| Flow cytometric detection of perforin upregulation in human CD8 T cells. | |
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MedLine Citation:
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PMID: 18615597 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Perforin and granzymes work synergistically to induce apoptosis in target cells recognized by cytotoxic T lymphocytes. While perforin is readily detectable by flow cytometry in resting CD8 T cells, upregulation of perforin in activated cells is thought to require proliferation. However, perforin undergoes numerous conformational changes during its maturation, which may affect the ability of conventional antibodies to recognize newly synthesized perforin. Polychromatic flow cytometry was used to detect perforin and cytokine production following stimulation of ex vivo human CD8 T cells. Two different anti-perforin antibodies, clones B-D48 and deltaG9, were used to discriminate various forms of perforin after cellular activation. We provide evidence for the rapid upregulation of perforin protein, which may contribute to the ability of CD8 T cells to kill multiple targets over time. The deltaG9 clone recognizes the granule-associated conformation of perforin, while the B-D48 clone is able to detect perforin in multiple forms. Finally, we show there is variability in the ability of CD8 T cells to upregulate perforin. Human CD8 T cells are capable of new perforin production immediately following activation. This work defines a novel flow cytometric procedure that can be used to more completely assess the cytotoxic capacity of human CD8 T cells. |
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Authors:
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Adam R Hersperger; George Makedonas; Michael R Betts |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Cytometry. Part A : the journal of the International Society for Analytical Cytology Volume: 73 ISSN: 1552-4930 ISO Abbreviation: Cytometry A Publication Date: 2008 Nov |
Date Detail:
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Created Date: 2008-10-29 Completed Date: 2008-12-09 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101235694 Medline TA: Cytometry A Country: United States |
Other Details:
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Languages: eng Pagination: 1050-7 Citation Subset: IM |
Affiliation:
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Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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CD8-Positive T-Lymphocytes
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metabolism* Clone Cells Flow Cytometry / methods* Granzymes / genetics, metabolism Humans Kinetics Lymphocyte Activation Perforin / chemistry, genetics* Protein Conformation RNA, Messenger / genetics, metabolism Staining and Labeling Up-Regulation / genetics* |
| Chemical | |
Reg. No./Substance:
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0/RNA, Messenger; 126465-35-8/Perforin; EC 3.4.21.-/Granzymes |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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