In vivo cytotoxicity assay for assessing immunity

De Chu Tang, Adi F. Gazdar, David P. Carbone

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

A rapid, quantitative, in vivo assay of cytotoxic responses would facilitate experimental evaluation of the potency of novel vaccine strategies. We have developed an in vivo cytotoxicity assay in which target cells expressing a luciferase reporter gene are implanted as monolayers on polystyrene disks onto the muscle tissue of mice. The luciferase activity retrievable from the adjacent tissue is used as an index of cytotoxicity. Implantation of B16 or NIH/3T3 cells expressing the β-galactosidase gene indicated that the target cells migrated to the muscle tissue from plastic within 4 h and then remained localized in the area of the disk. The amounts of luciferase retrievable from the adjacent tissue a few days post implantation readily detected the immune response induced by allo-immunization of fibroblasts or by production of interleukin-4 by tumor cells co-mixed with implanted reporter cells. Histologic analysis showed a correlation between the amount of luciferase retrieved and the number of viable target cells at the implantation site. Recruitment of immune effector cells which may be responsible for target cell death and luciferase elimination could be readily visualized. This simple cytotoxicity assay can be used as an in vivo assay of the net effect of cytotoxic immune responses.

Original languageEnglish (US)
Pages (from-to)173-182
Number of pages10
JournalJournal of Immunological Methods
Volume189
Issue number2
DOIs
StatePublished - Feb 5 1996

Keywords

  • Cytotoxicity
  • Immunity
  • Luciferase
  • Transplantation
  • Tumor

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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