Zhong L, Granelli-Piperno A, Pope M, Ignatius R, Lewis M G, Frankel S S, Steinman R M
Laboratory of Cellular Physiology and Immunology, The Rockefeller University, New York, NY 10021-6399, USA.
Eur J Immunol. 2000 Nov;30(11):3281-90. doi: 10.1002/1521-4141(200011)30:11<3281::AID-IMMU3281>3.0.CO;2-4.
To pursue the capacity of monkey dendritic cells (DC) to be modified by adenoviral vectors and present the encoded antigens, we generated DC from blood monocytes and infected them with recombinant adenoviruses encoding GFP reporter and SIVgag or nef genes. Recombinant, E1- and E3-deleted, adenoviruses could transfect immature DC to >90% efficiency. When differentiated in the presence of a maturation stimulus, the infected cells were identical to control uninfected DC in surface markers and potent stimulatory activity for the mixed leukocyte reaction. Recombinant adeno-SIVgag was comparable to vaccinia-gag in stimulating IFN-gamma-secreting CD8(+) T cells from PBMC of macaques vaccinated with SIV(mac239) Deltanef and challenged with pathogenic SIV or chimeric SIV/HIV. Small numbers of adeno-SIVgag-infected DC were sufficient to trigger specific ELISPOT responses by CD8(+) T cells from these animals. Some CD4(+) IFN-gamma-secreting cells were also found in the three of eight vaccinated animals with the highest CD8(+) responses. T cells from control animals did not respond to DC transfected with adeno-gag. Therefore recombinant adenoviruses efficiently transfect monkey DC in a nonperturbing fashion, and these DC efficiently present antigens to SIVgag immune CD8(+) T cells. These findings will allow autologous DC, expressing SIV genes with high efficiency, to be tested in vivo to achieve strong specific T cell immunity.
为了探究猴树突状细胞(DC)被腺病毒载体修饰并呈递编码抗原的能力,我们从血液单核细胞中生成DC,并将其用编码绿色荧光蛋白(GFP)报告基因以及猴免疫缺陷病毒(SIV)gag或nef基因的重组腺病毒进行感染。重组的、缺失E1和E3的腺病毒能够以大于90%的效率转染未成熟DC。当在成熟刺激物存在的情况下进行分化时,被感染的细胞在表面标志物以及对混合淋巴细胞反应的强大刺激活性方面与未感染的对照DC相同。重组腺病毒-SIVgag在刺激来自接种了SIV(mac239)Deltanef并受到致病性SIV或嵌合SIV/艾滋病毒攻击的猕猴外周血单核细胞(PBMC)中分泌γ干扰素的CD8(+) T细胞方面与痘苗病毒-gag相当。少量被腺病毒-SIVgag感染的DC足以引发这些动物的CD8(+) T细胞产生特异性酶联免疫斑点(ELISPOT)反应。在八只接种动物中,有三只具有最高的CD8(+)反应,在这些动物中也发现了一些分泌γ干扰素的CD4(+)细胞。来自对照动物的T细胞对用腺病毒-gag转染的DC没有反应。因此,重组腺病毒能够以非干扰的方式有效地转染猴DC,并且这些DC能够有效地将抗原呈递给针对SIVgag免疫的CD8(+) T细胞。这些发现将使高效表达SIV基因的自体DC能够在体内进行测试,以实现强大的特异性T细胞免疫。