Quantitative in situ analysis of IL-6 mRNA and its producing cells in transplant kidneys
Final Report Abstract
During my postdoctoral fellowship at the University of California, San Francisco, I optimized and developed an algorithm to perform whole slide high-resolution digital imaging and image analysis to quantitatively identify the precise cellular source of mRNA signals in transplanted kidneys at a single cell resolution. We showed that the mIFISH assay provides unique information on spatial context of cytokine producing cells that can be acquired and analyzed. Such information may help to improve in-situ assessment of immunological processes in kidney transplants and other transplanted organs where similar rejection mechanisms apply, and may eventually contribute to improved diagnostics and patient care. In a second project, I studied whether the four commonly used immunosuppressive drugs - prednisolone, mycophenolate mofetil, tacrolimus and rapamycin - have an impact on in vitro functional T-cell assays. We showed that T-cell function could be restored by removing residual immunosuppressive drugs during sample processing by a simple freezing/thawing cycle, followed by overnight incubation prior to the functional T cell assay. Our findings may help to further optimize and standardize functional T cell assays in the posttransplant period. I contributed to another research project where we studied the impact of belatacept on the phenotypic heterogeneity of renal T-cell mediated alloimmune responses. Our results might suggest differential involvement of the innate immune system, and an altered B cell engagement during T cell mediated rejection in belatacept-treated patients, relative to CNI-treated patients.
Publications
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Clinical Transplantation – Clinical Transplantation e14084. The impact of belatacept on the phenotypic heterogeneity of renal T-cell mediated alloimmune response: the critical role of maintenance treatment and inflammatory load
Dobi D, Vincenti F, Chandran S, Greenland J, Bowman C, Chen A, Junger H, Laszik Z
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J Histochem Cytochem. Novel in-situ hybridization and multiplex immunofluorescence technology combined with whole slide digital image analysis in kidney transplantation
Junger H, Dobi D, Chen A, Lee J, Vasquez JJ, Tang Q, Laszik ZG
