INHIBITION OF HUMAN-IMMUNODEFICIENCY-VIRUS REPLICATION IN A HUMAN T-CELL LINE BY ANTISENSE RNA EXPRESSED IN THE CELL

被引:15
作者
GYOTOKU, JI
ELFARRASH, MA
FUJIMOTO, S
GERMERAAD, WTV
WATANABE, Y
TESHIGAWARA, K
HARADA, S
KATSURA, Y
机构
[1] KYOTO UNIV, CHEST DIS RES INST, DEPT IMMUNOL, KYOTO 606, JAPAN
[2] KUMAMOTO UNIV, FAC MED, DEPT MED VIROL, KUMAMOTO 860, JAPAN
[3] KYOTO UNIV, FAC PHARMACEUT SCI, DEPT MOLEC MICROBIOL, KYOTO 606, JAPAN
关键词
HUMAN IMMUNODEFICIENCY VIRUS; ANTISENSE RNA; GENE TRANSDUCTION; RETROVIRAL VECTOR; HEMATOPOIETIC STEM-CELLS; ANTI-SENSE RNA; GENE-EXPRESSION; MESSENGER-RNA; HTLV-III; AIDS; RETROVIRUS; TYPE-1; HIV-1; COMPLEMENTARY;
D O I
10.1007/BF00568969
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The effect of the expression of antisense RNA against the human immunodeficiency virus (HIV) genome in a human T-cell line CEM on HIV replication was investigated. A 2.7 kilobase (kb) fragment of the HIV genome, including tat and a part of rev and env, was cloned into the retroviral vector WB in the antisense orientation under the SV40 or H-2K promoter. CEM cells transduced with this antisense gene via recombinant retrovirus expressed the RNAs of three different molecular sizes containing the antisense construct. CEM cells and these transduced cells were infected with HIV. HIV replication was evaluated 4-10 days later by an immunofluorescense assay and by determining the reverse transcriptase activity in the culture supernatant. The results indicate that although the recombinant retrovirus WB strongly enhanced the HIV replication in CEM cells. the expression of antisense RNA in the cells was highly effective in impeding the replication of HIV. The inhibitory effect was especially high in CEM cells transduced with the antisense gene under the control of SV40 promoter. In this case, HIV antigen-positive cells and reverse transcriptase activity in the culture supernatant of transduced cells were reduced to 30-50% and 5-10% of those in CEM cells and in the CEM cells transduced with WB, respectively.
引用
收藏
页码:189 / 202
页数:14
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