Rationale: Lupus nephritis (LuN) is the most common severe manifestation of systemic lupus erythematosus. Up to 50% of SLE patients develop LuN which usually requires toxic immunosuppressive therapies. Despite such aggressive treatments, up to 50% of LuN patients progress to renal failure within 5 years of diagnosis. We propose that these therapeutic failures reflect an incomplete knowledge of LuN pathogenesis. Goals: The goals of this Career Development Award (K08) application are to validate and iteratively develop a novel algorithm (Cell Distance Mapping - CDM) that is able to identify cognate cellular interactions between lymphocytes by means of computerized analysis, performed on images acquired from multi-channel immunofluorescent confocal laser scanning microscopy (CLSM), as recently published in Science Translational Medicine. We plan to extend this algorithm to the analysis of cognate T:dendritic cell (DC) interactions and validate it by means of using several unique systems with the first comprised of a triple adoptive transfer mouse model of cell tracker-labeled, antigen-pulsed, activated DCs, wild-type T cells, and antigen-specific T cells in the context of lymph node germinal centers. The second model is comprised of mice with sex- mismatched renal allografts combined with adoptive lymphocyte transfer, wherein we are able to restrict the response of T cells based on the gender origin of the source graft antigen(s). Finally, we propose to utilize human sex-mismatched renal allografts, wherein fluorescent in-situ hybridization techniques will allow us to determine the source of antigen-presenting cells and compare their interactions with cognate and non-cognate inflammatory cells. After iterative development of the above method, we propose to apply it to examine the characteristics and make-up of the inflammatory cell infiltrate in human LuN tubulointerstitial inflammation and renal allograft rejection, specifically focusing on the role(s) of DCs, their subtypes, and relation to T cells. We also plan to relate this information to renal disease severity and disease pathogenesis in patients. The experimental methods used for the above consist of multi-channel immunofluorescent CLSM, image analysis techniques, quantitative PCR performed on mRNA from Laser Capture Microscopy samples, as well as the use of super-resolution techniques such as Ground State Depletion Microscopy. Scientific Disciplines, Encompassed by Proposed Research: Immunology, Pathology, Medical Physics. Applicable Research Areas: Autoimmunity, inflammation, transplant immunology, image analysis, molecular and cellular biology. Trainee information: The proposed 4-year project will be performed by Dr. Vladimir Liarski, M.D., a junior faculty member at the level of Instructor and a recent Institutional Career Development Award (K12) recipient.
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, in which the immune response is misdirected at normal tissues and sites of the body. One of the most frequently and severally affected sites are the kidneys, in a process termed tubulointerstitial nephritis, where multiple inflammatory cells are arranged within parts of the kidney and appear similar to other sites of organ inflammation in related autoimmune diseases. We aim to confirm these findings and explore the ways in which this inflammation begins, how it alters the course of disease in human patients, and which types of cells are involved so that we may potentially target them for treatment in the future.
| Trotter, Kimberly; Clark, Marcus R; Liarski, Vladimir M (2016) Overview of pathophysiology and treatment of human lupus nephritis. Curr Opin Rheumatol 28:460-7 |
