Seroprevalence of neutralizing antibodies to human mastadenovirus serotypes 3 and 7 in healthy children from guangdong province.

Severe adenovirus pneumonia is becoming more common in children infected with human mastadenovirus (HAdV)-3 and HAdV-7 than in those infected with other types of adenoviruses. Recently, there has been a trend toward an increasing prevalence of pneumonia caused by HAdV-7, an important viral pathogen in Pediatric Intensive Care Unit infections. Children infected with HAdV-7 have more serious symptoms of acute respiratory infections and other complications than those infected with HAdV-3. No specific anti-adenovirus drugs or vaccines are available for treatment or prevention. Therefore, we investigated the seroprevalence and titer levels of neutralizing antibodies (NAbs) against HAdV-3 and HAdV-7 in healthy children in Guangdong Province. We found that the seropositivity rates and antibody titers for HAdV-3 NAb were higher than those for HAdV-7 NAb. In children between 6 and 12 months of age, the seropositivity rates and titers were significantly low against HAdV-3 and HAdV-7. The HAdV-7-positive rate was significantly higher in the HAdV-3-positive samples than in the HAdV-3-negative samples. The HAdV-7 NAbs carried by the 0-6-month age group were dominated by low titers. These results reveal a low level of herd immunity against HAdV-3 and HAdV-7 in children, clarifying the importance of monitoring these two highly virulent adenoviruses, developing prophylactic vaccines, and predicting potential outbreaks.

Erratum: Two engineered site-specific antibody-drug conjugates, HLmD4 and HLvM4, have potent therapeutic activity in two DLL4-positive tumour xenograft models.

[This corrects the article on p. 2387 in vol. 10, PMID: 32905508.].

Two engineered site-specific antibody-drug conjugates, HLmD4 and HLvM4, have potent therapeutic activity in two DLL4-positive tumour xenograft models.

The humanized Delta-like 4 (DLL4) monoclonal antibody H3L2 with a quite high affinity for hrDLL4 inhibits the DLL4-mediated human umbilical vein endothelial cell (HUVEC) phenotype, inducing dysfunctional angiogenesis and tumour cell apoptosis, which effectively arrests breast cancer cell growth in vivo. To develop a more effective therapy, an engineered cysteine residue at alanine 121 (Kabat numbering) on each H3L2 heavy chain or at valine 207 (Kabat numbering) on each H3L2 light chain was established by site-directed mutagenesis. Three engineered antibodies, THL4, TH2 and TL2, were identified, and the specific-site antibody-drug conjugates (ADCs) THL4-mpeoDM1 (named HLmD4), TH2-mpeoDM1 (named HmD2), TL2-mpeoDM1 (named LmD2) and THL4-vcMMAE (named HLvM4), were produced, which exhibit much more potent antitumour activity than the naked antibody. The engineered ADCs can be directed against DLL4 and effectively internalized, followed by the release of small molecule cytotoxic agents, e.g., DM1 or MMAE, into the cytosol, which inhibit the synthesis of microtubules and induce G2/M phase growth arrest and cell death through the induction of apoptosis. ADC-conjugated DM1 was highly potent against DLL4-expressing cells in vitro. We systematically compared the in vitro potency and the in vivo preclinical efficacy and safety profiles of the heterogeneous conventional ADC, H3L2-mpeoDM1 (named JmD4) with that of the homogeneous engineered conjugate HLmD4. The engineered anti-DLL4 ADCs, particularly HLmD4, showed more potent antitumour activity than Docetaxel and superior safety compared with JmD4 in two xenograft tumour models. Our findings indicate that engineered ADCs have promising potential as effective preclinical therapies for cancers.

Eupatilin induces human renal cancer cell apoptosis via ROS-mediated MAPK and PI3K/AKT signaling pathways.

Phosphatidylinositol 3-kinase (PI3K)/AKT and mitogen activated protein kinase (MAPK) signaling cascades have significant roles in cell proliferation, survival, angiogenesis and metastasis of tumor cells. Eupatilin, one of the major compounds present in Artemisia species, has been demonstrated to have antitumor properties. However, the effect of eupatilin in renal cell carcinoma (RCC) remains to be elucidated. Therefore, the present study investigated the biological effects and mechanisms of eupatilin in RCC cell apoptosis. The results of the present study demonstrated that eupatilin significantly induced cell apoptosis and enhanced the production of reactive oxygen species (ROS) in 786-O cells. In addition, eupatilin induced phosphorylation of p38α (Thr180/Tyr182), extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase 1/2 (Thr183/Tyr185), and decreased the phosphorylation of PI3K and AKT in 786-O cells in a concentration-dependent manner. Furthermore, the ROS inhibitor N-acetyl-L-cysteine was able to rescue the MAPK activation and PI3K/AKT inhibition induced by eupatilin. Taken together, the results of the present study provide evidence that inhibition of eupatilin induces apoptosis in human RCC via ROS-mediated activation of the MAPK signaling pathway and inhibition of the PI3K/AKT signaling pathway. Thus, eupatilin may serve as a potential therapeutic agent for the treatment of human RCC.

The Seroprevalence of Some Pathogen Specific IgM in Children with Acute Respiratory Tract Infections in Guangzhou Region, 2011 - 2012.

BACKGROUND: Acute respiratory tract infections (ARTIs) are the leading cause of morbidity and death in children < 5 years worldwide. The aim of this study is to analyze the seroprevalence of nine pathogen specific IgMs in children with ARTIs with respect to gender, age, and seasonality in the Guangzhou region. METHODS: Serum samples were collected from 20160 children with ARTIs admitted to the Guangzhou Women and Children's Medical Center between 2011 and 2012. Serum-specific IgM antibodies to nine respiratory pathogens, Mycoplasma pneumonia (MP), Legionella pneumophila (LP), Coxiella burnetii (C. burnetii), Chlamydophila pneumonia (CP), adenovirus (ADV), respiratory syncytial virus (RSV), type A and type B influenza virus (IVA and IVB), and parainfluenza virus (PIV), were detected using immunofluorescence assay. RESULTS: The male-to-female ratio of all patients was 1.9:1. The median age was 3 years and 8 months with a significant difference in seropositivity to respiratory tract pathogens between children from different age groups. Seropositivity was detected in 43.53% of the children with the top three pathogens being MP (33.15%), RSV (10.27%), and ADV (6.63%), followed by IVB (2.63%), LP (2.25%), IVA (1.59%), PIV (1.57%), CP (0.27%), and C. burnetii (0.13%). The prevalence of single, double, and triple seropositivity was 70.20% (6160/8775), 25.22% (2213/8775), and 4.57% (401/8775), respectively. The total IgM seropositivity for any kind of pathogen in the nine kinds of pathogens peaked in winter (46.53%), while the nadir was observed in summer (41.97%). CONCLUSIONS: The top three seroprevalence of nine kinds of pathogen specific IgM was MP, followed by RSV and ADV. The epidemic pathogen specific IgM had a season-specific seropositivity distribution. Seroprevalence of the pathogen should be a focus of attention.

Analysis and solution of false-positives when testing CVA16 sera using an antibody assay against the EV71 virus.

Hand, foot and mouth disease (HFMD) in humans is caused mainly by Enterovirus 71(EV71) and Coxsackievirus A16 (CVA16). EV71 is associated with severe HFMD cases but not CVA16. Use of IgM-capture enzyme-linked immunosorbent assay (ELISA) is important for the early diagnosis of EV71 infection, but cross-reactivity of the anti-CVA16 IgM antibody with EV71 produces false-positive results. In this report, we designed a new EV71 IgM-capture ELISA method using the EV71 VP1 peptide instead of the EV71 virion as the detectable antigen, and tested sera from patients infected with EV71 or CVA16. The results showed that acute sera from 76 EV71-infected patients had similar sensitivity for virus detection (98.68%) or VP1 detection (97.37%). When acute sera from patients infected with CVA16 were used, significant differences between the two methods were observed. The cross-reactivity rate of the virus detection method was 29.4% (5/17), but no cross-reactivity was observed using the VP1 detection method. Western immunoblotting demonstrated that EV71 VP3 cross-reacted with part of the CVA16 IgM antibody. The results demonstrate that EV71 VP3 is the cross-reactive antigen in the EV71 IgM-capture ELISA when testing CVA16 sera using the virus-antibody detection method. The problem of false-positive results was resolved by using the VP1 peptide as the detectable antigen.

[Complete genomic sequence analysis on human enterovirus 71 strain in Guangzhou, in 2008 and 2010].

OBJECTIVE: To study the genomic genotypes and variation of human enterovirus 71 (EV71) infected infants in Guangzhou city, in 2008 and 2010. METHODS: Primers were designed on the basis of the genomic sequence of EV71 SHZH03 strain (AY465356) in the GenBank, and EV71 genome amplified by RT-PCR. PCR-products were directly sequenced and the genomic nucleotide sequences were analyzed with the programs of Clustal W/X, DNASTAR and MEGA 4.1. RESULTS: 9 strains of EV71 genome appeared to be 7405 bp in length. The genomic sequences of EV71 Guangzhou strains were compared with those of EV71 in GenBank, which revealed that the homology with EV71 genotype C4a Fuyang strains ranged between 98% - 99%. Homology with genotype C4b were 92% - 94%, with genotypes C1, C2, C3 as 82% - 83%, with genotypes B3, B4, B5 as 81% - 83% and the homology with genotype A was 80%. When compared the VP1 genes of EV71 Guangzhou strains with genotypes A, B, C virus, we revealed that the highest homology was also with genotype C4a. When compared the VP1 amino acid sequences of EV71 Guangzhou strains with genotype A, B, C virus by Clustal W program, the results revealed that the amino acid residue Q at position 22 in VP1 gene was transformed to H, while 213 (S→T) and 1764 (V→I) mutations in polyprotein were discovered. CONCLUSION: Data from the sequences and phylogenetics analysis on those Guangzhou strains in 2008 and 2010 revealed that those isolates belong to genotype C4a, with the homology with Fuyang strains as 98%-99%. Mutation of amino acid residue H at position 22 in VP1 gene was discovered and the neutralizing antibody of EV71 might have been conversed by this residue. 213 (S→T) and 1764 (V→I) mutations in polyprotein were also discovered.