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首页> 外文期刊>AIDS Research and Human Retroviruses >Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus
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Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus

机译：使用人类阴道乳杆菌表达人类免疫缺陷病毒1型中和抗体片段

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摘要

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, 3g/ml. All L. jensenii-expressed antibody fragments bound to gp120/CD4 complex; Lactobacillus-produced m36.4 inhibited HIV-1(BaL) 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.
机译：通过接种能产生广泛中和抗体的表位来消除人类1型免疫缺陷病毒（HIV-1）是HIV预防的最终目标。然而，证明产生适当的免疫应答是困难的。通过阴道定殖的乳杆菌表达广泛中和的抗体提供了一种将这些抗体被动地靶向粘膜的方法。我们测试了在乳杆菌中表达单链和单域抗体（dAbs）用作局部杀菌剂/活体生物治疗剂的可行性。乳杆菌提供了表达抗HIV蛋白的绝佳平台。已经鉴定出针对HIV-1包膜上的表位的广泛中和抗体，并已制成活性抗体片段。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。我们制备了表达dAb-m36.4的重组詹氏乳杆菌1153-1128。乳杆菌表达所有抗体片段m9，m36和m36.4。但是，我们注意到较小的m36 / m36.4被表达为更高的水平，即3g / ml。所有表达詹森氏乳杆菌的抗体片段均与gp120 / CD4复合物结合;在中和试验中，乳酸菌产生的m36.4抑制了HIV-1（BaL）。使用TZM-bl分析，我们表征了m36.4的中和广度。乳杆菌传递的dAb可以使这些抗体被动转移到粘膜，并在HIV-1传播的地方长寿。

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