Immunotherapy with peptide MHC tetramer isolated T cells

描述（由申请人提供）：T细胞表型和 高速排序已经结合起来，允许直接测试一个重要的 免疫治疗的基本原则-特别是过继免疫治疗， 抗原特异性T细胞将在体内根除抗原表达细胞。 尽管用体外活化和扩增的T细胞进行免疫治疗已经显示出 有希望，一种替代的广泛适用的策略可能允许治疗， 抗原特异性T细胞在显著活化之前。具体而言，肽 主要组织相容性复合体（MHC）四聚体允许鉴定T 细胞特异性结合独特的纳米肽的背景下， 人类白细胞抗原（HLA）等位基因。这项技术允许 分选和分离高度纯化的抗原特异性T细胞群， 来自外周血单核细胞（PBMC）的细胞。我们假设 用纯化抗原特异性T细胞群进行过继免疫治疗 细胞将在骨移植后免疫抑制的患者中具有临床益处。 骨髓/外周血干细胞移植。相关模型系统测试 这一假设存在于巨细胞病毒免疫功能低下的患者中 (CMV)感染或爱泼斯坦巴尔病毒（EBV）相关的LPD（LPD）。临床 通过施用CMV特异性克隆扩增， 体外和用自体EBV体外活化的少至10（7）个细胞/M2 PBMC 转化的B细胞。虽然这些策略在临床上有效，但它们 不能广泛应用，因为它们是复杂和耗时的， 长至3至4个月以产生用于施用的适当细胞。 一个令人兴奋的替代这个费力的过程将是直接分离 通过高速分选来自外周血样品的CMV或EBV特异性T细胞 CMV或EBV肽MHC四聚体。这一战略将使快速 分离高达10-40 × 10（6）抗原特异性T细胞。这些细胞可以 然后直接给药，或在短暂的离体给药后给药 激活和扩展。本申请建议进行临床前研究， 肽类MHC四聚体过继免疫治疗的初步临床试验 分选的T细胞。预计这些研究将提供重要的 这一一般概念的原理证明，将具有广泛的应用 用于病毒性疾病和癌症的抗原特异性过继免疫治疗。

DESCRIPTION (provided by applicant): Recent advances in T cell phenotyping and high speed sorting have coalesced to allow the direct testing of an important basic tenet in immunotherapy-specifically that adoptive immunotherapy with antigen specific T cells will eradicate antigen expressing cells in vivo. Although immunotherapy with T cells activated and expanded in vitro have shown promise, an alternative broadly applicable strategy may allow therapy with antigen specific T cells prior to significant activation. Specifically, peptide major histocompatibility complex (MHC)-tetramers allow the identification of T cells that specifically bind to unique nanopeptides in the context of a specific human leukocyte antigen (HLA) allele. This technology allows for the sorting and isolation of highly purified populations of antigens specific T cells from peripheral blood mononuclear cells (PBMC). We have hypothesized that the adoptive immunotherapy with purified populations of antigen specific T cells will have clinical benefits in patients immunosuppressed following bone marrow/peripheral blood stem cell transplant. A relevant model system to test this hypothesis exists in immuno-compromised patients with cytomegalovirus (CMV) infection or Epstein Barr virus (EBV) associated LPD (LPD). Clinical benefit has been achieved with administration CMV specific clones expanded in vitro and as few as 10(7) cells/M2PBMC activated in vitro with autologous EBV transformed B cells. While these strategies are clinically effective, they cannot be widely applied because they are complex and time-consuming, taking as long as to 3 to 4 months to generate the appropriate cells for administration. An exciting alternative to this laborious process would be to directly isolate CMV or EBV specific T cells from peripheral blood samples by high speed sorting with CMV or EBV peptide MHC tetramers. This strategy would allow for the rapid isolation of up to 10-40 x 10(6) antigen specific T cells. These cells could then be directly administered, or administered after a brief period of ex vivo activation and expansion. This application proposes pre-clinical studies and pilot clinical trials of adoptive immunotherapy with peptide MHC tetramer sorted T cells. It is anticipated that these studies would provide an important proof of principle for this general concept which would have wide application for antigen specific adoptive immunotherapy of vial disorders and cancer.