DNA encoding an HIV-1 Gag/human lysosome-associated membrane protein-1 chimera elicits a broad cellular and humoral immune response in Rhesus macaques.
DNA encoding an HIV-1 Gag/human lysosome-associated membrane protein-1 chimera elicits a broad cellular and humoral immune response in Rhesus macaques.
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DOI：
10.1371/journal.pone.0000135
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发表时间：
2006-12-27
期刊：
影响因子：
3.7
通讯作者：
Marques ET
中科院分区：
文献类型：
--
作者：
Chikhlikar P;Barros de Arruda L;Maciel M;Silvera P;Lewis MG;August JT;Marques ET
先前的小鼠HIV-1 p55 Gag免疫的研究已经证明了通过使用编码Gag的DNA抗原制剂作为与小鼠溶酶体相关膜蛋白（mLAMP/gag）的嵌合体，将抗原靶向含有主要组织相容性复合体II型（MHC II）复合物分子的细胞区室的有用性。在本研究中，我们分析了恒河猴免疫编码人LAMP/gag（hLAMP/gag）嵌合体的DNA后引起的GAG特异性T淋巴细胞和抗体反应的幅度和广度。ELISPOT分析表明，由hLAMP/gag嵌合体引发的平均Gag特异性IFN-γ应答在所有五只免疫的猕猴中仅在两次或三次裸DNA免疫后就可检测到，并且达到平均1000个斑点形成细胞（SFC）/106个PBMC。在CD 8+耗尽的细胞中检测到高IFN-γ ELISPOT应答，表明CD 4 + T细胞在这些应答中起主要作用。还通过使用ELISPOT测试了四只猕猴对12个重叠的15个氨基酸（aa）肽池的T细胞应答，每个肽池含有10个肽，包括完整的Gag蛋白序列。两个Mamu 08免疫的猕猴对8个和12个库有应答，Mamu B 01对6个库有应答，而另一个猕猴对5个库有应答，这表明hLAMP/gag DNA抗原制剂激发了针对Gag的广泛T细胞应答。此外，有强烈的HIV-1特异性IgG应答。每次DNA注射后IgG抗体滴度增加，表明强烈的遗忘B细胞反应，并且在三次免疫后在所有猕猴中高度升高。此外，每只猕猴的血清识别覆盖完整Gag氨基酸序列的20-aa肽文库的49种肽中的13种。此外，HIV-1特异性伊加抗体存在于血浆和外部分泌物中，包括鼻洗液。这些数据支持编码为LAMP嵌合体的基因疫苗的免疫原性增加的发现，包括非人灵长类动物对DNA疫苗的应答。
Previous studies of HIV-1 p55Gag immunization of mice have demonstrated the usefulness of targeting antigens to the cellular compartment containing the major histocompatibility complex type II (MHC II) complex molecules by use of a DNA antigen formulation encoding Gag as a chimera with the mouse lysosome-associated membrane protein (mLAMP/gag). In the present study, we have analyzed the magnitude and breadth of Gag-specific T-lymphocyte and antibody responses elicited in Rhesus macaques after immunization with DNA encoding a human LAMP/gag (hLAMP/gag) chimera. ELISPOT analyses indicated that the average Gag-specific IFN-γ response elicited by the hLAMP/gag chimera was detectable after only two or three naked DNA immunizations in all five immunized macaques and reached an average of 1000 spot-forming cells (SFC)/106 PBMCs. High IFN-γ ELISPOT responses were detected in CD8+-depleted cells, indicating that CD4+ T-cells play a major role in these responses. The T-cell responses of four of the macaques were also tested by use of ELISPOT to 12 overlapping 15-amino acids (aa) peptide pools containing ten peptides each, encompassing the complete Gag protein sequence. The two Mamu 08 immunized macaques responded to eight and twelve of the pools, the Mamu B01 to six, and the other macaque to five pools indicating that the hLAMP/gag DNA antigen formulation elicits a broad T-cell response against Gag. Additionally, there was a strong HIV-1-specific IgG response. The IgG antibody titers increased after each DNA injection, indicating a strong amnestic B-cell response, and were highly elevated in all the macaques after three immunizations. Moreover, the serum of each macaque recognized 13 of the 49 peptides of a 20-aa peptide library covering the complete Gag amino acid sequence. In addition, HIV-1-specific IgA antibodies were present in the plasma and external secretions, including nasal washes. These data support the findings of increased immunogenicity of genetic vaccines encoded as LAMP chimeras, including the response to DNA vaccines by non-human primates.
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影响因子：
4.4
作者：
Arruda, Luciana B.;Sim, Del;Marques, Ernesto T. A.
通讯作者：
Marques, Ernesto T. A.
DOI：
10.1073/pnas.93.24.14082
发表时间：
1996-11-26
影响因子：
11.1
作者：
Kessler, PD;Podsakoff, GM;Byrne, BJ
通讯作者：
Byrne, BJ
影响因子：
5.4
作者：
Casimiro, DR;Tang, AM;Shiver, JW
通讯作者：
Shiver, JW
影响因子：
3.7
作者：
Anwar, A;Chandrasekaran, A;August, JT
通讯作者：
August, JT
影响因子：
6.4
作者：
De Arruda, LB;Chikhlikar, PR;Marques, ETA
通讯作者：
Marques, ETA