P1 and cosmid clones define the organization of 280 kb of the mouse H-2 complex containing the Cps-1 and Hsp70 loci.

A 280 kilobase (kb) contig was isolated from mouse genomic P1 and cosmid libraries, using as probes human cDNA and genomic DNA fragments that map in the interval between the second component of complement and tumor necrosis factor genes of the HLA complex. The clone contig demonstrates synteny of eleven mouse genes that are homologous to genes initially mapped within the human major histocompatibility complex. These include the mouse homologs of BAT2 (HLA-B-associated transcript 2) through BAT9 and also three HSP70-related genes. Five P1 clones form a contig of 240 kb that spans from BAT9 through BAT3. Twelve cosmid clones are arranged in three contigs that confirm most of the structure of the P1 contig and link the mouse BAT3 homolog to the BAT2 homolog approximately 15 kb farther telomeric. Polymorphic DNA markers within the cloned region were used to map the cleft palate susceptibility-1 (Cps-1) locus to the interval between Hsp70.1 and BAT6 (valyl-tRNA synthetase). This refines the location of the Cps-1 locus to a 45 kb region contained in the H2-124 P1 insert.

Association between alleles of the transforming growth factor-alpha locus and the occurrence of cleft lip.

DNA samples from 100 patients with cleft lip with or without cleft palate (CL/P) were compared with those of 98 unaffected control individuals with respect to transforming growth factor alpha (TGFA) genotypes. Among the Caucasians in this population (83 CL/P, 84 controls), there was a significant difference in the restriction fragment length polymorphisms (RFLPs) observed after digestion with TaqI (chi 2 = 4.68, P = 0.03). The frequency of the C2 allele in the Caucasian CL/P population was 0.169, whereas that in the control group was 0.089. When the data for Caucasians, African-Americans, and Asians were examined jointly, the chi 2 value for the pooled sample was 5.08 (P = 0.02). This confirms the hypothesis of Ardinger et al. [1989, Am J Hum Genet, 45:348-353] that TFGA itself or a closely linked gene contributes to the development of CL/P in humans.

Restriction fragment length polymorphisms, glucocorticoid receptors, and phenytoin-induced cleft palate in congenic strains of mice with steroid susceptibility differences.

A gene that affects susceptibility to cortisone-induced cleft palate maps between H-2S and H-2D on mouse chromosome 17. Congenic mouse strains that differ at this locus, designated Cps-1 (cleft palate susceptibility-1), have been tested for the presence of several closely linked markers. All data obtained so far are consistent with a gene order of H-2S-Cps-1-BAT-5-BAT-2-TNF-H-2D. The Cps-1 gene does not appear to affect the level of glucocorticoid receptors or the susceptibility of mice to phenytoin-induced cleft palate.

Chromosome assignments of the genes for glucocorticoid receptor, myelin basic protein, leukocyte common antigen, and TRPM2 in the rat.

We have utilized rat-mouse somatic cell hybrids to make chromosomal assignments for the glucocorticoid receptor (GR), myelin basic protein (MBP), leukocyte common antigen (LCA), and testosterone-repressed prostate message-2 (TRPM2) genes in the rat. The genes for GR and MBP both map on chromosome 18 of the rat, which corresponds to the mapping of both genes on chromosome 18 of the mouse. The gene for LCA maps on chromosome 13, which is where C4b-binding protein beta-chain (C4BPB), coagulation factor V (F5), and renin have previously been assigned. This linkage group appears to be homologous to a substantial portion of mouse chromosome 1 and human chromosome 1q. Finally, the TRPM2 gene has been assigned to rat chromosome 15.

Genetic control of resistance to clinical EAE accompanied by histological symptoms.

The susceptibility of rats to experimental allergic encephalomyelitis (EAE) induced by myelin basic protein (MBP) was studied in a variety of genetic crosses. Rats were evaluated according to weight loss, neurological symptoms, and histological criteria. The results demonstrate that three different types of genes are involved in susceptibility. An RT1-linked gene is necessary but not sufficient for full expression of EAE induced by MBP in complete Freund's adjuvant (CFA). Additional genes are required for the occurrence of histological EAE, but a full-blown inflammatory reaction is not sufficient for the expression of clinical EAE. A third type of gene, which can be demonstrated in appropriate crosses, is required for the consistent expression of clinical symptoms. Dominant genes for resistance to clinical symptoms were transferred to the Lewis (LEW) background from the BN.B1 strain through two generations of backcrossing. Thus, there are genetically controlled mechanisms involved in the neurological expression of EAE which are independent of the inflammatory reaction as observed in central nervous system (CNS) histology.

Class I and II major histocompatibility complex gene product expression by a rat insulinoma cell line in vitro following exposure to gamma interferon.

A study of Class I and II major histocompatibility complex gene product expression by a rat insulinoma cell line (RINm5F) was performed using monoclonal antibodies and immunoperoxidase techniques. RINm5F cells were incubated with different concentrations of gamma interferon. RINm5F cells exhibit low levels of Class I molecules and are normally devoid of Class II gene products. Upon exposure to gamma interferon, RINm5F cells showed a dramatic increase in Class I expression. This expression was homogenous and could be detected on all cells after 18 h of incubation with as little as 1 unit/ml of interferon. In contrast, de novo Class II expression was not homogeneous and required 36 h of incubation with 10 units/ml of interferon. The number of RINm5F cells expressing Class II antigens was dose- and time-dependent. Interferon treatment did not affect the morphology of RINm5F cells as determined by ultrastructural analysis. Withdrawal of interferon from the culture medium for as long as 78 h diminished but did not abolish the expression of Class I and Class II molecules already induced. The ability of interferon to enhance expression of Class I gene products and induce de novo expression of Class II molecules on B-cell-derived RINm5F cells supports the hypothesis that aberrant expression of major histocompatibility complex gene products on pancreatic B cells may be an important factor in triggering the immune response in Type 1 (insulin dependent) diabetes mellitus.

Structural studies of the class II histocompatibility antigens of the ACI rat.

The class II antigens of the ACI rat were studied using both conventional alloantisera and monoclonal antibodies. By sequential immunoprecipitation experiments and cell binding studies, alloantisera were shown to contain antibodies to both the RT1.B and the RT1.D gene products. Using one- and two-dimensional gel electrophoresis, the structures of these gene products were shown to be distinguishable. The importance of these differences for the immune response and antigen presentation is discussed.

A Class II monoclonal antibody specific for the RT1.B, rather than the RT1.D, product of the rat major histocompatibility complex.

An allospecific monoclonal antibody, 79.7.5., has been shown to be specific for a Class II histocompatibility product of the ACI (RT1.AaBaDa) rat. To further specify the reaction of this antibody to the B or D locus Class II products of RT1, we examined the binding of this antibody to peripheral blood lymphocytes (PBLs) of the WRC rat (haplotype RT1.AnBnDa). Radiolabelled monoclonal antibody 79.7.5. did not bind to PBLs from the WRC rat, but it did bind to PBLs from the WRA rat (RT1.AdBaDa) and the DA (RT1.AaBaDa) rat. These results were confirmed using radiolabelled Staphylococcus protein A in an indirect binding assay. In addition, binding of 79.7.5 could be inhibited by alloantiserum BN anti-BN.1A (DA) (directed against AaBaDa) but not by BN anti WRC (directed against Da). These data demonstrate that monoclonal antibody 79.7.5. reacts with the a allele product of RT1.B rather than RT1.D. This antibody can be used to probe the structures and functional roles of different Class II products of the rat major histocompatibility complex.