T Cell Trafficking in Adoptive Cellular Immunotherapy

长期目标是评价和重新设计 基于淋巴细胞基础的过继免疫治疗（AI）方案 贩卖人口最初的计划被重新聚焦在最新颖和最有前途的 目标（目标2）根据3年奖励期。初步工作显示， 肿瘤反应性Ti前效应细胞集中在肿瘤引流淋巴液中 淋巴结（TDLN）细胞合成血管的结合位点（配体） 粘附受体P-和E-选择素。 选择素-配体的合成发生在一小部分活化的T细胞上 这意味着选择性培养这些细胞（Plighgh细胞）可能会改善 体内抗肿瘤活性。顺磁分馏程序，使用 设计了重组人IgM：鼠P-选择素嵌合体（IgM set HM）， 测试假设。处理5-10所需的大量IgMPselHM x 108个细胞/实验促进了高容量 瞬时转染系统。高密度细胞的选择性培养 肺微转移抑制（3天）增加10-20倍， MCA-205肉瘤和B16黑色素瘤分别大于70倍。50 MCA肉瘤转移的抑制百分比需要0.1-0.3 x 106培养 高，2-4 x 106未分级培养物和大于9 x 106培养物 Pliglow TDLN/动物。B16黑色素瘤转移的50%抑制需要1 每种培养基中，10 × 106培养的Plighhigh与70 × 106培养的未分级TDLN 动物因此，TDLN分馏使用的粘附受体参与Ti 贩运显著增强了Al的治疗益处。 在这些有希望的结果的基础上，我们有以下目标。(1)识别 TDLN中在培养期间产生抗肿瘤活性的高浓度亚群。的 从TDLN分离的高密度细胞是异质性的。因此，这一目标 确定细胞类型（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)识别效应器机制， 提高培养的Plighigh TDLN的效力。T细胞的贡献 亚群，血管粘附受体和趋化因子将被研究， 在不同的位置有反应的转移。(3)确定治疗是否 与促炎细胞因子一起改善了募集和抗肿瘤 培养的Plighigh细胞的活性。换句话说，是否可以增加 培养的Plighgh细胞的递送和临床益处， 在肿瘤承载器官或个别巨转移的炎症反应？ (4)使用来自患者的标本评估人类中基于选择素的分选 接受过继性免疫疗法治疗播散性肾细胞癌

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.