Generation of equine TSLP-specific antibodies and their use for detection of TSLP produced by equine keratinocytes and leukocytes

Veterinary Immunology and Immunopathology
J JandaE Marti

Abstract

Allergic horses react to innocuous environmental substances by activation of Th2 cells and production of allergen-specific IgE antibodies. The mechanisms leading to Th2 differentiation are not well understood. In humans and mice, epithelial cell-derived thymic stromal lymphopoietin (TSLP) plays a central role in this process. Little is known about equine TSLP (eqTSLP) and its role in allergic diseases and our current knowledge is limited to the assessment of TSLP mRNA expression. In order to be able to study eqTSLP at the protein level, the aim of the present study was to produce recombinant eqTSLP protein and generate TSLP-specific antibodies. EqTSLP was cloned from a skin biopsy sample from a horse with chronic urticaria and eqTSLP protein was expressed in E.coli and in mammalian cells. Recombinant proteins were designed to include C-terminal Histag, which allowed subsequent purification and detection by Histag-specific Ab. Polyclonal and monoclonal eqTSLP-specific Ab were generated after immunization of mice with E.coli-expressed TSLP. Their specificity was tested by western blotting and ELISA. In addition, a commercially available polyclonal human TSLP-specific antibody was tested for cross-reactivity with eqTSLP. Expressio...Continue Reading

References

Apr 22, 2005·Veterinary Dermatology·Silvia RüfenachtPetra J Roosje
Sep 6, 2005·Nature Immunology·Baohua ZhouSteven F Ziegler
Jan 25, 2006·The Journal of Experimental Medicine·Yong-Jun Liu
Feb 27, 2008·Nature Immunology·Caroline L SokolRuslan Medzhitov
Apr 27, 2010·Veterinary Immunology and Immunopathology·J Klukowska-RötzlerJ Janda
Aug 10, 2011·Respirology : Official Journal of the Asian Pacific Society of Respirology·Mathilde LeclereJean-Pierre Lavoie
Feb 22, 2012·Veterinary Immunology and Immunopathology·Jolanta Klukowska-RötzlerJozef Janda

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Citations

Nov 26, 2013·The Veterinary Clinics of North America. Equine Practice·Valerie A Fadok

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