Functional genetic variants in DC-SIGNR are associated with mother-to-child transmission of HIV-1

Geneviève Boily-Larouche, Anne-Laure Iscache, Lynn S Zijenah, Jean H Humphrey, Andrew J Mouland, Brian J Ward, Michel Roger, Geneviève Boily-Larouche, Anne-Laure Iscache, Lynn S Zijenah, Jean H Humphrey, Andrew J Mouland, Brian J Ward, Michel Roger

Abstract

Background: Mother-to-child transmission (MTCT) is the main cause of HIV-1 infection in children worldwide. Given that the C-type lectin receptor, dendritic cell-specific ICAM-grabbing non-integrin-related (DC-SIGNR, also known as CD209L or liver/lymph node-specific ICAM-grabbing non-integrin (L-SIGN)), can interact with pathogens including HIV-1 and is expressed at the maternal-fetal interface, we hypothesized that it could influence MTCT of HIV-1.

Methods and findings: To investigate the potential role of DC-SIGNR in MTCT of HIV-1, we carried out a genetic association study of DC-SIGNR in a well-characterized cohort of 197 HIV-infected mothers and their infants recruited in Harare, Zimbabwe. Infants harbouring two copies of DC-SIGNR H1 and/or H3 haplotypes (H1-H1, H1-H3, H3-H3) had a 3.6-fold increased risk of in utero (IU) (P = 0.013) HIV-1 infection and a 5.7-fold increased risk of intrapartum (IP) (P = 0.025) HIV-1 infection after adjusting for a number of maternal factors. The implicated H1 and H3 haplotypes share two single nucleotide polymorphisms (SNPs) in promoter region (p-198A) and intron 2 (int2-180A) that were associated with increased risk of both IU (P = 0.045 and P = 0.003, respectively) and IP (P = 0.025, for int2-180A) HIV-1 infection. The promoter variant reduced transcriptional activity in vitro. In homozygous H1 infants bearing both the p-198A and int2-180A mutations, we observed a 4-fold decrease in the level of placental DC-SIGNR transcripts, disproportionately affecting the expression of membrane-bound isoforms compared to infant noncarriers (P = 0.011).

Conclusion: These results suggest that DC-SIGNR plays a crucial role in MTCT of HIV-1 and that impaired placental DC-SIGNR expression increases risk of transmission.

Conflict of interest statement

Competing Interests: J. H. received an operating grant from BASF for the ZVITAMBO project.

Figures

Figure 1. DC-SIGNR haplotypes reconstruction with minor…
Figure 1. DC-SIGNR haplotypes reconstruction with minor allele frequency >5% (MAF) and selected htSNPs in Zimbabwean population.
Figure 2. Effect of the int2-180A variant…
Figure 2. Effect of the int2-180A variant on placental DC-SIGNR isoform expression.
(a) Schematic representation of the int2-180A variant location within the DC-SIGNR gene. (b) Proportion of DC-SIGNR membrane-bound and soluble isoforms in homozygous H1 (int2-180AA) and homozygous WT (int2-180GG) placental samples estimated by qRT-PCR assays in three independent experiments performed in triplicate. Data are presented as percentage of total transcripts. Differences between isoform proportions among the homozygous H1 and homozygous WT were calculated by the χ2 test.
Figure 3. Transcriptional activity of DC-SIGNR promoters.
Figure 3. Transcriptional activity of DC-SIGNR promoters.
(a, b) Effect of the p-198A variant on transcriptional activity in luciferase reporter assay in vitro in transfected HeLa cells. (a) Schematic representation of reporter gene constructs corresponding to the DC-SIGNR promoter region from –715 to –1, spanning with either C (WT) or A at position −198. (b) Relative luciferase expression from pGL2-Basic, the parental vector without the promoter. Expression of the DC-SIGNR promoter constructs were calculated relatively to this value. Data are presented in mean±S.E.M. values of three independent experiments performed in triplicate and difference in relative luciferase expression between the p-198 variants was examined with the Student's t test. (c) Total number of DC-SIGNR isoforms in homozygous H1 (p-198AA, int2-180AA) and homozygous WT (p-198CC, int2-180GG) placental samples measured by qRT-PCR assays in three independent experiments performed in triplicate. Data are shown in mean±S.E.M. copies number normalized by 105 copies number of GAPDH. Student's t test was used to calculate differences in the number of isoforms between the H1 and WT groups.

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