Functional impact of IgA nephropathy-associated selectin gene haplotype on leukocyte-endothelial interaction.
Previously, we discovered single-nucleotide polymorphisms (SNPs) associated with Immunoglobulin A (IgA) nephropathy in selectin genes, which were 712C>T(P238S) in L selectin, -642A>G in the promoter region of L selectin, and 1402C>T(H468Y) in E selectin. Interestingly, these SNPs were in nearly complete linkage disequilibrium, thus two haplotypes, disease-associated TGT and wild-type (Wt) CAC, were constructed. To investigate the functional significance of TGT haplotype, a stable CHO transfectant expressing a P238S-L-selectin variant (CHO-varL) and a recombinant adenovirus vector containing an H468Y-E-selectin variant (Ad-varE) were established and compared to their Wt counterparts. Under flow, CHO-varL exhibited significantly less adhesion over IL-1beta-activated HUVEC monolayers compared to CHO-wtL. Furthermore, a luciferase reporter construct, containing a promoter region of the L-selectin variant (luc-varL), exhibited significantly less transcription activity compared to Wt (luc-wtL). These results suggest that the adhesive interactions and expression level of L selectin in disease-associated haplotypes are significantly compromised, indicating a potential role of these SNPs in the pathogenesis of inflammatory diseases, including IgA nephropathy.
Takei
Takashi
T
Department of Medicine, Kidney Center, Tokyo Women's Medical University, Tokyo, Japan.
Hiraoka
Megumi
M
Nitta
Kosaku
K
Uchida
Keiko
K
Deushi
Michiyo
M
Yu
Tao
T
Nitta
Noriko
N
Tsuchiya
Ken
K
Yumura
Wako
W
Nihei
Hiroshi
H
Nakamura
Yusuke
Y
Yoshida
Masayuki
M
D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000235:genetics* / Q000276:immunology; D006224:Cricetinae; D003412:Cricetulus; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000276:immunology; D005922:Glomerulonephritis, IGA / Q000235:genetics* / Q000276:immunology; D018922:HL-60 Cells; D006239:Haplotypes; D006801:Humans; D019041:L-Selectin / Q000235:genetics / Q000378:metabolism*; D007962:Leukocytes / Q000276:immunology*; D009154:Mutation; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011401:Promoter Regions, Genetic
D019040:E-Selectin; D019041:L-Selectin