Responses of pokeweed mitogen-stimulated peripheral mononuclear cells to human recombinant interleukins 3 and 4.

Lymphokine research Pub Date : 1990-01-01
M Kato, M Fujii, T Ito, K Kumagai
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Abstract

Peripheral blood mononuclear cells (PBMC) that had been stimulated with a polyclonal B cell activator, pokeweed mitogen (PWM), were examined for mitogenic responsiveness to interleukins (IL) and colony stimulating factors (CSF) or other cytokines using recombinant preparations from E. coli or Cos cells expressing their cloned cDNAs. PWM-stimulated PBMC (PWM-blasts) were found to be responsive to IL-3 depending upon the concentrations of the agent. Mitogenic response of PWM-blasts in the reaction to IL-3 was suppressed by the addition of a specific monoclonal antibody against IL-3 but not by anti-IL-2 serum, excluding the possibility that the DNA replication induced by IL-3 is a result of the proliferative response of IL-2 responder cells to IL-2, which might be produced during the culture of PWM-blasts in the presence of IL-3. The PWM-blasts were also responsive to IL-2 and IL-4 but not to other interleukins, namely, IL-1, IL-5 and IL-6, nor to granulocyte/macrophage (GM)-, granulocyte (G)- nor macrophage (M)-CSFs. They were also not responsive to interferon gamma. Similar responsiveness to IL-3 and IL-4 was found in the PBMC stimulated with a B-cell mitogen, Staphylococcus aureus Cowan 1 (Sac-1), but not in the PBMC stimulated with T cell mitogens, PHA or ConA. These results suggest that the human PBMC stimulated with B cell mitogens such as PWM or Sac-1 contain activated IL-3- and IL-4- responsive cells in sufficient number to detect these lymphokine activities.(ABSTRACT TRUNCATED AT 250 WORDS)

商陆有丝分裂原刺激的外周单核细胞对人重组白细胞介素3和4的反应。
用多克隆B细胞激活剂美洲商陆有丝分裂原(PWM)刺激外周血单核细胞(PBMC),用表达克隆cdna的大肠杆菌或Cos细胞重组制剂检测有丝分裂对白细胞介素(IL)和集落刺激因子(CSF)或其他细胞因子的反应性。pwm刺激的PBMC (pwm -blast)被发现对IL-3有反应,这取决于介质的浓度。加入针对IL-3的特异性单克隆抗体可抑制pwm母细胞对IL-3的有丝分裂反应,但抗IL-2血清不能抑制,排除了IL-3诱导的DNA复制是IL-2应答细胞对IL-2的增殖反应的结果,这种增殖反应可能是在IL-3存在的pwm母细胞培养过程中产生的。pwm母细胞对IL-2和IL-4也有反应,但对其他白细胞介素,即IL-1、IL-5和IL-6没有反应,对粒细胞/巨噬细胞(GM)-、粒细胞(G)-和巨噬细胞(M)- csf也没有反应。它们对干扰素也没有反应。在b细胞有丝分裂原金黄色葡萄球菌1 (sac1)刺激的PBMC中发现了对IL-3和IL-4的类似反应,而在T细胞有丝分裂原PHA或ConA刺激的PBMC中则没有。这些结果表明,受B细胞有丝分裂原(如PWM或Sac-1)刺激的人PBMC中含有足够数量的激活的IL-3和IL-4应答细胞,以检测这些淋巴因子的活性。(摘要删节250字)
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