Immunogenicities of Env glycoproteins from circulating HIV-1 isolates in China focusing on the strategy of "DNA prime plus protein boost"
Immunogenicities of Env glycoproteins from circulating HIV-1 isolates in China focusing on the strategy of "DNA prime plus protein boost"
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DOI：
10.3760/cma.j.issn.0366-6999.2009.19.028
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发表时间：
2009-10-05
影响因子：
6.1
通讯作者：
Lu Shan
中科院分区：
文献类型：
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作者：
Wang Zheng;Wang Shi-xia;Lu Shan
2007年9月，默克公司开发的基于腺病毒的HIV-1疫苗意外失败。这使艾滋病毒疫苗开发战略发生了重大转变，重新将重点放在诱导中和抗体上。开发HIV-1疫苗的一个主要挑战是确定免疫原，并采用能够引发针对不同遗传亚型初级分离株的广泛中和抗体的递送方法。方法中国流行的HIV-1分离株主要由Thai-B、CRF_BC和CRF01_AE组成。为了构建针对这3种HIV-1亚型的DNA疫苗，从HIV-1分离株GX48（AE）、GX79（AE）、NX22（BC）、GS22（BC）、HN24（Thai-B）中构建了携带gp120区域的DNA疫苗。在瞬时转染的293T细胞中，用Western blotting检测这些DNA疫苗中gp120的表达。采用“DNA引物+蛋白增强”的方法对新西兰大白兔进行了预免疫，并在基于Tzm-bl细胞的实验中检测了对不同HIV-1毒株的中和抗体反应。结果DNA引物后，gp120特异性抗体反应较低（平均滴度=10(4.72)）；而gp120特异性抗体的滴度随着2次蛋白增强而升高（平均滴度=10(6.81)）。最重要的是，这种联合方法诱导的中和抗体（Nab）滴度比单独使用DNA或蛋白质诱导的抗体滴度要好得多(P
Background The adenovirus-based HIV-1 vaccine developed by Merck Company suffered from an unexpected failure in September 2007. This generated a big shift in the strategy of HIV vaccine development with renewed focus on the induction of neutralizing antibodies. A major challenge in developing an HIV-1 vaccine is to identify immunogens and adopt delivery methods that can elicit broadly neutralizing antibodies against primary isolates of different genetic subtypes.Methods Most circulating HIV-1 isolates in China are composed of clades Thai-B, CRF_BC and CRF01_AE. In order to construct DNA vaccines against these 3 HIV-1 subtypes, DNA vaccines carrying the gp120 regions from HIV-1 isolates of GX48(AE), GX79(AE), NX22(BC), GS22(BC), HN24(Thai-B) were constructed. Expression of gp120 from these DNA vaccines was detected by Western blotting in transiently transfected 293T cells. Pilot immunizations of New Zealand white rabbits were performed using the strategy of "DNA prime plus protein boost" and the neutralizing antibody response was detected in a Tzm-bl cell based assay against different HIV-1 strains.Results Response of gp120-specific antibody was relatively low after DNA primes (mean titer=10(4.72)); however, the titer of gp120-specific antibody went up with 2 protein boosts (mean titer=10(6.81)). Above all, neutralizing antibody (Nab) titers induced by this combined approach were much better than those elicited by DNA or protein used alone (P