Bartal A H, Feit C, Hirshaut Y
Nat Immun Cell Growth Regul. 1986;5(2):107-12.
In an effort to determine the effect of dexamethasone on hybridoma formation, spleen cells from BALB/c mice hyperimmunized with sheep red blood cells (SRBC) were fused with mouse plasmacytoma cells (P3U1) in the presence of polyethylene glycol (PEG). Dexamethasone was added in decreasing doses (10(-3) to 10(-9) mM) to the hypoxanthine-aminopterin-thymide (HAT) medium immediately after the PEG-mediated cell fusion. 10(-3) mM of this steroid was found to inhibit markedly the number and size of hybridoma clones generated, while 10(-5) mM dexamethasone was shown to enhance hybridoma formation. The effect of 10(-3) mM dexamethasone was most pronounced when added immediately after fusion. When this dose was given 48 or 120 h after cell fusion, the extent of the inhibitory effect was less pronounced. High concentration of dexamethasone may also inhibit monoclonal antibody production by hybridomas once generated. An increase in the number of clones formed was observed when 10(-5) mM dexamethasone was added to HAT medium as well as an increase in the average colony size. Large clones were also observed with lower dexamethasone doses ranging from 10(-7) to 10(-9) mM. Possible mechanisms on the effect of dexamethasone on hybridoma formation are discussed.
为了确定地塞米松对杂交瘤形成的影响,将用绵羊红细胞(SRBC)高度免疫的BALB/c小鼠的脾细胞与小鼠浆细胞瘤细胞(P3U1)在聚乙二醇(PEG)存在的情况下进行融合。在PEG介导的细胞融合后,立即将地塞米松以递减剂量(10^(-3)至10^(-9) mM)添加到次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷(HAT)培养基中。发现10^(-3) mM的这种类固醇显著抑制产生的杂交瘤克隆的数量和大小,而10^(-5) mM地塞米松则显示可增强杂交瘤的形成。10^(-3) mM地塞米松在融合后立即添加时效果最为明显。当在细胞融合后48或120小时给予该剂量时,抑制作用的程度不太明显。高浓度的地塞米松一旦产生,也可能抑制杂交瘤产生单克隆抗体。当将10^(-5) mM地塞米松添加到HAT培养基中时,观察到形成的克隆数量增加,并且平均集落大小也增加。在10^(-7)至10^(-9) mM的较低地塞米松剂量下也观察到了大克隆。讨论了地塞米松对杂交瘤形成影响的可能机制。