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questions about PBMCs study

PBMC cytokine proliferation

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4 replies to this topic

#1 joy123

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Posted 22 July 2013 - 07:37 AM

Hi, I want to analyze if a stimulant could stimulate immune response from patients' peripheral blood mononuclear cells (PBMCs). I have some questions here:

1. How should I testify if the immune response is activated? What parameters should I choose?

2. I see papers test cytokines. So I want to ask how should I choose which cytokines to test?

3. Besides cytoknies, some publications also include Cell proliferation assay. I tried BrdU kit but it doesn't work well. Is cell proliferation assay necessary?

I look forward for your suggestions. THANKS VERY MUCH!Posted Image

#2 bob1

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Posted 23 July 2013 - 02:01 AM

(Warning - I'm not an immunologist, so may give not such good advice here)

Yes, cytokines (e.g. interleukins) would be appropriate to measure, as would T-cell or B-cell activation.

How to choose depends on what you are looking at - bacteria will produce a different response to viruses etc.

Cell proliferation could be a marker, I would go with MTT/MTS assays or a simple cell count (maybe by Flow cytometry) rather than BrdU.

#3 joy123

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Posted 23 July 2013 - 06:36 AM

(Warning - I'm not an immunologist, so may give not such good advice here)

Yes, cytokines (e.g. interleukins) would be appropriate to measure, as would T-cell or B-cell activation.

How to choose depends on what you are looking at - bacteria will produce a different response to viruses etc.

Cell proliferation could be a marker, I would go with MTT/MTS assays or a simple cell count (maybe by Flow cytometry) rather than BrdU.


Thanks! But cell proliferation is not quite realistic for me. Can I skip it? If so, should I do something else to supplement the cytokine analysis?

Edited by joy123, 23 July 2013 - 06:36 AM.


#4 bob1

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Posted 23 July 2013 - 12:38 PM

No idea, sorry. There is bound to be something else that can be done (migration assays perhaps?).

#5 waldomedina

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Posted 26 August 2013 - 04:23 PM

Hello:
If you want to get the percentage of activated cells you have to know that markers used, for example CD25 and / or CD69 in T cells.
The proliferation assay is also a basic tool: BrdU could be good, I usually use CFSE.
And finally if you want to evaluate cytokines, you have to know what cytokines are involved in the process that you are evaluating.
 
Good Luck.

Edited by waldomedina, 26 August 2013 - 04:24 PM.






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