De Vries E, Van der Weij J P, Van der Veen C J, Doekes G, Steven M M, Cats A
Eur J Immunol. 1986 Feb;16(2):122-8. doi: 10.1002/eji.1830160203.
In this study we report on the preparation and application of pokeweed mitogen (PWM) conjugated to latex particles. This conjugate (PWM-latex) was prepared by incubation of PWM with latex particles in the presence of glutaraldehyde. The effect of the addition of PWM-latex to human peripheral blood mononuclear cells (PBMC) was fully comparable to the addition of PWM alone i.e. differentiation of lymphocytes into blast cells followed by proliferation of these blast cells as measured by DNA synthesis and total cell number. Electronmicroscopically PWM-latex was found to be taken up by monocytes within the first 24 h after addition. Although no direct interaction could be observed between PWM-latex and lymphocytes, the latter were found to differentiate into blast cells. Due to interactions of these blast cells with latex-containing monocytes, latex particles were obviously released from the phagocytes and close contacts between latex particles and blast cells were regularly seen. In addition, it was found that blast cells of T cell origin, as judged by their positive reaction with anti-T cell monoclonal antibodies, had taken up latex particles. Based on enzyme-cytochemical, functional and light microscopical studies, monocytes could not be detected in the PWM-latex-driven PBMC stimulation after 6 days. At the electronmicroscopical level evidence was found that the inability to demonstrate macrophages could be due to the release of the lysosomal enzyme content and of parts of the cytoplasm.
在本研究中,我们报告了与乳胶颗粒偶联的商陆有丝分裂原(PWM)的制备及应用。这种偶联物(PWM-乳胶)是通过在戊二醛存在下将PWM与乳胶颗粒孵育制备而成。将PWM-乳胶添加到人外周血单个核细胞(PBMC)中的效果与单独添加PWM完全相当,即淋巴细胞分化为母细胞,随后这些母细胞增殖,这可通过DNA合成和总细胞数来衡量。电子显微镜观察发现,添加后24小时内单核细胞摄取了PWM-乳胶。虽然未观察到PWM-乳胶与淋巴细胞之间有直接相互作用,但发现后者分化为母细胞。由于这些母细胞与含乳胶的单核细胞相互作用,乳胶颗粒明显从吞噬细胞中释放出来,并且经常能看到乳胶颗粒与母细胞之间的紧密接触。此外,发现经抗T细胞单克隆抗体阳性反应判断为T细胞来源的母细胞摄取了乳胶颗粒。基于酶细胞化学、功能和光学显微镜研究,6天后在PWM-乳胶驱动的PBMC刺激中未检测到单核细胞。在电子显微镜水平上发现,无法证明存在巨噬细胞可能是由于溶酶体酶含量和部分细胞质的释放。