On the role of the transmembrane anchor sequence of influenza hemagglutinin in target cell recognition by class I MHC-restricted, hemagglutinin-specific cytolytic T lymphocytes.
We have examined the requirement for the transmembrane hydrophobic anchor sequence of the influenza hemagglutinin (HA) in the formation of the antigenic moiety on the surface of target cells recognized by class I MHC-restricted murine CTL. For this analysis we have used a line of CV-1 monkey epithelial cells that express the transfected murine H-2Kd gene product as target cells and have used recombinant SV40-based late replacement vectors to achieve expression of genes encoding wild-type and mutant forms of HA. We have found that the majority of Kd-restricted HA-specific CTL clones recognize target cells that express a secreted HA molecule that lacks the transmembrane and cytoplasmic domains of the parent glycoprotein. Several Kd-restricted CTL clones that recognize subtype-specific and crossreactive epitopes on HA fail to recognize the anchor-negative, secreted HA or chimeric HA molecules containing the transmembrane and cytoplasmic domains of unrelated glycoproteins. These CTL clones appear to be directed to antigenic epitopes located within the transmembrane domain of HA, as defined by their capacity to recognize target cells sensitized with a synthetic 23-amino-acid peptide corresponding to sequences within this domain. The implications of these results for class I MHC-restricted CTL recognition are discussed.
D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D000956:Antigens, Viral; D002460:Cell Line; D002999:Clone Cells / Q000276:immunology; D004274:DNA, Recombinant; D000939:Epitopes / Q000276:immunology; D006183:H-2 Antigens / Q000276:immunology; D006388:Hemagglutinins / Q000235:genetics / Q000276:immunology*; D008285:Major Histocompatibility Complex; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D009975:Orthomyxoviridae / Q000276:immunology*; D013539:Simian virus 40 / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection
D000954:Antigens, Surface; D000956:Antigens, Viral; D004274:DNA, Recombinant; D000939:Epitopes; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D006388:Hemagglutinins; D008565:Membrane Proteins