Antigen Presentation and Cytotoxic T Lymphocyte Killing Studied in Individual, Living Cells
| Autor: | D. Lansing Taylor, R L DeBiasio, Antoinette Tishon, Michael B. A. Oldstone, Hanna Lewicki, Greg LaRocca, Jean Edouard Gairin, Klaus M. Hahn |
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| Rok vydání: | 1994 |
| Předmět: |
Microinjections
Molecular Sequence Data Antigen presentation Cell chemical and pharmacologic phenomena Biology Major histocompatibility complex Lymphocytic choriomeningitis Viral Proteins Virology medicine Lymphocytic choriomeningitis virus Cytotoxic T cell Amino Acid Sequence Cells Cultured Antigen Presentation Cell Death Rhodamines Histocompatibility Antigens Class I Videotape Recording hemic and immune systems T lymphocyte medicine.disease Molecular biology In vitro CTL medicine.anatomical_structure Microscopy Fluorescence biology.protein T-Lymphocytes Cytotoxic |
| Zdroj: | Virology. 201:330-340 |
| ISSN: | 0042-6822 |
| Popis: | 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 51 Cr 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 led 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. |
| Databáze: | OpenAIRE |
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