Gene mapping in a murine cell line by immunoselection with cytotoxic T lymphocytes.
Minor histocompatibility (H) loci encode alloantigens that are recognized by cytotoxic T (Tc) lymphocytes. A (C57BL/10 x 129)F1-derived transformed lymphocyte cell line was immunoselected in vitro with cloned Tc cells that were specific for H-3aa, a Chromosome 2-encoded minor H antigen. This cell line is heterozygous at H-3a (former symbol, Cd-1) and other loci. Three groups of antigen-loss variants were identified. One group contained mutations affecting only the antigen-encoding gene. Another group probably arose through a single homologous interchromosomal exchange, resulting in extensive regions of loss of heterozygosity (LOH). The third group of variants contained an interstitial LOH, one of which was shown to be a significant deletion. Several deletion boundaries were identified, one of which ordered the closely linked H-3a and beta 2-microglobulin (B2m) genes. We suggest that Tc immunoselection against minor H antigens is a promising approach for targeting negative selection to specified chromosomal regions and can provide high-resolution genetic map information.
Zuberi
A R
AR
Jackson Laboratory, Bar Harbor, Maine 04609.
Dudley
M E
ME
Christianson
G J
GJ
Roopenian
D C
DC
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics*; D018949:Antigens, CD1; D002461:Cell Line, Transformed; D002874:Chromosome Mapping / Q000379:methods*; D003602:Cytotoxicity, Immunologic; D005260:Female; D005796:Genes; D006579:Heterozygote; D007621:Karyotyping; D008297:Male; D051379:Mice / Q000235:genetics*; D008810:Mice, Inbred C57BL; D009154:Mutation; D012641:Selection, Genetic; D017384:Sequence Deletion; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D001613:beta 2-Microglobulin / Q000235:genetics
D015703:Antigens, CD; D018949:Antigens, CD1; D001613:beta 2-Microglobulin