IVIG中IgG片段对巨噬细胞吞噬致敏红细胞功能的影响  

Effect of IgG fragments in IVIG(pH4) on phagocytosis of sensitized erythrocytes by macrophage

作　　者：朱丽媛 张巍 侯明霞 刘卿 覃余燕 马莉[1] 李长清[1] ZHU Liyuan;ZHANG Wei;HOU Mingxia;LIU Qin;QIN Yuyan;Ma Li;LI Changqing(Institute of Blood Transfusion,Chinese Academy of Medical Sciences&Peking Union Medical College,Chengdu 610052,China)

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052 [2]深圳市卫光生物制品股份有限公司

出　　处：《中国输血杂志》2022年第12期1199-1203,共5页Chinese Journal of Blood Transfusion

基　　金：中国医学科学院医学与健康科技创新工程(2021-12MI-060)、(2021-12M-1-042);国家药品监督管理局血液制品质量控制重点实验室(依托单位:广东省药品检验所)开放课题(KF2021011)。

摘　　要：目的 研究静注人免疫球蛋白(pH4)(human immunoglobulin for intravenous injection, IVIG)中主要成分免疫球蛋白G(immunoglobulin G,IgG) Fab段、以及F(ab′)2段及Fc段对THP-1来源的巨噬细胞吞噬致敏红细胞吞噬功能的影响。方法 首先利用蛋白酶切技术和分离纯化技术,从IVIG中制备IgG Fc段、Fab段以及F(ab′)2段,并采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE)进行鉴定;其次,使用佛波酯诱导THP-1细胞分化为M0型巨噬细胞;使荧光染料羟基荧光素二醋酸盐琥珀酰亚胺脂标记致敏红细胞,采用流式细胞术检测上述各组分对M0型巨噬细胞吞噬致敏红细胞的吞噬能力的影响。结果 SDS-PAGE结果显示,所制备的IgG各片段均满足后续试验要求。流式结果表明,已成功建立M0型巨噬细胞吞噬致敏红细胞的吞噬模型。将IVIG中IgG各片段分别作用于M0型巨噬细胞吞噬致敏红细胞,当IgG Fc段蛋白浓度从0.1μg/mL增加至10μg/mL时,其对M0型巨噬细胞吞噬致敏红细胞的吞噬率从(24.21±0.58)%降至(12.27±0.19)%;当IVIG蛋白浓度从0.1μg/mL增加至10μg/mL,其吞噬率从(20.57±0.39)%下降至(0.20±0.03)%。当蛋白浓度相同时(10μg/mL),IgG Fc段对吞噬作用的抑制作用仅为IVIG的一半。此外,IgG Fab片段、F(ab′)2以及人血清白蛋白均不能抑制M0型巨噬细胞的吞噬作用。结论 IVIG可以有效地抑制THP-1来源的M0型巨噬细胞的吞噬作用,这种抑制作用主要依赖于IgG Fc段,而与IgG Fab段和F(ab′)2无关。Objective To research the effect of the Fc, Fab and F(ab′)2fragments of immunoglobulin G, the main components of Human Immunoglobulin(pH4) for Intravenous Injection(IVIG), on the phagocytic function of macrophages derived from THP-1 cells. Methods First of all, IVIG was digested with papain and pepsin to obtain Fc, Fab and F(ab′)2, and these components were then identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). Afterwards, propylene glycol monomethyl ether acetate(PMA) was used to induce THP-1 cells to differentiate into M0 macrophages. Finally, the sensitized erythrocytes were labeled with carboxy fluorescein succinimidyl ester(CFSE), and the effect of the above components on the phagocytic ability of M0 macrophages to engulf sensitized erythrocytes was detected by flow cytometry. Results The identification results of SDS-PAGE showed that the prepared IgG fragments met the requirements of subsequent experiments. Flow cytometry performs showed that the phagocytosis model of M0 macrophages had been successfully established. When the concentration of Fc increased from 0.1μg/mL to 10μg/mL, the phagocytosis rate of erythrocytes sensitized by M0 macrophages decreased from(24.21±0.58) % to(12.27±0.19) %. When the concentration of IVIG protein increased from 0.1 μg/ml to 10 μg/ml, the phagocytosis rate decreased from(20.57±0.39) % to(0.20±0.03) %. Meanwhile, at the same protein concentration(10 μg/ml), the inhibitory effect of Fc on phagocytosis was only half that of IVIG. In addition, Fab, F(ab′)2, and human serum albumin could not inhibit phagocytosis of M0 macrophages. Conclusion IVIG can effectively inhibit the phagocytosis of THP-1 derived M0 macrophages, which is mainly dependent on the Fc, but not related to the Fab of IgG and F(ab′)2.

关 键 词：静注人免疫球蛋白 FC段 THP-1细胞 吞噬 

分 类 号：R457.11[医药卫生—治疗学]