Warncke Max, Vogt Birgit, Ulrich Jacqueline, von Laer Meike Dorothee, Beyer Winfried, Klump Hannes, Micheel Burkhard, Sheriff Ahmed
Genethor GmbH, Goltzstr. 52, 10781 Berlin, Germany.
Biochem Biophys Res Commun. 2004 Jun 4;318(3):673-9. doi: 10.1016/j.bbrc.2004.04.057.
The aim of this study was to determine the impact of lentiviral transduction on primary murine B cells. Studying B cell activities in vivo or using them for tolerance induction requires that the cells remain unaltered in their biological behavior except for expression of the transgene. As we show here, murine B cells can efficiently be transduced by lentiviral, VSV-G-pseudotyped vectors without the necessity of prior activation. Culture with LPS gave enhanced transduction efficiencies but led to the upregulation of CD86 and proliferation of the cells. Transduction of naive B cells by lentiviral vectors was dependent on multiplicity of infection and did not lead to a concomitant activation. Furthermore, the transduced cells could be used for studies in the NOD mouse system without altering the onset of diabetes. We conclude that lentiviral gene transfer into naive B cells is a powerful tool for manipulation of B cells for therapeutic applications.
本研究的目的是确定慢病毒转导对原代小鼠B细胞的影响。在体内研究B细胞活性或将其用于诱导耐受性要求细胞除转基因表达外,其生物学行为保持不变。正如我们在此所展示的,小鼠B细胞可被慢病毒、VSV-G假型载体有效转导,而无需预先激活。用LPS培养可提高转导效率,但会导致CD86上调和细胞增殖。慢病毒载体对幼稚B细胞的转导取决于感染复数,且不会导致伴随的激活。此外,转导后的细胞可用于非肥胖糖尿病(NOD)小鼠系统的研究,而不会改变糖尿病的发病时间。我们得出结论,将慢病毒基因导入幼稚B细胞是一种用于治疗应用中操纵B细胞的强大工具。