Chimpanzee Fab fragments and a derived humanized immunoglobulin G1 antibody that efficiently cross-neutralize dengue type 1 and type 2 viruses
Chimpanzee Fab fragments and a derived humanized immunoglobulin G1 antibody that efficiently cross-neutralize dengue type 1 and type 2 viruses
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DOI：
10.1128/jvi.78.23.12910-12918.2004
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发表时间：
2004-12-01
影响因子：
5.4
通讯作者：
Lai, CJ
中科院分区：
文献类型：
--
作者：
Goncalvez, AP;Men, R;Lai, CJ
使用来自人类或非人灵长类动物的单克隆抗体进行被动免疫，是预防登革病毒（DENV）和其他黄病毒（包括西尼罗河病毒）所致疾病的一种有吸引力的疫苗替代方法。在先前的一项研究中，描述了从感染所有四种登革病毒血清型（登革病毒血清型1[DENV - 1]至DENV - 4）的黑猩猩骨髓mRNA中进行抗体库克隆以回收Fab片段。在该研究中，回收并鉴定了一种能有效中和DENV - 4的人源化免疫球蛋白G1（IgG1）抗体。在本研究中，利用从黑猩猩构建的噬菌体文库来回收针对其他三种登革病毒血清型的Fab抗体。未鉴定出血清型特异性中和Fab。相反，我们回收了能特异性沉淀包膜蛋白且对所有四种登革病毒血清型都有交叉反应的登革病毒中和Fab。其中三种Fab在与DENV - 1和DENV - 2结合时相互竞争，尽管这些Fab各自包含不同的互补决定区3（CDR3）-H序列。具有相同或几乎相同CDR3 - H序列的Fab以相似的高50%蚀斑减少中和试验（PRNT50）滴度交叉中和DENV - 1和DENV - 2，滴度范围从0.26到1.33微克/毫升，并且能中和DENV - 3和DENV - 4，但滴度低10到20倍。其中一种Fab，1A5，在测试的其他黄病毒中也能最有效地中和西尼罗河病毒。Fab 1A5与人类序列结合转化为全长抗体，以便在哺乳动物CHO细胞中生产。人源化IgG1 1A5在分别以0.48和0.95微克/毫升的滴度交叉中和DENV - 1和DENV - 2时，被证明与Fab 1A5一样有效。IgG1 1A5还以约3.2到4.2微克/毫升的PRNT50滴度中和DENV - 3、DENV - 4和西尼罗河病毒。这种人源化抗体是进一步开发针对登革病毒以及可能针对其他黄病毒相关疾病的免疫预防的有吸引力的候选者。
Passive immunization with monoclonal antibodies from humans or nonhuman primates represents an attractive alternative to vaccines for prevention of illness caused by dengue viruses (DENV) and other flaviviruses, including the West Nile virus. In a previous study, repertoire cloning to recover Fab fragments from bone marrow mRNA of chimpanzees infected with all four DENV serotypes (dengue virus serotype 1 [DENV-1] to DENV-4) was described. In that study, a humanized immunoglobulin G1 (IgG1) antibody that efficiently neutralized DENV-4 was recovered and characterized. In this study, the phage library constructed from the chimpanzees was used to recover Fab antibodies against the other three DENV serotypes. Serotypespecific neutralizing Fabs were not identified. Instead, we recovered DENV-neutralizing Fabs that specifically precipitated the envelope protein and were cross-reactive with all four DENV serotypes. Three of the Fabs competed with each other for binding to DENV-1 and DENV-2, although each of these Fabs contained a distinct complementarity determining region 3 (CDR3)-H sequence. Fabs that shared an identical or nearly identical CDR3-H sequences cross-neutralized DENV-1 and DENV-2 at a similar high 50% plaque reduction neutralization test (PRNT50) titer, ranging from 0.26 to 1.33 mu/ml, and neutralized DENV-3 and DENV-4 but at a titer 10- to 20-fold lower. One of these Fabs, 1A5, also neutralized the West Nile virus most efficiently among other flaviviruses tested. Fab 1A5 was converted to a full-length antibody in combination with human sequences for production in mammalian CHO cells. Humanized IgG1 1A5 proved to be as efficient as Fab 1A5 for cross-neutralization of DENV-1 and DENV-2 at a titer of 0.48 and 0.95 mug/ml, respectively. IgG1 1A5 also neutralized DENV-3, DENV-4, and the West Nile virus at a PRNT50 titer of approximately 3.2 to 4.2 mug/ml. This humanized antibody represents an attractive candidate for further development of immunoprophylaxis against DENV and perhaps other flavivirus-associated diseases.