CAJ | 학술논문

目的：研究经加热处理的人黑色素瘤细胞A375对外周血单个核细胞来源的自然杀伤(NK)细胞及树突状细胞(DC)免疫活性的影响并探讨其作用机制。方法：体外培养人外周血单个核细胞来源的NK和DC细胞，并使其与43℃水浴加热后的A375细胞共孵育24 h。应用CCK-8法测定加热和非加热组效靶比(NK∶DC∶A375)分别为1∶2∶1、3∶6∶1、6∶12∶1时NK/DC细胞的杀伤率；应用酶联免疫吸附法(ELISA)检测上述各效靶比组NK细胞培养液中γ-干扰素(γ-INF)的释放情况。结果：在每个效靶比，与非加热组比较，加热组NK/DC的杀伤率均明显提高(P<0.05)，且其杀伤率随NK/DC细胞的浓度升高而升高(<0.05)。在加热组和非加热组，含有DC较不含DC组杀伤率均明显提高(P<0.05)。加热组NK细胞γ-INF释放均较非加热组明显增加(P<0.05)。结论：经加热处理后的黑色素瘤细胞A375能够刺激NK细胞分泌更多的γ-INF，明显提高NK细胞的杀伤活性，且DC在其中起着重要作用。
목적：연구경가열처리적인흑색소류세포A375대외주혈단개핵세포래원적자연살상(NK)세포급수돌상세포(DC)면역활성적영향병탐토기작용궤제。방법：체외배양인외주혈단개핵세포래원적NK화DC세포，병사기여43℃수욕가열후적A375세포공부육24 h。응용CCK-8법측정가열화비가열조효파비(NK∶DC∶A375)분별위1∶2∶1、3∶6∶1、6∶12∶1시NK/DC세포적살상솔；응용매련면역흡부법(ELISA)검측상술각효파비조NK세포배양액중γ-간우소(γ-INF)적석방정황。결과：재매개효파비，여비가열조비교，가열조NK/DC적살상솔균명현제고(P<0.05)，차기살상솔수NK/DC세포적농도승고이승고(<0.05)。재가열조화비가열조，함유DC교불함DC조살상솔균명현제고(P<0.05)。가열조NK세포γ-INF석방균교비가열조명현증가(P<0.05)。결론：경가열처리후적흑색소류세포A375능구자격NK세포분비경다적γ-INF，명현제고NK세포적살상활성，차DC재기중기착중요작용。
To study the effects of the heated A375 melanoma cells on natural killer (NK) cells and dendritic cells (DC) generated from human cultured peripheral blood mononuclear cells (PBMCs) in vitro,and to explore the mechanism. METHODS:NK cells and DCs cultured from PBNCs in vitro and reacted with A375 melanoma cells in a water-bath at 43℃. Using CCK-8 kit to test the killing rate of NK cells and DCs against A375 melanoma cells at the effector∶ratio of efficacy to target(E/T) (NK∶DC∶A375) of 1∶2∶1,3∶6∶1,and 6∶12∶1. ELISA was used to measure concentrations ofγ-INF released by NK cells at the effector∶E/T (NK∶DC∶A375) of 1∶2∶1,3∶6∶1 and 6∶12∶1. RESULTS:At each E/T ,the killing rate of the heated group against A375 melanoma cells were stronger than the unheated group (P<0.05),and the killing rate increased as the concentrations of NK and DC increased (P<0.05). The concentration of γ-INF released by the heated group was also more than the unheated group(P<0.05). CONCLUSION:Heated A375 melanoma cells could induce NK to release moreγ-INF and significantly increased the activity of NK cells ,in which DC played an important role.