Stimulation of camel polyclonal antibody against human T cell immunoglobulin and mucin 3


Homayouni V., Khanahmad H., Ganjalikhani-Hakemi M., Behdani M., Ghasemi P., Rezaei A.

Iranian Journal of Biotechnology, vol.15, no.3, pp.166-171, 2017 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 15 Issue: 3
  • Publication Date: 2017
  • Doi Number: 10.15171/ijb.1427
  • Journal Name: Iranian Journal of Biotechnology
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.166-171
  • Keywords: Camel antibody, Gal9, HcAb, HEK 293, TIM-3
  • Istanbul Medipol University Affiliated: No

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.