Insulin receptor based lymphocyte trafficking in the progression of type 1 diabetes


  • Michael P. Morran University of Toledo
  • Ali G. Al-Dieri University of Toledo
  • Andrea L. Nestor?Kalinoski University of Toledo
  • Richard K. Jordan University of Toledo
  • Nirdesh K. Gupta
  • Marcia F McInerney University of Toledo



chemotaxis, Cre-recombinase, diabetes, insulin receptor, insulitis


The insulin receptor (IR) is a transmembrane receptor which recognizes and binds the hormone insulin. We describe two models that were devised to explore the role of IR over-expression on T-lymphocytes and their chemotactic motility in the progression of type 1 diabetes. FVB/NJ-CD3-3×FLAG-mIR/MFM mice were generated to selectively over-express 3×FLAG tagged murine IR in T-lymphocytes via an engineered CD3 enhancer and promoter construct. Insertion of the 3×FLAG-mIR transgene into FVB/NJ mice, a known non-autoimmune prone strain, lead to a minor population of detectable 3×FLAG-mIR tagged T-lymphocytes in peripheral blood and the presence of a few lymphocytes in the pancreas of the Tg+/- compared to age matched Tg-/- control mice. In order to induce stronger murine IR over-expression then what was observed with the CD3 enhancer promoter construct, a second system utilizing the strong CAG viral promoter was generated. This system induces cell specific IR over-expression upon Cre-Lox recombination to afford functional 3×FLAG tagged murine IR with an internal eGFP reporter. The pPNTlox2-3×FLAG-mIR plasmid was constructed and validated in HEK-Cre-RFP cells to ensure selective Cre recombinase based 3×FLAG-mIR expression, receptor ligand affinity towards insulin, and functional initiation of signal transduction upon insulin stimulation.

Author Biography

Marcia F McInerney, University of Toledo

Distinguished University Professor


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How to Cite

Morran MP, Al-Dieri AG, Nestor?Kalinoski AL, Jordan RK, Gupta NK, McInerney MF. Insulin receptor based lymphocyte trafficking in the progression of type 1 diabetes. J Biol Methods [Internet]. 2018Jan.23 [cited 2021Oct.21];5(1):e85. Available from:




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