Production of mmLFA3-Ig with immunosuppressive activity by recombinant expression in Pichia pastoris
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1
Key Lab of Transplant Engineering and Immunology, Ministry of Health,SiChuan University, China
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2
Regenerative Medicine Research Center, West China Hospital, Sichuan University, China
Engagement of lymphocyte function antigen 3 (LFA3) and CD2 can increase the activation of T cells. Blockade of LFA3-CD2 pathway efficiently down-regulated the abnormal immune activation, thus resulting in the development of human LFA3-Ig (hLFA3-Ig) as a treatment for psoriasis. Nevertheless, it is far from to unravel the role of LFA3-CD2 in regulation of immune system. Rhesus monkey macaca mulatta has been widely used as animal models for human immune disorders. Due to specificity of the immune system, M. mulatta LFA3-Ig (mmLFA3-Ig) might be required for pathogenesis of immune disorders in rhesus monkey models. The gene encoding mmLFA3 were amplified from peripheral blood mononuclear cells (PBMCs) by RT-PCR. Amino acid sequence analysis demonstrated that the identity between the predicted extracellular domain (ECD) of mmLFA3 and that of hLFA3 is about 86%. To prepare mmLFA3-Ig, the ECD of mmLFA3 was fused to the N-terminus of human IgG1 Fc fragment. The gene encoding mmLFA3 ECD was firstly optimized according to the codon bias of P. pastoris. MmLFA3-Ig produced by P. pastoris specifically bound CD2-positive cells. Positive staining was detected by flow cytometer in approximately 90% CD2-positive Jurkat cells, compared to that in 5% CD2-negative Raji cells. Moreover, mmLFA3-Ig showed dose-dependent suppression on monkey and human lymphocyte proliferation in both Con A- and allogenic antigen-stimulated systems. Interestingly, mmLFA3-Ig showed stronger immunosuppression on monkey lymphocyte proliferation than that of human. These results suggest that mmLFA3-Ig produced by P. pastoris might be a used for pathogenesis of immune disorder in rhesus monkey model.
Keywords:
LFA3,
CD2/CD58,
Immunosuppression,
Immunotherapy,
Pichia pastoris
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Translational immunology and immune intervention
Citation:
Zhu
J,
Zhu
S,
Yang
H,
Cheng
J,
Lu
X and
Wan
L
(2013). Production of mmLFA3-Ig with immunosuppressive activity by recombinant expression in Pichia pastoris.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00467
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Received:
13 Apr 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Dr. Lin Wan, Key Lab of Transplant Engineering and Immunology, Ministry of Health,SiChuan University, Chengdu, China, scuwan@163.com