[Polyclonal B cell activation induced by Actinomyces viscosus T14V. Responsiveness of human peripheral blood B cells]

We have previously reported that the sonicate extrast from Actinomyces viscosus T 14 V (Av. sup) causes polyclonal B cell activation (PBA) in murine splenic B cells without any T cell assistance. However, there are some reports describing T cell dependency of PBA induced by A. viscosus in human peripheral blood B cells, and it is still controversial whether or not A. viscosus has the capability of exhibiting PBA without the help of T cells. In this report, we examined PBA in human peripheral blood B cells using the same Av. sup which induced T cell independent PBA in murine splenic B cells. The PBA was evaluated in terms of immunoglobulin (Ig) production by means of micro-ELISA method and cell proliferation by incorporation of 3H-TdR after incubation of cells for 5 or 7 days with Av. sup. Human peripheral blood B cells were highly purified by the following methods; The peripheral blood was collected by venipuncture and the peripheral blood mononuclear cells were harvested using gradient centrifugations. These cells were passed through a Sephadex G-10 column, and T cells were eliminated by rosette formation with sheep red blood cells. B cells were highly purified from these T cell-depleted cell populations by panning method with anti-human Ig antibody coated dishes followed by complement-dependent cytotoxisity using monoclonal antibody cocktails consisting of OKT 3, OKT 4 and OKT 8. These highly purified B cells could not respond to T cell mitogens such as PHA.(ABSTRACT TRUNCATED AT 250 WORDS)