Cloning, expression analyses, and chromosomal location of porcine interleukin-18 receptor alpha chain (IL-18Ralpha).
We cloned and sequenced a cDNA that contains the coding sequence of the porcine interleukin-18 receptor alpha chain (PoIL-18Ralpha). Based on the conserved nucleotide sequences between human (HuIL-18Ralpha) and murine IL-18Ralpha (MuIL-18Ralpha), we performed reverse transcription-polymerase chain reaction (RT-PCR) with total RNA prepared from porcine peripheral blood lymphocytes (PBLs) stimulated with PoIL-12 to clone the cDNA of PoIL-18Ralpha. The open reading frame (ORF) of the PoIL-18Ralpha cDNA is 1620 base pairs (bp) in length and encodes 539 amino acids. The predicted amino acid sequence showed 68.2% and 50.2% identity to the human and murine amino acid sequences, respectively. Stimulation with concanavalin A (ConA) and IL-12, but not with IL-4, was shown to upregulate the expression of IL-18Ralpha mRNA in pig PBLs by RT-PCR analysis. Flow cytometric analysis also demonstrated that IL-18Ralpha was constitutively expressed on PoPBLs, and this expression was augmented by ConA stimulation. Furthermore, the PoIL-18Ralpha gene was mapped by fluorescence in situ hybridization (FISH) to porcine chromosome 3 (3q13-q14), near the location at which the IL-1beta gene had already been mapped. The present results will be helpful for understanding PoIL-18 and interferon gamma (IFN-gamma)-mediated T helper 1 (Th1) cell development.
Muneta
Yoshihiro
Y
Department of Immunology, National Institute of Animal Health, Tsukuba 305-0856, Japan. ymuneta@affrc.go.jp
Uenishi
Hirohide
H
Yamamoto
Ryuji
R
Yoshihara
Kazuhiro
K
Yasue
Hiroshi
H
Awata
Takashi
T
Mori
Yasuyuki
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping / Q000662:veterinary*; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018664:Interleukin-12 / Q000494:pharmacology; D053727:Interleukin-18 Receptor alpha Subunit; D015847:Interleukin-4 / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D021122:Protein Subunits / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D021101:Radiation Hybrid Mapping; D018123:Receptors, Interleukin / Q000096:biosynthesis / Q000235:genetics*; D053724:Receptors, Interleukin-18; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013552:Swine / Q000235:genetics*
D018076:DNA, Complementary; C508572:IL18R1 protein, human; C508573:Il18r1 protein, mouse; D053727:Interleukin-18 Receptor alpha Subunit; D021122:Protein Subunits; D012333:RNA, Messenger; D018123:Receptors, Interleukin; D053724:Receptors, Interleukin-18; D003208:Concanavalin A; D018664:Interleukin-12; D015847:Interleukin-4