INDUCTION OF TNF- and IFN (WHOLE BLOOD METHOD)

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Tony Rook
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INDUCTION OF TNF- and IFN (WHOLE BLOOD METHOD)

Please find the link for the following protocol:

INDUCTION OF TNF- and IFN (WHOLE BLOOD METHOD)

http://aactg.s-3.com/pub/download/imm/iminducy.pdf

Principle:

The capacity for production of cytokines is an important functional measure of immunologic status. Production of IFN( in response to soluble antigens is mainly a reflection of CD4 cell function, and is progressively lost in HIV infection. Production of TNF" is mainly a reflection of monocyte and macrophage activation. The capacity for production of TNF" is generally maintained in HIV infection, and may be increased during
certain opportunistic infections. Production of both these cytokines may be affected by immunomodulatory drugs such as prednisone. RBCs and PMNs neither contribute to nor inhibit production of these cytokines. Thus, it is not necessary to remove these cells prior to stimulation with antigens or mitogens in culture. This is the basis for the "whole blood" method. Other cytokines can potentially be studied by this method, including IL-2, IL-6, IL-10, IL-12, RANTES, and TGF$. These have not yet been studied systematically by the ACTG, however. It may also be possible to study CD4 and CD8 responses to HIV gp120 with this method, by stimulating,
respectively, with soluble gp120 protein, or by infection with a gp120 vaccine construct. These methods are under development.