Gzyl Jaroslaw, Bolesta Elizabeth, Wierzbicki Andrew, Kmieciak Dariusz, Naito Toshio, Honda Mitsuo, Komuro Katsutoshi, Kaneko Yutaro, Kozbor Danuta
Department of Immunology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA.
Virology. 2004 Jan 20;318(2):493-506. doi: 10.1016/j.virol.2003.10.009.
Induction of cross-reactive cellular and humoral responses to the HIV-1 envelope (env) glycoprotein was examined after DNA immunization of BALB/c mice with gp140(89.6)-derived constructs exhibiting partial or complete deletions of the V1, V2, and V3 domains. It was demonstrated that specific modification of the V3 loop (mV3) in combination with the V2-modified (mV2) or V1/V2-deleted (DeltaV1/V2) region elicited increased levels of cross-reactive CD8(+) T cell responses. Mice immunized with the mV2/mV3 or DeltaV1/V2/mV3 gp140(89.6) plasmid DNA were greater than 50-fold more resistant to challenge with recombinant vaccinia virus (rVV) expressing heterologous env gene products than animals immunized with the wild-type (WT) counterpart. Sera from mV2/mV3- and DeltaV1/V2/mV3-immunized mice exhibited the highest cross-neutralizing activity and displayed intermediate antibody avidity values which were further enhanced by challenge with rVV expressing the homologous gp160 glycoprotein. In contrast, complete deletion of the variable regions had little or no effect on the cross-reactive antibody responses. The results of these experiments indicate that the breadth of antibody responses to the HIV-1 env glycoprotein may not be increased by removal of the variable domains. Instead, partial deletions within these regions may redirect specific responses toward conserved epitopes and facilitate approaches for boosting cross-reactive cellular and antibody responses to the env glycoprotein.
在用展示V1、V2和V3结构域部分或完全缺失的gp140(89.6)衍生构建体对BALB/c小鼠进行DNA免疫后,检测了对HIV-1包膜(env)糖蛋白的交叉反应性细胞和体液反应的诱导情况。结果表明,V3环(mV3)与V2修饰(mV2)或V1/V2缺失(DeltaV1/V2)区域的特异性修饰引发了更高水平的交叉反应性CD8(+) T细胞反应。用mV2/mV3或DeltaV1/V2/mV3 gp140(89.6)质粒DNA免疫的小鼠,比用野生型(WT)对应物免疫的动物对表达异源env基因产物的重组痘苗病毒(rVV)攻击的抵抗力高50倍以上。来自mV2/mV3和DeltaV1/V2/mV3免疫小鼠的血清表现出最高的交叉中和活性,并显示出中等抗体亲和力值,在用表达同源gp160糖蛋白的rVV攻击后,这些值进一步增强。相比之下,可变区的完全缺失对交叉反应性抗体反应几乎没有影响。这些实验结果表明,去除可变结构域可能不会增加对HIV-1 env糖蛋白的抗体反应广度。相反,这些区域内的部分缺失可能会使特异性反应转向保守表位,并有助于增强对env糖蛋白的交叉反应性细胞和抗体反应的方法。