This one is for the B cell immunologists. In the absence of an animal model or antigen-experienced samples, is it possible to robustly induce antigen-specific antibodies from naive PBMCs in vitro over a given period of time? NB: the PBMCs/donors would not have "seen" the antigen before.
For example, if I took 1e6 PBMCs and stimulated them with a bacterial peptide pool (and perhaps additional co-stimulatory molecules like IL-2/6) for 5-7 days could I detect antigen-specific responses via indirect ELISA?
All the major players would present in the well; however, there's obviously no defined germinal center, etc so I have my doubts as to the validity of the whole exercise. Have any of you tried something similar?
Thanks in advance for your input!
Hi Nathan,
Considering a restricted number of cells of a cell culture assay as you mentioned, it is unlikely that you stimulate the specific B cell clone to expand and differentiate into antibody-secreting cells.
However, other reagents (Pokeweed mitogen + Staphylococcus aureus cowan + CpG ODN) may present a better result if combined with a peptide pool for incubation with PBMCs.
The description of these reagents as an optimal stimulum for B cell expansion and diferentiation into antibody-secreting cells in vitro can be found in the paper below:
Tracking human antigen-specific memory B cells: a sensitive and generalized ELISPOT system. Journal of Immunological Methods
Volume 286, Issues 1–2, March 2004, Pages 111-122
Shane Crotty, Rachael D. Auberta, John Glidewell, Rafi Ahmed
Good luck,
Eduardo
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts