Tuttle T M, Inge T H, Wirt C P, Frank J L, McCrady C M, Bear H D
Department of Surgery, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298.
J Immunother (1991). 1992 Aug;12(2):75-81. doi: 10.1097/00002371-199208000-00001.
Several strategies have been used to stimulate the growth of tumor-specific T cells in place of tumor antigen. One approach is to use pharmacologic agents to activate the second messenger pathways of T-cell activation. In the present study, we examined the ability of the protein kinase C activator bryostatin 1 (B) plus the calcium ionophore ionomycin (I) to stimulate the growth of lymphocytes obtained from the axillary lymph nodes (DLN) draining a progressively growing intradermal plasmacytoma tumor. Draining lymph node cells were initially cultured with autologous tumor cells and 20 U/ml of interleukin-2 (IL-2) for 7 days. The lymphocytes were then incubated with various concentrations of bryostatin 1 plus 1 microM ionomycin and cultured for an additional 14 days in IL-2. DLN cells initially cultured with autologous tumor and then restimulated with 5 nM bryostatin 1 and 1 microM ionomycin exhibited marked in vitro proliferation and 15-fold expansion of cell numbers over 2 weeks. The cells expanded with B/I were predominantly CD8+ T cells and retained specific in vitro cytotoxicity against autologous tumor. When adoptively transferred to mice with established liver metastases, DLN cells restimulated with B/I-mediated specific tumor regression.
已经采用了几种策略来替代肿瘤抗原刺激肿瘤特异性T细胞的生长。一种方法是使用药物激活T细胞活化的第二信使途径。在本研究中，我们检测了蛋白激酶C激活剂苔藓抑素1（B）加钙离子载体离子霉素（I）刺激从引流逐渐生长的皮内浆细胞瘤肿瘤的腋窝淋巴结（DLN）获得的淋巴细胞生长的能力。引流淋巴结细胞最初与自体肿瘤细胞和20 U/ml白细胞介素-2（IL-2）一起培养7天。然后将淋巴细胞与各种浓度的苔藓抑素1加1 μM离子霉素一起孵育，并在IL-2中再培养14天。最初用自体肿瘤培养然后用5 nM苔藓抑素1和1 μM离子霉素再刺激的DLN细胞在体外表现出明显的增殖，并且在2周内细胞数量扩增了15倍。用B/I扩增的细胞主要是CD8 + T细胞，并保留了对自体肿瘤的特异性体外细胞毒性。当将其过继转移到已建立肝转移的小鼠中时，用B/I再刺激的DLN细胞介导了特异性肿瘤消退。
