In Vivo Assessment of NK Cell-Mediated Cytotoxicity by Adoptively Transferred Splenocyte Rejection

  • Nathan J. Schloemer
  • Alex M. Abel
  • Monica S. Thakar
  • Subramaniam MalarkannanEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2097)


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.”


NK 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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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Nathan J. Schloemer
    • 1
    • 2
  • Alex M. Abel
    • 1
    • 2
    • 3
  • Monica S. Thakar
    • 1
    • 2
  • Subramaniam Malarkannan
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Laboratory of Molecular Immunology and Immunotherapy, Blood Research InstituteBlood Center of WisconsinMilwaukeeUSA
  2. 2.Department of PediatricsMedical College of WisconsinMilwaukeeUSA
  3. 3.Department of Microbiology and ImmunologyMedical College of WisconsinMilwaukeeUSA
  4. 4.Department of MedicineMedical College of WisconsinMilwaukeeUSA

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