Objective To observe the protective immunity induced by the nucleic acid vaccine of 21.^7 kDa membrane protein molecule of Schistosoma japonicum Chinese mainland strain (SjC 21.^7) in BALB/c mice. . Methods. A pair of primers (P1 and P2) was synthesized according to the DNA sequence of the SjC21.^7. The ORF sequence of SjC21.^7 was amplified by PCR, and the Kozark sequence was added to the position of initiator. The gene fragment was inserted into the eukaryotic expression plasmid pcDNA3.^1 to form the recombinant plasmid SjC21.^7-pcDNA3.^1. Forty-eight BALB/c mice were divided into three groups: control, test and boost. Each mouse was injected in quadriceps femoris with plasmid pcDNA3.^1 (control) or recombinant plasmid SjC21.^7-pcDNA3.^1 (test, boost); for the boost group, with additional P35-pcDNA3.^1 and P40-pcDNA3.^1. All mice were immunized three times with an interval of 2 weeks, challenged each with 45 cercariae of S.^japonicum at the 30th day after final immunization. At day 45 after challenge,all mice were sacrificed, the numbers of worms and hepatic eggs were counted. Antibody level in the sera of mice before and two weeks after immunization was determined with ELISA. The expression of the target gene in quadriceps femoris was observed with immunohistochemistry. . Results . The immunohistochemistry analysis showed that there were specific antigens expressed in the local tissue of the test group mice. There was specific IgG in the serum of partial mice in test and boost groups. Compared with the control group, the worm reduction rate was 29.^9% and its egg reduction rate 13.^8% in the test group; 31.^9% and 28.^0% respectively in the boost group. The egg reduction rate in the boost group was higher than that of the test group (P

Macrophages acquire distinct phenotypes during tissue stress and inflammatory responses. Macrophages are roughly categorized into two different subsets named inflammatory M1 and anti-inflammatory M2 macrophages. We herein identified a unique pathogenic macrophage subpopulation driven by IL-23 with a distinct gene expression profile including defined types of cytokines. The freshly isolated resting mouse peritoneal macrophages were stimulated with different cytokines in vitro, the expression of cytokines and chemokines were detected by microarray, real-time PCR, ELISA and multiple colors flow cytometry. Adoptive transfer of macrophages and imiquimod-induced psoriasis mice were used. In contrast to M1- and M2-polarized macrophages, IL-23-treated macrophages produce large amounts of IL-17A, IL-22 and IFN-γ. Biochemical and molecular studies showed that IL-23 induces IL-17A expression in macrophages through the signal transducer and activator of transcription 3 (STAT3)-retinoid related orphan receptor-γ T (RORγT) pathway. T-bet mediates the IFN-γ production in IL-23-treated macrophages. Importantly, IL-23-treated macrophages significantly promote the dermatitis pathogenesis in a psoriasis-like mouse model. IL-23-treated resting macrophages express a distinctive gene expression prolife compared with M1 and M2 macrophages. The identification of IL-23-induced macrophage polarization may help us to understand the contribution of macrophage subpopulation in Th17-cytokines-related pathogenesis.
Animals ; Cell Polarity ; Imiquimod ; Interleukin-23 ; metabolism ; Macrophages ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Psoriasis ; chemically induced ; metabolism ; pathology