特异性DNA探针检测细胞因子诱导的杀伤细胞在患者体内的生存时间

目的 观察应用荧光原位杂交技术(FISH)研究细胞因子诱导的杀伤细胞(CIK)在性别错配的女性受者体内生存时间.方法 通过细胞采集仪采集男性供者的单个核细胞,经多种细胞因子体外诱导培养成CIK细胞,输注给女性受者.治疗数月后,用Y染色体特异性DNA探针测定女性受者体内淋巴细胞上Y染色体的表达.结果 流式细胞仪检测第14天时CIK细胞高表达CD3+ CD56+为55.4％.CSP X探针定位于Xp11.1-q11.1,杂交信号为绿色,CSP Y探针定位于Yp11.1-q11.1,杂交信号为红色,女性细胞染色体XX显示两个绿色荧光信号,男性细胞染色体XY 显示1个绿色信号,1个红色荧光信号.在女性受者体内淋巴细胞上有Y染色体的表达,表达比例为0.5％ ～3.0％.结论 Y染色体特异性DNA探针可以确定CIK细胞在患者体内的生存时间.

Detection of survival time of cytokine-induced killer cells in vivo by specific DNA probes

Objective To observe the survival time of cytokine-induced killer cells (CIK) in vivo in gender-mismatched female recipients by using fluorescence in situ hybridization (FISH).Methods Mononuclear cells of male donors were collected with cell collection device,and induced into CIK cells by several kinds of cytokines in vitro,and then were infused to the female recipients.Several months after CIK treatment,the expression of Y chromosome in lymphocytes of the female recipients was detected by using Y chromosome specific DNA probes.Results CIK cells were highly expressed after 14 days,and CD3 +CD56 + was 55.4％ by flow cytometry.CSP X probe was located at Xp11.1-q11.1,and the hybridization signal was green; CSP Y probe was located at Yp11.1-q11.1,and the hybridization signal was red.Female cell chromosome was XX,showing two green fluorescent signals; Male cell chromosome was XY,showing one green fluorescent signal and one red fluorescent signal.Y chromosome was expressed in the lymphocytes of the female recipients,and the expression rate was 0.5％ to 3.0％.Conclusion The survival time of CIK cells in vivo can be determined by using Y chromosome specific DNA probes.