应用流式细胞仪研究抗IL-8单抗对IL-8激活人颗粒细胞活力的中和作用(英文)

FLOW CYTOMETRY ANALYSIS OF THE NEUTRALIZATION EFFECT OF ANTI-IL-8 MCABS ON IL-8- ACTIVATED HUMAN GRANULOCYTES

以IL-8免疫的BALB/C小鼠脾细胞与Sp2/0或653小鼠骨髓瘤细胞融合构建了淋巴细胞杂交瘤克隆I8-S2和I8-63。ELISA叠加试验(ELISA Additivity Test)表明这两杂交瘤克隆分泌的单抗分别识别IL-8分子的不同表位。IL-8能激活人颗粒细胞,引起细胞内Ca^(2+)浓度([Ca^(2+)]_i)上升。通过流式细胞仪分析[Ca^(2+)]_i的变化,发现两个克隆单抗对IL8激活细胞的活力具有不同的中和作用。克隆I8-S2具有很强的中和作用,而克隆18-63则不然。上述结果提示IL-8的激活细胞活力局限于该分子的某表位。

Two hybridoma clones,18-S2 and I 8-63. secreting anti- IL-g antibodies were constructed from fusion of immunized spleen cells of BALB/c mice with either Sp 2/0 or 653 myeloma cells. They recognized different epitopes on IL-8 molecule.IL-8 could activate human granulocytes and induce the elevation of [Ca2+]1. Through flow cytometry analysis the two clones displayed different neutralizing effects on the cell activation function of IL-8. The neutralizing effect of clone I 8-S 2 was apparently higher than that of clone I 8-63. It is suggested that the cell activation function of IL-8 is restricted to a certain epitope of IL-8 molecule,