粘胶放线菌T14V诱导多克隆B细胞活化。人外周血B细胞的反应性[j]。

T Okada, Y Harada, H Ito, T Nozaki, S Ebisu, S Kimura, H Okada
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引用次数: 0

摘要

我们之前报道过来自粘胶放线菌t14v (Av. sup)的声波提取物在没有T细胞辅助的情况下引起小鼠脾B细胞的多克隆B细胞活化(PBA)。然而,有一些报道描述了粘胶原在人外周血B细胞中诱导PBA的T细胞依赖性,粘胶原是否能够在没有T细胞帮助的情况下表现出PBA的能力仍然存在争议。在本报告中,我们使用同样的Av. sup在小鼠脾B细胞中诱导T细胞非依赖性PBA,检测了人外周血B细胞中的PBA。用微酶联免疫吸附试验(elisa)和3H-TdR与Av. sup共孵育5天或7天后,对PBA进行免疫球蛋白(Ig)产生和细胞增殖的评价。静脉穿刺法采集外周血,梯度离心法采集外周血单个核细胞。将这些细胞通过Sephadex G-10柱,用羊红细胞形成玫瑰花结消除T细胞。B细胞从这些T细胞耗尽的细胞群中通过抗人Ig抗体包被培养皿的淘洗方法高度纯化,然后使用由okt3、okt4和okt8组成的单克隆抗体鸡尾酒进行补体依赖性细胞毒性。这些高度纯化的B细胞不能对T细胞裂丝原如PHA产生反应。(摘要删节250字)
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[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)

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