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Stimulation of Camel Polyclonal Antibody Against Human T Cell Immunoglobulin and Mucin 3 Publisher



Homayouni V1 ; Khanahmad H2 ; Ganjalikhanihakemi M1 ; Behdani M3 ; Ghasemi P1 ; Rezaei A1
Authors
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Authors Affiliations
  1. 1. Immunology Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  2. 2. Genetic Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  3. 3. Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, P.O.Box: 1316543551, Tehran, Iran

Source: Iranian Journal of Biotechnology Published:2017


Abstract

Background: T cell Immunoglobulin, Mucin (TIM)-3, is a type I transmembrane glycoprotein belonging to TIM family. This receptor expresses on T helper type 1 (Th1) cells that binds to galectin-9 (Gal9); inducing an inhibitory signal. As a result, apoptosis of Th1 cells occurs and cytotoxicity of CD8 T cells becomes evident in vitro. Therefore, this immunomodulatory molecule may be used as a novel target for clinical purposes. The production of camel polyclonal antibodies against TIM-3-expressing cell line was the purpose of this study. Objectives: In this study, we aimed to use HEK 293 cells expressing human TIM-3 to obtain camel polyclonal antibody against TIM-3 by immunization. Materials and Methods: A pre-synthesized human TIM-3cDNA was inserted into pcDNA3.1 plasmid and the new construct was transfected in HEK cell. TIM-3 expression was confirmed by qRT-PCR and flow cytometry. A camel (6 months old) was immunized with the lysate prepared from rTIM-3 expressing HEK cells 4 times. The anti-TIM-3 antibody level was evaluated using ELISA method. Results: TIM-3 was successfully cloned in HEK cells with 88% success rate. High level of anti-TIM-3 antibody was detected in the serum of the camel immunized with the recombinant cell lysate, after final injection. Conclusions: Our rhTIM-3 cell display system can be useful for future diagnostic or therapeutic approaches. © 2017 The Author(s).
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