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. 1973 Aug;70(8):2401–2405. doi: 10.1073/pnas.70.8.2401

Lambda-Chain Production in Human Lymphoblast-Mouse Fibroblast Hybrids

Stuart H Orkin †,*, Philip D Buchanan ‡, William J Yount †,§,, Howard Reisner †,§,, John W Littlefield †,
PMCID: PMC433744  PMID: 4599625

Abstract

Mutant human lymphoblast cells deficient in hypoxanthine phosphoribosyltransferase (EC 2.4.2.8) activity were hybridized with thymidine kinase (EC 2.7.1.21)-deficient mouse fibroblasts. Hybrid cells were readily selected, as both parental lines were nonreverting and eliminated by hypoxanthine-amethopterinthymidine medium. Human lambda (λ) chain was the only immunoglobulin chain produced by the lymphoblast parent, as determined by immunofluorescent techniques. Two independent hybrid clones chosen for detailed study synthesized human λ chain, and continued to do so after prolonged culture.

As in both parental lines, no human immunoglobulin heavy chains, complements C3 or C4, or α1-antitrypsin, or mouse immunoglobulin chains or complement C5 were detectable in the hybrids. Selection against thymidine kinase-containing hybrid cells with 5-bromodeoxyuridine did not eliminate positive λ-chain reactivity, suggesting that the kinase and λ-chain loci are not linked.

The continued production of an immunoglobulin chain by human lymphoblast-mouse fibroblast hybrids contrasts with the extinction of other differentiated functions in several hybrid systems, and indicates that gene localization and linkage analysis for human immunoglobulin chains should be feasible with this system.

Keywords: immunoglobulins, gene localization, chromosome E-17, HAT selection, differentiation

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Selected References

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