Teresa Woźniakowska-Gęsicka

Distribution of cytomegalovirus gN variants and associated clinical sequelae in infants

BACKGROUND Human cytomegalovirus (HCMV) is the most widespread cause of congenital infection. The effects of various viral strains and viral loads on the infection outcome have been under debate. OBJECTIVES To determine the distribution of gN variants in HCMV strains isolated from children with congenital or postnatal infection and to establish the relationship between the viral genotype, the viral load, and the sequelae. STUDY DESIGN The study population included congenitally HCMV-infected newborns and children with postnatal or unproven congenital HCMV infection. The genotyping was performed by RFLP analysis of PCR-amplified fragments, and the viral load was measured by quantitative real-time PCR. RESULTS Our results demonstrated that the HCMV genotypes gN3b, gN4b, and gN4c were prevalent in the patients examined. There were no differences in the distributions of gN genotypes in the congenitally and postnatally infected children. Multiple HCMV strains were detected in both groups of children. A significant association between the HCMV gN4 genotype and the incidence of neurological disorders was observed (p=0.045). Our results suggest that the detection of the gN2 or the gN4 genotype may be indicative of serious manifestations in children. In contrast, the gN3b and the gN1 genotypes represent less pathogenic HCMV strains. The HCMV load in urine was significantly higher in children with congenital infection compared with children with postnatal infection. No correlation was found between the viral load and the genotype. CONCLUSION Our results suggest that the gN genotype may be a virological marker of symptomatic HCMV infection in newborns.

Relationship between toll-like receptor 2 Arg677Trp and Arg753Gln and toll-like receptor 4 Asp299Gly polymorphisms and cytomegalovirus infection

OBJECTIVES The association among specific single-nucleotide polymorphisms (SNPs) in TLR2 (Arg677Trp, Arg753Gln) and TLR4 (Asp299Gln) and human cytomegalovirus (CMV) infection was studied in infants and adults. METHODS The TLR2 and TLR4 polymorphisms were genotyped in 151 patients with CMV infections and in 78 unrelated healthy individuals. Genotyping was performed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified fragments. The viral load was measured by quantitative real-time PCR. RESULTS Almost all of the patients with CMV infections were wild-type homozygotes without TLR2 and TLR4 polymorphisms. No significant differences in TLR2 and TLR4 polymorphisms were observed between infants with or without CMV infection. Compared with adults with CMV infections, heterozygosity for the TLR2 Arg677Trp and TLR4 Asp299Gly SNPs was detected more frequently in healthy individuals (p<0.05). Logistic regression analysis showed that the wild-type TLR2 genotype was associated with an increased risk of CMV infection and that heterozygosity for TLR2 and TLR4 SNPs diminished the risk of CMV infection in adult patients. An association between CMV load and the TLR4 SNP was found. CONCLUSION Our results suggest that the wild-type TLR2 genotype may be a risk factor for CMV replication in adult patients.

TLR9 -1486T/C and 2848C/T SNPs Are Associated with Human Cytomegalovirus Infection in Infants

Toll-like receptor 9 (TLR9) recognizes non-methylated viral CpG-containing DNA and serves as a pattern recognition receptor that signals the presence of human cytomegalovirus (HCMV). Here, we present the genotype distribution of single-nucleotide polymorphisms (SNPs) of the TLR9 gene in infants and the relationship between TLR9 polymorphisms and HCMV infection. Four polymorphisms (-1237T/C, rs5743836; -1486T/C, rs187084; 1174G/A, rs352139; and 2848C/T, rs352140) in the TLR9 gene were genotyped in 72 infants with symptomatic HCMV infection and 70 healthy individuals. SNP genotyping was performed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Digested fragments were separated and identified by capillary electrophoresis. The HCMV DNA copy number was measured by a quantitative real-time PCR assay. We found an increased frequency of heterozygous genotypes TLR9 -1486T/C and 2848C/T in infants with HCMV infection compared with uninfected cases. Heterozygous variants of these two SNPs increased the risk of HCMV disease in children (P = 0.044 and P = 0.029, respectively). In infants with a mutation present in at least one allele of -1486T/C and 2848C/T SNPs, a trend towards increased risk of cytomegaly was confirmed after Bonferroni’s correction for multiple testing (Pc = 0.063). The rs352139 GG genotype showed a significantly reduced relative risk for HCMV infection (Pc = 0.006). In contrast, the -1237T/C SNP was not related to viral infection. We found no evidence for linkage disequilibrium with the four examined TLR9 SNPs. The findings suggest that the TLR9 -1486T/C and 2848C/T polymorphisms could be a genetic risk factor for the development of HCMV disease.

Cytomegalovirus alpha-chemokine genotypes are associated with clinical manifestations in children with congenital or postnatal infections.

Human cytomegalovirus (HCMV) is the leading cause of congenital infections. The aim of our study was to determine the prevalence of genotypes based on the highly polymorphic UL146 and UL147 HCMV genes and the relationship between the genotype and symptoms or viral load. We analyzed samples from 121 infants with symptomatic HCMV infection, including 32 congenitally infected newborns. The G7 and G5 genotypes were predominant in postnatal infection, whereas the G1 genotype was prevalent in congenital infection. Central nervous system (CNS) damage and hepatomegaly were detected more frequently among children infected with the G1 genotype than in those infected by other genotypes. An association between the viral genotype and viruria level was found. There was a strong correlation between HCMV genotypes determined through the UL146 and UL147 sequences (ĸ=0.794). In conclusion, we found that certain vCXCL genotypes are associated with clinical sequelae following HCMV infection.

Cytomegalovirus glycoprotein H genotype distribution and the relationship with hearing loss in children

Cytomegalovirus (CMV) is a leading cause of congenital infection and a leading infectious cause of hearing loss in children. The ORF UL75 gene encodes envelope glycoprotein H (gH), which is essential for CMV entry into host cells and the target of the immune response in humans. However, the distribution of gH variants and the relationship between the viral genotype, viral load, and sequelae in children infected with CMV is debated. The UL75 genetic variation of CMV isolates from 42 newborns infected congenitally with CMV and 93 infants with postnatal or unproven congenital CMV infection was analyzed. Genotyping was performed by analysis of PCR‐amplified fragments, and the viral load was measured by quantitative real‐time PCR. There were no differences in the distribution of gH genotypes in the children infected congenitally and postnatally. Mixed‐genotype infections with both gH1 and gH2 variants were detected in approximately 25% of the examined patients. No relationship between UL75 gene polymorphisms and the symptoms at birth was observed. The results suggest that the infection with gH2 genotype diminishes the risk of hearing loss in children (P = 0.010). In addition, sensorineural hearing loss was associated with CMV gH1 genotype infection in infants (P = 0.032) and a high viral load in urine (P = 0.005). In conclusion, it was found that the gH genotype does not predict clinical sequelae in newborn infants following congenital CMV infection. However, these results suggest that the gH genotype might be associated with hearing loss in children. J. Med. Virol. 86:1421–1427, 2014.

Efficacy of pegylated interferon α-2b and ribavirin in chronic hepatitis C virus (genotypes 1 and 4) infection.

Background and Objective: The course of chronic hepatitis C in children is often mild or asymptomatic, but may lead to liver cirrhosis and neoplasm. The aim of our study was retrospective evaluation of treatment efficacy using pegylated interferon (IFN)-&agr;2b with ribavirin in children and adolescents with chronic hepatitis C, both treatment naïve and re-treated. Methods: The study comprised 79 patients with chronic hepatitis C ages 8 to 18 years (43 patients re-treated; 54 infected with genotype 1 hepatitis C virus and 25 with genotype 4), treated with pegylated IFN-&agr;2b (1.5 &mgr;g · kg−1 · week−1) plus ribavirin (15 mg · kg−1 · day−1) for 48 weeks. The primary endpoint was sustained virologic response (SVR). Results: Early viral response (EVR) was observed in 43.1% and end-of-treatment response in 47.9% of patients. In 44.3% of patients, SVR was achieved, which was maintained for at least the next 6 months. Patients not treated before significantly more frequently attained EVR, end-of-treatment response, and SVR (64%, 65.6%, and 63.9%, respectively) as compared with re-treated patients (30%, 33.3%, and 27.9%, respectively). Among 28 patients who attained EVR, 23 achieved SVR. In 2 patients, despite lack of EVR, SVR was observed. There were numerous adverse effects. They were not so severe as to discontinue therapy. Conclusions: Combined therapy with pegylated IFN-&agr;2b and ribavirin in patients with chronic hepatitis C, infected with hepatitis C virus genotypes 1 and 4, was more effective in treatment-naïve patients (63.9%) as compared with re-therapy cases (27.9%). SVR was maintained for at least the next 6 months in all of the patients. The applied treatment has limited efficacy and evokes numerous adverse effects; thus, search for new methods of treatment is mandatory.

Association of TLR3 L412F Polymorphism with Cytomegalovirus Infection in Children

Intracellular Toll-like receptor 3 (TLR3) recognizes viral double-stranded RNA (dsRNA) and activates antiviral immune responses through the production of type I interferons (IFNs) and inflammatory cytokines. This receptor binds to dsRNA molecules produced during human cytomegalovirus (HCMV) replication. TLR7 senses viral single-stranded RNA (ssRNA) in endosomes, and it can interact with endogenous RNAs. We determined the genotype distribution of single-nucleotide polymorphisms (SNPs) within the TLR3 and TLR7 genes in children with HCMV infection and the relationship between TLR polymorphisms and viral infection. We genotyped 59 children with symptomatic HCMV infection and 78 healthy individuals for SNPs in the TLR3 (rs3775290, c.1377C>T, F459F; rs3775291, c.1234C>T, L412F; rs3775296, c.-7C>A) and TLR7 (rs179008, c.32A>T, Q11L; rs5741880, c.3+1716G>T) genes. SNP genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and capillary electrophoresis. The HCMV DNA load was quantified by real-time PCR. We found an increased frequency of the heterozygous genotype TLR3 L412F in children with HCMV infection compared with uninfected cases. In individuals with a mutation present in at least one allele of the L412F SNP, an increased risk of HCMV disease was found, and this result remained highly significant after Bonferroni’s correction for multiple testing (Pc < 0.001). The heterozygous genotype of this SNP was associated with the increased risk of HCMV disease in an adjusted model that included the HCMV DNA copy number in whole blood and urine (P < 0.001 and P = 0.008, respectively). Moreover, those with a heterozygous genotype of rs3775296 showed an increased relative risk of HCMV infection (P = 0.042), but this association did not reach statistical significance after correction for multiple testing. In contrast, the rs3775290 SNP of TLR3 and TLR7 SNPs were not related to viral infection. A moderate linkage disequilibrium (LD) was observed between the SNPs rs3775291 and rs3775296 (r2 = 0.514). We suggest that the L412F polymorphism in the TLR3 gene could be a genetic risk factor for the development of HCMV disease.

Ostre rozsiane zapalenie mózgu i rdzenia kręgowego u dzieci jako następstwo jałowego zapalenia opon mózgowo-rdzeniowych – opis dwóch przypadków

Streszczenie Ostre rozsiane zapalenie mozgu i rdzenia kregowego ( acute disseminated encephalomyelitis – ADEM) charakteryzuje sie odczynem zapalnym i demielinizacją w obrebie ośrodkowego ukladu nerwowego, ktore są nastepstwem chorob zakaźnych lub szczepien. Wystepuje bardzo rzadko, cześciej u dzieci niz u doroslych, zwykle konczy sie pelnym wyzdrowieniem, ale moze prowadzic do zgonu lub powodowac odlegle nastepstwa. Diagnoza choroby jest trudna i wymaga glownie roznicowania ze stwardnieniem rozsianym; najbardziej pomocne dla rozpoznania jest badanie za pomocą rezonansu magnetycznego. W ostrym okresie choroby postepowanie z wyboru stanowi steroidoterapia. W pracy przedstawiono dwa przypadki ADEM (u 6-letniego chlopca i 5-letniej dziewczynki) o niejasnej etiologii, bedące nastepstwem jalowego zapalenia opon mozgowo-rdzeniowych.

Częstość mykoplazmatycznych zapaleń płuc u dzieci w Łodzi i makroregionie na podstawie obserwacji własnych

Wstep Zakazenia ukladu oddechowego u dzieci stanowią czesty problem kliniczny. Wśrod czynnikow etiologicznych istotne znaczenie ma Mycoplasma pneumoniae . Cel pracy Autorzy ocenili czestośc zakazenia Mycoplasma pneumoniae jako czynnika przyczynowego zapalen pluc u dzieci. Material i metody Analizą objeto lącznie 23 779 dzieci w wieku od 1 miesiąca do 18 roku zycia leczonych na wybranych oddzialach dzieciecych w Łodzi i makroregionu lodzkiego w okresie od 2003 do 2007 r. Wyniki Wykazano wystepowanie zakazenia Mycoplasma pneumoniae u 1313 dzieci (5,5%). Zaobserwowano stopniowy wzrost czestości mykoplazmatycznych zapalen pluc w poszczegolnych latach obserwacji, z najwiekszym nasileniem w ostatnich dwoch latach. Stwierdzono rowniez, ze zakazenie Mycoplasma pneumoniae wystepowalo w kazdej grupie wiekowej, przy czym w najwyzszym odsetku (43,7%) u dzieci powyzej siodmego roku zycia, a w najnizszym u dzieci 2–3-letnich (16,2%) i u niemowląt (17,9%). Wnioski Stwierdzenie u 5,5% dzieci zakazenia Mycoplasma pneumoniae wskazuje na istotny udzial tego czynnika w etiologii zapalen pluc u dzieci. Wykazano, ze Mycoplasma pneumoniae jest rowniez przyczyną zapalenia pluc u niemowląt i malych dzieci. Wczesne rozpoznanie ma istotne znaczenie, poniewaz umozliwia wdrozenie prawidlowego leczenia.

Bakteryjne powikłania w przebiegu ospy wietrznej – opis przypadków

Streszczenie Ospa wietrzna nadal jest jedną z najczestszych chorob zakaźnych w Polsce. Zakazenie wirusem ospy wietrznej, wbrew powszechnym opiniom, nie zawsze przebiega w postaci lagodnej choroby. Nadal utrzymują sie dośc wysokie wskaźniki hospitalizacji z powodu powiklan w przebiegu choroby. W pracy przedstawiono 2 przypadki ospy wietrznej powiklanej zakazeniami bakteryjnymi wymagającymi interwencji chirurgicznej.