cDNA cloning and functional analysis of goose interleukin-2.
cDNA encoding goose IL-2 (GoIL-2) was cloned from Con A-stimulated goose splenic mononuclear cells (SMC) using oligonucleotide primers based on the conserved sequence of duck (DuIL-2), chicken (ChIL-2) and turkey IL-2s (TuIL-2). The GoIL-2 cDNA is 718nt long, which contains an open reading frame (ORF) of 423 base pairs encoding a protein of 141 aa. The GoIL-2 shows, respectively, 79%, 82-85%, and 91-92% identities with TuIL-2, ChIL-2 and DuIL-2 in cDNA, and also shows, respectively, 63%, 63-64%, and 82-85% identities with TuIL-2, ChIL-2 and DuIL-2 in amino acid sequence. Recombinant GoIL-2 (rGoIL-2) protein expressed in Escherichia coli has an approximate molecular weight of 18kDa. The rGoIL-2 has biological effect on goose and duck as well as chicken lymphocytes in a dose-dependent manner, though the effect on duck and chicken lymphocytes has been found to be relatively weak. In addition, rGoIL-2 also strengthens goose immune responses induced by vaccinating the inactivated oil emulsion vaccine against avian influenza virus. The monoclonal antibodies (mAb) to rGoIL-2 recognized the binding epitopes of nature GoIL-2 protein expressed in vero cells. Antiserum and mAb 5B10 to rGoIL-2 can inhibit the biological activity of rGoIL-2 and endogenous GoIL-2. The results, at the first time, indicated that goose IL-2 reserves species-specialties in the biological functions and can be used as a potential immunoadjuvant for goose vaccination and immunotherapeutic purposes. Finally, the mAbs to rGoIL-2 also provide a useful tool for further immunobiological studies of IL-2 in avian immune systems.
Zhou
Ji-Yong
JY
Laboratory of Virology and Immunology, Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, 268 Kaixuan Road, Hangzhou, Zhejiang Province 310029, PR China. jyzhou@zju.edu.cn
Chen
Ji-Gang
JG
Wang
Jin-Yong
JY
Wu
Jian-Xiang
JX
Gong
Hui
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001681:Biological Assay; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005777:Geese; D006801:Humans; D007150:Immunohistochemistry; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism*; D008214:Lymphocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010802:Phylogeny; D012313:RNA / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014162:Transfection
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012313:RNA