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Nucleic Acids Research, 1987, Vol. 15, No. 24 10551-10567
© 1987

Articles

The complete sequence of the MHC class II chain RT1D{alpha}u of the diabetic BB rat: mRNA levels of RT1.D{alpha} in lymphocytes

Eugene W. Holowachuk, Mary-Kay Greer and Diego R. Martin

Banting and Best Department of Medical Research, The C.H.Best Institute, University of Toronto 112 College Street, Toronto, Ontario M5G 1L6, Canada

Received June 8, 1987. Revised October 21, 1987. Accepted October 21, 1987.

The major histocompatibility complex of the rat (RT1 complex) encodes two sets of class II molecules referred to as RT1.B and RT1.D. The complete structure of the RT1.D{alpha}u chain of the diabetes prone BB rat was determined by the isolation and characterization of a full size cDNA. Comparisons of the nucleotide and protein sequences of RT1 D{alpha} with the analogous molecules, H-2 I-E{alpha} and HLA DR{alpha}, revealed that these alpha chains have been highly conserved during evolution. Southern blot analysis indicated an association of the RT1 haplotypes, ‘u’ and ‘l’, with Bam H1 DNA bands of 9.8 kb and 11.7 kb, respectively.

The BB rat develops insulin dependent diabetes as an autoimmune abnormality Accumulating evidence suggests a cellular mediated etiology and the involvement of class II molecules. The steady state levels of RT1.D{alpha} mRNA were measured in splenic lymphocytes of diabetes prone BB rats and age matched histocompatible normal nondiabetic WF rats by a RNase protection assay. Compared to WF rats, elevated transcripts of RT1.D{alpha} were found in lymphocytes of young BB rats (~4x and ~2.5x greater at 20–40 d and 40–75 d, respectively). In lymphocytes of older diabetic and nondiabetic BB rats (>75 d) the levels of RT1.D{alpha} mRNA were lower than in the young BB rats and were found at the WF control levels. The increased steady state RT1.D{alpha} mRNA levels in the young BB rats may reflect differences in the proportion of splenic lymphocytes expressing this gene (activated lymphocytes), and thus differences in splenic lymphocyte populations. The steady state RT1.D{alpha} mRNA levels in lymphocytes of the normal rats were found to be relatively similar at all ages examined. The increased class II gene transcripts found in lymphocytes of young BB rats indicates that they possess a highly activated immune system.


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