IL-2/HSVtk Gene Expression in Neuroblastoma Cell Line / 대한소아혈액종양학회지

PURPOSE: We investigated the efficacy of in vitro transduction of neuro 2a cells with interleukin-2 (IL-2) and herpes simplex virus-thymidine kinase (HSVtk) gene followed by assay of IL-2 and administration of the antiviral drug ganciclovir (GCV). METHPDS: The LNC/IL-2/IRES/HSVtk vectors were transferred in vitro into the murine neuro 2a-cell lines and the transduced cell lines were selected in G-418, 500 mug/mL, for 14 days. The neuro-2a cells transduced with 1x10(6) LNC/IL-2/IRES/HSVtk vector (neuro- 2a/IL-2 /HSVtk) were cultured in 6 well culture plates for 24 hours and we measured IL-2 concentrations of 1 x10(5) neuro-2a/IL-2/HSVtk by ELISA method. And also, 1x10(5) neuro- 2a/IL-2/HSVtk were cultured in 96 well culture plates in 10 mug/mL concentrations of GCV for 72 hours and then we measured the surviving cells 24 hours, 48 hours and 72 hours after GCV administration. RESULTS: We selected 5 colonies of the neuro-2a/IL-2/HSVtk and expanded in G-418 contained media. The IL-2 concentration of the neuro-2a/ IL-2/HSVtk was from 78.0+/-8.7 IU/day to 92.1+/-7.5 IU/day. The cell numbers of neuro-2a/IL-2/HSVtk survived after GCV administration were 3.7+/-1.5x10(2), 4.4+/-0.4x10(3), 5.0+/-1.0x10(2), 1.0+/-0.2x10(4), and 1.5+/-0.5x10(2), respectively. And the cell numbers of LNCX transduced neuro-2a cells survived were 7.5+/-0.5x10(5). CONCLUSION: In vitro transduction of neuro-2a cells with the LNC/IL-2/IRES/HSVtk vector expressed effectively IL-2 and HSVtk gene simultaneously.

Role of Adenovirus in Diarrheal Children

In order to investigate the role of adenovirus in diarrheal children, we evaluated 907 children with diarrhea and 193 children without diarrhea for a 22-month. Stools were tested for group A rotavirus antigen and for adenovirus types 40/41 (Ad 40/41) by using ELISA, cell technique and indirect immunofluorescent method. Adenovirus was detected in 10.1% of the diarrheal children and 3.1% of the non-diarrheal children. Ad40/41 was detected in 4.85% of the diarrheal children. Other nonenteric adenovirus was detected in 5.29% of the diarrheal children. In addition, 24% of the adenovirusinfected children excreted rotavirus simultaneously. Rotavirus was found in 57.1% of the diarrheal patients. Among the diarrheal children. 88% of those with adenovirus were younger than 24 months. Although peaks of adenovirus infection were detected in July and Autumn in the study, there is no apparent seasonal trend with adenovirus. The average duration of admission caused by adenoviral infection was 4.9 days and 88% of the cases accompanied by fever. Stool occult blood test revealed positive reaction in 66.3% of the cases and serum aminotransferase activities transiently elevated in 5.4% of the cases. Diarrhea with more than 10 stools per day, vomiting, or fever in adenovirusinfected children were similar with ratavirus, where as the first two manifestions were associated with confection of rotavirus and adenovirus. Nine out of 92 cases(9.8%) of the gastroenteritis caused by adenovirus revealed intussusception. We conclude that Ad40/41 is an important cause of diarrhea among infants and children, and non-enteric adenovirus is considered to be a cause of diarrhea, but their role in diarrheal children should be further studied.