Most DC-SIGNR transcripts at mucosal HIV transmission sites are alternatively spliced isoforms.
The repeat region of DC-SIGNR (CD209L) is polymorphic on the genomic level, and, in a separate study, we observed a correlation between the DC-SIGNR genotype and HIV-1 susceptibility during sexual contact. However, previous investigations using immunohistochemistry failed to detect membrane-bound DC-SIGNR on cells in the genital and rectal mucosa. We therefore explored the presence of DC-SIGNR in these compartments with a more sensitive limiting dilution RT-PCR, which also allowed for quantification of alternatively spliced mRNA isoforms. DC-SIGN (CD209) and DC-SIGNR mRNA transcript isoforms were found in all 12 vaginal and two rectal biopsies obtained from 14 healthy individuals. For DC-SIGNR, we detected significantly more isoform than full-length transcripts (mean copy numbers/mug RNA: 602 vs 26; P=0.0009). Four mucosal samples lacked full-length DC-SIGNR transcripts entirely. Cloning and sequencing of DC-SIGNR mRNA in three additional individuals revealed a diverse repertoire of DC-SIGNR isoforms, many of which encoded for proteins predicted to be soluble and secreted. Indeed, in one vaginal sample, we detected only soluble isoforms. In conjunction with our prior observation that the DC-SIGNR genotype has an effect on HIV-1 transmission in vivo, these findings emphasize that DC-SIGNR, in addition to DC-SIGN, should be considered as a cofactor in sexual HIV-1 transmission. Soluble isoforms, in particular, may modulate the efficiency of viral transmission and dissemination.
Liu
Huanliang
H
Department of Laboratory Medicine, University of Washington School of Medicine, Seattle, WA 98195, USA.
Hladik
Florian
F
Andrus
Thomas
T
Sakchalathorn
Polachai
P
Lentz
Gretchen M
GM
Fialkow
Michael F
MF
Corey
Lawrence
L
McElrath
M Juliana
MJ
Zhu
Tuofu
T
D000595:Amino Acid Sequence; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D005838:Genotype; D015658:HIV Infections / Q000635:transmission*; D015497:HIV-1; D006801:Humans; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000737:chemistry; D037181:Lectins, C-Type / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D009092:Mucous Membrane / Q000737:chemistry*; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D012007:Rectum; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014621:Vagina; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D020033:Protein Isoforms; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C433029:CLEC4M protein, human