Robust vaginal colonization of macaques with a novel vaginally disintegrating tablet containing a live biotherapeutic product to prevent HIV infection in women.
Robust vaginal colonization of macaques with a novel vaginally disintegrating tablet containing a live biotherapeutic product to prevent HIV infection in women.
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DOI：
10.1371/journal.pone.0122730
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Parks TP
中科院分区：
文献类型：
--
作者：
Lagenaur LA;Swedek I;Lee PP;Parks TP
MucoCept是一种用于预防女性HIV-1感染的生物疗法，它含有一种人类阴道延seni乳酸杆菌，该乳酸杆菌经过基因增强，可表达HIV-1进入抑制剂修饰的氰毒蛋白n （mCV-N）。本研究的目的是开发一种固体阴道剂型，支持MucoCept乳酸菌在先前显示的水平上持续阴道定植，使用新鲜制备的培养物，以保护猕猴免受SHIV感染，并在猕猴阴道定植模型中测试该制剂。将配制好的细菌在片剂型模具中冻干，然后用干燥剂包装在箔袋中，制成阴道崩解片。考察了崩解时间、药效和稳定性。为了进行定植，研究人员在五天内给非同步猕猴阴道服用一片或五片药片。分别在给药后3天、14天和21天采集阴道样本，并进行培养以测定乳酸菌定植水平。为了确认MucoCept乳酸菌菌株的身份，在第14天和第21天从样品中提取基因组DNA，并进行菌株特异性PCR。用Western blot检测细菌上清液中mCV-N蛋白的存在。该片剂易操作，2分钟崩解，药效（5.7 × 1011 CFU/g），在4℃和25℃下稳定。阴道给药后，66%的猕猴在给药后14天和83%的猕猴在给药后21天定植了MucoCept乳杆菌。1片和5片给药方案的定植水平无显著差异（第14天p=0.88，第21天p=0.99）。菌株特异性PCR证实，即使在培养阴性的猕猴中也存在这种细菌。最后，使用特异性抗mCV-N抗体，通过Western blot分析证实mCV-N蛋白的存在。
MucoCept is a biotherapeutic for prevention of HIV-1 infection in women and contains a human, vaginal Lactobacillus jensenii that has been genetically enhanced to express the HIV-1 entry inhibitor, modified cyanovirin-N (mCV-N). The objective of this study was to develop a solid vaginal dosage form that supports sustained vaginal colonization of the MucoCept Lactobacillus at levels previously shown, with freshly prepared cultures, to protect macaques from SHIV infection and to test this formulation in a macaque vaginal colonization model. Vaginally disintegrating tablets were prepared by lyophilizing the formulated bacteria in tablet-shaped molds, then packaging in foil pouches with desiccant. Disintegration time, potency and stability of the tablets were assessed. For colonization, non-synchronized macaques were dosed vaginally with either one tablet or five tablets delivered over five days. Vaginal samples were obtained at three, 14, and 21 days post-dosing and cultured to determine Lactobacillus colonization levels. To confirm identity of the MucoCept Lactobacillus strain, genomic DNA was extracted from samples on days 14 and 21 and a strain-specific PCR was performed. Supernatants from bacteria were tested for the presence of the mCV-N protein by Western blot. The tablets were easy to handle, disintegrated within two minutes, potent (5.7x1011 CFU/g), and stable at 4°C and 25°C. Vaginal administration of the tablets to macaques resulted in colonization of the MucoCept Lactobacillus in 66% of macaques at 14 days post-dosing and 83% after 21 days. There was no significant difference in colonization levels for the one or five tablet dosing regimens (p=0.88 Day 14, p=0.99 Day 21). Strain-specific PCR confirmed the presence of the bacteria even in culture-negative macaques. Finally, the presence of mCV-N protein was confirmed by Western blot analysis using a specific anti-mCV-N antibody.
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影响因子：
2.4
作者：
Gengiah, Tanuja N.;Moosa, Atika;Naidoo, Anushka;Mansoor, Leila E.
通讯作者：
Mansoor, Leila E.
影响因子：
3.7
作者：
Benning L;Golub ET;Anastos K;French AL;Cohen M;Gilbert D;Gillevet P;Munyazesa E;Landay AL;Sikaroodi M;Spear GT
通讯作者：
Spear GT
影响因子：
4.9
作者：
Liu, Xiaowen;Lagenaur, Laurel A.;Xu, Qiang
通讯作者：
Xu, Qiang
影响因子：
3.7
作者：
Brichacek, Beda;Lagenaur, Laurel A.;Hamer, Dean H.
通讯作者：
Hamer, Dean H.
DOI：
10.4103/2231-4040.90885
发表时间：
2011-10-01
影响因子：
1.4
作者：
Basu, Biswajit;Bagadiya, Abhishek;Dharamsi, Abhay
通讯作者：
Dharamsi, Abhay