IL-2受体放射配基结合分析法的建立

Development of a radioligand binding assay for interlenkin 2 receptor

为建立一种稳定可靠的白细胞介素-2受体(IL-2R)放射配基结合分析(RBA)法,对6例健康男性取血分离并培养外周血单个核细胞(PBMC),进行 IL-2R 多点法 RBA;10例健康男性进行 PBMC IL-2R 单点法 RBA。结果:多点法 RBA测得每个细胞高亲和力 IL-2R 的数量为1125.7±180.4个,其 Kd_(高)=(7.060±2.928)×10^(-11)mol/L;低亲和力 IL-2R 的数量为12 524.5±3 816.0个,其 Kd_(低)=(2.374±0.924)×10~9mol/L。单点法 RBA 测得每个细胞表面高亲和力 IL-2R 的数量为1180.4±291.8个;低亲和力 IL-2R 的数量为12430.1±3161.2个。两种方法所测数据无明显差异。因此,单点法RBA 是一种简单有效的测定 IL-2R 方法。

PURPOSE To develop a stable and reliable interleukin 2 receptor (IL-2R) radioligand binding assay.METH- ODS The periphreal blood mononuclear cells (PBMC) were separated from healthy human blood and cultured in RPMI-1640 with PHA-M for 72h.The IL-2R in PBMC was analyzed using either multipoint RBA method or single point RBA method.RE- SULTS The results of multipoint RBA (n=6) were:for higb affinity IL-2R,RT=1125.7±180.4((?)±s)per cell,Kd= (7.060±2.928)×10^(-11)mol/L,and for low affinity IL-2R,RT=12524.5±3816((?)±s) per cell,Kd=(2.374±0.924)× 10^(-9)mol/L.The results of single point RBA method (n=10) were:for high affinity IL-2R,RT=1180.40±291.8((?)±s) per cell,and for low affinity IL-2R,RT=12430.1±3161.2((?)±s) per cell.There was no statistical significance between the results of the two methods.CONCLUSIONS Single point IL-2R RBA is a practical method for estimation of receptor density or PBMC.