Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus.
Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus.
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DOI：
10.1089/aid.2015.0378
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发表时间：
2016
影响因子：
1.5
通讯作者：
Lagenaur LA
中科院分区：
文献类型：
--
作者：
Marcobal A;Liu X;Zhang W;Dimitrov AS;Jia L;Lee PP;Fouts TR;Parks TP;Lagenaur LA
通过接种产生广泛中和抗体的表位来根除人类免疫缺陷病毒1型（HIV-1）是HIV预防的最终目标。然而，产生适当的免疫反应已被证明是困难的。通过阴道定殖乳酸杆菌表达广泛中和抗体提供了将这些抗体被动靶向粘膜的方法。我们测试了在乳杆菌中表达单链和单结构域抗体（dAb）以用作局部杀微生物剂/活生物素的可行性。乳酸杆菌为表达抗HIV蛋白提供了极好的平台。已经鉴定了针对HIV-1包膜上的表位的广泛中和抗体，并将其制成活性抗体片段。我们测试了单链可变片段m9和dAb-m36及其衍生物m36.4作为原型抗体。我们在詹氏乳杆菌-1153中克隆并表达了抗体片段m9、m36和m36.4，并测试了表达水平和功能性。我们构建了重组L.表达dAb-m36的jensenii 1153-1128。所有抗体片段m9、m36和m36.4均由乳杆菌表达。然而，我们注意到较小的m36/m36.4表达水平较高，≥3 μg/ml。阿尔湖jensenii表达的抗体片段与gp 120/CD 4复合物结合;乳酸杆菌产生的m36.4在中和试验中抑制HIV-1BaL。使用TZM-bl测定，我们表征了m36.4的中和宽度。通过乳酸杆菌递送dAb可以将这些抗体被动转移到粘膜，并在HIV-1传播部位延长寿命。
Eradication of human immunodeficiency virus type 1 (HIV-1) by vaccination with epitopes that produce broadly neutralizing antibodies is the ultimate goal for HIV prevention. However, generating appropriate immune responses has proven difficult. Expression of broadly neutralizing antibodies by vaginal colonizing lactobacilli provides an approach to passively target these antibodies to the mucosa. We tested the feasibility of expressing single-chain and single-domain antibodies (dAbs) in Lactobacillus to be used as a topical microbicide/live biotherapeutic. Lactobacilli provide an excellent platform to express anti-HIV proteins. Broadly neutralizing antibodies have been identified against epitopes on the HIV-1 envelope and have been made into active antibody fragments. We tested single-chain variable fragment m9 and dAb-m36 and its derivative m36.4 as prototype antibodies. We cloned and expressed the antibody fragments m9, m36, and m36.4 in Lactobacillus jensenii-1153 and tested the expression levels and functionality. We made a recombinant L. jensenii 1153-1128 that expresses dAb-m36.4. All antibody fragments m9, m36, and m36.4 were expressed by lactobacilli. However, we noted the smaller m36/m36.4 were expressed to higher levels, ≥3 μg/ml. All L. jensenii-expressed antibody fragments bound to gp120/CD4 complex; Lactobacillus-produced m36.4 inhibited HIV-1BaL in a neutralization assay. Using a TZM-bl assay, we characterized the breadth of neutralization of the m36.4. Delivery of dAbs by Lactobacillus could provide passive transfer of these antibodies to the mucosa and longevity at the site of HIV-1 transmission.