Down-regulation of CD53 expression in Epinephelus coioides under LPS, poly (I:C), and cytokine stimulation.

Tetraspanins are a group of cell surface molecules involved in cell adhesion, motility, metastasis, signal transduction, and immune cell activation. Members of the tetraspanin family include CD9, CD37, CD63, CD53, and others. However, few tetraspanins have been investigated in teleosts. In this study, we obtained the open reading frame of CD53 cDNA from orange spotted grouper (Epinephelus coioices), an economically important fish. The predicted amino acid structure contains four membrane-spanning domains and a conserved CCG motif. The amino acid identity between human and grouper CD53 was only 38%; however, both CD53 proteins share the same structure. Quantitative real-time PCR revealed that mRNA is abundant in immune organs, including the head and trunk kidneys, spleen, thymus, gill, and blood. Immunochemistry and immunofluorescence analyses further revealed that CD53 was majorly expressed in the leukocytes of various organs. Finally, mRNA and protein expression for CD53 was down-regulated in fish treated with immune stimulators, including LPS, Poly (I:C), Vibrio, recombinant grouper IL-6, and CCL4. Our results indicate that the expression of CD53 may play important roles in pathogen invasion and inflammation reaction.

The biofunction of orange-spotted grouper (Epinephelus coioides) CC chemokine ligand 4 (CCL4) in innate and adaptive immunity.

CC chemokine (motif) ligand 4 (CCL4) is indispensable to the chemoattraction of macrophages, natural killer cells, and lymphocytes in mammals; however, it has only been cloned in a limited number of fish species and information related to its biofunction remains ambiguous with regard to teleosts. To explore the role of teleost CCL4, we first evaluated the mRNA expression of the Epinephelus coioides CCL4 (gCCL4) gene in various organs under LPS and poly (I:C) stimulated; secondary, we evaluated the immune-related genes expression of fish under the recombinant gCCL4 protein stimulated. Our results revealed an increase in the mRNA of gCCL4 in immune organs immediately following stimulation by poly (I:C); however, in LPS stimulated fish, the expression did not increase until nearly 24 h after induction. In biofunction assays, recombinant gCCL4 was found to induce chemotactic activity in the peripheral blood leukocytes of groupers and up-regulate the gene expressions of grouper TNFA1 (TNF-α1), TNFA2 (TNF-α2), IFNG (IFN-γ), MX, TBX21 (T-bet), CD8 (α and ß chain). These findings indicate that grouper CCL4 attracts leukocytes, induces an inflammatory response, and drives lymphocyte differentiation into the Th1 pathway.

Comparison of polycyclic aromatic hydrocarbon emissions on gasoline- and diesel-dominated routes.

Three diesel-dominated routes (DDRs) and three gasoline-dominated routes (GDRs) were chosen as the study sites. The total number of vehicles on GDRs (47,200) was much higher than that on DDRs (14,500). The concentration of polycyclic aromatic hydrocarbons (PAHs), elemental carbon, organic carbon, and metals from GDR roadsides was higher than that for DDRs. The diagnostic ratios (ANTHR/PHE + ANTHR, FLT/FLT + PYR, BaA/BaA + CHR, and IND/IND + BghiP + ANTHN) all indicated that the major PAH source on DDR and GDR was emissions from vehicle engine combustion. The marked diesel ratios of low molecular weight PAH2.5/T-PAH2.5, methyl-PAH2.5/T-PAH2.5, methyl-PHE/PHE, and Mo/PM2.5 on DDRs were higher than those on GDRs. Significant correlations were found between the number of vehicles and the concentration of T-PAH2.5, Car-PAHs2.5, and BaPeq2.5 on DDRs and GDRs. The increase in the levels of T-PAH2.5, Car-PAHs2.5, and BaPeq2.5 per 100 vehicles on DDRs was about 3.3, 3.5, and 4.2 times higher than that on GDRs, respectively. The higher percentage of high-exhaust volume from the larger amount of diesel vehicles on DDRs than that on GDRs was the main factor leading to these results. The diagnostic ratios BaA2.5/CHR2.5 and (BbF + BkF)2.5/BghiP2.5 showed significant differences between the fine PAH sources emitted on DDRs and GDRs, whereas the diagnostic ratios Me-PAH2.5/T-PAH2.5 and (BbF + BkF)2.5/BghiP2.5 showed good correlations with the percentages of diesel exhaust volume in the total exhaust volume (E(diesel)/E(total)) on DDRs.