Abacioglu Y H, Fouts T R, Laman J D, Claassen E, Pincus S H, Moore J P, Roby C A, Kamin-Lewis R, Lewis G K
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore 21201.
AIDS Res Hum Retroviruses. 1994 Apr;10(4):371-81. doi: 10.1089/aid.1994.10.371.
To define protein folding patterns of HIV-1 Env subunit vaccines, we have isolated a set of 30 monoclonal antibodies (MAbs) from BALB/c mice immunized with a recombinant gp160 vaccine (rgp160) expressed in a baculovirus system. This article describes epitope mapping for the MAb panel and topology of the epitopes for rgp160 and a recombinant gp120 (rgp120) also expressed in a baculovirus system. The following results are reported: (1) rgp160 harbors a minimum of 4 antigenic domains, 3 mapping to the C1, C2, and C3/V4 regions of gp120 and 1 mapping to the cytoplasmic tail of gp41; (2) there are at least 3 adjacent or overlapping epitopes in each antigenic domain; (3) a minimum of 14 independent epitopes were mapped, all of which are continuous sites; (4) each of the epitopes is exposed on rgp160 without prior manipulation of the protein; and (5) by contrast, 6 of the 8 epitopes mapping to the C1, C2, and C3/V4 regions are not exposed on rgp120, but become exposed when the protein is denatured. Taken together, these results show that rgp160 and rgp120 are folded differently, illustrating the use of this MAb panel to compare epitope topographies of recombination HIV-1 Env proteins. This MAb panel may aid in the refinement of HIV-1 Env subunit vaccines.
为了确定HIV-1包膜亚基疫苗的蛋白质折叠模式，我们从用杆状病毒系统表达的重组gp160疫苗（rgp160）免疫的BALB/c小鼠中分离出一组30种单克隆抗体（MAb）。本文描述了该单克隆抗体组的表位作图以及rgp160和同样在杆状病毒系统中表达的重组gp120（rgp120）的表位拓扑结构。报告了以下结果：（1）rgp160至少含有4个抗原结构域，3个定位于gp120的C1、C2和C3/V4区域，1个定位于gp41的胞质尾；（2）每个抗原结构域中至少有3个相邻或重叠的表位；（3）至少定位了14个独立表位，所有这些表位都是连续位点；（4）每个表位在未经预先处理的rgp160上都是暴露的；（5）相比之下，定位于C1、C2和C3/V4区域的8个表位中的6个在rgp120上不暴露，但在蛋白质变性时会暴露。综上所述，这些结果表明rgp160和rgp120的折叠方式不同，说明了利用该单克隆抗体组比较重组HIV-1包膜蛋白的表位拓扑结构。该单克隆抗体组可能有助于优化HIV-1包膜亚基疫苗。
