Cloning,in vitroexpression, and bioactivity of interleukin-18 isolated from a domestic porcine breed found in Henan

Abstract

To evaluate the effects of recombinant porcine interleukin-18 (rpIL-18) on the replication of viruses in host cells and proliferation of lymphocytes, porcine IL-18 (pIL-18) isolated from a domestic big-white porcine breed found in the Henan province (HN) was cloned using a reverse transcriptase-PCR. The cloned HN pIL-18 contained an ORF of 579 base pairs encoding a 192-amino-acid precursor protein. The amino acid sequence of HN pIL-18 was compared with all the other pIL-18 amino acid sequences and varied by at least one amino acid to the consensus of all the others available. HN pIL-18 mature protein gene was inserted into a prokaryotic vector pGEX-4T-1 and expressed in Escherichia coli BL21. The expression of glutathione-S-transferase-pIL18 fusion protein was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. The rpIL-18 induced in vitro proliferation of concanavalin-A-stimulated porcine splenocytes, as revealed by the MTT assay. We studied the antiviral activities of the rpIL-18 on the replication of porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), and porcine parvovirus (PPV) cultured in two homologous cell lines. The results suggested that rpIL-18 can stimulate the proliferation of lymphocytes and inhibit viral pathogens infecting the porcine population.

IL-22 suppresses the infection of porcine enteric coronaviruses and rotavirus by activating STAT3 signal pathway

“…IL-22 has been demonstrated to combat a broad range of infectious agents in the gut through the up-regulation of antimicrobial peptides or cytokines Sonnenberg et al, 2011). Beta-defensin-2 (BD-2), IL-18 and IFN-l are among the antimicrobials and antiviral cytokines in small intestinal epithelial cells that are well studied (Chen et al, 2009;Lazear et al, 2015;Schroder and Harder, 1999;Zhang et al, 2014). We initially investigated whether mpIL-22 induced the expression of BD-2, IL- ).…”

Immune Responses of Piglets Immunized by a Recombinant Plasmid Containing Porcine Circovirus Type 2 and Porcine Interleukin-18 Genes

“…Three days after transfection, the total cellular lysates were collected and electrophoresed through an SDS-12% polyacrylamide gel. Afterwards, proteins were then transferred onto a nitrocellulose membrane (Bio-Rad, Hercules, CA) as described previously (7). The blots were probed with mouse anti-PCV2 mAb (Rural Technologies, Inc., Brookings, SD) or mouse anti-porcine IL-18 mAb (produced by our laboratory, unpublished data), washed, and exposed to horseradish peroxidase (HRP)-labeled anti-mouse IgG antibody (Southern Biotechnology Associates, Inc., Birmingham, AL).…”

Co-Formulation of Recombinant Porcine IL-18 Enhances the Onset of Immune Response in a New Lawsonia intracellularis Vaccine

“…The epIL-18 and ppIL-18 biological activity was evaluated in pig peripheral blood lymphocyte cultures using the MTT colorimetric assay [ 57 , 58 ]. The increased protein concentrations were lower than those reported in porcine IL-18 expressed in E. coli [ 57 ].…”