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Journal of Virology, May 2000, p. 3948-3952, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Generation of Cytomegalovirus-Specific Human T-Lymphocyte Clones by Using Autologous B-Lymphoblastoid Cells with Stable Expression of pp65 or IE1 Proteins: a Tool To Study the Fine Specificity of the Antiviral Response

Christelle Retière,1 Virginie Prod'homme,1 Berthe-Marie Imbert-Marcille,2 Marc Bonneville,1 Henri Vié,1 and Marie-Martine Hallet1,*

Institut National de la Santé et de la Recherche Médicale U4631 and Laboratoire de Virologie,2 Institut de Biologie, 44093 Nantes Cedex 1, France

Received 10 November 1999/Accepted 29 January 2000

Cytotoxic T lymphocytes (CTLs) play a central role in the control of persistent human cytomegalovirus (HCMV) infection in healthy virus carriers. Previous analyses of the specificity of HCMV-reactive CD8+ CTLs drawn from in vitro models in which antigen-presenting cells were autologous fibroblasts infected with laboratory HCMV strains have shown focusing of CTL responses against the major tegument protein, pp65. By contrast, the 72-kDa major immediate-early protein (IE1) was identified as a minor target for this response. Here we have studied the fine specificity and T-cell-receptor features of T-cell clones generated against autologous B lymphoblastoid cell lines stably transfected with HCMV cDNA coding for either pp65 or a natural variant of IE1. This strategy allowed efficient generation of T-cell clones against IE1 and pp65 and led to the identification of several new IE1 and pp65 epitopes, including some located in polymorphic regions of IE1. Such an approach may provide relevant information about the characteristics of the CTL response to IE1 and the effect of viral polymorphism on the immune response against HCMV.

* Corresponding author. Mailing address: INSERM U463, Institut de Biologie, 9 quai Moncousu, 44093 Nantes Cedex 1, France. Phone: (33) 02-40-08-47-12. Fax: (33) 02-40-35-66-97. E-mail: mmhallet{at}nantes.inserm.fr.

Journal of Virology, May 2000, p. 3948-3952, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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