Plasma cells in circulating blood.

[CD19]

I don't use LPS. I did few tests and I found in some people's supernatant IgG. When I finish I will have to confront the antibodies titres with MFI of surface antigens (among others I evaluated CD40 and CD40L). This is just crazy.

[Zagami]

Good luck

[CD19]

Is this possible that I after culture there is higher concentration of IgG than IgM? The difference is not significant, but I found it.

[CD19]

After 72 hours I didn't find IgG, but after 120h I found and it some cases it was higher than IgM, but maybe this is normal, coz IgM is fast response and IgG is after switch?

[luisillo]

I've never done it but, according to literature (I don't have the article right now, though), IgG appears in long term cultures: 5 or more days. The authors mentioned something about not changing media enhances IgG over IgM production. I will try to get at least the reference and post it.

Good luck

[luisillo]

Might help you:

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[CD19]

O, thanks.

[CD19]

I am reading now article about PBMC stimulation with Pokeweed (PWM) + S. aureus + CpG. They are tracking in these research memory B cells and counting IgG titres, so as I understand such a stimulation activated memory B cells. Since 40% of B cells in circulating blood are memory B cells then I also have them in my culture, BUT I stimulate T cells with anti-CD3 and due to cytokine production and also CD40-CD40L (etc.) interaction I activate B cells. Is anti-CD3 stimulation able to stimulate memory B cells? i dont add any specific antigen to provoke memory B cells. I activate only T cells.

[CD19]

I read at mirorao.com that memory B cells can be activated only by protein antigen which I dont have in my culture. So IgM and IgG titres I received appeared from newly formed plasma cells which were generated after anti-cd3 stimulation of T cells. There are almost no plasmablasts and plasmocytes in blood of healthy humans so they were created de novo in my culture. Am I finally right?