Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
Glycoform and net charge heterogeneity in gp120 immunogens used in HIV vaccine trials.
复制标题
DOI：
10.1371/journal.pone.0043903
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Berman PW
中科院分区：
文献类型：
--
作者：
Yu B;Morales JF;O'Rourke SM;Tatsuno GP;Berman PW
RV 144临床试验首次表明，疫苗接种可以提供适度但重要的保护，免受HIV-1感染。为了了解保护性应答，并提高疫苗的效力，重要的是要确定在初免/加强方案中使用的免疫原的结构。在这里，我们研究了AIDSVAX B/E疫苗中所含gp 120免疫原的净电荷异质性，这归因于糖型变异。等电聚焦和糖苷酶消化用于评估RV 144试验中使用的AIDSVAX B/E疫苗中所含gp 120净电荷的变化。我们观察到MN-rgp 120的16个变体和A244-rgp 120的24个变体。将中国仓鼠卵巢细胞中产生的gp 120的糖型变异与293 F人胚肾细胞系中产生的gp 120的糖型变异进行比较，后者通常用于中和试验。我们发现CHO细胞中产生的gp 120变体明显比293细胞中产生的gp 120变体酸性更强。使用单克隆抗体评估糖型异质性对抗原性的影响。无论是在CHO中还是在293中产生，广泛中和的PG 9 MAb与A244-rgp 120结合，但不与MN-rgp 120结合。然而，PG 9能够以高亲和力结合在N-乙酰葡糖胺基转移酶I缺陷的293细胞中产生的MN-rgp 120和A244-rgp 120。RV 144试验中使用的MN-和A244-rgp 120由于含唾液酸糖型的变化而在净电荷方面表现出广泛的异质性。这些差异是细胞系依赖性的，影响重组包膜蛋白的抗原性，并可能影响用于测量中和的测定。这些研究，以及最近的报告记录广泛中和抗体针对糖蛋白120的碳水化合物表位，表明糖型变异是一个关键变量，要考虑在生产和评估亚单位疫苗，旨在预防艾滋病毒感染。
The RV144 clinical trial showed for the first time that vaccination could provide modest but significant protection from HIV-1 infection. To understand the protective response, and to improve upon the vaccine's efficacy, it is important to define the structure of the immunogens used in the prime/boost regimen. Here we examined the heterogeneity in net charge, attributable to glycoform variation, of the gp120 immunogens contained in the AIDSVAX B/E vaccine. Isoelectric focusing and glycosidase digestion were used to assess variation in net charge of the gp120s contained in the AIDSVAX B/E vaccine used in the RV144 trial. We observed 16 variants of MN-rgp120 and 24 variants of A244-rgp120. Glycoform variation in gp120 produced in Chinese hamster ovary cells was compared to glycoform variation in gp120 produced in the 293F human embryonic kidney cell line, often used for neutralization assays. We found that gp120 variants produced in CHO cells were distinctly more acidic than gp120 variants produced in 293 cells. The effect of glycoform heterogeneity on antigenicity was assessed using monoclonal antibodies. The broadly neutralizing PG9 MAb bound to A244-rgp120, but not to MN-rgp120, whether produced in CHO or in 293. However, PG9 was able to bind with high affinity to MN-rgp120 and A244-rgp120 produced in 293 cells deficient in N-acetylglucosaminyltransferase I. MN- and A244-rgp120 used in the RV144 trial exhibited extensive heterogeneity in net charge due to variation in sialic acid-containing glycoforms. These differences were cell line-dependent, affected the antigenicity of recombinant envelope proteins, and may affect assays used to measure neutralization. These studies, together with recent reports documenting broadly neutralizing antibodies directed against carbohydrate epitopes of gp120, suggest that glycoform variation is a key variable to be considered in the production and evaluation of subunit vaccines designed to prevent HIV infection.
登录
查看更多内容
影响因子：
3.7
作者：
Eggink, Dirk;Melchers, Mark;Sanders, Rogier W.
通讯作者：
Sanders, Rogier W.
影响因子：
3.7
作者：
Kang, SM;Quan, FS;Compans, RW
通讯作者：
Compans, RW
HIV-1 疫苗功效试验的免疫相关分析。
DOI：
10.1056/nejmoa1113425
发表时间：
2012-04-05
期刊：
The New England journal of medicine
影响因子：
--
作者：
Haynes BF;Gilbert PB;McElrath MJ;Zolla-Pazner S;Tomaras GD;Alam SM;Evans DT;Montefiori DC;Karnasuta C;Sutthent R;Liao HX;DeVico AL;Lewis GK;Williams C;Pinter A;Fong Y;Janes H;DeCamp A;Huang Y;Rao M;Billings E;Karasavvas N;Robb ML;Ngauy V;de Souza MS;Paris R;Ferrari G;Bailer RT;Soderberg KA;Andrews C;Berman PW;Frahm N;De Rosa SC;Alpert MD;Yates NL;Shen X;Koup RA;Pitisuttithum P;Kaewkungwal J;Nitayaphan S;Rerks-Ngarm S;Michael NL;Kim JH
通讯作者：
Kim JH
影响因子：
56.9
作者：
LASKY, LA;GROOPMAN, JE;BERMAN, PW
通讯作者：
BERMAN, PW
影响因子：
3.7
作者：
Berman, PW;Huang, W;Gregory, TJ
通讯作者：
Gregory, TJ