Stimulation of Camel Polyclonal Antibody against Human T cell Immunoglobulin and Mucin 3

Document Type: Research Paper

Authors

1 Immunology Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

2 Genetic Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

3 Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, P.O.Box: 1316543551, Iran

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 confi rmed by qRT-PCR and fl ow 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 fi nal injection.
Conclusions: Our rhTIM-3 cell display system can be useful for future diagnostic or therapeutic approaches.

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