ANTIGEN PRESENTATION AND CYTOTOXIC T-LYMPHOCYTE KILLING STUDIED IN INDIVIDUAL, LIVING CELLS

被引:19
|
作者
HAHN, K [1 ]
DEBIASIO, R [1 ]
TISHON, A [1 ]
LEWICKI, H [1 ]
GAIRIN, JE [1 ]
LAROCCA, G [1 ]
TAYLOR, DL [1 ]
OLDSTONE, M [1 ]
机构
[1] CARNEGIE MELLON UNIV, CTR LIGHT MICROSCOPE IMAGING BIOMED SCI, PITTSBURGH, PA 15213 USA
关键词
D O I
10.1006/viro.1994.1298
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Interactions between individual, living fibroblasts and cytotoxic T lymphocyte (CTL) clones were analyzed by using video-enhanced differential interference contrast and fluorescence microscopy in a multimode configuration. Fibroblasts expressing known major histocompatibility complex I alleles (MC57: H-2b; Balb: H-2d) were sensitized for killing by incubating or microinjecting them with peptide fragments of lymphocytic choriomeningitis virus. Previous determination of the CTL clones' specificity for these peptides and MHC-I alleles enabled us to study CTL killing of fibroblasts, and nonlethal CTL interaction with targets due to ''mismatches'' of the CTL, target, and/or peptide. During viral peptide-specific MHC-restricted CTL killing, distinct morphological alterations were observed (CTL shape changes, movements of granules in CTL cytoplasm, and target cell contraction and blebbing). When no killing occurred, CTL engaged in prolonged, nonrandom movement on the target cells. Alloreactive and virus-specific CTL displayed the same morphology during killing. To study antigen presentation further within individual, living cells, a LCMV glycoprotein peptide (aa 272-286, LSDSSGVENPGGYCL) was covalently labeled with tetramethylrhodamine. In Cr-51 release assays, the labeled peptide specifically induced potent CTL killing, but neither labeled nor unlabeled peptide proved toxic for unsensitized targets. Microinjection of the labeled peptide into the cytoplasm of fibroblast cells]ed to CTL killing of those cells, yet nearby uninjected cells contacted by CTL were not killed, indicating that killing was due to presentation of microinjected peptide rather than binding of extracellular peptide to cell surface MHC. Peptide-injected target cells were killed only when combined with CTL specific for the peptide and for the MHC allele of the injected cell. (C) 1994 Academic Press, Inc.
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页码:330 / 340
页数:11
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