NK Cell Education in Recipients of Allogeneic HCT

项目总结(见说明)： 在目前的研究中，我们发现，虽然NK细胞在异基因移植后早期大量重建，但它们表现出杀伤免疫球蛋白受体(KIR)减少，它们是低反应性和低杀伤肿瘤靶点。利用体外发育模型，我们还发现IL-15可以诱导KIR的获得和完全效应功能。该项目的主要假设是，造血移植后早期的NK细胞没有受过教育，NK细胞的教育受到IL-15、激活受体(2B4Tim-3)和抑制性受体(KIR、NKG2A、LIR-1)连接的发育调节，所有这些都在决定AML靶点是否被杀伤方面发挥作用。目前资助的项目1的有希望的结果表明，与有利的KIR B单倍型供者进行异基因造血细胞移植(通过较高的B含量评分和Cen-B/B模式而增强)可以防止AML复发。在第二个战略(项目3)中，建立了自然杀伤细胞过继转移平台。为了改进现有的方法，NCI最近开发了一种生产rhLL-15的cGMP工艺，将向校外社区提供。SAI将测试IL-15，作为扩大过继转移的成人NK细胞的新策略的一部分。第二个疗程的IL-15将在第+42天进行，以培养来自同一捐赠者的CD34+干细胞重建的NK细胞。在SA2中，我们将研究KIR免疫遗传学是否决定NK细胞功能(临床上发现的有利的Cen-B/B模式)，以及IL-15响应的KIR启动子元件是否控制转录调控，以形成KIR谱系。SA3将评估一种在大多数NK细胞上表达的新的激活受体TIM-3，该受体可被IL-15上调。我们推测它通过上调2B4的SAP适配器(也是对IL-15的反应)在NK细胞的培养中发挥作用。TIM-3和2B4可以直接识别表达AML靶点的Galecfin-9(Gal-9)和CD48，也可以间接接受表达相同配体的树突状细胞的激活信号。随着这些研究的完成，我们希望改变使用IL-15转移NK细胞的做法，并向我们介绍IL-15诱导NK细胞教育和识别AML靶点的机制。

PROJECT SUMMARY (See instructions): In the current funding we found that although NK cells reconstitute in high numbers early after allogeneic transplant, they display diminished killer immunoglobulin receptors (KIR), they are hypo-responsive and poorly kill tumor targets. Using in vitro development models, we also found that the acquisifion of KIR and full effector function can be induced by IL-15. The overarching hypothesis of this Project is that NK cells eariy after hematopoetic transplantafion are uneducated and that NK cell education is developmentally regulated by IL-15, activafing receptors (2B4 Tim-3), and inhibitory receptor (KIR, NKG2A, LIR-1) ligation, all of which play a role in determining whether AML targets are killed. Promising results from Project 1 ofthe current funding show that allogeneic hematopoietic cell transplantafion with favorable KIR B haplotype donors (enhanced by a higher B content score and a Cen-B/B pattern) results in protecfion from AML relapse. In a second strategy (Project 3), a platform for NK cell adoptive transfer has been established. To improve on existing methods, the NCI has recently developed a cGMP process for production of rhlL-15 which will be made available to the Extramural community. SAI will test IL-15 as part of a novel strategy to expand adoptively transferred adult NK cells. A second course of IL-15 will be given on Day +42 to educate NK cells that reconstitute from CD34+ stem cells from the same donor. In SA2, we will invesfigate whether KIR immunogenetics determines NK cell function (the favorable Cen-B/B pattern found clinically) and whether IL-15 responsive KIR promoter elements control transcriptional regulation to form the KIR repertoire. SA3 will evaluate a novel activating receptor expressed on most NK cells, Tim-3, which is upregulated by IL-15. We hypothesize it plays a role in NK cell education.by upregulating the SAP adaptor (also IL-15 responsive) for 2B4. Tim-3 and 2B4 can directly recognize Galecfin-9 (Gal-9) and CD48 expressing AML targets or may indirectly receive activating signals from dendrific cells that express the same ligands. At complefion of these studies, we expect to change practice of NK cell adopfive transfer by use of IL-15 and to inform us about IL-15 mechanisms to induce NK cell education and recognifion of AML targets.