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Infection and Immunity, October 2008, p. 4609-4614, Vol. 76, No. 10
0019-9567/08/$08.00+0     doi:10.1128/IAI.00563-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Enumeration of Cytotoxic CD8 T Cells Ex Vivo during the Response to Listeria monocytogenes Infection{triangledown}

Dietmar M. W. Zaiss,{dagger} Alice J. A. M. Sijts,{dagger} and Tim R. Mosmann*

David H. Smith Center for Vaccine Biology and Immunology and Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York 14642

Received 8 May 2008/ Returned for modification 23 June 2008/ Accepted 30 July 2008

Cytotoxicity is a key effector function of CD8 T cells. However, what proportion of antigen-specific CD8 T cells in vivo exert cytotoxic activity during a functional CD8 T-cell response to infection still remains unknown. We used the Lysispot assay to directly enumerate cytotoxic CD8 T cells from the spleen ex vivo during the immune response to infection with the intracellular bacterium Listeria monocytogenes. We demonstrate that not all antigen-responsive gamma interferon (IFN-{gamma})-secreting T cells display cytotoxic activity. Most CD8 T cells detected at early time points of the response were cytotoxic. This percentage continuously declined during both the expansion and contraction phases to about 50% at the peak and to <10% of IFN-{gamma}-producing cells in the memory phase. As described for clonal expansion, this elaboration of a program of differentiation after an initial stimulus was not affected by antigen or CD4 help but, like proliferation, could be influenced by later reinfection. These data indicate that cytotoxic effector function during the response to infection is regulated independently from IFN-{gamma} secretion or expansion or contraction of the overall CD8 T-cell response.

* Corresponding author. Mailing address: David H. Smith Center for Vaccine Biology and Immunology, University of Rochester Medical Center, 601 Elmwood Avenue, Box 609, Rochester, NY 14642. Phone: (585) 275-9120. Fax: (585) 273-2452. E-mail: Tim_Mosmann{at}urmc.rochester.edu

{triangledown} Published ahead of print on 4 August 2008.

Editor: J. L. Flynn

{dagger} Present address: Division of Immunology, Faculty of Veterinary Medicine, University Utrecht, Utrecht, The Netherlands.

Infection and Immunity, October 2008, p. 4609-4614, Vol. 76, No. 10
0019-9567/08/$08.00+0     doi:10.1128/IAI.00563-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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