Molecular cloning and expression analysis of pig lymphocyte activation gene-3 (LAG-3; CD223).
Lymphocyte activation gene-3 (LAG-3; CD223) is a co-receptor that may function as a down-modulator of T cell activity via an interaction with MHC class II molecules. In this study, we cloned and sequenced the complete cDNA sequence of pig Lag3. Pig Lag3 cDNA contains an open reading frame (1524bp) encoding 507 amino acids. The putative amino acid identity of pig LAG-3 with those of the human, rat, and mouse is 78%, 66%, and 67%, respectively. Pig Lag3 mRNA transcript was mainly expressed in lymphoid tissues, and was inducible upon stimulation of phytohemagglutinin (PHA). Next, we developed mouse antiserum against the extracellular domain (D1 and D2 domains) of pig LAG-3. Pig LAG-3 is detected as an approximately 67kDa of a single species by Western blot analysis. Moreover, flow cytometry analysis confirmed that LAG-3 on pig T cells (CD3(+) cells) is inducible upon PHA stimulation. These results indicated that the structure and expression pattern of pig LAG-3 are conserved among mammalian species.
Kim
Seon-Soo
SS
School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Republic of Korea.
Kim
Sang-Hoon
SH
Kang
Hyung-Sik
HS
Chung
Hee Yong
HY
Choi
Inho
I
Cheon
Yong-Pil
YP
Lee
Ki Ho
KH
Lee
Dong-Mok
DM
Park
Jongsun
J
Lee
Sang Yeol
SY
Chun
Taehoon
T
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D003001:Cloning, Molecular; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D008213:Lymphocyte Activation; D008297:Male; D008969:Molecular Sequence Data; D012313:RNA / Q000737:chemistry / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D016415:Sequence Alignment; D013552:Swine / Q000235:genetics* / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D000093322:Lymphocyte Activation Gene 3 Protein
D015703:Antigens, CD; D012313:RNA; D000093322:Lymphocyte Activation Gene 3 Protein