2C-TCR viewed by Vyse and Kotzin, 1998). (anti-H-2L d ) transgenic mice of the H-2 b/d background Under such circumstances, genetically manipulated resulted in the chronic and systemic graft-versusmurine models that develop various aspects of lupushost-like disease. Furthermore, CD8 ؉ 2C-TCR ؉ PD-1 Ϫ/Ϫ like disease would be quite helpful in providing insights T cells exhibited markedly augmented proliferation in into the pathogenesis of lupus. Although the overexvitro in response to H-2 d allogenic cells. Collectively, pression of the bcl-2 (Strasser et al., 1991) or fli (Zhang it is suggested that PD-1 is involved in the maintenance et al., 1995) oncogene was reported to develop glomeruof peripheral self-tolerance by serving as a negative lonephritis, these transgenic mouse models may not regulator of immune responses. be suitable for elucidation of etiological mechanisms of natural autoimmune diseases because this pathogene-Introduction sis is primarily due to proliferation and/or survival of lymphocytes expressing the transgenes rather than to Self-tolerance is maintained in distinct immunocompespecific defects in the immune regulation. Glomerulonetent cells at different stages (reviewed by Mondino et phritis but not arthritis was also reported in lyn-deficient al., 1996). In T cells, for instance, autoreactive T cell mice (Hibbs et al., 1995; Nishizumi et al., 1995). Unlike clones can be clonally eliminated in the thymus during the natural lupus-prone mice and human SLE patients the differentiation process (negative selection or centhat produce elevated levels of IgG, however, lyn-defitral tolerance) (Kappler et al., 1987; reviewed by von cient mice exhibit a rather selective increase in IgM. So Boehmer, 1990, 1991; Nossal, 1994). T cells that have far, none of the known genes have been shown to be managed to escape from negative selection and are responsible for both glomerulonephritis and arthritis, the released in the periphery are still constrained to avoid hallmarks of systemic autoimmune diseases. response to autoantigens (peripheral tolerance) (re-The PD-1 gene, which belongs to the immunoglobulin viewed by Miller and Morahan, 1992; Mondino et al., gene superfamily, encodes a 55 kDa type I transmem-1996). Several distinct mechanisms for peripheral tolerbrane protein (Agata et al., 1996) containing the immuance have been proposed based upon various animal noreceptor tyrosine-based inhibitory motif (ITIM), V/Ixmodels, including clonal anergy, downregulation of YxxL, (reviewed by Thomas, 1995; Vivier and Daeron, antigen receptor/coreceptor complex (Rocha and von 1997) in its cytoplasmic tail. The structure of PD-1 in-Boehmer, 1991), and involvement of active suppression cluding the ITIM motif is conserved between mouse and human (Shinohara et al., 1994; Finger et al., 1997). Although PD-1 is expressed only on a tiny population To whom correspondence should be addressed (e-mail: honjo@ mfour.med.kyoto-u.ac.jp). in the thymus of normal mice, it has been found to be cryosections were pretreated with 2% BSA in PBS/0.05% Tween (1995). Genetic contributions to lupus-like disease in (NZB ϫ NZW)F1 mice. Immunol. Rev. 144, 51-74. (PBS/T) for 30 min and incubated with biotinylated CD8a or 1B2 overnight. After washing twice with PBS/T, sections were incubated Finger, L.R., Pu, J., Wasserman, R., Vibhakar, R., Louie, E., Hardy, with avidin and biotinylated-HRP (VECTASTAIN ABC kit, Vector Lab-R.R., Burrows, P.D., and Billips, L.G. (1997). The human PD-1 gene: oratory) and developed with 0.02% H 2 O 2 and diaminobenzidine tetcomplete cDNA, genomic organization, and developmentally regurahydrochloride (DAB) (Wako, Osaka, Japan) according to the prolated expression in B cell progenitors. Gene 197, 177-187. cedures of the manufacturer. The sections were counterstained with Goodnow, C.C., Cyster, J.G., Hartley, S.B., Bell, S.E., Cooke, M.P., hematoxylin by standard methods. Healy, J.I., Akkaraju, S., Rathmell, J.C., Pogue, S.L., and Shokat, K.P. (1995). Self-tolerance checkpoints in B lymphocyte develop-Flow Cytometric Analysis ment. Adv. Immunol. 59, 279-368. The following antibodies were purchased from PharMingen: FITC-Hibbs, M.L., Tarlinton, D.M., Armes, J., Grail, D., Hodgson, G., Maglabeled anti-mouse CD8a (53-6.7), phycoerythrin-R (PE)-conjugated litto, R., Stacker, S.A., and Dunn, A.R. (1995). Multiple defects in the anti-CD4 (RM4-5), FITC-labeled anti-mouse Thy-1 (30-H12), PE-conimmune system of Lyn-deficient mice, culminating in autoimmune jugated anti-mouse CD45R/B220 (RA3-6B2), biotinylated antidisease. Cell 83, 301-311. mouse CD25 (7D4), biotinylated anti-mouse CD44(1M7), biotinylated Ishida, Y., Agata, Y., Shibahara, K., and Honjo, T. (1992). Induced anti-mouse CD45RB (16A), biotinylated anti-mouse CD62L (MELexpression of PD-1, a novel member of the immunoglobulin gene 14), and biotinylated anti-mouse CD69 (H1.2F3). Anti-2C TCR␣ superfamily, upon programmed cell death. EMBO J. 11, 3887-3895. clonotypic mAb, 1B2 (Kranz et al., 1984), and anti-mouse PD-1 mAb Izui, S., Kelley, V.E., Masuda, K., Yoshida, H., Roths, J.B., and Mur-(J43) were biotinylated. Cells were preincubated with 2.4G2 (antiphy, E.D. (1984). Induction of various autoantibodies by mutant gene Fc receptor mAb) and then stained with the indicated antibodies, lpr in several strains of mice. J. Immunol. 133, 227-233. followed by flow cytometric analysis. Izui, S., Iwamoto, M., Fossati, L., Merino, R., Takahashi, S., and Ibnou Zekri, N. (1995). The Yaa gene model of systemic lupus erythe-Proliferative Assay matosus. Immunol. Rev. 144, 137-156. Splenocytes from 2C and 2C ϫ PD-1 Ϫ/Ϫ mice on the H-2 b/b background were stimulated with mitomycin C (Sigma Chemical)-treated Kappler, J.W., Roehm, N., and Marrack, P. (1987). T cell tolerance splenocytes from BALB/c mice in the culture for 1 week. After reby clonal elimination in the thymus. Cell 49, 273-280. peated stimulation under the same conditions, the cells were Kariyone, A., Takiguchi, M., Igarashi, S., and Kano, K. (1988). Ontogharvested. Ten thousand viable cells were cultured in U-bottom eny and function of B220 ϩ L3T4 ϩ T-cell subset of MRL/Mplpr/lpr 96-well culture dishes (Iwaki Glass, Chiba, Japan) with indicated mice. Cell. Immunol. 115, 112-120. stimulators for 72 hr, pulsed with 0.5 Ci of methyl-3 H thymidine Kawai, K., Matsumoto, Y., Watanabe, H., Ito, M., and Fujiwara, M. per well (Amersham Pharmacia Biotech, Tokyo, Japan) for the last (1991). Induction of cutaneous graft-versus-host disease by local 12 hr, and then harvested with Ready Filter for Skatron (Beckman) injection of unprimed T cells. Clin. Exp. Immunol. 84, 359-366. followed by scintillation counting. When splenocytes or cell lines Kotzin, B.L. (1996). Systemic lupus erythematosus. Cell 85, 303-306. were used as stimulators, they were treated with 50 g/ml of mito-Kranz, D.M., Sherman, D.H., Sitkovsky, M.V., Pasternack, M.S., and mycin C (Sigma) for 30 min followed by three washings. Eisen, H.N. (1984). Immunoprecipitation of cell surface structures of cloned cytotoxic T lymphocytes by clone-specific antisera. Proc.
T cell receptor alpha-chain repertoire of pathogenic autoantibody-inducing T cells in lupus mice
The Journal of Immunology
The production of pathogenic anti-DNA autoantibodies in mice with lupus nephritis is dependent on special autoimmune Th cells that can also transfer the disease into preautoimmune mice. In previous work, these pathogenic Th cells were cloned and their TCR beta-chains were sequenced to reveal a recurrent motif of anionic residues in their CDR3 loops. Accordingly, approximately half of the Th clones were found to be specific for nucleosomal Ag that contain cationic residues. Herein, we analyzed the TCR alpha-chain repertoire of 15 of these pathogenic Th clones and found them to be heterogeneous, even among the nucleosome-specific Th clones. Most of these autoimmune TCR alpha-chains contained anionic residues in their CDR3 in addition to cationic residues. Therefore, these pathogenic Th clones of lupus probably recognize epitopes with mixed charge runs that are derived from autoantigens, such as histone-DNA complexes. Interestingly, the V alpha gene segments used by 10 of these Th clon...
Treatment of murine lupus with cDNA encoding IFN-γR/Fc
Journal of Clinical Investigation, 2000
Introduction Lupus, the prototypic systemic autoimmune disease, is characterized by a high female predominance, multiorgan pathology, and a broad spectrum of autoantibodies, of which those against nuclear antigens typically predominate (1). The etiology of this disease is still unknown, but a strong genetic predilection appears to be a dominant factor (reviewed in ref. 2). Despite considerable advances in the management of this disease, morbidity and mortality remain high, and intense efforts are ongoing to develop less toxic and more efficacious treatments. Among the many avenues pursued in experimental models, those related to immunointervention with blocking peptides (3), antibodies, and other agents that inhibit coreceptor or costimulatory molecules (4-6), and those using agonist and antagonists of cytokines (reviewed in ref. 7 and 8), appear to be promising. Many cytokine abnormalities have been identified in lupus-predisposed mice and humans (reviewed in ref. 7 and 8), the most prominent of which is increased levels of IFN-γ in serum, lymphoid organs, and afflicted tissues. The importance of this Th1 type inflammatory cytokine in lupus pathogenesis was suggested by the initial demonstration of Jacob et al. (9) that (NZBxW)F 1 lupus mice treated with IFN-γ or its inducers showed accelerated disease, whereas those treated with anti-IFN-γ Ab beginning at the early stage showed significant delay in disease onset. The most compelling evidence for the deleterious effects of IFN-γ in lupus was obtained recently by us (10) and
Arthritis research, 2001
The classification of T helper cells into type 1 (Th1) and type 2 (Th2) led to the hypothesis that Th1 cells and their cytokines (interleukin [IL]-2, interferon [IFN]-gamma) are involved in cell-mediated autoimmune diseases, and that Th2 cells and their cytokines (IL-4, IL-5, IL-10, IL-13) are involved in autoantibody(humoral)-mediated autoimmune diseases. However, this paradigm has been refuted by recent studies in several induced and spontaneous mouse models of systemic lupus erythematosus, which showed that IFN-gamma is a major effector molecule in this disease. These and additional findings, reviewed here, suggest that these two cross-talking classes of cytokines can exert autoimmune disease-promoting or disease-inhibiting effects without predictability or strict adherence to the Th1-versus-Th2 dualism.
Genetic Complementation Results in Augmented Autoantibody Responses to Lupus-Associated Antigens
The Journal of Immunology, 2009
Lupus-prone female NZM2328 mice develop high titers of anti-nuclear and anti-dsDNA autoantibodies. Despite high expression of type I IFNs, these mice do not develop autoantibodies to the small nuclear ribonucleoprotein (snRNP) complex. Thus, additional genetic factors must regulate the generation of anti-snRNP autoantibodies. In contrast, despite much lower expression of type 1 IFNs, the diabetes-prone NOD mice spontaneously make anti-snRNP autoantibodies, albeit at a low incidence. To determine whether combination of high type I IFN response of NZM mice with appropriate susceptibility genes of NOD mice would result in anti-snRNP antibody response, cohorts of (NZM2328 × NOD) F1 mice were generated and characterized for development of autoimmunity.
Journal of Experimental Medicine, 1993
Males from the BXSB murine strain (H-2b) spontaneously develop an autoimmune syndrome with features of systemic lupus erythematosus (SLE), which results in part from the action of a mutant gene (Yaa) located on the Y chromosome. Like other H-2b mice, the BXSB strain does not express the class II major histocompatibility complex antigen, I-E. Here we report that the expression of I-E (E alpha dE beta b) in BXSB males bearing an E alpha d transgene prevents hypergammaglobulinemia, autoantibody production, and subsequent autoimmune glomerulonephritis. These transgenic mice bear on the majority of their B cells not only I-E molecules, but also an I-E alpha chain-derived peptide presented by a higher number of I-Ab molecules, as recognized by the Y-Ae monoclonal antibody. The I-E+ B cells appear less activated in vivo than the I-E- B cells, a minor population. This limited activation of the I-E+ B cells does not reflect a functional deficiency of this cell population, since it can be sti...