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

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

Journal Name: Protein & Peptide Letters

Volume 24 , Issue 6 , 2017

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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.

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

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Article Details

VOLUME: 24
ISSUE: 6
Year: 2017
Page: [545 - 550]
Pages: 6
DOI: 10.2174/0929866524666170404161649
Price: $58

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