Identification of novel CTL epitopes of CMV-pp65 presented by a variety of HLA alleles
Identification of novel CTL epitopes of CMV-pp65 presented by a variety of HLA alleles
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DOI：
10.1182/blood-2003-03-0824
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发表时间：
2004-01-15
期刊：
影响因子：
20.3
通讯作者：
Takahashi, T
中科院分区：
文献类型：
--
作者：
Kondo, E;Akatsuka, Y;Takahashi, T
巨细胞病毒（CMV）特异性t细胞免疫在免疫功能低下患者的巨细胞病毒疾病保护中起重要作用。细胞毒性t淋巴细胞（CTL）表位的鉴定是监测t细胞免疫和免疫治疗的必要条件。在本研究和之前的研究中，使用pp65转导的cd40活化B （CD40-B）细胞作为抗原呈递细胞，成功地从所有11名cmv血清阳性的健康供体中生成了cmv -pp65特异性CTL系。通过酶联免疫斑点（ELISPOT）检测，单个CTL表位可以与pp65基因的截短形式定位。对于具有已知结合基序的人白细胞抗原（HLA）等位基因，通过计算机算法预测确定区域内的CTL表位。对于没有已知结合基序的HLA等位基因（HLA-Cw*0801， -Cw*1202和-Cw*1502），通过逐步截断pp65基因来确定表位。本研究共鉴定出14个新的CMV-pp65 CTL表位。有趣的是，3个肽被发现由2个不同的HLA I类等位基因或亚型呈现。此外，使用CD40-B细胞与合成肽的混合物脉冲导致从一些血清阴性供体产生pp65特异性CTL系。因此，该研究证明了一种有效的策略来识别由多种HLA等位基因呈现的CTL表位。
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 identifled. 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.