Development of an ELISA for bovine IL-10

被引：117

作者：

Kwong, LS ^{[1
]}

Hope, JC ^{[1
]}

Thom, ML ^{[1
]}

Sopp, P ^{[1
]}

Duggan, S ^{[1
]}

Bembridge, GP ^{[1
]}

Howard, CJ ^{[1
]}

机构：

[1] Inst Anim Hlth, Compton RG20 7NN, Berks, England

来源：

VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY | 2002年 / 85卷 / 3-4期

基金：

英国生物技术与生命科学研究理事会;

关键词：

interleukins; bovine IL-10; ELISA; cattle; cytokines;

D O I：

10.1016/S0165-2427(02)00007-7

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

The objective of the study was to develop an assay for bovine IL-10 that could be applied to analyses of immune responses and advance understanding of a variety of diseases of cattle. Recombinant bovine IL-10 (rbo IL-10) was transiently expressed in Cos-7 cells and shown to inhibit the synthesis of IFNgamma by bovine cells stimulated with antigen in vitro. Mice were immunised with a plasmid containing a cDNA insert encoding rbo IL-10 and inoculated with rbo IL-10. A number of monoclonal antibodies (mAb) were generated that reacted with rbo IL-10 in an ELISA. Some of these mAb neutralised the ability of rbo IL-10 to inhibit IFNgamma synthesis by antigen-stimulated bovine cells. A pair of mAb was identified that together could be used to detect both recombinant and natural bovine IL-10 present in supernatant of PBMC stimulated with ConA. A luminescent detection method was applied to the ELISA making it more sensitive. Using this method native IL-10 was detected in supernatants of PBMC, diluted blood and undiluted blood from cattle immunised with Mycobacterium bovis BCG or ovalbumin and incubated in vitro with antigen indicating the applicability of the assay to a number of in vitro culture systems. (C) 2002 Elsevier Science B.V. All rights reserved.

引用

页码：213 / 223

页数：11

共 17 条

[1] Bovine interleukin-12 and modulation of IFNγ production [J].