targeting CD154 (CD40L) to clone T cells is probably a good indication as CD154, in its active form and trimeric, is mainly present on activated T cells.

But all CD154 antibodies are not suitable: for example, LL2 and LL48 inhibit the binding between CD40 and CD154, and can have an effect on proliferation, whereas LL55 has no inhibitory activity.

Other clones, as 115H5, are more specific for CD154 present on T regs.

Finally the use of CD40 can also be a solution, but this interaction is mainly described for the activation of B cells

Thanks Dr Jean It surly helps a lot about selecting the best clone.

Harish

Dear Dr Harish Chandra, you can try to do T hybridomas. This is much more efficient way to obtain big number of T cell clones in comparison with classical procedures of T cell cloning.

Thanks Dr Kazansky I am looking for  Antigen specific T cell clones  of Mycobacterium by challenging  autologous dendtritic cells  with whole cell antigens and subsequently interacting these with naive T cells isolated from healthy subjects in DC-T cell assay in vitro. Do have a detailed protocol to generate these T cell hybridomas?

You can get it here. http://www.springerprotocols.com/Pdf/doi/10.1385/1-59259-682-7:185?encCode=TU1JOjU4MTo3LTI4Ni05NTI5NS0x&tokenString=L7+H9++gEJQfYnC2byjq8w==

Thanks Dr. Kazansky