Mathew A, Kurane I, Rothman A L, Zeng L L, Brinton M A, Ennis F A
Division of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical Center, Worcester 01655, USA.
J Clin Invest. 1996 Oct 1;98(7):1684-91. doi: 10.1172/JCI118964.
A severe complication of dengue virus infection, dengue hemorrhagic fever (DHF), is hypothesized to be immunologically mediated and virus-specific cytotoxic T lymphocytes (CTLs) may trigger DHF. It is also likely that dengue virus-specific CTLs are important for recovery from dengue virus infections. There is little available information on the human CD8+ T cell responses to dengue viruses. Memory CD8+CTL responses were analyzed to determine the diversity of the T cell response to dengue virus and to identify immunodominant proteins using PBMC from eight healthy adult volunteers who had received monovalent, live-attenuated candidate vaccines of the four dengue serotypes. All the donors had specific T cell proliferation to dengue and to other flaviviruses that we tested. CTLs were generated from the stimulated PBMC of all donors, and in the seven donors tested, dengue virus-specific CD8+CTL activity was demonstrated. The nonstructural (NS3 and NS1.2a) and envelope (E) proteins were recognized by CD8+CTLs from six, five, and three donors, respectively. All donors recognized either NS3 or NS1.2a. In one donor who received a dengue 4 vaccine, CTL killing was seen in bulk culture against the premembrane protein (prM). This is the first demonstration of a CTL response against the prM protein. The CTL responses using the PBMC of two donors were serotype specific, whereas all other donors had serotype-cross-reactive responses. For one donor, CTLs specific for E, NS1.2a, and NS3 proteins were all HLA-B44 restricted. For three other donors tested, the potential restricting alleles for recognition of NS3 were B38, A24, and/or B62 and B35. These results indicate that the CD8+CTL responses of humans after immunization with one serotype of dengue virus are diverse and directed against a variety of proteins. The NS3 and NS1.2a proteins should be considered when designing subunit vaccines for dengue.
登革出血热（DHF）是登革病毒感染的一种严重并发症，据推测是由免疫介导的，病毒特异性细胞毒性T淋巴细胞（CTL）可能引发DHF。登革病毒特异性CTL对于从登革病毒感染中恢复也可能很重要。关于人类CD8 + T细胞对登革病毒的反应，现有信息很少。我们分析了记忆性CD8 + CTL反应，以确定T细胞对登革病毒反应的多样性，并使用来自八名健康成年志愿者的外周血单个核细胞（PBMC）来鉴定免疫显性蛋白，这些志愿者接种了四种登革血清型的单价减毒活候选疫苗。所有供体对登革病毒和我们测试的其他黄病毒都有特异性T细胞增殖。从所有供体受刺激的PBMC中产生CTL，并且在测试的七名供体中，证明了登革病毒特异性CD8 + CTL活性。非结构蛋白（NS3和NS1.2a）和包膜蛋白（E）分别被六名、五名和三名供体的CD8 + CTL识别。所有供体都识别NS3或NS1.2a。在一名接种登革4疫苗的供体中，在大量培养中观察到针对前膜蛋白（prM）的CTL杀伤作用。这是首次证明针对prM蛋白的CTL反应。使用两名供体的PBMC进行的CTL反应具有血清型特异性，而所有其他供体具有血清型交叉反应。对于一名供体，针对E、NS1.2a和NS3蛋白的CTL均受HLA - B44限制。对于测试的其他三名供体，识别NS3的潜在限制等位基因是B38、A24和/或B62以及B35。这些结果表明，人类接种一种血清型登革病毒后的CD8 + CTL反应是多样的，并且针对多种蛋白。在设计登革亚单位疫苗时应考虑NS3和NS1.2a蛋白。
