猪补体研究：猪CH50、ACH50及补体成分的测定

Studies of Pig Complement: Measurement of Pig CH50, ACH50, and Components.

作者信息

Sakai R, Kitano E, Hatanaka M, Lo P, Matsuura R, Deguchi K, Eguchi H, Maeda A, Watanabe M, Matsunari H, Nagashima H, Okuyama H, Miyagawa S

机构信息

Department of Pediatric Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.

Department of Medical Technology Faculty of Health Sciences, Kobe Tokiwa University, Kobe, Japan.

出版信息

Transplant Proc. 2016 May;48(4):1282-4. doi: 10.1016/j.transproceed.2015.10.066.

DOI:10.1016/j.transproceed.2015.10.066

PMID:27320604

Abstract

BACKGROUND

On the basis of a comparison of the hemolytic complement titer in pigs with that in humans, the complement system of pigs was investigated. The response of innate immunity, such as the natural antibodies, against humans was also examined.

METHODS

Hemolytic complement activity of pig serum was measured with the use of a microtitration technique. CH50 was determined according to the method of Mayer. ACH50 was assayed according to the methods of Platts-Milles and Ishizaka. Hemolytic activities of C1, C4, C2, C3, C5, C8, and C9 were estimated through the use of intermediate cells and reagents, as described previously. In addition, the pig natural anti-human antibody was studied with the use of human peripheral blood mononuclear cells (PBMCs). Human PBMCs were stained with 5% pig serum, followed by staining with fluorescein isothiocyanate-labeled goat anti-pig IgG and IgM. The resulting stained cells were quantified by use of a FACScalibur system. The alternative pathway of pig complement was also measured with the use of human erythrocytes and normal pooled pig serum with or without Mg(++)EGTA.

RESULTS

Both the CH50 and ACH50 titers were lower than those of humans. Concerning the components, except for C3, each component, that is, C1, C4, C2, C5, C8, and C9, was also lower than that of humans, based on measured values for human complement components. Pig serum clearly contains natural antibodies, IgG and IgM, to human PBMCs. The alternative pathway of pig complement reacted with human erythrocytes.

CONCLUSIONS

As a whole, pig innate immunity, the complement system and natural antibody, recognizes the surfaces of human cells.

摘要

背景

通过比较猪与人的溶血补体滴度，对猪的补体系统进行了研究。还检测了猪的天然抗体等固有免疫对人的反应。

方法

采用微量滴定技术测定猪血清的溶血补体活性。根据迈耶方法测定CH50。根据普拉茨 - 米尔斯和石坂的方法测定ACH50。如前所述，通过使用中间细胞和试剂估计C1、C4、C2、C3、C5、C8和C9的溶血活性。此外，利用人外周血单个核细胞（PBMC）研究猪天然抗人抗体。人PBMC用5%猪血清染色，然后用异硫氰酸荧光素标记的山羊抗猪IgG和IgM染色。使用FACScalibur系统对所得染色细胞进行定量。还使用人红细胞和含或不含Mg(++)EGTA的正常混合猪血清测量猪补体的替代途径。