Investigation of alloreactive NK cells in mixed lymphocyte reactions using paraformaldehyde-silenced target cells.
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.
Zenhaeusern
Gabriela
G
Immunobiology Laboratory, University Hospital Basel, Switzerland.
Gasser
Olivier
O
Saleh
Lanja
L
Villard
Jean
J
Tiercy
Jean-Marie
JM
Hess
Christoph
C
D016207:Cytokines / Q000378:metabolism*; D004307:Dose-Response Relationship, Radiation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005240:Feasibility Studies; D005404:Fixatives / Q000494:pharmacology*; D005557:Formaldehyde / Q000494:pharmacology*; D005720:Gamma Rays; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000528:radiation effects; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000528:radiation effects; D008213:Lymphocyte Activation / Q000187:drug effects* / Q000528:radiation effects; D007959:Lymphocyte Culture Test, Mixed / Q000379:methods*; D011108:Polymers / Q000494:pharmacology*
D016207:Cytokines; D005404:Fixatives; D011108:Polymers; D005557:Formaldehyde; D007371:Interferon-gamma; C003043:paraform