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ISSN 0582-9879 Acta Biochim et Biophysica Sinica 2004, 36(6):430-436 CN
31-1300/Q
Human Recombinant B7-H3 Expressed in E. coli Enhances T
Lymphocyte Proliferation and IL-10 Secretion in Vitro
Guang-Bo ZHANG#, Yong-Jing CHEN#, Qin SHI,
Hong-Bing MA, Yan GE, Qin WANG, Zhi JIANG, Ying XU, and Xue-Guang ZHANG*
Institute of Biotechnology, Soochow
University, Key Laboratory of Clinical Immunology of Jiangsu Province, Suzhou
215007, China
Abstract To explore the
biofunctions of human B7-H3 on activated T lymphocyte, the gene of human B7-H3
encoding the extracellular region (IgV-like and IgC-like domains) was obtained
by RT-PCR from human lung cells and subcloned into the prokaryotic expression
vector pGEX-5X-3 to express glutathione S-transferase (GST) fusion protein. A
49 kD fusion protein (named as GST/hB7-H3 hereafter) was induced by IPTG and
purified by standard methods reported in prokaryotic system. In the presence of
the first signal imitated by anti-CD3 monoclonal antibody, T lymphocyte
proliferation was observed by incubating purified T cells with soluble
GST/hB7-H3 fusion protein by MTT assay. The concentrations of IFN-g and IL-10 in
the supernatants of T cells were determined by ELISA. The results showed that
the GST/hB7-H3 protein produced in bacteria had modest biological activities to
proliferate the T lymphocyte and enhance IFN-g as well as IL-10
secretion.
Key words B7-H3; GST-fusion
protein; T lymphocyte proliferation
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Received: February 24, 2004
Accepted: March 26, 2004
This work was supported by the grants from the National Basic
Research Program (No. 2001CB51003) and the National Natural Science Foundation
of China (300330540)
#These two authors contributed equally
to this work
*Corresponding author: Tel/Fax, 86-512-65104908; E-mail,
[email protected]