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B Lymphoblastoid Cell Lines as Efficient APC to Elicit CD8⁺ T Cell Responses Against a Cytomegalovirus Antigen¹

Qi Sun^*, Robert L. Burton^*, Li-Jun Dai^*, William J. Britt and Kenneth G. Lucas^2,*

^* Hematology and Oncology, School of Medicine, University of Alabama, and Department of Pediatrics, The Children’s Hospital of Alabama, Birmingham, AL 35294

Potent and readily accessible APC are critical for development of immunotherapy protocols to treat viral disease and cancer. We have shown that B lymphoblastoid cell lines (BLCL) that stably express CMV phosphoprotein 65 (BLCLpp65), as a result of retroviral transduction, can be used to generate ex vivo CTL cultures that possess cytotoxicity against CMV and EBV. In this report, we demonstrate that the EBV-specific cytotoxicity in the BLCLpp65-primed culture had a spectrum of EBV-Ag recognition similar to that of the BLCL-primed counterpart, suggesting that retroviral transduction and expression of the CMV Ag would not compromise the Ag-presenting capacity of BLCL. In addition, BLCLpp65 appeared to present multiple natural pp65 epitopes, because pp65-specific CTL, which recognized different CMV clinical isolates, were generated in BLCLpp65-primed cultures from individuals with various HLA backgrounds. Consistent with a polyclonal expansion of virus-specific CTL, T cell lines established from the BLCLpp65-primed CTL cultures expressed different TCR-V_ß. Although most of the virus-specific T cell isolates were CD8⁺, EBV-specific CD4⁺ lines were also established from BLCLpp65-primed cultures. Western blot analysis revealed that the CD8⁺ lines, but not the CD4⁺ line, expressed granzyme B, consistent with features of classic CTL. Thus, our results suggested that BLCL stably expressing a foreign Ag might be used as a practical APC to elicit CD8⁺ T cell responses.

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