Pan Hua, Sun Li-Rong
Women and Children's Medical and Health Care Center of Qingdao, Qingdao, Shandong 266011, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2012 Mar;14(3):184-7.
To study the effect of bacillus Callmette-Guérin (BCG) on cytotxicity of cytotoxic T lymphocyte (CTL) from human peripheral blood of children with acute lymphoblastic leukemia (ALL) for killing HL-60 cells in vitro.
The mononuclear cells were isolated from peripheral blood of ALL children and healthy children, and were cultured with RPMI1640, interleukin-2 (IL-2), phytohemagglutinin (PHA) and BCG.The growth of CTLs was observed by light microscopy. The proportions of CD3, CD3+CD4+ and CD3+CD8+ were determined by flow cytometry 10 days after culture. MTT method was performed to detect the cytotoxicity of CTLs for killing HL-60 cells.
Neither the cell number nor the volume of CTLs changed significantly within 2 days of culture, but both began increasing on the 3rd day of culture and reached a peak on the 6-10th days. On the 10th day of culture, the cell number of CTLs in the BCG treatment group was much higher than in the group without BCG treatment. The CD3+CD8+ proportion in the leukemia group was much higher than in the control group. With the effect of BCG, the CD3+CD8+ proportion of the two groups became much higher. The cytotoxicity of CTLs for killing HL-60 cells in the leukemia group was weaker than in the control group.
BCG along with IL-2 and PHA promotes the proliferation of CTLs and enhances the ability of CTLs in killing HL-60 cells.
研究卡介苗（BCG）对急性淋巴细胞白血病（ALL）患儿外周血细胞毒性T淋巴细胞（CTL）体外杀伤HL-60细胞的细胞毒性的影响。
从ALL患儿和健康儿童外周血中分离单个核细胞，用RPMI1640、白细胞介素-2（IL-2）、植物血凝素（PHA）和BCG进行培养。通过光学显微镜观察CTL的生长情况。培养10天后，用流式细胞术检测CD3、CD3+CD4+和CD3+CD8+的比例。采用MTT法检测CTL对HL-60细胞的杀伤细胞毒性。
培养2天内，CTL的细胞数量和体积均无明显变化，但在培养第3天开始增加，并在第6 - 10天达到峰值。培养第10天，BCG治疗组的CTL细胞数量远高于未用BCG治疗组。白血病组的CD3+CD8+比例远高于对照组。在BCG的作用下，两组的CD3+CD8+比例均显著升高。白血病组CTL对HL-60细胞的杀伤细胞毒性弱于对照组。
BCG联合IL-2和PHA可促进CTL的增殖，并增强CTL杀伤HL-60细胞的能力。
