CLINICAL TRIAL: IMMUNOLOGIC STUDIES FROM BONE MARROW DONORS AND VOLUNTEERS FOR T

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 来自几个实验室的先前结果已经表明，来自CMV的pp 65片段蛋白是由对该病毒呈血清阳性的个体识别的主要免疫原性蛋白。 我们的实验室已经鉴定了CTL克隆，其在10种不同的HLA I类等位基因的背景下识别pp 65。 已经为这些克隆定义的表位库代表了美国约95%的种族人口。 评价这些CTL表位是否主要用于一般人群需要分析刺激来自CMV血清阳性的HLA型个体的外周血淋巴细胞的方法的免疫原性，该方法采用一种技术，其中将游离肽表位以高浓度加入微孔培养物中的PBMC中。 我们已经将这种方法用于对pp 65具有特异性并受HLA A*0201、A*1101、A*2402、A*6901和B*0702限制的CTL表位。 在每种情况下，我们已经检查了不到5名健康志愿者，他们是血清反应阳性，并已被证明对他们表达的HLA等位基因特异性的CTL表位有反应。识别肽负载和CMV感染的靶标的CD 8 +CTL在两周内扩增100倍。 体外刺激程序的使用允许灵敏地确定个体是否对CMV产生CTL应答，以及pp 65是否是该应答的组分。 我们希望证明至少5个，如果可能的话，10个随机选择的个体将对单个表位产生应答，这表明表达相同限制性I类等位基因的11个个体对该表位产生普遍应答的潜力。 这为生产疫苗分子的方法提供了理论基础，这些分子含有一个或多个这些表位，以使高危个体免受CMV感染。 获得外周血和活检的临床程序是临床方案的核心部分，最好在GCRC的主持下进行。 我们已经开发了表达HLA等位基因的个体的队列，我们已经从pp 65和最近的pp 150获得了这些等位基因的表位。 我们希望进行这些研究，以表征CMV pp 65和pp 150的所有主要表位，这些表位在人类对病毒的免疫应答中很重要。 进行这些研究的目的是确定一种适用于所有高危个体的基于肽的疫苗。 我们还将继续推导新的CMV表位，以完善我们的表位库，从而确保尽可能完整地代表构成美国人口的不同种族的个体。 本方案有几份知情同意书（A-D）。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Previous results from several laboratories have indicated that the pp65 segament protein from CMV is the major immunogenic protein recognized by individuals who are seropositive for the virus. CTL clones, which recognize pp65 in the context of ten different HLA Class I alleles, have been identified by our laboratory. The repertoire of epitopes that have been defined for these clones are representative of approximately 95% of the ethnic population of the United States. Evaluation of whether these CTL epitopes are predominantly used in the general population requires an analysis of the immunogenecity of the approach of stimulating peripheral blood lymphocytes from HLA-typed individuals who are seropositive for CMV, in a technique in which the free peptide epitope is added at a high concentration to PBMC in microwell cultures. We have used this approach with CTL epitopes specific for pp65 and restricted by HLA A*0201, A*1101,A*2402,A*6901 and B*0702. In each case, we have examined less than five healthy volunteers who are seropositive and have been shown to respond to the CTL epitopes that are specific for the HLA allele, which they express. CD8+CTL, which recognize both peptide loaded and CMV infected targets, are amplified 100-fold in a two-week period. The use of the in vitro stimulation procedure allows a sensitive determination of whether an individual is making a CTL response to CMV, and whether pp65 is a component of that response. We wish to demonstrate that at least five, and if possible, ten randomly chosen individuals will respond to a single epitope, suggesting the potential for a universal response to that epitope from ll individuals who express the same restricting Class I allele. This provides the rationale for an approach to producing vaccine molecules, containing one or more of these epitopes to immunize at-risk individuals against CMV infection. The clinical procedures of obtaining both peripheral blood and biopsy are the central part of the clinical protocol and could be best carried out under the auspices of the GCRC. We have developed cohorts of individuals who express the HLA alleles for which we have epitopes from pp65, and more recently, pp150. We wish to carry out these studies, to characterize all of the major epitopes of CMV pp65 and pp150, which are important in the human immune response to the virus. The purpose for carrying on these studies is to define a peptide-based vaccine that would be applicable to all at-risk individuals. We will also continue to derive new CMV epitopes, to complete our repertoire of epitopes that would ensure as complete a representation as possible of individuals of different ethnicities that make up the American population. There are several consent forms (A-D) which are active for the protocol.