Department of Surgery, Hebei Medical University, Shijiazhuang, Hebei 050017, China.
Chin Med J (Engl). 2012 Mar;125(5):794-800.
Adoptive transfer of allogeneic tumor-specific T cells often results in severe graft-versus-host disease (GVHD). Here, we sought to maximize graft-versus-tumor and minimize GVHD by using haploidentical T cells in pre-irradiated B16-melanoma bearing mice.
C57BL/6 mice bearing B16-melanoma tumors were irradiated with 0, 5, or 7 Gy total body irradiation (TBI), or 7 Gy TBI plus bone marrow transplantation. Tumor areas were measured every 3 days to assess the influence of irradiation treatment on tumor regression. B16-melanoma bearing mice were irradiated with 7 Gy TBI; sera and spleens were harvested at days 1, 3, 5, 7, 9, 11, and 13 after irradiation. White blood cell levels were measured and transforming growth factor β1 (TGF-b1) and interleukin 10 (IL-10) levels in serum were detected using enzyme-linked immunosorbent assay (ELISA) kits. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry were performed to test TGF-b1, IL-10 and Foxp3 mRNA levels and the proportion of CD4+CD25+Foxp3+ T-regulatory cells (Tregs) in spleens. B16-melanoma bearing C57BL/6 mice were irradiated with 7 Gy TBI followed by syngeneic (Syn1/Syn2) or haploidentical (Hap1/Hap2), dendritic cell-induced cytotoxic T lymphocytes (DC-CTLs) treatment, tumor areas and system GVHD were observed every 3 days. Mice were killed 21 days after the DC-CTLs adoptive transfer; histologic analyses of eyes, skin, liver, lungs, and intestine were then performed.
Irradiation with 7 Gy TBI on the B16-melanoma-bearing mice did not influence tumor regression compared to the control group; however, it down-regulated the proportion of Tregs in spleens and the TGF-b1 and IL-10 levels in sera and spleens, suggesting inhibition of autoimmunity and intervention of tumor microenvironment. Adoptive transfer of haploidentical DC-CTLs significantly inhibited B16-melanoma growth. GVHD assessment and histology analysis showed no significant difference among the groups.
Adoptive transfer of haploidentical tumor-specific T cells in irradiation-pretreated B16-melanoma bearing mice preserved antitumor capacity without causing a GVHD response.
过继输注同种异体肿瘤特异性 T 细胞常导致严重的移植物抗宿主病（GVHD）。在这里，我们试图通过使用半相合 T 细胞在预先照射的 B16-黑色素瘤荷瘤小鼠中最大限度地发挥移植物抗肿瘤作用，同时最小化 GVHD。
用 0、5 或 7 Gy 全身照射（TBI）或 7 Gy TBI 加骨髓移植照射携带 B16-黑色素瘤肿瘤的 C57BL/6 小鼠。每 3 天测量肿瘤面积以评估照射治疗对肿瘤消退的影响。用 7 Gy TBI 照射携带 B16-黑色素瘤的小鼠；在照射后第 1、3、5、7、9、11 和 13 天采集血清和脾脏。用酶联免疫吸附试验（ELISA）试剂盒测量白细胞水平，并检测血清中转化生长因子β1（TGF-b1）和白细胞介素 10（IL-10）水平。用实时逆转录-聚合酶链反应（RT-PCR）和流式细胞术检测脾脏中 TGF-b1、IL-10 和 Foxp3 mRNA 水平以及 CD4+CD25+Foxp3+T 调节细胞（Tregs）的比例。用 7 Gy TBI 照射携带 B16-黑色素瘤的 C57BL/6 小鼠，然后用同种（Syn1/Syn2）或半相合（Hap1/Hap2）、树突状细胞诱导的细胞毒性 T 淋巴细胞（DC-CTLs）治疗，每 3 天观察肿瘤面积和系统 GVHD。在 DC-CTL 过继转移后 21 天处死小鼠，然后对眼睛、皮肤、肝脏、肺和肠进行组织学分析。
与对照组相比，用 7 Gy TBI 照射携带 B16-黑色素瘤的小鼠并不影响肿瘤的消退；然而，它下调了脾脏中 Tregs 的比例以及血清和脾脏中 TGF-b1 和 IL-10 的水平，提示抑制自身免疫和干预肿瘤微环境。过继输注半相合 DC-CTLs 可显著抑制 B16-黑色素瘤的生长。GVHD 评估和组织学分析显示各组间无显著差异。
在预先照射的携带 B16-黑色素瘤的小鼠中过继输注半相合肿瘤特异性 T 细胞可保留抗肿瘤能力而不引起 GVHD 反应。
