利用包膜和GAG基因产物制备基于iscom技术的HIV实验疫苗。

Biotechnology therapeutics Pub Date : 1993-01-01
L Akerblom, P Nara, N Dunlop, S Putney, B Morein
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引用次数: 0

摘要

在之前的实验中,将gp160掺入iscom可以诱导对HIV-1同源和异源分离株的中和抗体(Akerblom et al., AIDS Res., 1991)。在目前的工作中,我们将HIV-1的三种重组DNA产品纳入iscoms。gp120的羧基末端部分在大肠杆菌- pb -1中表达;含有大肠杆菌gag中表达的p24和p15部分的嵌合体;杆状病毒gp160在杆状病毒中克隆并在昆虫细胞中产生。iscom制剂对同源抗原、确定的重组产物和含有中和表位的合成肽RP135 (aa 304-328)诱导免疫应答。用PB1-iscoms和gp160 (baculo) iscoms免疫小鼠血清进行合胞抑制试验。PB1 iscoms免疫小鼠血清与合成肽RP135反应强烈,并能中和同源分离HIV-1/IIIB,中和效价为1/64。检测了3种gp160 (baculo) iscom抗血清,其中2种与合成肽RP135反应强烈,但不能中和同源分离HIV-1/IIIB。用2微克的gp160对同源抗原和覆盖部分gp41的重组DNA大肠杆菌构建体p121诱导小鼠高血清滴度。两次免疫后抗体反应达到上限。PB1-和GAG-iscoms需要三次免疫才能达到抗体应答的上限。
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HIV experimental vaccines based on the iscom technology using envelope and GAG gene products.

In previous experiments gp160 incorporated into iscom was shown to induce neutralizing antibodies to the homologous as well as the heterologous isolates of HIV-1 (Akerblom et al., AIDS Res., 1991). In the present work we have incorporated into iscoms three defined recombinant DNA products of HIV-1. The carboxy-terminal part of gp120 expressed in E. Coli-PB-1; a chimera containing parts of both p24 and p15 expressed in E. coli-GAG; and baculovirus gp160 cloned in baculovirus and produced in insect cells. Immune responses were induced by the iscom preparations to the homologous antigen as well as to defined recombinant products and to the synthetic peptide RP135 (aa 304-328) harboring a neutralizing epitope. Sera from mice immunized with PB1-iscoms and gp160 (baculo) iscoms were tested in a syncytie inhibition assay. The serum from a mouse immunized with PB1 iscoms reacted strongly with the synthetic peptide RP135 and also neutralized the homologous isolate HIV-1/IIIB with a neutralization titer of 1/64. Three gp160 (baculo) iscom antisera were tested, of which two reacted strongly with the synthetic peptide RP135 but did not neutralize the homologous isolate HIV-1/IIIB. High serum titers were induced in mice by the gp160 iscoms (2 micrograms) to homologous antigen and the recombinant DNA E. coli construct p121 covering part of gp41. The ceilings of the antibody responses were reached after two immunizations. The PB1- and GAG-iscoms required three immunizations to reach the ceiling of the antibody response.

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