To W C, Seeley B M, Barthel S W, Shu S
Department of Otolaryngology and Communicative Disorders, Cleveland Clinic Foundation, Ohio, USA.
Laryngoscope. 2000 Oct;110(10 Pt 1):1648-54. doi: 10.1097/00005537-200010000-00015.
To evaluate the cytokine secretion profile and therapeutic efficacy of Th1 CD4+ L-selectin-tumor-draining lymph node lymphocytes in the treatment of murine pulmonary metastases.
Prospective, murine in vivo and in vitro study.
B6 mice were injected bilaterally subcutaneously with MCA 205 sarcoma cells to initiate tumor growth. Eleven days later, tumor-draining inguinal lymph nodes were harvested. Single-cell suspensions were prepared and fractionated using magnetically activated cell sorting. Sorted CD4+ L-selectin-lymphocytes were activated with anti-CD3 monoclonal antibody for 48 hours either alone to give a Th1 phenotype, or in the presence of interleukin (IL)-4 and anti-interferon-gamma (alpha-IFN-gamma) monoclonal antibody to elicit a Th2 phenotype. Activated cells were then expanded for 3 days in IL-2. Resulting cells were used to treat 3-day pulmonary metastases. Enzyme-linked immunosorbent assay and intracellular fluorescent-activated cell-sorter (FACS) scanning were used to evaluate the cytokine secretion profiles of these cells.
Activated and expanded L-selectin- CD4+ T cells demonstrated a Th1 cytokine profile and excellent antitumor efficacy. In contrast, L-selectin- CD4+ lymphocytes activated in the presence of IL-4 and alpha-IFN-gamma monoclonal antibody demonstrated a Th2-like profile and significantly (P < .05) poorer antitumor efficacy.
The cytokine environment during the activation of tumor-draining lymph nodes can influence the therapeutic efficacy of activated L-selectin-, CD4+ T cells. Cell mediated, Th1-dependent immunity appears to play an important role in mediating tumor regression. Culture conditions promoting Th2 cells resulted in T cells associated with diminished antitumor efficacy.
评估Th1 CD4+ L选择素-肿瘤引流淋巴结淋巴细胞在治疗小鼠肺转移中的细胞因子分泌谱及治疗效果。
前瞻性小鼠体内和体外研究。
双侧皮下注射MCA 205肉瘤细胞至B6小鼠以启动肿瘤生长。11天后，收集肿瘤引流腹股沟淋巴结。制备单细胞悬液并使用磁珠分选法进行分离。分选的CD4+ L选择素淋巴细胞用抗CD3单克隆抗体单独激活48小时以获得Th1表型，或在白细胞介素（IL）-4和抗干扰素-γ（α-IFN-γ）单克隆抗体存在的情况下激活以诱导Th2表型。然后将激活后的细胞在IL-2中扩增3天。所得细胞用于治疗3天的肺转移。采用酶联免疫吸附测定和细胞内荧光激活细胞分选仪（FACS）扫描评估这些细胞的细胞因子分泌谱。
激活并扩增的L选择素-CD4+ T细胞表现出Th1细胞因子谱及出色的抗肿瘤效果。相比之下，在IL-4和α-IFN-γ单克隆抗体存在下激活的L选择素-CD4+淋巴细胞表现出类似Th2的谱，且抗肿瘤效果显著较差（P <.05）。
肿瘤引流淋巴结激活过程中的细胞因子环境可影响激活的L选择素-CD4+ T细胞的治疗效果。细胞介导的、Th1依赖的免疫似乎在介导肿瘤消退中起重要作用。促进Th2细胞的培养条件导致T细胞抗肿瘤效果降低。
