[Cloning, prokaryotic expression of LAIR and identification of their immunological reactivities].
OBJECTIVE
To clone and express the leukocyte-associated immunoglobulin-like receptor (LAIR) and to identify the immune reactivity of LAIR-2 to anti-LAIR-1 specific monoclonal antibodies(mAb).


METHODS
Genes encoding LAIR-1 and LAIR-2 were cloned by RT-PCR from human peripheral blood mononuclear cells(PBMC) and two leukemia cell lines Jurkat and HL-60. The extracellular region gene of LAIR1 and LAIR-2 were inserted into vector pGEX-4T-3 expressing GST fusion protein, expressed on IPTG induction and purified through glutathione-sepharose 4B column. The immunological reactivity of expressed LAIR-2 to anti-LAIR-1 mAb was identified by indirect ELISA.


RESULTS
LAIR-1 and LAIR-2 cDNAs had been cloned and expressed. Five new LAIR-1 cDNA isoforms were cloned. Among them, two isoforms from HL-60 included LAIR-1 open reading frames (ORF) and three isoforms from Jurkat were LAIR-1 cDNA segments. The LAIR-1 and LAIR-2 showed different immunological reactivities.


CONCLUSIONS
The transcription, processing after transcription and expression of LAIRs may be related to disparities in individuals and disease status. The difference in immunological reactivity may be involved in their structure.
Xie
Xin
X
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Ouyang
Wei-Ming
WM
Xue
Jiang-Nan
JN
Zhang
Xin-Hai
XH
Liu
Fei
F
Zhuang
Ran
R
Jin
Bo-Quan
BQ
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007963:Leukocytes, Mononuclear; D011993:Recombinant Fusion Proteins / Q000235:genetics
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins