Flow cytometric detection of degranulation reveals phenotypic heterogeneity of degranulating CMV-specific CD8+ T lymphocytes in rhesus macaques
In: Journal of Immunological Methods, Jg. 325 (2007-08-01), S. 20-34
Online
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Zugriff:
Flow-cytometric conditions for detection of lysosomal-associated membrane proteins (LAMPs) on the surface of recently degranulated cells were optimized for rhesus macaques and used to investigate the functional properties of rhesus cytomegalovirus (rhCMV)-specific CD8+ T lymphocytes with regards to cytotoxicity and interferon (IFN)-gamma secretion in six asymptomatic CMV-seropositive rhesus macaques. Unlike humans, the rhesus macaque LAMP-1 protein CD107a underwent little or no endocytosis over a six to 18 h stimulation period. Following in vitro stimulation, rhCMV-specific CD8+ T lymphocytes were heterogeneous with regards to the composition of cells positive for CD107a and/or IFN-gamma, time to reach peak degranulation, and kinetics of IFN-gamma secretion relative to degranulation. Responder CD8+ T lymphocytes that underwent degranulation without IFN-gamma production (CD107a+IFN-gamma-) were predominantly composed of terminally differentiated effectors (CD28-CD45RA+). Moreover, they had significantly lower frequencies of effector memory (CD28-CD45RA-) cells compared to the IFN-gamma-secreting cells that did or did not undergo degranulation (CD107a+IFN-gamma+ or CD107a-IFN-gamma+). The perforin content of effector CD8+ T lymphocytes was significantly greater than that of effector memory CD8+ T lymphocytes in rhesus macaques, suggesting that they were more cytolytic. Our findings suggest that the composition of rhCMV-specific CD8+ T lymphocytes with regards to CD107a+IFN-gamma- responders may be an important determinant of their ability to control CMV replication.
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Flow cytometric detection of degranulation reveals phenotypic heterogeneity of degranulating CMV-specific CD8+ T lymphocytes in rhesus macaques
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Autor/in / Beteiligte Person: | Kaur, Amitinder ; Chan, Kenneth S. |
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Zeitschrift: | Journal of Immunological Methods, Jg. 325 (2007-08-01), S. 20-34 |
Veröffentlichung: | Elsevier BV, 2007 |
Medientyp: | unknown |
ISSN: | 0022-1759 (print) |
DOI: | 10.1016/j.jim.2007.05.011 |
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