Experimental allergic encephalomyelitis (EAE)' is an autoimmune disease of the central nervous system (CNS) that is induced by immunization of susceptible animal strains with myelin basic protein (MBP). The disease is characterized by lymphocyte infiltration leading to CNS lesions, demyelination, and chronic relapsing paralysis, which are symptoms similar to those seen for multiple sclerosis (MS) in humans (1-6). Prevention of EAE by treatment with CD4-specific antibodies (7,8) and adoptive transfer of disease symptoms with MBP-specific T helper (Th) cells (9-16) have implicated CD4+ Th cells as the primary disease-inducing component of EAE.The dominant T cell response to MPB in mice of particular MHC haplotypes is directed towards distinct MBP determinants (16)(17)(18)(19)(20). For example, B10.PL and PL/J mice (H-2°) respond primarily to an acetylated NH2-terminal epitope, while SJL/J mice (H-2g) respond primarily to a more COOH-terminal epitope. Recent studies have indicated that in H-2u mice the T cell response to the dominant NH2-terminal epitope of MBP is of limited heterogeneity (21-23). Molecular characterization of the TCRs used by B10.PL-derived Th cells revealed the use of only two distinct V/3 gene segments, V/38.2 and VS13, and two distinct Va gene segments, Va2.3 and Va4.3 (21). The gene products of either V/3 segment can pair with the gene products ofeither Va segment to yield a total of four discrete types ofTh cells. The majority of Th cells examined expressed the V08.2 gene segment (84%) while the remainder expressed the V013 gene segment (16%). The distribution ofVa gene segments was less skewed among these MBP-specific Th cells with 60% expressing Va2 .3 and 40% expressing Va4.3 .The limited use of TCR V region genes in the response to MBP allows the application of unique strategies of specific immune intervention . Since the majority of MBP NH2-terminal reactive Th cells in H-2u mice use V08.2, mAbs to V08 (F23 .1,

SummaryCollagen type II-induced arthritis (CIA) is generated in susceptible rodent strains by intradermal injections of homologous or heterologous native type II collagen in complete Freund's adjuvant. Symptoms of CIA are analogous to those of the human autoimmune disease, rheumatoid arthritis. CIA is a model system for T cell-mediated autoimmune disease. To study the T cell receptor (TCR) repertoire of bovine type II-specific T cells that may be involved in the pathogenesis of CIA in DBA/1Lac.J (H-2q) mice, 13 clonally distinct T cell hybridomas specific for bovine type II collagen have been established and the ot and/3 chains of their TCRs have been analyzed. These T cell hybridomas recognize epitopes that are shared by type II collagens from distinct species and not by type I collagens, and exhibit a highly restricted TCP,.-ot/3 repertoire. The ot chains of the TCRs employ three Vot gene subfamilies (Votll, Vot8, and Vot22) and four Jot gene segments (Jot42, Jot24, Jot37, and Jot32). The Vot22 is a newly identified subfamily consisting of approximately four to six members, and exhibits a high degree of polymorphism among four mouse strains of distinct Vot haplotypes. In addition, the B chains of the TCRs employ three V/3 gene subfamilies (V~8, VB1, and VB6), however the V38.2 gene segment is preferentially utilized (58.3%). In contrast, the JB gene segment usage is more heterogeneous. On the basis of the highly limited TCR-ot/3 repertoire of the TCRs of the panel of bovine type II-specific T cell hybrid clones, a significant reduction (60%) of the incidence of arthritis in DBA/1Lac.J mice is accomplished by the use of anti-V38.2 antibody therapy. Collagen type II-induced arthritis (CIA) 1 in animals is an experimental animal model system of the human autoimmune disease, rheumatoid arthritis (1-3). CIA is induced in susceptible rodents by intradermal injections of homologous or heterologous native collagen type II (1, 2). In contrast, similar injections of other joint tissue proteins such as collagen type I and proteoglycans do not lead to arthritis. In addition, susceptibility to CIA in rodents is linked to MHC genes (4, 5). Among the inbred mouse strains, only mice of the H-2q and H-2 r haplotypes generally acquire an inflammatory polyarthritis upon immunization with collagen type II in CFA (5). However, SWR (H-2q) and RIII (H-2 r) inbred mouse strains are resistant to CIA (6, 7), suggesting that non-MHC genes are also crucial for the induction of the disease. Previous studies indicate that CIA is associated with a high level of both cellular and humoral responses to collagen type II. However, the role of Abs and T lymphocytes in the pathogenesis of the disease is ill-defined. It has been reported that transfer of anti-collagen type II Abs to naive animals results in transient synovitis with a histopathologic picture different from that seen in CIA (8). Hence, antibodies to collagen type II alone are not sufficient for the development of the prototypical lesions associated with arthritis. In contrast, adopti...

Studies ofIg gene structure and organization during the past decade have illuminated the central mechanisms of antibody diversification: the somatic rearrangement and reassortment of multiple gene segments, junctional flexibility, and point mutation . The inherited set of Ig gene segments provides a diversified genetic basis upon which these dynamic processes operate during ontogeny. Thus, the composition of these germline genes imposes a major influence on the development of the antibody repertoire.The present study examines the germline content and organization of the mouse heavy chain variable region genes (V genes) . We set out to analyze the locus in sufficient detail and resolution to provide the basis for determining the extent of inherited Vgene diversity, the evolution ofthe germline repertoire, and any functional consequences of the physical arrangement of the V gene segments .The mouse Igh locus consists of at least 100-200 V genes (1-3). V gene families, defined by nucleotide sequence relationships, comprise distinct sets of highly related V genes that can be identified by hybridization using prototypic V gene probes . This classification of V gene families (l, 4) has provided a useful framework for the study of germline V gene content, polymorphism, and utilization (1-7). Whereas the general organization of the Igh locus is 5'-V HDJ .-C -3' (8, 9), previous studies of V gene family organization have resulted in partial, relatively low resolution maps lacking consistency between reports (10-12)."Deletion mapping" takes advantage of the fact that V, gene rearrangements result in the deletion of DNA originally separating the rearranged V gene segment and the DJ .-CH region (13). We have constructed a panel of 32 pre-B cell lines, most of which have rearranged V, genes on both chromosomes. Since these cell lines were derived from Fi mice heterozygous at the Igh locus, V, gene deletions can be identified using RFLPs. V, gene analyses of 51 independently rearranged chromosomes are consistent with a single V, gene map order of nine V, gene families. The genomic stability of these cell lines and consistent deletion profiles of all 51 rearranged loci provide a high resolution V gene map that has compelling experimental support.

Animal models of autoimmune diseases have been instrumental in advancing our understanding of autoimmunity in humans. Collagen-induced arthritis in mice is an autoimmune disease model of rheumatoid arthritis, which is MHC class II restricted and CD4 T cell dependent. To better understand the fundamental role of T cells in arthritis, we have generated a transgenic mouse carrying the rearranged Valpha11.1 and Vbeta8.2 TCR chain genes isolated from a type II collagen (CII)-specific T cell hybridoma. Cell surface analysis indicated that Vbeta8.2 chain was expressed on the surface of nearly all peripheral T cells. Analysis of T cell subsets in transgenic mice revealed a profound skewing in peripheral T cells towards the CD4 population. Although peripheral T cells were not tolerant to CII and responded to CII stimulation in vitro, transgenic mice did not develop spontaneous arthritis. However, a rapid onset of arthritis with severe clinical signs was detected in transgenic mice after immunization with CII in complete Freund's adjuvant. Histological analysis of inflamed joints showed a great resemblance to arthritic joints in man. This unique transgenic mouse model provides valuable insights into the mechanism of arthritis and into potential specific immune interventions.

Animal models of autoimmune diseases have been instrumental in advancing our understanding of autoimmunity in humans. Collagen-induced arthritis (CIA) in mice is an autoimmune disease model of rheumatoid arthritis. Susceptibility to CIA in mice is linked to genes of the major histocompatibility complex (MHC). CD4(+) T cells that express the T-cell receptor (TCR) Tcra-V11.1 and/or Tcrb-V8.2 play a key role in the pathogenesis of arthritis in the DBA/1 mouse (H2(q)). We identified an inbred mouse strain, FVB/NJ (H2(q)), that is resistant to arthritis induction and exhibits a genomic deletion of certain Tcrb-V gene segments. We report a novel polymerase chain reaction-based method for the rapid identification of new mouse strains that exhibit germline Tcrb-V gene deletions. We mapped for the first time both the 5' and 3' breakpoints of the Tcrb-V deletion in the FVB/NJ, SWR, SJL, C57L, and C57BR strains to within 1.1 kilobases. Since there is an association between a particular Tcra-V allele (Tcra-V11.1(d)) and arthritis susceptibility in H2(q) mouse strains, we examined the allelic polymorphisms of the Tcra-V11 gene subfamily members between the arthritis-susceptible DBA/1 mouse and the arthritis-resistant FVB/NJ mouse strain. The amino acid sequences of the Tcra-V11.1 alleles differ at two positions (codons 18 and 68). Therefore, the resistance of FVB/NJ mouse to arthritis induction may be due in part to Tcra-V11.1 coding sequence polymorphism and Tcrb-V8.2 gene segment deletion, as we have recently demonstrated in the case of SWR mouse strain.