Previous studies in our laboratory using primed lymphocyte typing (PLT) in an informative family have led to the identification of four traits and two regions, separable by recombination within the HLA-DR system. In the present study, we analysed the heterogeneity of HLA-DR antigens among members of this family as a first step towards defining the molecular entities responsible for the observed cellular reactivities. Cell lines of various genotypes derived from the members of this family were labelled biosynthetically with 35S-methionine. The total glycoproteins and immunoprecipitates prepared with two monoclonal anti-HLA-DR antibodies, two monoclonal anti-mouse Ia antibodies cross-reactive with HLA-DR, and a rabbit anti-HLA-DR (anti-p27,33) antiserum were fractionated by two-dimensional gel electrophoresis. In agreement with previous results, HLA-DR light chains were clearly polymorphic whereas the heavy chains looked rather monomorphic. The light chain subunits either segregated with one haplotype, or were shared by some but not all haplotypes or were common to all haplotypes. Whereas the haplotype-specific polypeptides correlated best with HLA-DR3 and HLA-DR5 specificities, the shared subunits may have corresponded to the cross-reactivities observed by PLT. Comparison of the patterns obtained with the various monoclonal antibodies and with the rabbit antiserum revealed that each monoclonal antibody bound a subset of HLA-DR light chains, all of which were present in the anti-p27,33 precipitates. This rabbit antiserum and one of the monoclonal antibodies immunoprecipitated a set of heavy chains not bound by the other 3 antibodies. Since overlapping pools of light chains were present in all five immunoprecipitates, these results suggest that different heavy chains may associate with the same light chains.

Animals↗

PubMedDate not supplied

Structure of the Ki-ras gene of the human lung carcinoma cell line Calu-1.

The homologue of the viral Kirsten ras (v-Ki-ras) gene found in the human lung carcinoma cell line, Calu-1, has an intron-exon structure similar to that of the human homologue of the viral Harvey ras (v-Ha-ras) gene. A second, potential fourth coding exon is present in the human Ki-ras gene and similar sequences are found in the Kirsten murine sarcoma virus. Cysteine is encoded at the twelfth amino acid position, suggesting that the Calu-1 Ki-ras gene has undergone a mutational activation at the same position as the human Ha-ras gene of the bladder carcinoma cell line, T24. A comparison of their predicted amino acid sequences suggests that ras proteins have a 'constant' region and a 'variable' region. Here we propose a common modular structure for ras gene products in which the variable region forms a physiologically important combining site.

Amino Acid Sequence↗

PubMedDate not supplied

Hypothesis: more mutations to cure cancer?

Cancer results from the accumulation of selected mutations. We propose here that some combinations of mutations may be counterselected. To illustrate this, we show an analysis of allelotyping in human breast tumors in which we demonstrate that specific associations of genetic events are statistically under-represented. This emerging concept could be used in a new therapeutical approach of cancer.

Breast Neoplasms↗

PubMedDate not supplied