Jiao Yu-Meng, Fang Qiang, Xia Hui, Wang Xue-Mei, Tao Zhi-Yong, Chen Xing-Zhi, Shen Ji-Long
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2013 Apr;31(2):95-8.
To observe the role of CD8+ T cells in the tumor growth delay induced by Toxoplasma gondii excreted-secreted antigens (TgESA) in B16FI0 mouse melanoma model in the early stage.
TgESA were prepared by incubating T. gondii tachyzoites for 12 h in vitro. 15 C57BL/6 mice were randomly assigned to group A, B, and C (5 mice per group). Each mouse in group B and C was subcutaneously injected in right flank with 2 x 10(5) B16F10 cells. Mice in group C were intraperitoneally injected with TgESA (100 microl per mouse) at 7d after B16F10 cells injection. Mice of group A were only injected with PBS. On the 13th day after melanoma cell injection, the mice were sacrificed and spleen was removed. The percentage of CD8+ T cells in the spleen was analyzed by flow cytometry. CD8+ T cells were isolated from spleen cells by using immunomagnetic beads. The activity of CD8+ T cells against B16F10 melanoma cells was determined by LDH release assay at different effect-to-target cell ratios (2.5:1, 5:1, and 10:1). Other 30 C57BL/6 mice were randomly divided into group E, F, and G. Each mice were injected with 2 x 10(5) B16F10 cells. At the same time, mice in group F and G were simultaneously injected via the tail vein with CD8+ T cells isolated from mice in group B and C. Tumor growth, mortality and survival time of mice were observed and recorded during 35-d observation period.
The percentage of CD3+CD8+T cells in the spleen cells of group C [(15.74 +/- 0.28)%] was significantly higher than that of group B [(14.18 +/- 0.27)%] and A [(13.86 +/- 0.13)%] (P < 0.05). At different effect-to-target cell ratios, the activity of CD8+ T cells against B16F10 cells in group C was significantly higher than that of group B (P < 0.05). The average time of tumor formation in group G [(14.9 +/- 1.2) d] was longer than that in group F [(11.9 +/- 0.7) d] and E [(9.4 +/- 1.2) d] (P < 0.05). The tumor size in these groups increased, but there was no obvious difference in the tumor growth rate among the three groups. The tumor size of group G was significantly smaller than the other two groups (P < 0.05). In group E, F and G, mice began to die on the 26th day, the 29th day and the 30th day after tumor inoculation, and the number of survival mice was 3, 5 and 7, respectively, at the 35th day after injection.
TgESA may up-regulate the quantity and function of CD8+ T cell in B16F10 melanoma mouse model, which plays a role of delaying tumor growth in early stage.
观察CD8 + T细胞在早期弓形虫排泄分泌抗原（TgESA）诱导的B16F10小鼠黑色素瘤模型肿瘤生长延迟中的作用。
通过将弓形虫速殖子在体外孵育12小时制备TgESA。将15只C57BL / 6小鼠随机分为A、B、C组（每组5只）。B组和C组的每只小鼠右腹侧皮下注射2×10⁵个B16F10细胞。C组小鼠在注射B16F10细胞后7天腹腔注射TgESA（每只小鼠100μl）。A组小鼠仅注射PBS。黑色素瘤细胞注射后第13天，处死小鼠并取出脾脏。通过流式细胞术分析脾脏中CD8 + T细胞的百分比。使用免疫磁珠从脾细胞中分离CD8 + T细胞。在不同效靶细胞比（2.5:1、5:1和10:1）下，通过乳酸脱氢酶释放试验测定CD8 + T细胞对B16F10黑色素瘤细胞的活性。另外30只C57BL / 6小鼠随机分为E、F、G组。每只小鼠注射2×10⁵个B16F10细胞。同时，F组和G组小鼠经尾静脉同时注射从B组和C组小鼠分离的CD8 + T细胞。在35天的观察期内观察并记录小鼠的肿瘤生长、死亡率和存活时间。
C组脾细胞中CD3 + CD8 + T细胞的百分比[（15.74±0.28）%]显著高于B组[（14.18±0.27）%]和A组[（13.86±0.13）%]（P < 0.05）。在不同效靶细胞比下，C组CD8 + T细胞对B16F10细胞的活性显著高于B组（P < 0.05）。G组肿瘤形成的平均时间[（14.9±1.2）天]长于F组[（11.9±0.7）天]和E组[（9.4±1.2）天]（P < 0.05）。这些组的肿瘤大小均增加，但三组之间肿瘤生长速率无明显差异。G组的肿瘤大小明显小于其他两组（P < 0.05）。在E、F和G组中，小鼠在肿瘤接种后第26天、第29天和第30天开始死亡，注射后第35天存活小鼠数量分别为3只、5只和7只。
在B16F10黑色素瘤小鼠模型中，TgESA可能上调CD8 + T细胞的数量和功能，在早期发挥延迟肿瘤生长的作用。
