Identification of I-E alpha genes in H-2 recombinant mouse strains by F1 complementation

W. P. Lafuse, S. Savarirayan, J. F. McCormick, C. S. David

Research output: Contribution to journalArticle

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Abstract

Nine recombinant H-2 mouse strains with crossovers in the I region between I-E-negative haplotypes f,q and I-E-positive haplotypes p,k were examined for I-E expression by microtoxicity dye exclusion assay. These recombinants were found to be negative for I-E expression. There are two possible genotypes in these recombinant mouse strains that could result in lack of I-E expression. Recombinants with crossovers between the E alpha gene and the S region would have both nonexpressed I-E alpha and beta genes (E(β)(f)E(α)(f), E(β)(q)E(α)(q)) and recombinants with crossovers between the E(β) and E(α) genes would have a nonexpressed E(β) gene (E(β)(f) or E(α)(q)) and a functional E(α) gene (E(α)(k) or E(α)(p)). To distinguish between these possible genotypes these recombinants were crossed to B10.A(4R), which carries a functional E(α)(k) gene but is I-E-negative due to a nonexpressed E(α)(b) gene. F1 mice were examined for transcomplementing I-E molecules by immunoprecipitation of 3H-leucine-radiolabeled detergent lysates of spleen cells with a monoclonal I-E antibody (14-4-4). Detection of a transcomplementing I-E molecule was confirmed by immunoprecipitation with a monoclonal antibody (H9-14.8) specific for the I-E(k) beta polypeptide chain derived from B10.A(4R) and by tryptic peptide map comparisons. Five recombinant mouse strains were able to complement with B10.A(4R) in F1 mice to generate a transcomplementing I-E molecule, and thus have an expressed I-E alpha gene (E(α)(k) or E(α)(p)). Four recombinants did not complement with B10.A(4R) in the F1 expression of I-E molecules, and thus have nonexpressed I-E alpha genes (E(α)(f) or E(α)(q)).

Original languageEnglish (US)
Pages (from-to)297-301
Number of pages5
JournalTransplantation
Volume43
Issue number2
StatePublished - 1987

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Genes
Immunoprecipitation
Haplotypes
Genotype
Peptides
Leucine
Detergents
Coloring Agents
Spleen
Monoclonal Antibodies
Antibodies

ASJC Scopus subject areas

  • Immunology
  • Transplantation

Cite this

Lafuse, W. P., Savarirayan, S., McCormick, J. F., & David, C. S. (1987). Identification of I-E alpha genes in H-2 recombinant mouse strains by F1 complementation. Transplantation, 43(2), 297-301.

Identification of I-E alpha genes in H-2 recombinant mouse strains by F1 complementation. / Lafuse, W. P.; Savarirayan, S.; McCormick, J. F.; David, C. S.

In: Transplantation, Vol. 43, No. 2, 1987, p. 297-301.

Research output: Contribution to journalArticle

Lafuse, WP, Savarirayan, S, McCormick, JF & David, CS 1987, 'Identification of I-E alpha genes in H-2 recombinant mouse strains by F1 complementation', Transplantation, vol. 43, no. 2, pp. 297-301.
Lafuse WP, Savarirayan S, McCormick JF, David CS. Identification of I-E alpha genes in H-2 recombinant mouse strains by F1 complementation. Transplantation. 1987;43(2):297-301.
Lafuse, W. P. ; Savarirayan, S. ; McCormick, J. F. ; David, C. S. / Identification of I-E alpha genes in H-2 recombinant mouse strains by F1 complementation. In: Transplantation. 1987 ; Vol. 43, No. 2. pp. 297-301.
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abstract = "Nine recombinant H-2 mouse strains with crossovers in the I region between I-E-negative haplotypes f,q and I-E-positive haplotypes p,k were examined for I-E expression by microtoxicity dye exclusion assay. These recombinants were found to be negative for I-E expression. There are two possible genotypes in these recombinant mouse strains that could result in lack of I-E expression. Recombinants with crossovers between the E alpha gene and the S region would have both nonexpressed I-E alpha and beta genes (E(β)(f)E(α)(f), E(β)(q)E(α)(q)) and recombinants with crossovers between the E(β) and E(α) genes would have a nonexpressed E(β) gene (E(β)(f) or E(α)(q)) and a functional E(α) gene (E(α)(k) or E(α)(p)). To distinguish between these possible genotypes these recombinants were crossed to B10.A(4R), which carries a functional E(α)(k) gene but is I-E-negative due to a nonexpressed E(α)(b) gene. F1 mice were examined for transcomplementing I-E molecules by immunoprecipitation of 3H-leucine-radiolabeled detergent lysates of spleen cells with a monoclonal I-E antibody (14-4-4). Detection of a transcomplementing I-E molecule was confirmed by immunoprecipitation with a monoclonal antibody (H9-14.8) specific for the I-E(k) beta polypeptide chain derived from B10.A(4R) and by tryptic peptide map comparisons. Five recombinant mouse strains were able to complement with B10.A(4R) in F1 mice to generate a transcomplementing I-E molecule, and thus have an expressed I-E alpha gene (E(α)(k) or E(α)(p)). Four recombinants did not complement with B10.A(4R) in the F1 expression of I-E molecules, and thus have nonexpressed I-E alpha genes (E(α)(f) or E(α)(q)).",
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