Cloning, in vitro expression, and bioactivity of interleukin-18 isolated from a domestic porcine breed found in Henan.
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.
Chen
Hong-Ying
HY
Henan Agricultural University, Zhengzhou, China.
Zheng
Lan-Lan
LL
Li
Xin-Sheng
XS
Wei
Zhan-Yong
ZY
Cui
Bao-An
BA
Li
Xiao-Kang
XK
Liu
Jin-Peng
JP
Yin
Hong-Zheng
HZ
Meng
Jiang-Tao
JT
Zhang
Yan
Y
Li
Shi-Min
SM
D000818:Animals; D048708:Cell Growth Processes / Q000187:drug effects; D002460:Cell Line; D002681:China; D003001:Cloning, Molecular / Q000379:methods; D003588:Cytopathogenic Effect, Viral; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D011558:Herpesvirus 1, Suid / Q000502:physiology; D020382:Interleukin-18 / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology; D028323:Parvovirus, Porcine / Q000502:physiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D019316:Porcine respiratory and reproductive syndrome virus / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D020539:Sequence Analysis, Protein; D013552:Swine; D014779:Virus Replication / Q000187:drug effects
D020382:Interleukin-18; D011994:Recombinant Proteins