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Clinical and Diagnostic Laboratory Immunology, March 2002, p. 230-235, Vol. 9, No. 2
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.2.230-235.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Depletion of Human NK and CD8 Cells prior to In Vitro H1N1 Flu Vaccine Stimulation Increases the Number of Gamma Interferon-Secreting Cells Compared to the Initial Undepleted Population in an ELISPOT Assay

Christophe Dercamp,,{dagger} Violette Sanchez, Julie Barrier, Emanuelle Trannoy, and Bruno Guy*

Research Department, Aventis Pasteur, 69280 Marcy l'Etoile, France

Received 18 September 2001/ Returned for modification 29 October 2001/ Accepted 20 November 2001

In order to study the respective roles of CD4, CD8, and CD56 (NK) cells in gamma interferon (IFN-{gamma}) production after in vitro stimulation with flu vaccine in a healthy adult human population, we depleted these cellular subtypes before stimulation with antigen (inactivated split vaccine, A/Texas H1N1, or A/Sydney H3N2). We observed that while CD4 cells were required for IFN-{gamma} secretion in both conditions in vitro, CD56 (NK) cells and, to a lesser extent, CD8 cells had a negative effect on such synthesis upon H1N1 stimulation, as judged by an increased number of spots compared to the initial undepleted population. This regulation of IFN-{gamma} secretion was associated with an increase in ICAM-1 expression, in particular on T and B cells. This study points out the importance of evaluating in vitro immune responses on a whole-cell population in addition to isolated subtypes if one needs to address potential cellular interactions occurring in vivo in some situations (H1N1 stimulation in the present case). Such cross-regulations occur even in vitro during the antigenic stimulation step.

* Corresponding author. Mailing address: Research Department, Aventis Pasteur, Campus Merieux, 1541 Av. M. Merieux, 69280 Marcy l'Etoile, France. Phone: 33 (0)4 37-37-38-75. Fax: 33 (0)4 37-37-36-39. E-mail: bruno.guy{at}aventis.com.

{dagger} Present address: Schering Plough, 69571 Dardilly, France.

Clinical and Diagnostic Laboratory Immunology, March 2002, p. 230-235, Vol. 9, No. 2
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.2.230-235.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.





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