Ozols D Y, Harnish D G, Rawls W E, Rosenthal K L
Department of Pathology, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada.
Arch Virol. 1990;115(3-4):209-25. doi: 10.1007/BF01310531.
Pichinde virus (PV) infection of mice results in induction of a strong H-2 restricted, virus-specific cytotoxic T lymphocyte (CTL) response and rapid clearance of the virus. To define the specificities of CTL induced by PV infection, we constructed vaccinia virus recombinants containing cloned cDNAs corresponding to full-length (VVNP) and a truncated form (VVNP 51-561) of the nucleoprotein (NP) gene of PV. Radioimmunoprecipitation analysis of infected cell lysates indicated that VVNP expressed a PV-specific product identical in size to that of authentic NP, while vaccinia virus recombinants containing truncated NP produced a polypeptide consistent with the synthesis of amino acids 51-561 of Pichinde virus NP. Interestingly, cells infected with VVNP synthesized easily detectable, but much lower levels of nucleoprotein relative to both PV and VVNP51-561. Primary virus-specific CTL induced in three different strains of inbred mice following intravenous infection with PV were able to lyse syngeneic target cells infected with PV but did not markedly lyse syngeneic targets expressing full-length or truncated NP following recombinant vaccinia virus infection. Similarly, secondary anti-PV specific CTL generated following in vitro restimulation by PV or selectively restimulated with vaccinia recombinants did not significantly lyse target cells expressing NP. Further, infection of mice with VVNP and VVNP51-561 did not induce CTLs specific for PV and did not prime mice for the generation of memory anti-PV CTL in vivo. These results suggest that PV gene products other than NP, such as the GPC or L protein, contain the major target epitope(s) recognized by PV-specific CTL.
小鼠感染皮钦德病毒（PV）会诱导产生强烈的H - 2限制的、病毒特异性细胞毒性T淋巴细胞（CTL）反应，并使病毒快速清除。为了确定由PV感染诱导的CTL的特异性，我们构建了痘苗病毒重组体，其包含与PV核蛋白（NP）基因的全长（VVNP）和截短形式（VVNP 51 - 561）相对应的克隆cDNA。对感染细胞裂解物的放射免疫沉淀分析表明，VVNP表达一种PV特异性产物，其大小与天然NP相同，而含有截短NP的痘苗病毒重组体产生一种与皮钦德病毒NP的氨基酸51 - 561合成一致的多肽。有趣的是，感染VVNP的细胞合成了易于检测到的核蛋白，但相对于PV和VVNP51 - 561，其水平要低得多。在三种不同品系的近交小鼠经静脉感染PV后诱导产生的原代病毒特异性CTL能够裂解感染PV的同基因靶细胞，但在重组痘苗病毒感染后，对表达全长或截短NP的同基因靶细胞没有明显的裂解作用。同样，在体外经PV再刺激或用痘苗重组体选择性再刺激后产生的继发性抗PV特异性CTL，对表达NP的靶细胞也没有明显的裂解作用。此外，用VVNP和VVNP51 - 561感染小鼠不会诱导产生针对PV的CTL，也不会在体内使小鼠产生记忆性抗PV CTL。这些结果表明，除NP外的PV基因产物，如GPC或L蛋白，包含PV特异性CTL识别的主要靶表位。
