Hi and happy new year to all of you. If I haven't said so before, it bears repeating... I thank you all for your support, past (and future as I have a query!!!) I have a MoAb from B-D to CD40L and want to use it to screen possible Hyper IgM cases. I am experimenting with different methods of stimulating my cells; I gather from reading thus far that one could stimulate with a) PHA for 24 hours but that PHA + PMA was more successful. Alternatively b) many have used immobilised antiCD3; however c) one paper (Eur J Immunol 1996 26;4;846-50 suggested that as opening of the Ca chanel unduced expression one could get induction in 2hours with Ionomycin. They also said that ficol prep PBL were more successful that whole blood..but I'm not sure why that would be. So, I attempted to stimulate PBL for 2hrs with PMA+Ionomycin, Ionomycin alone and Ca Ionophore with.... no success. I'm sure some of you have had experience here..any comments Many thanks Jane Hughes Chief, Institute of Child Health Lab, Red Cross Children's Hospital Rondebosch, Cape Town 7700. South Africa Tel 021 658 5315 Fax 021 689 1287
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