In vitro T-lymphocyte proliferative response to mouse thyroglobulin in experimental autoimmune thyroiditis

The in vitro lymphocyte proliferative response to mouse thyroglobulin (MTg) was studied in good and poor responder mice in relationship to in vivo antibody formation and thyroid infiltration. CBA(H-2^k) and BALB/c(H-2^d) mice were immunized in the hind footpads with MTg incorporated into complete Freund's adjuvant (CFA). At weekly intervals up to 28 days, groups of mice were sacrificed. Their popliteal lymph nodes were cultured in vitro for proliferative response to MTg and their antibody levels and thyroid involvement determined. In good responder CBA mice, the proliferative responses to MTg were strongest on Days 8 to 14, where they were 9- to 14-fold over control levels, depending on the day of harvest. The response declined to 2- to 4-fold over background on Days 21 to 28, although high antibody levels were present throughout this period. The proliferative response was abrogated by anti-Thy-1 treatment, indicating its dependence on T cells. In poor responder BALB/c mice, no significant proliferative responses to MTg were observed at any time, although the animals displayed moderate levels of MTg antibody. The responses to PPD, in contrast, were similar in both strains, usually being 4- to 7-fold above background. Thyroid infiltration, like the proliferative response to MTg, was observed only in CBA mice. Thus lymphocyte proliferation at 8 to 14 days represents a reliable, early in vitro correlate of autoimmune thyroiditis induced with CFA as adjuvant.     

Genetic study of mouse sensitivity to MHV3 infection: influence of the H-2 complex.

Genetic study of acute and chronic mouse hepatitis virus type 3 disease was carried out in segregating generations of a cross involving a susceptible (C57BL/6) and a resistant (A/J) mouse strain. The data obtained indicate that one or two recessive genes may be involved in resistance of acute and chronic diseases but suggest that the genes involved in both diseases are different. In this cross, no correlation was observed between H-2 and acute or chronic disease. In mice of congenic lines, however, A/Sn (H-2a), A.SW (H-2s), A.BY (H-2b), and A.CA (H-2f), it appeared that the presence of the H-2f allele conferred to heterozygote as well as to homozygote animals the capacity to resist the development of chronic disease. It seems, therefore, that MHV3 sensitivity in mice is under the influence of at least two major genes: one for the acute disease and the other, H-2 linked, for the chronic disease.     

Family organization of mouse H-2 class I genes

The H-2 complex of the mouse contains numerous class I genes of unknown function. These genes are here classified into families according to homology in the exons encoding the variable domains. There are one major and at least five minor families, whose members are partly clustered and partly interspersed on the mouse chromosome. DNA sequences show that not only Tla and Qa-2 but also other minor-family genes have intact coding domains. These may be expressible genes of novel types.     

DNA sequence analysis of the C3H H-2Kk and H-2Dk loci. Evolutionary relationships to H-2 genes from four other mouse strains

We generated nucleotide sequences for H-2Kk and H-2Dk from the C3H mouse, as well as for a genomic clone of H-2Db, in order to conduct an evolutionary analysis of the H-2 genes from three haplotypes, k, d, and b. H-2Kk from both the C3H and AKR strains, H-2Kd, H-2Kb, H-2Dk, H-2Ld, H-2Dd, H-2Db, and H-2Dp DNA sequences were aligned, and the alignments used to construct phylogenetic trees inferring the evolutionary relationships among the nine genes by two independent methods. Both approaches yielded trees with similar topologies. In addition, the sequence alignments revealed patterns of nucleotide substitutions which implicate both point mutation and recombination in the divergence of the H-2 genes. Future considerations for evolutionary analysis of class I genes are discussed.     

Non-H-2 and H-2-Linked immune response genes control the cytotoxic T-cell response to H-Y

The immunoregulation of cytotoxic T-cell responses to the male-specific antigen H-Y in mice has been found to be genetically controlled by genes of the major histocompatibility complex (H-2). Responsiveness was mainly confined to H-2 ^b strains, but it has also been found in recombinant strains, F₁ hybrids, and chimeras that carry at least part of the H-2 ^b haplotype. By using a different immunization procedure it has been shown recently that an H-2 ^k mouse strain (CBA) is also able to mount an equivalent H-Y-specific response. We investigate here, by applying this immunization technique, the responsiveness of other H-2 ^k strains and of strains of other independent H-2 haplotypes. Both responders and nonresponders are found in three haplotypes: k, s, and d. The strain distribution pattern of responsiveness shows a combined influence of non-H-2 and H-2 genes. In certain strains there is a high variability in responsiveness between genetically indentical individual animals. We discuss a model of immune response (Ir) gene function which could account for these observations.     

Nematospiroides dubius: two H-2-linked genes influence levels of resistance to infection in mice

Strains of mice sharing common H-2 haplotypes but different genetic backgrounds, and H-2 congenic strains of mice differing only at H-2 genes were studied to assess the role of H-2 and non-H-2 genes in immunity to challenge infections with the nematode parasite Nematospiroides dubius. Strains of mice sharing the H-2k haplotype were uniformly more susceptible to challenge than strains expressing H-2q alleles, regardless of genetic background. However, in some cases strains of mice sharing the k or q haplotypes differed significantly in levels of resistance. Therefore, non-H-2 genes must influence the response observed. H-2 cogenic strains of mice differed markedly in their ability to resist challenge infections. Mice sharing the C57BL/10 background but expressing k alleles were very susceptible to challenge, while the H-2q, H-2f, and H-2s, haplotypes were associated with resistance. Studies of H-2 congenic recombinant strains of mice suggested that two H-2 genes influence the antiparasite response. One of these genes maps to the left of E alpha and the other to the D-end of the H-2 complex. It is concluded also that the unique configuration of H-2 genes in F1 hybrids contributes to increased resistance to challange.     

The IgG2a antibody response to thyroglobulin is linked to the Igh locus in mouse

The IgG-subclass usage by several strains of mice in the response to immunization with mouse thyroglobulin (mTg) was examined in the experimental autoimmune thyroiditis model. While the subclass usage by most mouse strains was similar, the Igh^b allotype-bearing mice consistently produced lower IgG2a levels to mTg. Using CBA-Igh ^b congenic and recombinant inbred strains of mice, the lower level of IgG2a in the Igh^b mouse was mapped to the Igh locus. The regulation of IgG2a appeared to be cis controlled, as the CBA x C57BL/6F₁ mouse also produced reduced IgG2a of the Igh^b (B6) allotype but not Igh^j (CBA) allotype.     

Genetic control of the immune response to the H-2 associated Slp alloantigen in the mouse.

A survey of sixteen standard inbred and congenic resistant strains of mice reveals that the ability to mount an immune response to the Slp allotype is associated with the H-2 type of the recipient. Strains carrying the H-2f, H-2k, and H-2q haplotypes are able to produce specific antibody whereas strains of the H-2b haplotype are non-responders. Analysis of F1-hybrids and four informative intra-H-2-recombinants demonstrates that the ability to respond to the Slp allotype is controlled by a dominant gene associated with the K end of the H-2 complex.     

The male antigen. II. Regulation of the primary and secondary responses to H-Y by H-2 associated genes

The primary response of females of ten inbred mouse strains to the male antigen (H-Y) was investigated by transfer of peritoneal exudate cells (PEC). Three distinct classes of reactivity were seen. Early primary response to H-Y was associated with H-2 haplotypes b and s; intermediate response with H-2 haplotypes, k, d, i, and h; and late or absent response with H-2 haplotypes a and f. The failure of A.CA (H-2^f) females to mount a detectable primary response against syngeneic male cells was not due to the lack of the antigen from A.CA male cells. The ability of (A.CA × B10)F₁ hybrid females to respond to the male antigen demonstrated the dominant nature of the B10 (H-2^b) response and excluded the possibility that A.CA females possess a dominant self-antigen cross-reactive with H-Y. The secondary response of eight inbred strains was investigated; at least three distinct levels of reactivity were apparent. The speed of the secondary response was associated with the various H-2 haplotypes in the same way as the primary response. The implications of differential strain reactivity, background effect, and the association of Ir-1 with response to H-Y are discussed.     

Regulation of the hapten-specific T cell response. I. Preferential induction of hyporesponsiveness to the D-end of the major histocompatibility complex in the hapten-specific cytotoxic T cell response

In this paper we have examined the phenomenon of hapten-specific tolerance in the cytolytic T lymphocyte (CTL), using the trinitrophenyl (TNP) and azobenzenearsonate haptens. We found that the H-2 K and H-2 D-end restricted CTL in H-2a mice are differentiable in the ease with which they are tolerized to the TNP hapten. With TNP modified syngeneic spleen cells (TNP-SC), or low amounts of trinitrobenzylsulfonic acid as tolerogen, preferential hyporesponsiveness of D-end restricted CTL can be observed. Larger doses of hapten, e.g. a higher amount of trinitrobenzylsulfonic acid, will tolerize both K- and D-end restricted TNP-specific CTL in H-2a mice. The phenomenon of preferential D-end restricted CTL hyporesponsiveness is not observed in H-2d, H-2k, or H-2b mice, nor is it observed in H-2a mice with respect to the azobenzenearsonate hapten. We have also shown that the clones of CTL responsible for lysis of TNP-modified allogeneic targets (cross-reactive lysis) in H-2a mice probably overlap with the D-end restricted TNP-specific CTL since D-end restricted hyporesponsiveness induced by intravenous injection of TNP spleen cells also results in the elimination of cross-reactive lysis of TNP-modified allogeneic targets. The possible mechanisms of preferential D-end hyporesponsiveness to the TNP hapten in the H-2a mice as well as its significance and relationship to previous work in this area are discussed in this paper.     

Regulation of antitubercular immunity in mice by genes of the H-2 complex

Development of DTH reaction and survival time after M. tuberculosis H37Rv infection have been studied in H-2 congenic and recombinant mice pretreated with high doses of BCG vaccine. In addition, in vitro proliferation of lymphocytes from infected CBA, B6 and 4R mice to PPD was studied in the presence of anti-I-A and anti-I-E mAbs. High doses of BCG vaccination (1 mg/mouse) have led to a significant inhibition of DTH and diminution of survival time in B10.M (H-2f) mice only, and to opposite effects in all other strains tested (H-2a, b, d, k, h4). In I-A+, I-E- 4R mice anti-I-Ak mAbs abrogated lymphocyte proliferation to PPD completely, while in I-A+, I-E- CBA mice only the mixture of anti-I-Ak and anti-I-Ek mAbs was effective.     

Immune response to the Slp alloantigen in the mouse:H-2-linked qualitative and quantitative control

Immunization of inbred mouse strains lacking the Slp allotype results in the production of Slp antibodies in some strains but elicits no detectable response in other strains. Analysis of standard inbred and congenic resistant strains reveals that both the qualitative and quantitative ability to respond to the Slp allotype is associated with theH-2 haplotype of the recipient. Three different response phenotypes can be identified utilizing complement fixation and quantitative immunodiffusion tests. Strains which carry theH-2 ^q haplotype are high responders,H-2 ^k strains are intermediate in response, whileH-2 ^b andH-2 ^v strains produce no detectable antibody. The characteristic response patterns of high and intermediate responders were manifest by day 35 of immunization and continued as discrete response types after a second booster. Quantitative data in the immune response of the intra-H-2 recombinant B10.A(4R) suggest that the recombination event which established theH-2 ^h4 chromosome disturbs the proper function of the genetic determinant controlling response to Slp.