Effect of bacillus Callmette-Guérin on cytotxicity of cytotoxic T lymphocyte from children with acute lymphoblastic leukemia for killing HL-60 cells in vitro
PAN Hua, SUN Li-Rong
Women and Children's Medical and Health Care Center of Qingdao, Qingdao, Shandong 266011, China. Sunlr@vip.sina.com
Abstract OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: BCG along with IL-2 and PHA promotes the proliferation of CTLs and enhances the ability of CTLs in killing HL-60 cells.
PAN Hua,SUN Li-Rong. Effect of bacillus Callmette-Guérin on cytotxicity of cytotoxic T lymphocyte from children with acute lymphoblastic leukemia for killing HL-60 cells in vitro[J]. 中国当代儿科杂志, 2012, 14(3): 184-187.
PAN Hua,SUN Li-Rong. Effect of bacillus Callmette-Guérin on cytotxicity of cytotoxic T lymphocyte from children with acute lymphoblastic leukemia for killing HL-60 cells in vitro[J]. CJCP, 2012, 14(3): 184-187.
［1］Bequm NA, Ishii K, Kurita-Taniquchi M, Tanabe M, Kobayashi M, Moriwaki Y. Mycobacterium bovis BCG cell wall-specific differentially expressed genes identified by differential display and cDNA subtraction in human macrophages［J］. Infect Immun, 2004, 72(2):937-948.
［2］Fan XL, Yu TH, Gao Q, Yao W. Immunological properties of recombinant Mycobacterium bovis bacillus Calmette-Guérin strain expressing fusion protein IL-2-ESAT-6［J］. Acta Biochim Biophys Sin(Shanghai), 2006, 38(10): 683-690.
［3］Shin DM, Jeon BY, Lee HM, Jin HS, Yuk JM, Song CH, et al. Mycobacterium tuberculosis eis regulates autophagy, inflammation, and cell death through redox-dependent signaling［J］. PloS Pathog, 2010, 6(12): e1001230.
［4］Yuk JM, Shin DM, Yang CS, Kim KH, An SJ, Rho J, et al. Role of apoptosisregulating signal kinase 1 in innate immune responses by Mycobacterium bovis bacillus CalmetteGuérin［J］. Immunol Cell Biol, 2009, 87(1): 100-107.
［5］Yu DS, Lee CF, Hsieh DS, Chang SY. Antitumor effects of recombinant BCG and uinterleukin-12 DNA vaccines on xenografted murine bladder cancer［J］. Urology, 2004, 63(3): 596-601.
［6］Giri PK, Schorey JS. Exosomes Derived from M. Bovis BCG infected macrophages activate antigen-specific CD4+ and CD8+T cells in vitro and in vivo［J］. PLoS One, 2008, 3(16): e2461.
［7］Mathé G, Amiel JL. The roles of adoptive and active forms of immunotherapy in the cure of children suffering from acute lymphoid leukemia: a) underestimation of active immunotherapy benefit, b)its immunogenetic indications to select sensitive patients,hence prevent chemotherapy's late effects［J］. Biomed Pharmacother, 2001, 55(9-10): 531-542.
