Production and characterization of two monoclonal antibodies to bovine tumour necrosis factor alpha (TNF-α.) and their cross-reactivity with ovine TNF-α

被引：18

作者：

Kwong, L. S. ^{[1
]}

Thom, M. ^{[1
]}

Sopp, P. ^{[1
]}

Rocchi, M. ^{[2
]}

Wattegedera, S. ^{[2
]}

Entrican, G. ^{[2
]}

Hope, J. C. ^{[1
]}

机构：

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

[2] Int Res Ctr, Moredun Res Inst, Edinburgh EH26 0PZ, Midlothian, Scotland

来源：

VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY | 2010年 / 135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：

Tumour necrosis factor alpha; ELISA; Intracytoplasmic expression; MYCOBACTERIUM-BOVIS; TUBERCULOSIS; CELLS; ELISA;

D O I：

10.1016/j.vetimm.2010.01.001

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Tumour necrosis factor alpha (TNF-alpha) is an innate pro-inflammatory cytokine involved in protection against intracellular pathogens. Existing methods for measuring TNF-alpha production and function in ruminants are limited to ELISA and many rely on polyclonal antisera. With a view to developing improved detection methods for bovine (boy) TNF-alpha, monoclonal antibodies (mAb) were produced by immunising mice with a plasmid encoding boy TNF-alpha. Two of the resulting mAb, termed CC327 and CC328, were used to develop a sandwich ELISA capable of detecting both native and recombinant boy TNF-alpha. This ELISA did not detect recombinant ovine (ov) TNF-alpha. A luminometric method was applied to the ELISA to improve sensitivity for detection of native boy TNF-alpha in culture supernatants derived from bovine monocyte-derived dendritic cells (DC) infected with Mycobacterium bovis. Both CC327 and CC328 detected intracytoplasmic expression of TNF-alpha in mitogen-activated bovine T lymphocytes. However, only CC328 detected intracytoplasmic ovine TNF-alpha in transfected cells, explaining the failure of the sandwich ELISA to detect recombinant ov TNF-alpha. These mAbs have generated the capability to study the role of TNF-alpha in host immune protection and disease pathogenesis in ruminants. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：320 / 324

页数：5

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