Document Type: Research Paper
Immunology Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Genetic Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, P.O.Box: 1316543551, Iran
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.