Generation of the Fluorescent HMGB1-GFP Fusion Protein in Insect Cells and Evaluation of its Immunogenicity in Two Mice Models

Title:Generation of the Fluorescent HMGB1-GFP Fusion Protein in Insect Cells and Evaluation of its Immunogenicity in Two Mice Models

Volume: 24 Issue: 6

Author(s): Ali Anvar, Rouhollah Vahabpour *, Iman Salahshourifar and Azam Bolhassani *

Affiliation:

• Department of Medical Lab Technology, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences; Tehran,Iran

• Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran,Iran

Keywords: HMGB1, GFP, Immunogenicity, Baculovirus-insect cells, fusion protein, adjuvant.

Abstract: Background: High mobility group box 1 (HMGB1) is a highly conserved protein present in the nuclei and cytoplasm of cells which has an important role as a mediator of inflammation in the extracellular environment. HMGB1 was identified as an innate adjuvant that induces immune responses against soluble antigens in vivo.

Objective: Our goal is the generation of recombinant HMGB1-GFP fusion protein in insect cells for evaluation of immune responses in mouse model.

Method: In the current study, we used a baculovirus expression system for insect cells that was based on expression of HMGB1 with target gene (GFP), and purified the recombinant HMGB1- GFP fusion protein. We then demonstrated whether immunogenicity of GFP changes in the presence or absence of recombinant HMGB1 acting as an adjuvant in C57BL/6 and BALB/c mice.

Results: Our data showed that HMGB1 had a major influence on antibody immune responses induced by GFP in both animal models. The groups receiving HMGB1-GFP fusion protein showed total IgG and IgG2a responses significantly higher than IgG1 in BALB/c mice. Indeed, a mixed IgG1/IgG2a response was observed with high intensity toward IgG2a. In contrast, C57BL/6 mice immunized by HMGB1-GFP protein elicited the same levels of IgG1 and IgG2a. However, the levels of IgG2a and total IgG against the recombinant GFP (rGFP) in C57BL/6 mice were lower than those in BALB/c mice.

Conclusion: We concluded that fusion of HMGB1 with GFP was immunologically more effective than GFP alone.

Cite this article as:

Anvar Ali , Vahabpour Rouhollah*, Salahshourifar Iman and Bolhassani Azam*, Generation of the Fluorescent HMGB1-GFP Fusion Protein in Insect Cells and Evaluation of its Immunogenicity in Two Mice Models, Protein & Peptide Letters 2017; 24 (6) . https://dx.doi.org/10.2174/0929866524666170404161649

DOI https://dx.doi.org/10.2174/0929866524666170404161649	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305