Hello,
Can anyone tell me or provide a publication that can explain the effects of ammonium chloride on NK or T cells cytokine production? I am getting ready to perform some functional studies and was going to incorporate an RBC lysis into my cell preparation protocol of human whole blood. I am going to Ficoll the blood, isolate the PBMCs and lyse residual RBCs with ammonium chloride (150mM). RBC lysis would take place on ice for about 15 minutes. I could always run a study to see if it does or doesn't inhibit but thought I'd ask first.
Thank you!
Whatever we use for RBC lysis is washed out in the subsequent steps. Any effect in that 15 minutes should be minimal compared to culture.
The last thing you want to do when studying cytokine production is to introduce artefactual changes in the system. Ammonium chloride is harsh on cells (that's why it is used to lyse red cells). It may not kill the leucocytes, but may significantly affect the surface membrane proteins and cytoplasmic machinery both of which are, as you know, involved in cytokine production in response to external stimuli.
You may want to have a look at these articles.
Best wishes
Siva
Temporary Inhibition of Antibody-Dependent, Cell-Mediated Cytotoxicity by Pretreatment of Human Attacking Cells with Ammonium Chloride
Israel Yust, Richard W. Smith, John R. Wunderlich and Dean L. Mann J Immunol April 1, 1976, 116 (4) 1170-1172;
Article Antibody-dependent cellular cytotoxicity and natural cytotox...
Article Effect of red cell lysis protocols on the expression of rat ...
Red cell lysis with Ammonium chloride buffer inhibits NK killing of K562 target cells, so I would assume may significantly distort the interpretation of functional studies. I had to use hypotonic shock when I was working with PBMC to evaluate NK cell versus T-cell killing.
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