Hello,
After scanning the available literature on memory cells i found that, as many papers have indicated now that Memory b cell pool is usually generated early at GC reaction, and later some memory B cells join it but not with extensive SHM. So my question is, if MBC's develop so early, then why almost every other study on MBC's has time span of 2, 3 months or even longer. For my project all i am interested is to see the development of Memory B cells (or T cells) , what time point should i choose? how many immunization etc. Previously, i have to develop a stable model for memory B cells (or for T cells). I'm used OVA as an antigen to immunize the mouse , wait for nearly 2 months and give a boost, then 6-7 days later i looked for MBC's / MTC's in the spleen. I tried this protocol once, however i didn't get satisfactory result for either type. Central memory T cells (CD44+ CD62+ )even decreased and shifted to Effector memory (CD44+ CD62-). the time point is long enough to repeat multiple times. how should improve this protocol? kindly suggest me a Valid protocol to follow and get the stable memory cell model (For B and for T cell memory), which has already been established in the lab.
Also, regarding the flowcytometry panel, i have used B220+, CD38+, GL7-, OVA+, IgG /G1/ M+ . is this enough ? surprisingly , all my OVA+ cells seem to be around >90-95% IgG. which doesn't seem normal.
I would highly appreciate your worthy views and help on the subject. Thank You