I take fresh whole blood samples and spike them with bacteria. Within a short period (less than an hour) a noticeable amount of the bacteria has been killed by leukocyte activity. Due to the nature of the experiments, disrupting the cells' integrity would have a negative effect. I know that UVB can cause apoptosis or necrosis depending on intensity but I am unsure of a method that will leave the cells intact but inactive. Any suggestions or resources are welcome.
Did you try gamma-irradiation? Something between 30 to 50 Gy should inactivate the cells without killing them, maybe that could be a solution.
Dear Kim
Hope you found a method. Let us know.
If not, try this Table and the associated article
http://www.nature.com/bmt/journal/v33/n1/fig_tab/1704284t1.html
But any thing that inactivates may interfere with their function; will it result in interfering with the purpose of your study?
If they are not necessary why not remove the buffy coat?
Narayanan
Great chart! More things for me to look into for their mechanism of inactivation.
I don't want to remove the buffy coat because that would also take out platelets. We're studying septicemia, so it's most likely that the patients would have low immune cell populations (part of the cause of the infection) but that other blood components would be approximately the same as normal (though not always the case... aplastic anemia, for example, would be low across the board). Rupturing the white blood cells would change the consistency of the blood which is why I'd like to leave them intact, just inactive.
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