In Vivo Assessment of NK Cell-Mediated Cytotoxicity by Adoptively Transferred Splenocyte Rejection
NK cells are innate lymphocytes that are vital to clearance of virally infected or malignantly transformed cells. Assessment of the cytotoxic response is an important component of NK cell research and investigation of human disease. Standard assays of NK cell-mediated cytotoxicity of CD107a degranulation or 51Cr release assay utilize cultured or freshly isolated NK cell populations in vitro. In addition to requirements to maintain multiple target cell lines and radioactivity precautions in the case of 51Cr, these are in vitro evaluations of a complex in vivo function. Here, we describe the in vivo assessment of NK cell-mediated cytotoxicity through the adoptive transfer of splenocytes and their subsequent rejection. This protocol offers rapid, quantitative, and concurrent assessment of NK cell-mediated cytotoxicity against the prototypic NK stimulations of “missing-self” and “nonself.”
KeywordsNK cell Cytotoxicity Transplant Missing-self Nonself In vivo
Funding Support: We thank Lucia Sammarco and her Lulu’s Lemonade Stand for inspiration, motivation, and support. This work was supported in part by Ann’s Hope Melanoma Foundation (S.M. and M.S.T.); NIH R01 AI102893 (S.M.) and NCI R01 CA179363 (S.M. and M.S.T.); Alex Lemonade Stand Foundation (N.J.S); HRHM Program of MACC Fund/Children’s Hospital of Wisconsin (S.M.), Nicholas Family Foundation (S.M.); Gardetto Family (S.M.); MCW-Cancer Center-Large Seed Grant (S.M. & M.S.T.); and MACC Fund/Children’s Hospital of Wisconsin (M.S.T. and S.M.).
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