Regulation of human insulin gene expression in transgenic mice

Selden, Richard F.; Skośkiewicz, M; Howie, Kathleen Burke; Russell, Paul; Goodman, Howard M.

doi:10.1038/321525a0

Cited by 85 publications

(50 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…39). The specificity of these CD8 ϩ T cells has been unknown, but a clone isolated from the pancreas of a nonobese diabetic mouse has recently been reported to recognize insulin B-chain [15][16][17][18][19][20][21][22][23] (40). This raises the question of whether the CD8 ϩ CTL that recognize autologous insulin, which are described in this communication, could cause autoimmune diabetes.…”

Section: Discussionmentioning

confidence: 99%

“…The transfected cells (EL4-INS, M12.4.1-INS, and P815-INS) expressed the 0.55-kb mRNA (Fig. 1B, lanes 2, 4, and 6) that was expected for insulin (22), whereas the untransfected parental lines did not (Fig. 1B, lanes 1, 3, and 5).…”

Section: Generation Of Hins-transfected Cell Linesmentioning

confidence: 91%

“…F. Selden (Transkaryotic Therapies, Cambridge, MA) (22). For subcloning, a NcoI-HindIII fragment of the genomic human proinsulin gene (without the tissue-specific insulin promoter) was excised from pHINT1.…”

Section: Plasmid Construction and Transfectionmentioning

confidence: 99%

See 2 more Smart Citations

Bovine and Human Insulin Activate CD8+-Autoreactive CTL Expressing Both Type 1 and Type 2 Cytokines in C57BL/6 Mice

Ma¹,

Ke²,

et al. 2000

The Journal of Immunology

View full text Add to dashboard Cite

CD8+ T cells down-regulate a variety of immune responses. For example, porcine and human insulin do not stimulate Abs in C57BL/6 mice because CD8+ T cells inhibit CD4+ helper T cells. By contrast, bovine insulin induces Ab in C57BL/6 mice, and removal of CD8+ T cells does not alter this response. This raises the question of whether porcine, but not bovine, insulin activates CD8+ T cells or whether both insulins activate CD8+ T cells but CD4+ helper T cells are differentially inhibited by them. In this study, we show that insulin-specific CD8+ CTL can be cultured from C57BL/6 mice primed with either bovine or human insulin in CFA. Thus, exogenous Ags, besides OVA, induce CD8+ CTL when administered in an adjuvant, suggesting this is a typical response. These CTL are H-2Kb restricted and produce IL-5, IL-10, IFN-γ, and small amounts of IL-4, which is distinct from IFN-γ and TNF-α that are typically secreted by virus-specific CTL. Moreover, the CTL primed with either bovine or human insulin recognize an A-chain peptide that is identical to the mouse insulin sequence. That foreign proteins, which are closely related to self-proteins, activated autoreactive, CD8+ T cells in vivo is a novel finding. It raises the possibility that self-reactive CTL may be activated by cross-reacting Ags and once activated they might participate in autoimmunity. These results also suggest that down-regulation of insulin-specific responses by autoreactive CD8+ T cells is most likely due to the differential sensitivity of bovine and human insulin-specific CD4+ T cells.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Generation Of Hins-transfected Cell Linesmentioning

confidence: 91%

See 1 more Smart Citation

Bovine and Human Insulin Activate CD8+-Autoreactive CTL Expressing Both Type 1 and Type 2 Cytokines in C57BL/6 Mice

Ma¹,

Ke²,

et al. 2000

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…In the present study, we have investigated development of tolerance in transgenic mice that express the human preproinsulin gene. These mice, have been previously shown to express mRNA for human proinsulin, the precursor form of insulin, only in pancreatic islets (5). In the islets, human proinsulin is enzymatically cleaved into insulin and C-peptide, which are subsequently secreted in response to normal physiological signals (5).…”

Section: Introductionmentioning

confidence: 99%

Tolerance induced by physiological levels of secreted proteins in transgenic mice expressing human insulin.

Whiteley

Lake²,

Selden³

et al. 1989

J. Clin. Invest.

View full text Add to dashboard Cite

We have used transgenic mice to study immune tolerance to autologous, non-MHC encoded proteins that are expressed at physiological levels in the circulation. The transgenic mice used in these studies express the human preproinsulin gene and synthesize human proinsulin. Human and mouse insulin are secreted from the pancreatic islets of transgenic mice in response to normal physiological stimuli, such as glucose. Our data demonstrate that the transgenic mice have acquired tolerance to human insulin. The repertoire of T cells specific for exogenous antigens is shaped by the acquired tolerance to autologous proteins since pork but not beef or sheep insulin is also nonimmunogenic in the transgenic mice. We also found that the transgenic mice were tolerant to human proinsulin, the intracellular precursor of insulin. Unresponsiveness to human proinsulin most likely results from tolerance of insulin-specific and proinsulin-specific T cells that recognize the secreted enzymatic cleavage products of proinsulin, insulin and C-peptide.

show abstract

“…Multiple investigators have demonstrated functional insulin gene transfer both in vitro and in vivo. [2][3][4] However, attempts to regulate transgenic insulin production have proven inadequate. 5,6 Consequently, secretion of transgenic insulin has been either insufficient to normalize blood glucose, 4,[6][7][8][9] or has produced lethal hypoglycemia.…”

Section: Introductionmentioning

confidence: 99%

Regulated hepatic insulin gene therapy of STZ-diabetic rats

2000

View full text Add to dashboard Cite

Effective and safe insulin gene therapy will require regulation of transgenic insulin secretion. We have created a livertargeted insulin transgene by engineering glucose responsive elements into a hepatic promoter containing an inhibitory insulin response sequence. In this work, we demonstrate application of this transgene for the treatment of diabetes mellitus in vivo, by administering a recombinant adenovirus vector, Ad/(GlRE) 3 BP-1 2xfur, to rats made diabetic with streptozotocin. We verified hepatic expression of transgenic insulin by RT-PCR, and confirmed glucose responsive stimulation of transgenic insulin secretion in vivo by serum RIA. Following a portal system injection of either Ad/(GlRE) 3 BP-1 2xfur, or an empty adenoviral vector, animals made diabetic with either low (120 mg/kg), or high (290 mg/kg) dose streptozotocin (STZ) were monitored for changes in body weight, and blood glucose. Without subcutaneous insulin injections, blood glucose values of sham-

show abstract

Regulation of human insulin gene expression in transgenic mice

Cited by 85 publications

References 17 publications

Bovine and Human Insulin Activate CD8+-Autoreactive CTL Expressing Both Type 1 and Type 2 Cytokines in C57BL/6 Mice

Bovine and Human Insulin Activate CD8+-Autoreactive CTL Expressing Both Type 1 and Type 2 Cytokines in C57BL/6 Mice

Tolerance induced by physiological levels of secreted proteins in transgenic mice expressing human insulin.

Regulated hepatic insulin gene therapy of STZ-diabetic rats

Contact Info

Product

Resources

About