A Modified Peptide Stimulation Method for Efficient Amplification of Cytomegalovirus (CMV)-Specific CTLs  

作　　者：Guangping Ruan Li Ma Qian Wen Wei Luo Mingqian Zhou Xiaoning Wang 

机构地区：[1]Institute of Molecular Immunology, School of Biotechnology, SouthernMedical University, Guangzhou 510515, China [2]School of Biosciences ＆ Bioengineering, South China University ofTechnology, Guangzhou 510641, China

出　　处：《Cellular & Molecular Immunology》2008年第3期197-201,共5页中国免疫学杂志（英文版）

基　　金：This work was supported by grants from the National Natural Science Foundation of China(30771952,30771971);the Program for New Century Excellent Talents in University(NCET-07-0410);the Natural Science Foundation of Guangdong Province(07117783).

摘　　要：CMV-specific immunity is essential for control of human cytomegalovirus （HCMV） infection. Stem cell transplantation is used widely in the management of a range of diseases of the hemopoietic system. Patients are immunosuppressed profoundly in the early posttransplant period, and reactivation of cytomegalovirus （CMV） remains a significant cause of morbidity and mortality. Adoptive transfer of CMV-specific CD^8＋ T cell clones has been shown to reduce the rate of viral reactivation; however, the ex vivo production of cells for adoptive transfer is labor intensive and expensive. We report here a modified peptide stimulation method using CMV-specific epitope peptides to stimulate PBMCs for generation of CMV-specific CTLs. This method permits efficient amplification of CMV-specific CTLs and provides a large number of cells for FACS analysis from a single blood sample. Significantly, it achieves high frequencies of tetramer staining of CD^8＋ T cells allowing the data of different individuals to be easily compared and sequentially evaluated. Thus, this approach expands and selects HLArestricted CMV-pp65-reactive T-cell lines of high specificity for potential adoptive immunotherapy.CMV-specific immunity is essential for control of human cytomegalovirus （HCMV） infection. Stem cell transplantation is used widely in the management of a range of diseases of the hemopoietic system. Patients are immunosuppressed profoundly in the early posttransplant period, and reactivation of cytomegalovirus （CMV） remains a significant cause of morbidity and mortality. Adoptive transfer of CMV-specific CD^8＋ T cell clones has been shown to reduce the rate of viral reactivation; however, the ex vivo production of cells for adoptive transfer is labor intensive and expensive. We report here a modified peptide stimulation method using CMV-specific epitope peptides to stimulate PBMCs for generation of CMV-specific CTLs. This method permits efficient amplification of CMV-specific CTLs and provides a large number of cells for FACS analysis from a single blood sample. Significantly, it achieves high frequencies of tetramer staining of CD^8＋ T cells allowing the data of different individuals to be easily compared and sequentially evaluated. Thus, this approach expands and selects HLArestricted CMV-pp65-reactive T-cell lines of high specificity for potential adoptive immunotherapy.

关 键 词：cytotoxic T lymphocyte CYTOMEGALOVIRUS tetramer assay 

分 类 号：R36[医药卫生—病理学]