Details

Autor(en) / Beteiligte

• Long, Eric O.
• Wake, Claire T.
• Strubin, Michel
• Gross, Nicole
• Accolla, Roberto S.
• Carrel, Stefan
• Mach, Bernard

Titel

Isolation of Distinct cDNA Clones Encoding HLA-DR β Chains by Use of an Expression Assay

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 1982-12, Vol.79 (23), p.7465-7469

Ort / Verlag

United States: National Academy of Sciences of the United States of America

Erscheinungsjahr

1982

Quelle

MEDLINE

Beschreibungen/Notizen

• cDNA clones encoding different human Ia antigen β chains were isolated by use of a complementation-expression assay in Xenopus oocytes. The assay was based on two previous findings. First, oocytes injected with mRNA from a human B-cell line express HLA-DR antigen. The three intracellular DR chains are assembled in oocytes and can be immunoprecipitated with anti-DR monoclonal antibodies. Second, we have isolated cDNA clones encoding DR α and intermediate chains. In order to identify β -chain cDNA clones, mRNA was hybrid-selected with pools of cDNA clones, mixed with mRNA for the α and intermediate chains, and injected into oocytes. We isolated two distinct clones that could select DR β -chain mRNA as demonstrated by assembly of the translation product with DR α chains and immunoprecipitation with DR-specific monoclonal antibodies. One clone is specific for a β chain of the DR locus. The other clone, much weaker in its ability to select DR mRNA, encodes another Ia-like β chain. Full-length cDNA clones corresponding to the DR and Ia-like β chains were isolated and compared. Cross-hybridization was detectable in the coding regions but not in the 3′untranslated regions. Distinct RNAs homologous to the DR and the Ia-like β -chain clones were present in B cells but were undetectable in three T-cell lines.