Pcr based diagnosis of canine lymphoma and in vitro evaluation of chimeric antigen receptor expressing cells

Abstract

Cancer is a major health issue in companion animals with the incidence of 1 in 4 dogs which is more than the incidence rate in humans. Lymphoma accounts for about 7% - 24% of all neoplasia in dogs and it is the most prevalent form of hematopoietic neoplasm in dogs. In India, there are limited research reports pertaining to prevalence, incidence, clinical diagnosis, use of molecular techniques for phenotyping and grading as well as basic studies on cell based therapy. In this study, we have attempted different PCR based approaches including expression markers for diagnosis and differentiation of lymphoma. The study also performed in-vitro assessment of activated natural killer cells and also genetically modified NK cells that express the canine anti-CD20 CAR construct. newlinePreliminary studies were assessed with PBMC and NK enriched population. It could be observed that the CD3+ T cell depleted canine PBMC had higher cytotoxic potential to Raji cells indicating that a process to upscale the depletion of CD3+T cells and cytokine mediated activation can be a primary approach for clinical application in non-responsive cases. The transfected cells had higher cytotoxicity to Raji cells. With the use of 1/10 ratio of Raji cells the observed percentage of viability was 50.01% and 55.39% with canine PBMC and CD3+ T cell depleted canine PBMC expressing anti-CD20 CAR respectively. Both the canine PBMC and CD3+ T cell depleted canine PBMC expressing anti-CD20 CAR had a similar higher cytotoxic potential to Raji cells indicating the potential utility of the CAR construct. In future, the construct can be scaled up with viral approaches for adoptive cell therapy in canines. newline newlineKeywords: Canine lymphoma, PARR, IgH, TCRand#947;, PBMC, anti-CD20 CAR newline