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Blood, 15 January 2004, Vol. 103, No. 2, pp. 630-638.
Prepublished online as a Blood First Edition Paper on August 28, 2003; DOI 10.1182/blood-2003-03-0824.
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IMMUNOBIOLOGY
Identification of novel CTL epitopes of CMV-pp65 presented by a variety of HLA alleles
Eisei Kondo,
Yoshiki Akatsuka,
Kiyotaka Kuzushima,
Kunio Tsujimura,
Shoji Asakura,
Kohei Tajima,
Yoshitoyo Kagami,
Yoshihisa Kodera,
Mitsune Tanimoto,
Yasuo Morishima, and
Toshitada Takahashi
From the Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan; Department of Hematology and Chemotherapy, Aichi Cancer Center Hospital, Nagoya, Japan; Department of Hematology, Japanese Red Cross Nagoya First Hospital, Nagoya, Japan; and Department of Internal Medicine II, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan.
Cytomegalovirus (CMV)–specific T-cell immunity plays an important role in protection from CMV disease in immunocompromised patients. Identification of cytotoxic T-lymphocyte (CTL) epitopes is essential for monitoring T-cell immunity and also for immunotherapy. In this and previous studies, CMV-pp65–specific CTL lines were successfully generated from all of 11 CMV-seropositive healthy donors, using pp65-transduced CD40-activated B (CD40-B) cells as antigen-presenting cells. By use of enzyme-linked immunospot (ELISPOT) assays, individual CTL epitopes could be mapped with truncated forms of the pp65 gene. For human leukocyte antigen (HLA) alleles with a known binding motif, CTL epitopes within the defined regions were predicted by computer algorithm. For HLA alleles without a known binding motif (HLA-Cw*0801, -Cw*1202, and -Cw*1502), the epitopes were alternatively identified by step-by-step truncations of the pp65 gene. Through this study, a total of 14 novel CTL epitopes of CMV-pp65 were identified. Interestingly, 3 peptides were found to be presented by 2 different HLA class I alleles or subtypes. Moreover, use of CD40-B cells pulsed with a mixture of synthetic peptides led to generation of pp65-specific CTL lines from some of seronegative donors. The study thus demonstrated an efficient strategy for identifying CTL epitopes presented by a variety of HLA alleles.
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