Expression And Activity Identification Of Rat IFN-γ-inducible Protein-10 By Escherichia Coli Expression System

Objective:Chemokines have principal roles in immune surveillance and in disease. Chemokines are expressed by monocytes, endotheliocytes and lymphocytes in inflammation, which mediate immunologically competent cells activation and migration, and regulate immune responses. Recently, chemokines have been correlated with central nervous system (CNS) inflammation such as mutiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE). The pathological conditions of CNS inflammation is leukocytes trafficking into the CNS. Chemokines have principal roles in mediating leukocytes into CNS, in which interferon-y-inducible protein-10 (IP-10) is payed attention especially. IP-10 is three non-Glu-Leu-Arg CXC chemokine which can induce the migration of many inflammatory cells including T cells, monocytes and NK cells. Many studies have showed that during CNS inflammation, microglias were activated and expressed IP-10 that induced inflammatory cells infiltrating, which promoted CNS inflammation processes. For studying the bioactivity of IP-10 and the function of IP-10 in CNS inflammation. This study constructed prokaryotic expression plasmid pET-32a(+)-IP10 which was expressed in Escherichia coli BL21(DE3). Then the chemotactic activity of IP-10 fusion protein was detected after being purified with Ni²⁺ affinity chromatography. The study established foundation for studying new therapeutic methods for CNS inflammation, of which IP-10 acted as a target.