Graft-versus-leukemia

移植物抗白血病

• 批准号: 8149487
• 负责人: Austin John Barrett
• 金额: $ 327.4万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8149487
• 关键词: Aftercare; Age; Age-Years; Allogenic; Avidity; CD3 Antigens; CD34 gene; CD8B1 gene; CSF3 gene; Chronic; Clinical; Cyclophosphamide; Cyclosporine; Cyclosporins; Cytomegalovirus; DNA Viruses; Development; Disease; Donor Lymphocyte Infusion; Dose; Dyes; Environment; FCGR3B gene; Follow-Up Studies; Genotype; Goals; Growth Factor; Hemoglobin; Human; Immune system; Immunity; Immunosuppression; Immunosuppressive Agents; Immunotherapy; In Vitro; Incidence; Infection; K-562; Laboratories; Laboratory Research; Lymphoid; Malignant neoplasm of testis; Mesenchymal; Natural Killer Cells; Organ; Peripheral Blood Stem Cell; Photosensitizing Agents; Screening procedure; Stem cell transplant; Stromal Cells; Vaccinated; Variant; base; cytopenia; follow-up; graft vs host disease; immune function; improved; in vivo; killer immunoglobulin-like receptor; leukemia; preclinical study; safety study

(1) T cell interactions with leukemia cells: We have found that T cells recognizing all the leukemia antigens so far tested occue in normal individuals, and that after SCT T cells specific for several antigens increase in what appears to be a permissive environment for antigen-specific T cell expansion in the first weeks after transplant. This finding underpins development of new trials currently awaiting regulatory approval involving vaccinating donors with PR1 and WT1 nonapeptides and boosting patients with the same vaccines early after SCT. Two clinical trials - a safety study where patients with myeloid malignancies received a single dose of PR1 and WT1 vaccine and a second study where they received multiple doses of vaccine over a 16 week period demonstrated efficacy in increasing T cell frequencies to either or both vaccines in all 8 patents. However multiple vaccination resulted in loss of T cell responses and a loss of high avidity T cells recognizing the leukemia peptides suggesting the induction of tolerance. Currently our clinical grade vaccines are restricted to HLA A2 individuals and only stimulate CD8 Tcells. We are screening a peptide library of WT1 to identify peptides which induce other HLA class 1 molecules and also CD4 T cells so as to design a more widely applicable and more potent vaccine. (2) Natural Killer (NK) cell GVL: In a retospective analysis of over 270 SCT we found that stem cell donors with the KIR genotype 2DS1, 3DS1, 2DL5a conferred significantly less relapse in recipients with acute myelogenous leukemia (AML). A subsequent study of functional and phenotypic differences in NK cells between donors with and without this "favorable" genotype has not so far identified any differences. (3) GVL effect of mismatched T cells: We have shown that alloresponding T cells exist in both the naive and memory T cell compartments. memory alloreactive T cells often show cross reactivity with DNA virus antigens of CMV and EBV viruses. Using T cell receptor gene sequencing we confirmed that individual T cell clones can show dual reactivity against viral antigens and HLA mismatched target cells. To study whether cross-reactivity might present a limitation to the use of third party adoptive transfer of virus specific T cell clones we studied cross reactivity of virus specific T cells given to patients with post SCT viral reactivation and showed that in vivo such cross-reactivity did not result in GVHD. We are now exploring whether mismatched alloreactive T cell lines can be generated using an artificial genetically modified K562 antigen-presenting cell to provide GVL effects and also whether alloreacting T cell lines generated primarily against DNA viruses also have GVL effects and could be used to treat relapsed leukemia after SCT. (4) Immune recovery after SCT : We measured plasma cytokine profiles and lymphocyte repertoire to determine the optimum time to boost graft-versus-leukemia (GVL) effects after transplant and to characterize immune recovery in different transplant protocols. We found two patterns of cytokine expression - an early surge of lymphoid and myeolid growth factors in the first few days after transplant and a later surge of lymphokines released around 4 weeks after transplant. We are now extending cytokine profiling to compare patterns in autologous and T cell replete SCT. We will correlate recovery of different lymphocyte subsets with individual cytokine profiles to determine the factors which boost optimum recovery. (5) Selective depletion of alloreactive T cells: A clinical trial to selectively deplete T cells from the SCT that can cause GVHD and further translational work to develop improved techniques to selectively photodeplete activated T cells using rhodamine-based dyes has now been completed. We evaluated the ability of this photodepletion technique to selectively deplete graft-versus-host disease (GVHD) alloreacting T cells from stem cell transplants. Donor lymphocytes were stimulated with irradiated in-vitro expanded recipient T lymphocytes. Alloactivated T cells preferentially retaining the photosensitizer 4, 5-dibromorhodamine 123 (TH9402) (Kiadis Pharma, NL) were eliminated by light exposure. Twenty-four patients with hematological malignancies (16 high-risk) conditioned with fludarabine, cyclophosphamide, and total body irradiation received a CD34-selected stem cell allograft from an HLA identical sibling and 5x106/kg selectively depleted (SD) donor T cells. Low-dose cyclosporine was used as the only post-transplant immunosuppression. The overall probability of grade III-IV acute-GVHD was 13%. Fourteen patients developed (predominantly limited) chronic GVHD (c-GVHD). Five patients relapsed, 2 of whom remain alive in remission after further treatment. Thirteen patients survive at a median of 22 months. Overall survival and disease free survival actuarial probabilities (+ SEM) were 43 + 13% and 35 + 13% respectively. The SD technique resulted in a low incidence of relapse (24 + 10%), but was complicated by late non-relapse mortality associated with c-GVHD and infection of 50 + 14% at 4 years follow up. These results suggest that SD may effectively reduce severe a-GVHD while conserving graft-versus leukemia effects. (6) Mesenchymal stromal cells (MSC): Preclinical studies exploring the interaction of MSC with cytotoxic T cells showed that MSC can reduce cytotoxicity of T effector cells against leukemia lines. Further studies are warranted to explore a possible suppressive effect of MSC infusions on GVL reactivity. We have worked with the Clinical Center Cell Processing Section to develop clinical grade MSC for three trials covering the use of MSC to treat GVHD , poor marrow function and major organ damage. (7) A Clinical trial of T cell depleted SCT as a platform for immunotherapeutic approaches is ongoing. Thirty-six patients with hematologic malignancies underwent allogeneic HSCT from their HLA-identical siblings. The median age was 43 years (range 16-68). Transplant indications were acute leukemia (24), other myeloid malignancy (9), other lymphoid malignancy (3). Subjects received myeloablative conditioning regimen with cyclophosphamide, fludarabine and total body irradiation (12 Gy with lung shielding to 6 Gy). Subjects 55 years of age and older received 4 Gy. G-CSF mobilized peripheral blood stem cells from the donor were CD34+ selected utilizing the Miltenyi CliniMacs system, with infusion of a target CD34+ dose of 6 x 106/kg (range 3 to 10 x 106/kg) and a fixed CD3+ dose of 5 x 104 /kg. Patients received low-dose cyclosporine alone till day 21. Delayed lymphocyte add back (5 x 106 CD3+/kg) was given on day +90. Day 200 overall survival was 79%. Thirty-four subjects achieved complete donor (>95%) myeloid chimerism by day 14 and the median time to complete donor CD3+ chimerism was 45 days. The incidence of chronic GVHD was 34.3%. At a median follow up of 3.6 years, Kaplan-Meier estimates of relapse, nonrelapse mortality and overall survival were 35%, 33% and 44% respectively. These results represent acceptable engraftment and clinical outcomes. The trial is ongoing. A trial of alemtuzemab in LGL leukemia is ongoing Patients had chronic cytopenia, increased clonal CD3+CD8+ CD16+ or CD57+ T-LGL. After 1 mg test dose, alemtuzumab was administered at 10 mg/d for 10 days. Overall nine (56%; 95% CI 30%-80%) patients responded to alemtuzumab with five achieving a CR and four a PR. Of the 6 anemic patients who responded to therapy the median pre-treatment hemoglobin was 7.3 (range, 5.4-9.3) g/dL and the median post-treatment hemoglobin was 12.2 (range 10-13.7) g/dL. Three responders with an ANC < 500 /cu mm pre-treatment improved to an ANC > 1,000/cu mm after alemtuzumab. This study showed that alemtuzumab was safe and showed promising activity in pre-treated T-LGL patients.

(1) T细胞与白血病细胞的相互作用：我们发现正常个体中存在识别迄今为止所测试的所有白血病抗原的T细胞，并且在SCT后，对几种抗原具有特异性的T细胞在看似允许抗原的环境中增加- 移植后第一周内特异性 T 细胞扩增。这一发现支持了目前正在等待监管部门批准的新试验的开发，这些试验涉及为捐赠者接种 PR1 和 WT1 九肽疫苗，并在 SCT 后早期对患者加强接种相同的疫苗。 两项临床试验——一项安全性研究，骨髓恶性肿瘤患者接受单剂 PR1 和 WT1 疫苗；另一项研究，他们在 16 周内接受多剂疫苗，证明了其中一种或两种疫苗在增加 T 细胞频率方面的功效。全部8项专利。然而，多次疫苗接种导致 T 细胞反应丧失和识别白血病肽的高亲合力 T 细胞丧失，表明诱导了耐受性。目前，我们的临床级疫苗仅限于 HLA A2 个体，并且仅刺激 CD8 T 细胞。我们正在筛选WT1的肽库，以鉴定诱导其他HLA 1类分子和CD4 T细胞的肽，从而设计出更广泛适用和更有效的疫苗。 (2) 自然杀伤 (NK) 细胞 GVL：在对 270 多个 SCT 的回顾性分析中，我们发现具有 KIR 基因型 2DS1、3DS1、2DL5a 的干细胞供体使急性髓性白血病 (AML) 受者的复发率显着降低。随后对具有和不具有这种“有利”基因型的供体之间的 NK 细胞功能和表型差异进行的研究尚未发现任何差异。 (3) 不匹配T细胞的GVL效应：我们已经证明，幼稚T细胞区室和记忆T细胞区室中都存在同种异体反应T细胞。记忆同种反应性 T 细胞经常与 CMV 和 EBV 病毒的 DNA 病毒抗原表现出交叉反应。 通过 T 细胞受体基因测序，我们证实单个 T 细胞克隆可以表现出针对病毒抗原和 HLA 不匹配靶细胞的双重反应性。为了研究交叉反应性是否可能对病毒特异性 T 细胞克隆的第三方过继转移的使用产生限制，我们研究了给予 SCT 后病毒再激活患者的病毒特异性 T 细胞的交叉反应性，并表明在体内这种交叉反应性没有导致GVHD。我们现在正在探索是否可以使用人工基因修饰的 K562 抗原呈递细胞产生不匹配的同种反应性 T 细胞系以提供 GVL 效应，以及主要针对 DNA 病毒产生的同种反应性 T 细胞系是否也具有 GVL 效应并可用于治疗复发性癌症SCT后白血病。 (4) SCT 后的免疫恢复：我们测量了血浆细胞因子谱和淋巴细胞库，以确定移植后增强移植物抗白血病 (GVL) 效应的最佳时间，并表征不同移植方案中的免疫恢复特征。我们发现了细胞因子表达的两种模式——移植后最初几天淋巴和髓样生长因子的早期激增，以及移植后 4 周左右淋巴因子释放的后期激增。我们现在正在扩展细胞因子分析，以比较自体 SCT 和充满 T 细胞的 SCT 的模式。 我们将把不同淋巴细胞亚群的恢复与个体细胞因子谱相关联，以确定促进最佳恢复的因素。 (5) 选择性耗竭同种反应性 T 细胞：一项从 SCT 中选择性耗竭可导致 GVHD 的 T 细胞的临床试验，以及开发改进技术以使用基于罗丹明的染料选择性光耗竭活化 T 细胞的进一步转化工作现已完成。我们评估了这种光耗竭技术选择性耗竭干细胞移植物中移植物抗宿主病 (GVHD) 同种反应 T 细胞的能力。用经过辐射的体外扩增的受体 T 淋巴细胞刺激供体淋巴细胞。通过光照消除优先保留光敏剂 4, 5-二溴罗丹明 123 (TH9402) (Kiadis Pharma, NL) 的同种激活 T 细胞。接受氟达拉滨、环磷酰胺和全身照射治疗的 24 名血液恶性肿瘤患者（16 名高危患者）接受了来自 HLA 相同同胞的 CD34 选择干细胞同种异体移植物和 5x106/kg 选择性耗尽 (SD) 供体 T 细胞。低剂量环孢素被用作唯一的移植后免疫抑制剂。 III-IV 级急性 GVHD 的总体概率为 13%。 14 名患者出现（主要是有限的）慢性 GVHD (c-GVHD)。 5 名患者复发，其中 2 名患者在进一步治疗后仍处于缓解状态。 13 名患者的中位生存期为 22 个月。总生存率和无病生存精算概率 (+ SEM) 分别为 43 ± 13% 和 35 ± 13%。 SD 技术的复发率较低 (24 ± 10%)，但由于与 c-GVHD 相关的晚期非复发死亡率和 4 年随访时的感染率为 50 ± 14%，使情况变得复杂。这些结果表明，SD 可以有效减少严重的 a-GVHD，同时保留移植物抗白血病效应。 (6)间充质基质细胞(MSC)：探索MSC与细胞毒性T细胞相互作用的临床前研究表明，MSC可以降低效应T细胞对白血病细胞系的细胞毒性。需要进一步的研究来探索 MSC 输注对 GVL 反应性可能的抑制作用。我们与临床中心细胞处理部门合作，为三项试验开发临床级 MSC，涵盖使用 MSC 治疗 GVHD、骨髓功能不良和主要器官损伤。 (7) T 细胞耗尽 SCT 作为免疫治疗方法平台的临床试验正在进行中。 36 名患有血液系统恶性肿瘤的患者接受了来自其 HLA 相同兄弟姐妹的同种异体 HSCT。中位年龄为 43 岁（范围 16-68 岁）。移植适应症为急性白血病（24）、其他骨髓恶性肿瘤（9）、其他淋巴恶性肿瘤（3）。受试者接受环磷酰胺、氟达拉滨的清髓预处理方案和全身照射（12 Gy，肺屏蔽至 6 Gy）。 55 岁及以上的受试者接受了 4 Gy 的剂量。利用 Miltenyi CliniMacs 系统对来自供体的 G-CSF 动员的外周血干细胞进行 CD34+ 选择，输注目标 CD34+ 剂量为 6 x 106/kg（范围 3 至 10 x 106/kg），固定 CD3+ 剂量为 5 x 104 /公斤。患者仅接受低剂量环孢素直至第21天。在+90天给予延迟淋巴细胞加回（5 x 106 CD3+/kg）。第 200 天的总生存率为 79%。 34 名受试者在第 14 天时实现了完全供体 (>95%) 骨髓嵌合，完成供体 CD3+ 嵌合的中位时间为 45 天。慢性GVHD的发生率为34.3%。中位随访时间为 3.6 年，Kaplan-Meier 估计的复发率、非复发死亡率和总生存率分别为 35%、33% 和 44%。这些结果代表了可接受的移植和临床结果。审判正在进行中。 阿仑单抗治疗 LGL 白血病的试验正在进行中。患者患有慢性血细胞减少症，克隆性 CD3+CD8+CD16+ 或 CD57+T-LGL 增加。 1 mg 测试剂量后，阿仑单抗以 10 mg/d 给药，持续 10 天。总共有 9 名患者（56%；95% CI 30%-80%）对阿仑单抗有反应，其中 5 名患者达到 CR，4 名患者达到 PR。对治疗有反应的 6 名贫血患者中，治疗前血红蛋白中位数为 7.3（范围 5.4-9.3）g/dL，治疗后血红蛋白中位数为 12.2（范围 10-13.7）g/dL。治疗前 ANC < 500 /cu mm 的三名应答者在阿仑单抗治疗后改善至 ANC > 1,000/cu mm。 这项研究表明阿仑单抗是安全的，并且在接受过治疗的 T-LGL 患者中显示出有希望的活性。

项目成果

A clinical-scale selective allodepletion approach for the treatment of HLA-mismatched and matched donor-recipient pairs using expanded T lymphocytes as antigen-presenting cells and a TH9402-based photodepletion technique.

一种临床规模的选择性同种去除方法，使用扩增的 T 淋巴细胞作为抗原呈递细胞和基于 TH9402 的光去除技术，用于治疗 HLA 不匹配和匹配的供体-受体对。

• 发表时间: 2008-04-15
• 影响因子: 20.3
• 作者: Mielke, Stephan;Nunes, Raquel;Rezvani, Katayoun;Fellowes, Vicki S;Venne, Annie;Solomon, Scott R;Fan, Yong;Gostick, Emma;Price, David A;Scotto, Christian;Read, Elizabeth J;Barrett, A John
• 通讯作者: Barrett, A John

Leukemia-associated antigen-specific T-cell responses following combined PR1 and WT1 peptide vaccination in patients with myeloid malignancies.

骨髓恶性肿瘤患者联合 PR1 和 WT1 肽疫苗接种后白血病相关抗原特异性 T 细胞反应。

• 发表时间: 2008-01-01
• 影响因子: 20.3
• 作者: Rezvani, Katayoun;Yong, Agnes S M;Mielke, Stephan;Savani, Bipin N;Musse, Laura;Superata, Jeanine;Jafarpour, Behnam;Boss, Carol;Barrett, A John
• 通讯作者: Barrett, A John

Contribution of TCR-beta locus and HLA to the shape of the mature human Vbeta repertoire.

TCR-β 位点和 HLA 对成熟人类 Vbeta 库形状的贡献。

• 发表时间: 2008-05-15
• 作者: Melenhorst, J Joseph;Lay, Matthew D H;Price, David A;Adams, Sharon D;Zeilah, Josette;Sosa, Edgardo;Hensel, Nancy F;Follmann, Dean;Douek, Daniel C;Davenport, Miles P;Barrett, A John
• 通讯作者: Barrett, A John

Autoimmune mechanisms in the pathophysiology of myelodysplastic syndromes and their clinical relevance.

骨髓增生异常综合征病理生理学中的自身免疫机制及其临床相关性。

• 发表时间: 2009-04
• 影响因子: 10.1
• 作者: Barrett, A John;Sloand, Elaine
• 通讯作者: Sloand, Elaine

Pharmacokinetics of liposomal amphotericin B in pleural fluid.

脂质体两性霉素 B 在胸水中的药代动力学。

• 发表时间: 2010-04
• 影响因子: 4.9
• 作者: Moriyama, Brad;Torabi;Pratt, Alexandra K;Henning, Stacey A;Pennick, Gennethel;Shea, Yvonne R;Roy Chowdhuri, Sinchita;Rinaldi, Michael G;Barrett, A John;Walsh, Thomas J
• 通讯作者: Walsh, Thomas J