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01.12.2017 | Research | Ausgabe 1/2017 Open Access

Virology Journal 1/2017

Toll-like receptors genes polymorphisms and the occurrence of HCMV infection among pregnant women

Zeitschrift:
Virology Journal > Ausgabe 1/2017
Autoren:
Wioletta Wujcicka, Edyta Paradowska, Mirosława Studzińska, Jan Wilczyński, Dorota Nowakowska
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​s12985-017-0730-8) contains supplementary material, which is available to authorized users.

Abstract

Background

Human cytomegalovirus (HCMV) is the most common cause of intrauterine infections worldwide. The toll-like receptors (TLRs) have been reported as important factors in immune response against HCMV. Particularly, TLR2, TLR4 and TLR9 have been shown to be involved in antiviral immunity. Evaluation of the role of single nucleotide polymorphisms (SNPs), located within TLR2, TLR4 and TLR9 genes, in the development of human cytomegalovirus (HCMV) infection in pregnant women and their fetuses and neonates, was performed.

Methods

The study was performed for 131 pregnant women, including 66 patients infected with HCMV during pregnancy, and 65 age-matched control pregnant individuals. The patients were selected to the study, based on serological status of anti-HCMV IgG and IgM antibodies and on the presence of viral DNA in their body fluids. Genotypes in TLR2 2258 A > G, TLR4 896 G > A and 1196 C > T and TLR9 2848 G > A SNPs were determined by self-designed nested PCR-RFLP assays. Randomly selected PCR products, representative for distinct genotypes in TLR SNPs, were confirmed by sequencing. A relationship between the genotypes, alleles, haplotypes and multiple variants in the studied polymorphisms, and the occurrence of HCMV infection in pregnant women and their offsprings, was determined, using a logistic regression model.

Results

Genotypes in all the analyzed polymorphisms preserved the Hardy-Weinberg equilibrium in pregnant women, both infected and uninfected with HCMV (P > 0.050). GG homozygotic and GA heterozygotic status in TLR9 2848 G > A SNP decreased significantly the occurrence of HCMV infection (OR 0.44 95% CI 0.21–0.94 in the dominant model, P ≤ 0.050). The G allele in TLR9 SNP was significantly more frequent among the uninfected pregnant women than among the infected ones (χ2 = 4.14, P ≤ 0.050). Considering other polymorphisms, similar frequencies of distinct genotypes, haplotypes and multiple-SNP variants were observed between the studied groups of patients.

Conclusions

TLR9 2848 G > A SNP may be associated with HCMV infection in pregnant women.
Zusatzmaterial
Additional file 1: Figure S1. Exemplary PCR-RFLP products representative for various genotypes within TLR2 2258 G > A (A), TLR4 896 A > G (B), TLR4 1196 C > T (C) and TLR9 2848 G > A (D) SNPs. DNA fragments were resolved in 2% agarose gel, stained with ethidium bromide. Disparate lanes show restriction profiles for distinct genotypes in the range of studied TLR polymorphisms, determined in different pregnant women. The numbers on the right side of electropherograms show the size of separated DNA fragments. M—50 bp DNA marker; GG, GA, AA, AG, CT, CC—genotypes determined in studied TLR polymorphisms. (TIF 501 kb)
12985_2017_730_MOESM1_ESM.tif
Additional file 2: Figure S2. a and b Representative chromatograms of DNA fragments of various sequenced PCR products, encompassing TLR2 2258 G > A (A, B), TLR4 896 A > G (C, D), TLR4 1196 C > T (E, F) and TLR9 2848 G > A (G-I) SNPs. For TLR2 SNP, DNA forward strands were sequenced, and for TLR4 and TLR9—reverse strands were assayed. The numbers above some peaks of chromatograms indicate the following nucleotides determined in sequenced DNA fragments. Loci of the polymorphisms and genotypes analyzed in the study, are indicated with arrows. GG, GA, AA, AG, CT, CC—genotypes determined in studied TLR SNPs. (ZIP 1732 kb)
12985_2017_730_MOESM2_ESM.zip
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