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Destruction of conditional insulinoma cell lines in NOD mice: a role for autoimmunity
P Cattan1, D Rottembourg, S Cottet
1INSERM U561, Groupe hospitalier Cochin-Saint Vincent de Paul, Paris, France.
Aims/Hypothesis:
betaTC-tet (H2(k)) is a conditional insulinoma cell line derived from transgenic mice expressing a tetracycline-regulated oncogene. Transgenic expression of several proteins implicated in the apoptotic pathways increase the resistance of betaTC-tet cells in vitro. We tested in vivo the sensitivity of the cells to rejection and the protective effect of genetic alterations in NOD mice.
Methods:
betaTC-tet cells and genetically engineered lines expressing Bcl-2 (CDM3D), a dominant negative mutant of MyD88 or SOCS-1 were transplanted in diabetic female NOD mice or in male NOD mice with diabetes induced by high-dose streptozotocin. Survival of functional cell grafts in NOD-scid mice was also analyzed after transfer of splenocytes from diabetic NOD mice. Autoreactive T-cell hybridomas and splenocytes from diabetic NOD mice were stimulated by betaTC-tet cells.
Results:
betaTC-tet cells and genetically engineered cell lines were all similarly rejected in diabetic NOD mice and in NOD-scid mice after splenocyte transfer. In 3- to 6-week-old male NOD mice treated with high-dose streptozotocin, the cells temporarily survived, in contrast with C57BL/6 mice treated with high-dose streptozotocin (indefinite survival) and untreated 3- to 6-week-old male NOD mice (rejection). The protective effect of high-dose streptozotocin was lost in older male NOD mice. betaTC-tet cells did not stimulate autoreactive T-cell hybridomas, but induced IL-2 secretion by splenocytes from diabetic NOD mice.
Conclusion/Interpretation:
The autoimmune process seems to play an important role in the destruction of betaTC-tet cells in NOD mice. Genetic manipulations intended at increasing the resistance of beta cells were inefficient. Similar approaches should be tested in vivo as well as in vitro. High dose streptozotocin influences immune rejection and should be used with caution.
Insights
Genetic modifications did not protect betaTC-tet insulinoma cells from rejection in NOD mice. The autoimmune process is key in cell destruction, and streptozotocin
Area of Science:
- * Immunology and Diabetes Research
- * Cell Biology and Transplantation
Background:
- * BetaTC-tet cells are a conditional insulinoma cell line from transgenic mice.
- * In vitro studies show increased resistance with apoptotic pathway proteins.
- * NOD mice are a model for autoimmune diabetes.
Purpose of the Study:
- * To evaluate in vivo rejection sensitivity of betaTC-tet cells in NOD mice.
- * To assess the protective effect of genetic alterations against rejection.
- * To investigate the role of autoimmune processes in beta cell destruction.
Main Methods:
- * Transplantation of betaTC-tet cells and genetically engineered lines (Bcl-2, dominant-negative MyD88, SOCS-1) into diabetic NOD mice.
- * Analysis of cell graft survival in NOD-scid mice after splenocyte transfer from diabetic NOD mice.
- * Stimulation assays using autoreactive T-cell hybridomas and splenocytes from diabetic NOD mice.
Main Results:
- * BetaTC-tet cells and engineered lines were rejected in diabetic NOD mice and NOD-scid mice.
- * Temporary survival of cells was observed in young male NOD mice treated with high-dose streptozotocin, unlike in C57BL/6 mice.
- * This protective effect of streptozotocin diminished in older NOD mice; cells induced IL-2 secretion from diabetic NOD splenocytes.
Conclusions:
- * Autoimmune processes significantly contribute to betaTC-tet cell destruction in NOD mice.
- * Genetic modifications for enhanced beta cell resistance were ineffective in vivo.
- * High-dose streptozotocin impacts immune rejection, necessitating cautious use in experimental models.