1728-P: Inhibition of PKC-Theta in Prediabetic NOD Mice Prevents Diabetes Onset and Increases Regulatory CD4 T Cells in a Subset of Animals

Diabetes ◽  
2019 ◽  
Vol 68 (Supplement 1) ◽  
pp. 1728-P
Author(s):  
JAMES E. DILISIO ◽  
NOAH MISHKIN ◽  
BRENDAN PODELL
Keyword(s):  
T Cells ◽  
Nod Mice ◽  
Diabetes ◽  
2005 ◽  
Vol 54 (5) ◽  
pp. 1477-1486 ◽  
Author(s):  
D. A. Mahamed ◽  
A. Marleau ◽  
M. Alnaeeli ◽  
B. Singh ◽  
X. Zhang ◽  
...  

1993 ◽  
Vol 178 (1) ◽  
pp. 87-99 ◽  
Author(s):  
M J Rapoport ◽  
A Jaramillo ◽  
D Zipris ◽  
A H Lazarus ◽  
D V Serreze ◽  
...  

Beginning at the time of insulitis (7 wk of age), CD4+ and CD8+ mature thymocytes from nonobese diabetic (NOD) mice exhibit a proliferative unresponsiveness in vitro after T cell receptor (TCR) crosslinking. This unresponsiveness does not result from either insulitis or thymic involution and is long lasting, i.e., persists until diabetes onset (24 wk of age). We previously proposed that it represents a form of thymic T cell anergy that predisposes to diabetes onset. This hypothesis was tested in the present study by further investigating the mechanism responsible for NOD thymic T cell proliferative unresponsiveness and determining whether reversal of this unresponsiveness protects NOD mice from diabetes. Interleukin 4 (IL-4) secretion by thymocytes from > 7-wk-old NOD mice was virtually undetectable after treatment with either anti-TCR alpha/beta, anti-CD3, or Concanavalin A (Con A) compared with those by thymocytes from age- and sex-matched control BALB/c mice stimulated under identical conditions. NOD thymocytes stimulated by anti-TCR alpha/beta or anti-CD3 secreted less IL-2 than did similarly activated BALB/c thymocytes. However, since equivalent levels of IL-3 were secreted by Con A-activated NOD and BALB/c thymocytes, the unresponsiveness of NOD thymic T cells does not appear to be dependent on reduced IL-2 secretion. The surface density and dissociation constant of the high affinity IL-2 receptor of Con A-activated thymocytes from both strains are also similar. The patterns of unresponsiveness and lymphokine secretion seen in anti-TCR/CD3-activated NOD thymic T cells were also observed in activated NOD peripheral spleen T cells. Exogenous recombinant (r)IL-2 only partially reverses NOD thymocyte proliferative unresponsiveness to anti-CD3, and this is mediated by the inability of IL-2 to stimulate a complete IL-4 secretion response. In contrast, exogenous IL-4 reverses the unresponsiveness of both NOD thymic and peripheral T cells completely, and this is associated with the complete restoration of an IL-2 secretion response. Furthermore, the in vivo administration of rIL-4 to prediabetic NOD mice protects them from diabetes. Thus, the ability of rIL-4 to reverse completely the NOD thymic and peripheral T cell proliferative defect in vitro and protect against diabetes in vivo provides further support for a causal relationship between this T cell proliferative unresponsiveness and susceptibility to diabetes in NOD mice.


1999 ◽  
Vol 192 (2) ◽  
pp. 113-121 ◽  
Author(s):  
Patricia R. Hutchings ◽  
Suman Verma ◽  
Jenny M. Phillips ◽  
Silvia Zusman Harach ◽  
Sarah Howlett ◽  
...  
Keyword(s):  
T Cells ◽  

2011 ◽  
Vol 41 (5) ◽  
pp. 1480-1490 ◽  
Author(s):  
Kevin S. Goudy ◽  
Mark C. Johnson ◽  
Alaina Garland ◽  
Chengwen Li ◽  
Richard J. Samulski ◽  
...  

2011 ◽  
Vol 66 (5) ◽  
pp. 349-362 ◽  
Author(s):  
Yun Sun ◽  
Wenjing Wang ◽  
Bin Shan ◽  
Jingfang Di ◽  
Linlin Chen ◽  
...  

2020 ◽  
Vol 1 (1) ◽  
pp. 8-14
Author(s):  
Victor Tunje Jeza ◽  
Chen Jun ◽  
Wu Xiongwen

Currently, general immunosuppressive drugs are used to maintain tolerance to allografts. However, these drugs have a major drawback of rendering the patient susceptible to infections and other side effects like malignancy and drug related toxicities with an overall rejection of the organ at some point. Previous studies have shown that MHC-Ig dimers may suppress alloresponsive T cells in a donor specific manner in vitro. This work aimed to answer the question as to whether these dimers will surmount rejection through the direct mechanism of allorecognition by suppressing alloreactive CD8+ T cells. To do this, we first identified two mice models with a single mismatch at the MHC loci. We found and procured white albino NOD mice which happened to be transgenic for HLA-A2 and HLA-A24 molecules. We then constructed a human-mouse hybrid HLA-A2-Ig dimer by overlap-PCR to join parts of two different already cloned plasmids to form the full length HLA-A2β2α1α2murineα3 insert which was then cloned to pcDNA3.1 to form pcDNA3.1HLA-A2β2α1α2murineα3. The IgG2bFc region was added by restriction digestion and ligation to form the plasmid pcDNA3.1HLA-A2β2α1α2murineα3IgG2bFc. Sequencing was done and confirmed that the construction and cloning were successful. The plasmid pcDNA3.1HLA-A2β2α1α2murineα3IgG2bFc was then transfected by electroporation to J558L cells. Screening was done using G418 for 4 weeks in cell culture. We purified the dimer by affinity chromatography and then used ELISA to confirm expression of the dimer. The purified dimer was then used in 1-way MLC experiments where responder cells were mice cells expressing HLA-A24 molecules while stimulator cells were mice cells expressing HLA-A2 molecules. Cell samples were gated on anti-mouse CD3-PE/CY7, anti-mouse CD4-PE, and anti-mouse CD8-APC/CY7. Cell proliferation was analysed using CFSE. Our results showed that the proliferation of CD4+ T cells was inhibited in the presence of the dimer. This work is crucial for subsequent studies aiming to search for induction of donor specific tolerance.


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