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Updated: Feb 9, 2026

Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing
Published on: March 15, 2019
Stimulation of Camel Polyclonal Antibody against Human T cell Immunoglobulin and Mucin 3
Vida Homayouni1, Hossein Khanahmad2, Mazdak Ganjalikhani-Hakemi1
1Immunology Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, 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 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.
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