RESUMO
The innate immune response in the placenta depends on the ability of maternal immune cells and fetal trophoblast cells to detect and eliminate invading pathogens through germline-encoded pattern recognition receptors (PRRs). In the present study, we analysed the transcripts and protein expression of interferon (IFN)-inducible protein (IFI)16, melanoma differentiation-associated protein 5 (MDA5), RIG-I-like receptor (RIG-I) and Toll-like receptor (TLR)-3 in third-trimester human placentas and investigated cytokine profiles generated during herpes simplex type 1 (HSV-1) infection. Decidual and chorionic villous biopsies (38-42 weeks of gestation) were obtained from healthy women immediately after a caesarean section. The expression of the DDX58 (RIG-I), IFIH1 (MDA5), IFI16 and TLR3 transcripts was measured using quantitative real-time polymerase chain reaction (qRT-PCR). Extracellular cytokine and PRRs levels were then quantified by enzyme-linked immunosorbent assays (ELISAs). All examined PRRs genes, including DDX58, IFIH1, IFI16 and TLR3, were expressed constitutively at the mRNA and protein levels in the placental biopsies. The concentration of the IFI16 protein was increased in HSV-1-infected decidual and chorionic villous explants compared to those of mock-infected tissues (P = 0·029). Higher protein expression levels of RIG-I in both the maternal and fetal parts of the placenta were found (P = 0·009 and P = 0·004, respectively). In addition, increased production of IFN-ß by HSV-1-infected tissues was noticed (P = 0·004 for decidua, P = 0·032 for chorionic villi). No significant differences in the IFN-α, interleukin (IL)-6 and IL-8 levels were found. These results showed that HSV-1 infection can enhance the expression of IFI16 and RIG-I proteins in the human term placenta.
Assuntos
Proteína DEAD-box 58/metabolismo , Herpes Simples/imunologia , Herpesvirus Humano 1/fisiologia , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Placenta/fisiologia , Complicações Infecciosas na Gravidez/imunologia , Adulto , Biópsia , Células Cultivadas , Cesárea , Proteína DEAD-box 58/genética , Feminino , Humanos , Imunidade Inata , Proteínas Nucleares/genética , Técnicas de Cultura de Órgãos , Fosfoproteínas/genética , Gravidez , Terceiro Trimestre da Gravidez , Receptores Imunológicos , Regulação para CimaRESUMO
The purpose of this investigation was to describe a distribution of cytomegalovirus (CMV) single and multiple genotypes among infected pregnant women, their fetuses, and newborns coming from Central Poland, as well as congenital cytomegaly outcome. The study involved 278 CMV-seropositive pregnant women, of whom 192 were tested for viral DNAemia. Human cytomegalovirus (HCMV) genotyping was performed for 18 of 34 pregnant women carrying the viral DNA and for 12 of their 15 offspring with confirmed HCMV infections. Anti-HCMV antibodies levels were assessed by chemiluminescence immunoassay (CLIA) and enzyme-linked fluorescence assay (ELFA) tests. Viral DNA loads and genotypes were determined by real-time polymerase chain reaction (PCR) assays for the UL55 gene. In the pregnant women, we identified HCMV gB1, gB2, gB3, and gB4 genotypes. Single gB2, gB3, or gB4 genotypes were observed in 14 (77.8 %) women, while multiple gB1-gB2 or gB2-gB3 genotypes were observed in four (22.2 %). Maternal HCMV genotypes determined the genotypes identified in their fetuses and newborns (p ≤ 0.050). Half of them were infected with single HCMV gB1, gB2, or gB3 genotypes and the other half with multiple gB1-gB2 or gB2-gB3 genotypes. Single and multiple genotypes were observed in both asymptomatic and symptomatic congenital cytomegaly, although no gB3 genotype was identified among asymptomatic cases. In Central Poland, infections with single and multiple HCMV strains occur in pregnant women, as well as in their fetuses and neonates, with both asymptomatic and symptomatic infections. HCMV infections identified in mothers seem to be associated with the viral genotypes in their children.
Assuntos
Coinfecção/virologia , Infecções por Citomegalovirus/virologia , Citomegalovirus/classificação , Citomegalovirus/genética , Complicações Infecciosas na Gravidez/virologia , Proteínas do Envelope Viral/genética , Anticorpos Antivirais/sangue , Coinfecção/epidemiologia , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/epidemiologia , Feminino , Feto , Genótipo , Humanos , Imunoensaio , Recém-Nascido , Epidemiologia Molecular , Polônia/epidemiologia , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Human cytomegalovirus (HCMV) is the most common congenital infection. HCMV strains display genetic variability in different regions. Distribution of HCMV genotypes in the population of congenitally infected newborns from Central Poland and viral load in newborns' blood is described and discussed. HCMV isolates were analysed by sequencing at three sites on the genome: the UL144 tumour necrosis factor-alpha (TNFα)-like receptor gene, the US28 beta-chemokine receptor gene and the UL55 envelope glycoprotein B (gB) gene. The newborns' blood was examined for HCMV DNA with a nested (UL144, UL55) or heminested (US28) polymerase chain reaction, and the genotypes were determined by sequence analysis. HCMV DNA was detectable in 25 out of 55 examined newborns born by HCMV-infected mothers (45.5%). The blood viral load in mother-infant pairs was determined. Most of the newborns had identical virus genotype, gB2 (96%), UL144 B1 (88%) and US28 A2 (84%). These genotypes were detected in all newborns with asymptomatic congenital infection. The occurrence of UL144 B1 or US28 A2 genotypes in the babies examined was significant in comparison to other genotypes (p=0.0002 and p=0.040 respectively). There was no association between specific gB subtypes in all patients groups (p=0.463). There was no correlation between HCMV genotypes and the outcome.
