Interesting question. I believe that it does not exist, but I would be interested by the answers.
Hi Thierry,
I have plenty of EBV-transformed B cell clones, although they may not be suitable for your purposes. A few of them are growing slowly. I wonder whether they may be of help.
Best wishes.
Mario
Hi Thierry,
I have two suggestions for you: one is similar to Mario Mondelli's. Check a number of EBV-transformed cell lines. I could also offer more then 100 ;-)
Then, you may not be aware of the possibility to activate B cells poluclonal by CD40L or CD154. We use either the system described by Schultze et al. in 1997 (Nadler-Lab at that time point) basing on CD40L-transfected NIH3T3 cells or plates coated with rCD40L (antibodies against CD40 also work).
Best
Michael
Following on Michael's suggestion you can also try innate immune signals such as tickling TLR9 with CpG, although this would be a stronger signal compared with the CD40/CD40L pathway
I’m not an expert on this topic, but I recently saw a paper, may be of some interest to You:
(Human pre-B cell receptor signal transduction: evidence for distinct roles of PI3kinase and MAP-kinase signalling pathways. Immunity, Inflammation and Disease 1 (1), 26–36, October 2013 - Wiley http://onlinelibrary.wiley.com/doi/10.1002/iid3.4/full ),
In this paper, Anbazhagan et al. investigated pre-BCR signalling pathway to examine its regulation in normal primary pre-B lymphocytes and pre-B cell lines. Pre-BCR-dependent cell cycle entry and proliferation of 697 and Nalm6 pre-B cell lines following anti-m F(ab’)2 antibody stimulation are described.
Best wishes
Mauro
Indeed, you can use malignant B cell lines, like BL2 or DOHH2, with reduced % of FCS to have a limited growth, and stimulate them with a BCR / CD40L / CpG -(+/- IL2 and IL4) cocktail.
@ Mauro
Interesting paper. The thing is that, unlike the BCR, the pre-BCR is constitutively activated as a consequence of lambda-5 mediated pre-BCR aggregation. Then I am wondering what could be the effect of anti mu on it, i.e. superaggregation or disaggregation.
@ Paticia
Do you have a reference where proliferation is increased solely by adding anti-mu?
The human B cell line 2E2 was made by the Paolo Casali lab. It is derived from the Burkitt's lymphoma CL-01 line and is a model of activation and proliferation. It can be induced to undergo GC and plasma cell differentation from a variety of stimuli.
@ Van Luu : I am intrested in this cell line, do you know how to get it ? thanks
One of the numerous applications tested with the HuBBB Kit, sold by the company Dendritics, probably corresponds to the question.
The set (human blood B booster), which can stimulate and immortalize B lymphocytes from peripheral blood, has been used successfully to identify antibodies capable of activating B cells or on the contrary kill B cells specifically.
HuBBB is a process, turnkey, who can reactivate the B cells from peripheral blood, start to proliferate and secrete antibodies again, via the joint action of CD40 antibody and CD40L molecule.
The readout can be done either by following the proliferation or more simply by assay of secreted Igs.
A large number of molecules will be activated on the surface of the B cell (for example, CD20, CD21, HLA ...) and become the target of the antibody to be tested.
The procedure of the kit is described in www.dendritics.net
Hi Jean-Jacques
Do you have references where proliferation is stimulated by Fab'2 anti-IgM antibodies?
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