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

• Published in: Iranian journal of biotechnology Vol. 15; no. 3; pp. 166 - 171
• Main Authors: Homayouni, Vida, Khanahmad, Hossein, Ganjalikhani-Hakemi, Mazdak, Behdani, Mahdi, Ghasemi, Pouria, Rezaei, Abbas
• Format: Journal Article
• Language: English
• Published: National Institute of Genetic Engineering and Biotechnology 27-09-2017
• ISSN: 1728-3043; 2322-2921
• DOI: 10.15171/ijb.1427

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.