T Cell Trafficking in Adoptive Cellular Immunotherapy

长期目标是评估和重新设计 基于淋巴细胞基础的过继免疫治疗(Al)方案 贩卖人口。最初的计划被重新集中在最新颖和最有希望的 AIM(Aim 2)，考虑到3年的获奖期。初步工作表明， 肿瘤反应性钛前效应细胞集中在肿瘤引流淋巴中 合成血管结合部位(配体)的结节(TDLN)细胞 黏附受体P-和E-选择素。 选择素-配体的合成发生在激活的T细胞的一小部分上 这意味着对这些细胞(Plighigh细胞)的选择性培养可能会改善 体内抗肿瘤活性。一种顺磁分馏程序，使用 重组人1gm：小鼠P-选择素嵌合体(LgMPsetlHM) 检验这一假设。处理5-10个月需要大量的IgMPselHM X 108细胞/实验推动了高容量的发展 瞬时转染法。Plighigh细胞的选择性培养 肺微转移抑制(3天)增加10-20倍和 对于MCA-205肉瘤和B16黑色素瘤，分别超过70倍。50 抑制大脑中动脉肉瘤转移的百分比需要0.1-0.3×106培养 普通，2-4×106培养，大于9×106培养 每只动物提供Pliglow TDLN。需要抑制50%的B16-黑色素瘤Mets 1 培养的PlighHigh为106×106，培养的普通TDLN为70×106 动物。因此，利用与钛有关的黏附受体进行TDLN分级 贩运显著提高了铝的治疗效益。此次续订 在这些有希望的成果的基础上，实现以下目标。(1)识别 TDLN中在培养过程中产生抗肿瘤活性的高亚群。这个 从TDLN分离到的Plighigh细胞是异质性的。因此，这个目标 确定哪些细胞类型(a/b T细胞、NK、NK-T、13细胞、g/d T细胞APC) 影响抗肿瘤活性。(2)确定符合以下条件的执行机构 提高培养的Plighigh TDLN的效力。T细胞的贡献 将研究亚群、血管黏附受体和趋化因子 对不同部位的转移作出反应。(3)确定是否治疗 与促炎细胞因子一起提高募集和抗肿瘤作用 培养的Plighigh细胞的活性。换句话说，我们能不能增加 诱导分化培养的Plighigh细胞及其临床应用 是荷瘤器官的炎症反应还是单个大转移瘤？ (4)使用患者样本评估基于选择素的人类分选 接受播散性肾癌的过继免疫治疗。

The long-term objectives are the evaluation and redesign of adoptive immunotherapy (Al) protocols based on the fundamentals of lymphocyte trafficking. The original plan was refocused on the most novel and promising Aim (Aim 2) in light of the 3-year award period. Initial work revealed that tumor-reactive Ti pre-effector cells were concentrated in tumor draining lymph node (TDLN) cells synthesizing bindings sites (ligands) for the vascular adhesion receptors P- and E-selectin. Selectin-ligand synthesis occurred on a small subset of the activated T-cells implying that selective culture of these cells (Plighigh cells) might improve anti-tumor activity in vivo. A para-magnetic fractionation procedure, using recombinant human 1gM: murine P-selectin chimera (lgMPsetlHM), was devised to test the hypothesis. The large amounts of IgMPselHM needed for processing 5-10 x 108 cells/experiment fueled development of a high-capacity transient-transfection system. Selective culture of the Plighigh cells increased suppression of pulmonary micrometastases (3-day) by 10-20 fold and greater than 70 fold for MCA-205 sarcomas and B16 melanomas, respectively. 50 percent suppression of MCA-sarcoma metastases required 0.1-0.3 x 106 cultured Plighigh, 2-4 x 106 cultured unfractionated and greater than 9 x 106 cultured Pliglow TDLN per animal. 50 percent suppression of B16-melanoma mets required 1 x 106 cultured Plighhigh versus 70 x 106 cultured unfractionated TDLN per animal. Thus, TDLN fractionation using adhesion receptors involved in Ti trafficking markedly enhances the therapeutic benefit from Al. This renewal builds on these promising results with the following Aims. (1) Identify the PIighigh subsets in TDLN that produce anti-tumor activity during culture. The Plighigh cells isolated from TDLN are heterogeneous. Therefore, this aim determines which cell types (a/b T-cells, NK, NK-T, 13-cells, g/d T-cells APCs) influence anti-tumor activities. (2) Identify the effector mechanisms that increase the potency of cultured Plighigh TDLN. The contributions of T-cell subsets, vascular adhesion receptors and chemokines will be investigated for responding metastases in various locations. (3) Determine whether treatment with pro-inflammatory cytokines improves the recruitment and anti-tumor activities of cultured Plighigh cells. In other words, can one increase the delivery and clinical benefit of cultured Plighigh cells by inducing an inflammatory response in tumor-bearing organs or individual macrometastases? (4) Evaluate selectin-based sorting in humans using specimens from patients receiving adoptive immunotherapy for disseminated renal cell carcinoma.