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CD1 and TL were once thought to be genetic homologues because of their thymus-specific expression. We investigated their equivalents in the rat to clarify whether their structure and pattern of expression are conserved in rodents. Two rat class Ib genes, containing 3' sequences very similar to mouse TL, were identified and designated RT1.P. Neither of them, however, can encode ordinary class I molecules due to the accumulation of harmful mutations in the 5' regions that are unique to RT1.P, while the 3' TL-like regions still retain protein-coding capacity. Comparison of the structural organization of three types of TL family genes, which include mouse T3/T18-encoding TL antigens, mouse T1/T16, and rat RT1. P1/P2 pseudogenes, revealed the presence of a clear demarcation between the type-specific and TL-specific sequences at intron 3. This finding suggests that recombination plays an important role in creating the TL family genes in rodents. Characteristic features of TL, such as a low level of polymorphism and linkage to the major histocompatibility complex, were also observed in the rat. On the other hand, rat CD1 molecules were expressed at a high level on the surface of thymocytes. Absence of authentic TL antigens and thymic expression of CD1d molecules in the rat suggest the plasticity and conservation of class Ib genes in rodent evolution. Functions of TL may be substituted with CD1 or other class Ib molecules expressed by rat thymus.

SUMMARYExperimental autoimmune uveitis (EAU) is an organ-specific autoimmune disease and has served as a model of certain ocular inflammatory conditions in man. The present study was aimed at separating the effects of MHC and non-MHC genes on the development of EAU in the rat. EAU-susceptible LEW (RT1l) EAU-resistant WKAH (RTIk), and WKAH.1L (RTF) MHC congenic strain of WKAH background rats were immunized with retinal soluble antigen (S-Ag) in Freund's complete adjuvant (FCA). LEW rats showed typical EAU, while neither WKAH nor WKAH. 1L congenic rats developed EAU. However, when an additional i.v. injection of Bordetella pertussis was given, all rat strains developed EAU. Furthermore, when immunized with peptide M, an 18-mer synthetic peptide, which corresponds to amino acid positions 303–320 of bovine S-Ag, and given an additional i.v. injection of B. pertussis, LEW and WKAH. IL rats developed EAU, whereas WKAH did not. When ACI (RTIavl), BUF (RTIb), LEJ (RTIl), W(RTIk), F344(RTI1v1), BN (RTIq), NIG-III (RTIq), TO (RT1I), and SDJ (RT1u) rats were immunized with peptide M or S-Ag and then given B. pertussis, all strains developed EAU by immunization with S-Ag plus B. pertussis, but only F344 and NIG-III developed EAU by immunization with peptide M. These findings suggest that susceptibility to EAU in rats is controlled by both MHC and non-MHC genes; and that in the absence of B. pertussis adjuvant, the form of disease induced by native S-Ag in FCA is governed by non-MHC gene(s). However, this effect of non-MHC gene(s) could no longer be observed when the rats were also injected with B. pertussis adjuvant at sensitization.

The class II region of major histocompatibility complex of the rat. Rattus norvegicus (RT1) consists of RT1.B, RT1.D, and RT1.H subregions. The gene order around the H subregion was determined as RT1.A--H beta-H alpha--B by RFLP analysis of naturally occurring intra-RT1 recombinant rats with HLA DP probes. A unique recombinant strain, LEJ, was found to have its recombinational site between H beta and H alpha (RT1.AuH beta uH alpha bBbDb). Northern analysis of class II mRNAs showed that transcripts of RT1.D alpha, RT1.D beta, and RT1.B alpha shared identical sizes among various strains of rats, but RT1.B beta mRNA showed allele-specific size heterogeneities. Northern hybridization with HLA DP alpha probes detected possible RT1.H alpha transcripts. On the other hand, no clear signal of H beta was observed. BDIX whose RT1.B products had not been identified was found to transcribe B alpha and B beta mRNAs.

The RT1-B/D region-associated antigens which were serologically defined in the previous study (Ohhashi et al., 1981), were partially purified from membranes of a rat B cell leukaemia, KNL-14. Sequential immunoprecipitation test, with the partially purified 125I-B/Dak preparation using four different rat alloantisera, including a monoclonal antibody, disclosed three distinctive populations of beta units of the class II molecules. Highly purified beta units of three discriminable class II molecules were shown to have different structural properties in terms of molecular weights and of electrophoretic profiles on the isoelectric focusing. The beta units shifted to a position of higher molecular weight on SDS-PAGE under reducing condition, thus suggesting to carry intradisulfide bonds. Furthermore, the highly purified beta units cross-reacted with murine anti-Ia sera. The rebinding test revealed that at least two discriminable species of beta units cross-react with anti-I-Ak monoclonal antibody, whereas beta units purified by binding with the 1E4 monoclonal antibody cross-reacted with anti-I-Ab and/or anti-I-Ad antiserum. On the basis of structural and antigenic properties, we have postulated that the rat class II region can be divided into at least three subregions, each containing a locus which encodes a distinctive beta unit of the class II molecule.

Summary Seven different alloantisera absorbed with red blood cells (RBC) from appropriate strains of rats detected a series of B cell alloantigenic specificities that could be divided into two groups, presumably coded for by at least two different closely-linked loci in the rat major histocompatibility complex (MHC), RT1. The one locus had two allele codes for a broad specificity and the other locus codes for a unique specificity that was found only in the restricted strains of rats that shared the same mixed lymphocyte reaction (MLR) phenotype. RBC-absorbed alloantisera were monitored against a panel of B cell fractions obtained from sixteen inbred strains. Two alloantisera, ACI anti-W and W anti-TO detected two broad specificities, into either of which all inbred strains tested were classified. Two broad RT1-B region-associated specificities were thus designated provisionally as Ba-1.1 and -1.2. Another five alloantisera detected four respective specificities which have a narrower strain distribution. Sixteen inbred strains were classified into one of five specificities detected by W anti-F344, F344 anti-SDJ, WKA anti-ACI, W anti-BUF and ACI anti-W absorbed with LEJ lymph node cells. Each specificity was designated provisionally as Ba-2.1, -2.2, -2.4, -2.6, -2.7, respectively. A complete association of Ba-2 specificities with MLR phenotype was observed. Antigenic specificities of Ba-2.1, -2.2 and -2.4 were all classified in a group of Ba-1.1 specificity, whereas Ba-2.6 and -2.7 specificities were associated with Ba-1.2 specificity. This relationship suggested a linkage disequilibrium between the two loci for the Ba antigens.

The lymph node cells from 11 strains of rats, differing in the genotype of the major histocompatibility complex of the rat (RT1), were examined on the basis of their proliferative response to the cell wall antigen of Streptococcus mutans. The 11 rat strains fell into three groups: high, intermediate, and low responders. To demonstrate the influence of the major histocompatibility complex on immune responsiveness to S. mutans, further experiments were performed using the RT1-congenic rat strains WKAH.1L(LEW), WKAH. 1AV1(ACI), and WKAH.1J(LEJ), which differ only in the genotype of the RT1 region. Although the background genes of each strain were of WKAH origin, WKAH.1L(LEW) and WKAH.1AV1(ACI) rats showed a low response whereas WKAH.1J(LEJ) rats showed a moderate response to the S. mutans cell wall antigen. The results indicate that the immune response is controlled by the class II gene(s) in RT1. Furthermore, the RT1.D locus products were shown to play an important role in the restriction molecule, since a monoclonal antibody, HOK7, directed to the RT1.Dk locus products reduced the proliferative response of lymph node cells.

The immune response to bovine insulin (BI) in the rat is controlled by the major histocompatibility complex (Mhc)-linked immune response gene (Ir-BI) and immune suppression gene (Is-BI). In the present study, we investigated the low responsiveness to BI in the WKAH rat (RT1k) and attempted to explore the functional link between Is-BI and Mhc class II molecules. Lymph node cells (LNC) from the low responder (WKAH) rats responded well to BI when a large amount of antigen was added to the culture in vitro or after OX8-bearing (OX8+) T cells were eliminated. These LNC, after the elimination of OX8+ cells, could show the RT1.Dk-restricted proliferative response upon in vitro challenge with BI, BI-B chain, or pork insulin. In addition, OX8+ T cells, which were activated with BI and antigen-presenting cells (APC) in vitro, suppressed the anti-BI response of W3/25-bearing proliferating T cells from BI-immunized rats. The results have demonstrated that proliferating T-cell repertoires do exist to BI, which recognize BI-B chain in the context of RT1.Dk molecules in the WKAH rat, and that the state of low responsiveness is mediated to a great extent by antigen-specific OX8+ suppressor T (Ts) cells. Furthermore, the elimination of APC or the addition to RT1.Bk-specific monoclonal antibody in the in vitro secondary activation culture of Ts cells diminished the suppressive activity of OX8+ Ts cells. In the induction phase of Ts cells it therefore seems to be necessary for these cells to recognize BI together with RT1.Bk molecules on APC.

The immune response to bovine or pork insulin (BI or PI, respectively) was studied in the rat using the in vitro insulin-induced lymphocyte-proliferation assay. Results indicated that 11 inbred rat strains were divided into categories of high and low responders. Two high responders, SDJ (RT1u) and BN(RT1n) inbred rat strains, appeared to recognize different antigenic determinant(s) on the insulin molecule. The results of linkage and segregation analyses in F1, F2, backcross, and partially congenic rats showed that the Ir gene (Ir-BI), which encodes the high responsiveness in the SDJ rats, is inherited associated with RT1u, whereas the immune suppression gene (Is-BI), which encodes the low responsiveness in the WKA(RT1k) rats, is inherited together with RT1k. The Is-BI is the first major histocompatibility complex (MHC)-linked Is gene reported in the rat. The LEJ(RTI-AuBb) inbred rat strain showed a low response to BI, indicating that Ir-BI is closer to RTI-B/RTI-D region than to RTI-A.

Les differences concernant RT1-D ont un effet plus important que celles concernant RT1-B. L'incompatibilite concernant les 2 regions a la fois entraine un rejet de type aigu. Dans certains cas des greffons comportant une incompatibilite RT1 peuvent survivre indefiniment. Les foies des donneurs F1 survivent plus longtemps que dans les combinaisons parentales