Fluorospot IL-4 positive control

[Guest Leonie]

Hello,

I would like to perform the IFNg/IL-4 Fluorospot on PBMCs stimulated with a vaccine antigen in order to see if the vaccine induces predominantly a Th1 or Th2 response. Do you have a recommenation for a positive control for IL-4 secretion? E.g. PBMCs of a patient with a specific helminth infection, malaria,...? 

Thanks!

Leonie 

[Christian@mabtech.com]

Hello Leonie,

For human IFNg/IL4 I would recommend using either (AntiCD3+antiCD28) or PHA. For IL-4 secretion it is good to go up 100K-150K cells/well in order to get good response. 

Combining antiCD3 and antiCD28 will give the response a push. Works also without but spots do become stronger and often more plentiful. 

For PHA I would recommend using the same cell numer. 

best,
Christian