I think it is best to use isolated B cells. Otherwise other cells can produce cytokines that interfere with the skewing.

You should add something that crosslinks the BCR, such as anti-IgM. That together with CD40 ligation and IL-4 should give you IgE switching and proliferation.

Hi Jolien Suurmond,

I am very appreciated for your answer. I'll try once as you suggested, using isolated B cells. But for BCR crosslinking, I don't know how to choose antibody? Anti-IgM or/and anti-IgG? What is depends on? Looking forward to your professional answer.

Or maybe anti-IgM is enough, because only naive B cells occur CSR? Honestly, I am not sure if this is right?

I think it depends on what you want to do with the IgE+ cells. Anti-IgM alone is enough to induce switching in part of the B cells, but maybe adding anti-IgG could increase it further (I have never tried). Do you need a pure IgE+ B cell population?

There are some interesting papers out there about sequential switching of IgG+ B cells to IgE (mainly in mice):

Sequential class switching is required for the generation of high affinity IgE antibodies.Xiong H, Dolpady J, Wabl M, Curotto de Lafaille MA, Lafaille JJ. J Exp Med. 2012 Feb 13;209(2):353-64.

Unique maturation program of the IgE response in vivo. Erazo A, Kutchukhidze N, Leung M, Christ AP, Urban JF Jr, Curotto de Lafaille MA, Lafaille JJ. Immunity. 2007 Feb;26(2):191-203. Epub 2007 Feb 8.

Hi again,

Thanks a lot for your help. I am trying to see if my patients with critical high level of serum IgE (which has been tested is allergen specific) undergo more CSR to IgE than healthy controls. 

I'll post my further questions and progress of this topic. Hope that you will follow my topic and keep in touch  :)