en
Scientific article
English

Investigation of alloreactive NK cells in mixed lymphocyte reactions using paraformaldehyde-silenced target cells

Published inJournal of Immunological Methods, vol. 321, no. 1-2, p. 196-199
Publication date2007
Abstract

Mixed lymphocyte reactions (MLRs) remain central to the characterization of cellular allo-interactions. Here we show that irradiation, as used to 'silence' a given cell-population in unidirectional ('one-way') MLRs, is unable to abolish cytokine-production even at doses much higher than usually applied. By contrast, using target cells silenced via a formaldehyde-based fixation-protocol, we demonstrate feasibility to detect - in a true one-way reaction - secretion of IFNgamma by alloreactive NK cells. This simple, fixation-based protocol provides an accurate, robust and time-efficient means for assessing alloreactivity, avoiding cytokine-production by the MLR stimulator cells.

Keywords
  • Cytokines/metabolism
  • Dose-Response Relationship, Radiation
  • Enzyme-Linked Immunosorbent Assay/methods
  • Feasibility Studies
  • Fixatives/pharmacology
  • Formaldehyde/pharmacology
  • Gamma Rays
  • Humans
  • Interferon-gamma/metabolism
  • Killer Cells, Natural/drug effects/immunology/metabolism/radiation effects
  • Leukocytes, Mononuclear/drug effects/immunology/metabolism/radiation effects
  • Lymphocyte Activation/drug effects/radiation effects
  • Lymphocyte Culture Test, Mixed/methods
  • Polymers/pharmacology
Funding
  • Swiss National Science Foundation - SCORE-grant 3200B0/103253/1 & 3200B0/103254/1
  • Swiss National Science Foundation - 320000–11255
Citation (ISO format)
ZENHAEUSERN, Gabriela et al. Investigation of alloreactive NK cells in mixed lymphocyte reactions using paraformaldehyde-silenced target cells. In: Journal of Immunological Methods, 2007, vol. 321, n° 1-2, p. 196–199. doi: 10.1016/j.jim.2007.01.002
Main files (1)
Article (Published version)
accessLevelRestricted
Identifiers
ISSN of the journal0022-1759
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