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

Article Article scientifique Article Investigation of alloreactive NK cells in mixed lymphocyte reactions using paraformaldehyde-silenced target cells eng 2007 author Zenhaeusern Gabriela author Gasser Olivier 82983 author Saleh Lanja author Villard Jean 29595 author Tiercy Jean-Marie 26423 author Hess Christoph 196-199 10.1016/j.jim.2007.01.002 17306826 0022-1759 https://www.sciencedirect.com/science/article/pii/S0022175907000270 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. 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 Dewey 616 Swiss National Science Foundation SCORE-grant 3200B0/103253/1 & 3200B0/103254/1 CH Swiss National Science Foundation 320000–11255 CH 56 Département de médecine 221 false 298 Transplantation immunologie et osteoimmunologie 561 Journal of Immunological Methods 321 1-2 2ffa6ec1-9b3b-4220-8371-bfa07e56ce47 Article (Published version) Restricted 1-s2.0-S0022175907000270-main.pdf 1107179 application/pdf restrictedAccess Faculté de médecine / Section de médecine clinique / Département de médecine

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Investigation of alloreactive NK cells in mixed lymphocyte reactions using paraformaldehyde-silenced target cells	Journal of Immunological Methods			2007	205	0

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Gasser, Olivier