Three-dimensional Culture of Human Airway Epithelium in Matrigel for Evaluation of Human Rhinovirus C and Bocavirus Infections

Objective Newly identified human rhinovirus C （HRV-C） and human bocavirus （HBoV） cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses.Methods A platform for culturing human airway epithelia in a three-dimensional （3D） pattern using Matrigel as scaffold was developed. The features of 3D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3D cells at designated time points were quantitated by real-time polymerase chain reaction {PCR）. Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA.Results Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-2, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3D-cultured human airway epithelial （HAE） cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3D culture system.Conclusion Our data provide a preliminary indication that the 3D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.

Norovirus Infection and Histo-blood Group Antigens in Children Hospitalized with Diarrhea in Lulong and Chenzhou in China

Norovirus （NOV） is a pathogen that commonly causes viral diarrhea in children. Studies indicate that NoV recognizes human histo-blood group antigens （HBGAs） as cell attachment factors. In order to explore the correlation between of NoV infection and HBGAs, a cross-sectional study was conducted in children less than five years old who were hospitalized with diarrhea in two areas of China between November 2014 and February 2015. Of the paired stool and saliva samples taken from 424 children,

Gene Knockdown in Human Rhinovirus 1B Using 2′-OMe-modified si RNAs Results in the Reactivation of the Interferon Response

The aim of this study was to investigate the knockdown efficiency of 2＇-O-methylated （2＇-OMe）-modified small interfering RNAs （siRNAs） on human rhinovirus 1B （HRV1B） replication and the interferon response. Thus, 24 2＇-OMe-modified siRNAs were designed to target HRV1B. The RNA levels of HRV1B, Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and interferons were determined in HRV1B-infected HeLa and BEAS-2B epithelial cells transfected with 2＇-OMe-modified siRNAs. The results revealed that all 2＇-OMe-modified siRNAs interfered with the replication of HRVIB in a cell-specific and transfection efficiency-dependent manner. Viral activation of Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-1, and the interferon response was detected. In conclusion, the 2＇-OMe-modified siRNAs used in this study could interfere with HRV1B replication, possibly leading to the reactivation of the interferon response.

Detection of Human Bocavirus in Children with Acute Respiratory Tract Infections in Lanzhou and Nanjing,China

Objective The aim of this study was to explore the prevalent characteristics of HBoV1 and its co-infection.Methods PCR was used to detect HBoV1-DNA（HBoV1） and other viruses.A multivariate logistic regression model was used to explore possibility of co-detected for related viruses.Results The positivity rates in Nanjing and Lanzhou were 9.38%（74/789） and 11.62%（161/1386）,respectively（P〉0.05）.The HBoV1 positive group was younger than negative group（P〈0.05）.Seasonal differences were noted,with a higher frequency of infection in December and July.HBoV1-positive children [72.34%（169/235）] were co-infected with other respiratory viruses.Multifactorial analysis showed no correlations between HBoV1 and the clinical classification,region,gender,age,or treatment as an outpatient or in a hospital.Correlations were identified between HBoV1 infections with ADV（OR=1.53,95% CI 1.03-2.28）,RSV（OR=0.71,95% CI 0.52-0.98）,and IFVA（OR=1.77,95% CI 1.00-3.13）.Conclusions Presence of HBoV1 in nasopharyngeal aspirates did not correlate with region or gender,although the prevalence of HBoV1 was higher in younger children.There were no correlations between HBoV1 and other variables,except for the season and ADV,RSV,or IFVA infections.