CMV, ADV and EBV Viral Cytotoxic T-Lymphocytes Generated by a Novel Cytokine Capture System in Children, Adolescents and Young Adults with Refractory Viral Infection and T-Cell Immunodeficiency

项目摘要 儿童、青少年和青壮年(CAYA)，患有原发性T细胞免疫缺陷(PID)和/或 异基因干细胞移植后继发性T细胞免疫缺陷 难治性CMV、ADV和/或EBV感染和/或对抗病毒抗生素不耐受的人 预后(≥90%病死率)。CMV、ADV和EBV的难治性感染并不常见，这 代表美国的一种孤儿疾病(约250名患者)。令人惊讶的是，没有FDA 批准了对这种罕见患者群体的治疗。近年来，病毒特异性细胞毒的过继T细胞治疗 T淋巴细胞(CTL)目前处于早期临床研究阶段。这种方法利用了快速的 供者外周血单核细胞特异性刺激后病毒特异性CTL的分离技术 使用INF-g细胞因子捕获系统(Miltenyi)分离带有病毒衍生多肽的细胞和CTL 神童)。在单中心研究的少数患者中，这种方法是可行的、安全的和 很有效。这种新的细胞治疗方法在孤儿群体中的可行性、安全性和有效性 在更大的多中心，目前缺乏前瞻性研究。因此，我们假设部分人类白细胞抗原 利用Miltenyi生产的匹配(半相合)相关供者来源的CMV、ADV和EBV CTL CliniMACS Prodigy细胞因子捕获系统在任一PID的CAYA中都是可行、安全和有效的 有药物难治性病毒感染和/或对抗病毒药物不耐受的allSCT后和/或SID 抗生素。我们将通过三个大型多中心病毒CTL试验(IND17449)来研究这一假设 病毒CTL联盟(VIRCTLC)。具体的主要目标和次要目标包括(简要)：1)可行性 CMV、ADV和EBV CTL的生产和输注的安全性；2)病毒特异性CTL的有效性(消除 QRT-PCR法检测的病毒载量低于机构值下限；3)单倍体一致的持久性 供体病毒CTL输注；继发：4)发生AGVHD的可能性；5)无病毒和总体的可能性 存活；6)CMV、ADV和EBV的遗传、蛋白质组和免疫学特性的表征 CTL；以及7)特异性T细胞免疫重建、功能和与病毒细胞治疗反应的相关性。 实验设计包括(简要)：1)用病毒特异性抗原直接分离产生病毒CTL 多肽刺激和细胞因子捕获分离；2)三期II开放标记前瞻性研究；3)病毒CTL 有效性和功能性；4)CD3供体嵌合体的病毒CTL持久性；以及5)病毒CTL 通过流式细胞术、飞行时间(CyTOF)质量细胞术、单细胞细胞因子分析、 磷蛋白质组学分析，T细胞免疫图谱，T细胞受体(TCR)多样性和频率 免疫组织化学和单细胞mRNA测序。长期目标是获得可行性、安全性。 这种疗法的有效性将导致FDA批准由以下公司生产的CMV、ADV和EBV病毒CTL Miltenyi CliniMACS Prodigy细胞因子捕获系统在这一孤儿患者群体中的应用。

Project Summary Children, adolescents and young adults (CAYA) with a primary T-cell immunodeficiency (PID) and/or secondary T-cell immunodeficiency following allogeneic stem cell transplantation (AlloSCT) and a medically refractory CMV, ADV and/or EBV infection and/or who become intolerant to antiviral antibiotics have a dismal prognosis (≥90% mortality rate). Refractory infections with CMV, ADV and EBV are uncommon and this represents an Orphan Disease (approximately 250 patients) in the United States. Strikingly, there is no FDA approved therapy for this rare patient population. Recently, adoptive T-cell therapy with viral specific cytotoxic T-lymphocytes (CTLs) is currently under early phase clinical investigations. This approach utilizes a rapid technique of viral specific CTL isolation following specific stimulation of donor peripheral blood mononuclear cells with viral-derived peptides followed by CTL isolation utilizing an INF-g Cytokine Capture System (Miltenyi Prodigy). In a small number of patients in single center studies, this approach has been feasible, safe and efficacious. The feasibility, safety and efficacy of this novel cellular therapy approach in this orphan population in larger multicenter prospective studies is presently lacking. We therefore hypothesize that partially HLA matched (haploidentical) related donor derived CMV, ADV and EBV CTLs manufactured utilizing the Miltenyi CliniMACS Prodigy Cytokine Capture System will be feasible, safe and effective in CAYA with either PID and/or SID following AlloSCT with medically refractory viral infections and/or those intolerant to antiviral antibiotics. We will investigate this hypothesis through three large multicenter viral CTL trials (IND17449) via the Viral CTL Consortium (VIRCTLC). The specific primary and secondary aims include (Brief): 1) feasibility and safety of CMV, ADV and EBV CTL production and infusion; 2) efficacy of viral specific CTLs (elimination of specific viral load by qRT-PCR below the lower limit of institutional values; 3) persistence of haploidentical donor viral CTL infusion; secondary: 4) probability of developing AGVHD; 5) probability of viral free and overall survival; 6) characterization of the genetic, proteomic and immunological properties of CMV, ADV and EBV CTLs; and 7) specific T-cell immune reconstitution, function and correlation to response to viral cell therapy. The experimental design including (brief): 1) viral CTL production by direct isolation following viral specific peptide stimulation and cytokine capture isolation; 2) three phase II open label prospective studies; 3) viral CTL validation and functionality; 4) viral CTL persistence by CD3 donor chimerism; and 5) viral CTL characterization by flow cytometry, mass cytometry by time of flight (CYTOF), single cell cytokine analysis, phosphoproteomics analysis, T-cell immunoprofiling, T cell receptor (TCR) diversity and frequency by Immunoseq, and single cell mRNA sequencing. The long term objectives are to obtain the feasibility, safety and efficacy of this therapy that will lead to FDA approval of CMV, ADV and EBV viral CTLs manufactured by the Miltenyi CliniMACS Prodigy Cytokine Capture System in this orphan patient population.