Prolonged Survival of Adult Rat Pancreatic Islets Transfected with E1A-12s Adenovirus

Rutten, Michael J.; Lester, Linda R.; Quinlan, Margaret P.; Meshul, Charles K.; Deveney, Clifford W.; Rabkin, John M.

doi:10.1097/00006676-199908000-00012

Cited by 6 publications

(4 citation statements)

References 31 publications

(51 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transfection of pancreatic islets with an E1A-13S gene expressing only the 289-aa protein induced extensive islet mass apoptosis or necrosis compared with that seen with nontransfected islets, while transfection of pancreatic islets with an E1A-12S gene expressing only the 243-aa protein extended the life span and functionality of pancreatic islets with few apoptotic cells (44). The elevated expression of the proapoptosis cytokine TNF-␣ in A30 mice was in agreement with the results of another previous in vitro study demonstrating a significant increase in TNF-␣ expression in inflammatory cell lines by infection with the E1A gene expressing only the 289-aa protein but not by the E1A gene expressing only the 243-aa protein (34).…”

Section: Discussionmentioning

confidence: 96%

Transgenic Expression in Mouse Lung Reveals Distinct Biological Roles for the Adenovirus Type 5 E1A 243- and 289-Amino-Acid Proteins

Yang

McKerlie

Borenstein

et al. 2002

J Virol

View full text Add to dashboard Cite

Little is known about the biological significance of human adenovirus type 5 (Ad5) E1A in vivo. However, Ad5 E1A is well defined in vitro and can be detected frequently in the lungs of patients with pulmonary disease. Transgenic expression of the Ad5 E1A gene targeted to the mouse lung reveals distinct biological effects caused by two Ad5 E1A products. Either of two Ad5 E1A proteins was preferentially expressed in vivo in the transgenic lungs. The preferential expression of the Ad5 E1A 243-amino-acid (aa) protein at a moderate level was associated with cellular hyperplasia, nodular lesions of proliferating lymphocyte-like cells, and a low level of p53-dependent apoptosis in the lungs of transgenic mice. In contrast, the preferential expression of the Ad5 E1A 289-aa protein at a moderate level resulted in a proapoptotic injury and an acute pulmonary proinflammation in the lungs of transgenic mice, mediated by multiple apoptotic pathways, as well as an enhancement of the host immune cell response. Expression of the Ad5 E1A 243-aa protein resulted in proliferationstimulated p53 upregulation, while expression of the Ad5 E1A 289-aa protein led to DNA damage-induced p53 activation. These data suggest that the Ad5 E1A 243-and 289-aa proteins lead to distinct biological roles in vivo.

show abstract

Section: Discussionmentioning

confidence: 96%

Transgenic Expression in Mouse Lung Reveals Distinct Biological Roles for the Adenovirus Type 5 E1A 243- and 289-Amino-Acid Proteins

Yang

McKerlie

Borenstein

et al. 2002

J Virol

View full text Add to dashboard Cite

show abstract

“…Using various islet isolations and culturing techniques, human pancreatic islets have been maintained in vitro only for a limited time [3, 15, 34]. Although transfectional experiments have prolonged survival of rodent islets [27], no such studies have been successful in human islets. Recently, we have shown that hamster islet cells can be maintained in a defined medium, devoid of collagen, for more than a year [29], indicating that the quality of the culture medium is paramount for the islet cell maintenance.…”

Section: Discussionmentioning

confidence: 99%

Maintenance of human islets in long term culture

Batra¹,

Pour²,

Schmied³

et al. 2000

Differentiation

View full text Add to dashboard Cite

The long-term maintenance of human islets in culture has remained a challenge. Despite advancements in culture techniques, human islets proved to have a short life span in vitro. For the first time, we have succeeded in maintaining human islets in a defined culture medium for more than 12 months. Freshly isolated islets from a 38-year-old donor were cultured in M3:5 medium and placed on a rocker for 14 days to remove contaminated exocrine and mesenchymal cells which attached to the bottom. The floating islets were purified by daily hand-picking and transfer into fresh medium. After 14 days, purified islets were allowed to attach to the bottom of the flasks and to expand. At various time points, islets were examined immunohistochemically and electron microscopically, and the secretion of islet hormones and their mRNA were determined by radioimmunoassay and reverse transcriptase polymerase chain reaction, respectively. Within seven days of culture, ductular and acinar cells developed within the initially normal islets. With time, exocrine cell types expanded while the number of the endocrine cells and their secretion decreased. At day 60, only a few endocrine cells were identifiable, whereas most of the cells appeared undifferentiated and expressed cytokeratin 7 and 19, neuron specific enolase, tomato lectin, phaseolus leucoagglutinin, laminin, and vimentin. After 60 days, the culture consisted entirely of undifferentiated cells which could be maintained in culture for 270 days before they became senescent. This is the first report on the long-term maintenance of human islet cells in culture and allows an insight into the complex process of endocrine cell differentiation.

show abstract

“…In this regard several viral vectors have been tested for their efficacy to transduce islet cells. Adenovirus vectors [11][12][13] and lentivirus vectors [14][15][16] have been shown to transduce pancreatic islet cells efficiently. However, the efficacy of AAV vector transduction to islet cells has not been demonstrated to date.…”

Section: Introductionmentioning

confidence: 99%