Fu T M, Bonneau R H, Epler M, Tevethia M J, Alam S, Verner K, Tevethia S S
Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey 17033, USA.
Virology. 1996 Aug 1;222(1):269-74. doi: 10.1006/viro.1996.0419.
CD8+ cytotoxic T-lymphocytes recognize small epitope peptides in association with MHC class I molecules expressed on the cell surface. In this study, we have determined whether an 8 amino acid viral CTL epitope, when expressed in a cellular protein, can be appropriately processed, presented, and recognized by the corresponding epitope-specific CTL and whether it is capable of inducing a CTL response in vivo. An H-2Kb-restricted CTL epitope from herpes simplex virus type 1 (HSV-1) glycoprotein B (gB epitope, residues 498-505) was cloned into the mouse dihydrofolate reductase protein (DHFR) at amino acid position 87. The recombinant DHFRs were expressed in vaccinia virus recombinants. To distinguish the recombinant DHFR proteins from the endogenous DHFR, an antibody epitope, recognized by monoclonal antibody PAb 901 and derived from simian virus 40 (SV40) T antigen was tagged to the C-termini of recombinant DHFR proteins. In vivo expression of recombinant DHFR was demonstrated by immunoprecipitation with the monoclonal antibody PAb 901. The H-2b cells infected with recombinant vaccinia virus expressing the recombinant DHFR were specifically lysed by gB epitope-specific CTL. Furthermore, the recombinant DHFR was functional in inducing a long lasting HSV gB epitope-specific CTL response upon immunization of C57BL/6 (B6) mice. These results indicate that a viral epitope expressed in a cellular protein can be efficiently processed, presented, and recognized by epitope-specific CTL and show that cellular proteins expressing CTL epitopes can be used for induction of CD8+ T lymphocyte responses.
CD8 + 细胞毒性T淋巴细胞识别与细胞表面表达的MHC I类分子相关的小表位肽。在本研究中,我们确定了一个8氨基酸的病毒CTL表位,当在细胞蛋白中表达时,是否能被相应的表位特异性CTL适当地加工、呈递和识别,以及它是否能够在体内诱导CTL反应。来自单纯疱疹病毒1型(HSV - 1)糖蛋白B的H - 2Kb限制性CTL表位(gB表位,第498 - 505位氨基酸)被克隆到小鼠二氢叶酸还原酶蛋白(DHFR)的第87位氨基酸处。重组DHFR在痘苗病毒重组体中表达。为了将重组DHFR蛋白与内源性DHFR区分开来,一个由单克隆抗体PAb 901识别的、源自猿猴病毒40(SV40)T抗原的抗体表位被标记到重组DHFR蛋白的C末端。用单克隆抗体PAb 901进行免疫沉淀证明了重组DHFR在体内的表达。感染表达重组DHFR的重组痘苗病毒的H - 2b细胞被gB表位特异性CTL特异性裂解。此外,重组DHFR在免疫C57BL / 6(B6)小鼠时能诱导持久的HSV gB表位特异性CTL反应。这些结果表明,在细胞蛋白中表达的病毒表位可以被表位特异性CTL有效地加工、呈递和识别,并表明表达CTL表位的细胞蛋白可用于诱导CD8 + T淋巴细胞反应。