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Logo of bmcbiotBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Biotechnology
 
BMC Biotechnol. 2012; 12: 43.
Published online Jul 30, 2012. doi:  10.1186/1472-6750-12-43
PMCID: PMC3468374
Heterologous expression of human costimulatory molecule B7-2 and construction of B7-2 immobilized polyhydroxyalkanoate nanoparticles for use as an immune activation agent
Ming-Chuan Li,#1 Qian-Qian Liu,#1 Xiao-Yun Lu,corresponding author1 Ya-Li Zhang,1 and Lei-Lei Wang1
1Department of Biological Science and Bioengineering, Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an 710049, Shaanxi, P. R. China
corresponding authorCorresponding author.
#Contributed equally.
Ming-Chuan Li: daybreaklmc/at/gmail.com; Qian-Qian Liu: com9150/at/163.com; Xiao-Yun Lu: luxy05/at/mail.xjtu.edu.cn; Ya-Li Zhang: yarlee/at/163.com; Lei-Lei Wang: qiuyequmeng/at/qq.com
Received March 13, 2012; Accepted July 25, 2012.
Abstract
Background
Costimulation of T cells via costimulatory molecules such as B7 is important for eliciting cell-mediated antitumor immunity. Presenting costimulation molecules by immobilizing recombinant B7 on the surface of nanovectors is a novel strategy for complementary therapy. Polyhydroxyalkanoates (PHAs) are a family of biodegradable, non-toxic, biocompatible polyesters, which can be used as a nonspecific immobilizing matrix for protein presentation. Recombinant protein fusion with PHA granule binding protein phasin (PhaP) can be easily immobilized on the surface of PHA nanoparticles through hydrophobic interactions between PhaP and PHA, and therefore provides a low-cost protein presenting strategy.
Results
In this study, the extracellular domain of the B7-2 molecule (also named as CD86) was fused with PhaP at its N-terminal and heterogeneously expressed in recombinant Escherichia coli strain BL21 (DE3). The purified B7-2-PhaP protein was immobilized on the surface of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx)-based nanoparticles. Loading of 240 μg (3.2 pMol) of B7-2-PhaP protein per mg nanoparticles was achieved. Immobilized B7-2-PhaP on PHBHHx nanoparticles induced T cell activation and proliferation in vitro.
Conclusions
A PHA nanoparticle-based B7-2 costimulation molecule-presenting system was constructed. The PHA-based B7 presenting nanosystem provided costimulation signals to induce T cell activation and expansion in vitro. The B7-2-PhaP immobilized PHA nanosystem is a novel strategy for costimulation molecule presentation and may be used for costimulatory molecule complementary therapy.
Keywords: PHA nanoparticle, B7-2, Costimulation, PhaP, Immobilization
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