Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells

ABSTRACT Human Adenovirus 36 (HAdV-36) has been related to diverse effects on metabolism and may attenuate the lipid accumulation in kidneys with increased adiposity. Some of these effects would be related to viral persistence. However, until now, a model of persistent in vitro infection by HAdV-36 is unknown. In this study, we examined the cells of the Vero lineage to explore their permissiveness to long-term HAdV-36 infection. HAdV-36 was productively replicated in Vero cells and maintained long-term infection for up to 35 cell passages. A subculture was obtained from the cells that survived the primary infection at a low MOI (0.5). The production of the extracellular infectious virus with titers ranging from 104 to 106 TCID50/mL and DNA-bearing cells was detected. In long-term infected cells, the intracellular distribution of viral antigen was demonstrated by performing immunolocalization (IFI) and expression of cell-viral antigen in 50% of cells by flow cytometry, using anti-HAdV-36 hyperimmune rabbit serum. Furthermore, E1a and E4orf1 genes in long-term infected passages showed a decreasing trend. Our preliminary results reveal that renal epithelial monkey cells are permissive for the productive infection of HAdV-36. Vero cell culture long-term infection might be a promising model for addressing the fundamental aspects of the HAdV-36 biology that cannot reveal broadly-used cultures, which do not maintain long-term infection in primary or transformed cells.

Role of LncRNA ROR in Ad36-induced browning of human adipose-derived stem cells / 中华内分泌代谢杂志

Objective To investigate the role of LncRNA ROR in Ad36-induced browning of human adipose-derived stem cells(hADSC). Methods After hADSC was induced by cocktail and Ad36 for 2,4,6,and 8 days,Oil red O staining was performed for observing the adipogenic status. The mRNA expressions of LncRNA ROR, uncoupling protein 1(UCP1),and PRDM16 were detected by real-time PCR and the protein expressions of UCP1 and PRDM16 were detected by Western-blot. After LncRNA ROR was knocked down by siRNA, UCP1 and PRDM16 mRNA and protein expression levels in the process of Ad36-induced adipocyte differentiation were detected by real-time PCR and Western-blot. Results Oil red O staining showed that fat droplets in the cocktail-induced group were larger than those in the Ad36-induced group. Compared with the cocktail group,the mRNA expressions of LncRNA ROR, UCP1 and PRDM16, and the protein expressions of UCP1 and PRDM16 in Ad36 group were significantly increased(P<0.05). The mRNA and protein expressions of UCP1 and PRDM16 in LncRNA ROR knockdown group were significantly lower than those in control group(P<0.05). Conclusion In the process of Ad36-induced hADSC differentiation,the up-regulation of LncRNA ROR may stimulate UCP1 and PRDM16 expression,and thus promote the browning of hADSC.

The regulation of adenovirus type 36 infection and progranulin expression in Uygur obese patients / 西安交通大学学报(医学版)

ABSTRACT:Objective To investigate the regulation of adenovirus type 36 infection on precursor protein particles progranulin expression in Uygur obese patients.Methods Based on the diagnostic criteria of obesity,the samples were divided into obese group and non-obese group.Serum neutralization test was used to detect the antibody of Ad36.The progranulin mRNA expressions in abdominal omental and subcutaneous adipose tissues were detected by real-time quantitative PCR.ELISA method was used to determine serum progranulin protein levels. CD68 protein expression of macrophage was detected by immunohistochemistry. Results ① In the Uygur population of our study,compared with that in the non-obese group (38/1 1 7,32.5%),the number of obese patients (54/1 1 5,47.0%)infected with Ad36 was significantly higher than that in non-obese group (P <0.05 ).② Serum progranulin was significantly increased in the Ad36-infected obese group (408.45±1 56.92)than in non-obese group (326.1 1±1 58.60)(P <0.05).The mRNA expression of progranulin did not differ between the two groups.③ The macrophage infiltration was significantly higher in the Ad36-infected obese group (14 730.1 6 ± 2 227.39 )than in non-obese group (10 786.50 ± 2 772.80 )(P < 0.05 ).Conclusion Ad36 infection may be associated with the occurrence of obesity in Uygur population,and adenovirus type 36 infection may regulate the expression of serum progranulin at the protein level.