Antiviral lectins as microbicides

作为杀微生物剂的抗病毒凝集素

基本信息

批准号：
8056628
负责人：
KENNETH E PALMER
金额：
$ 38.33万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-15 至 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8056628
关键词：
Agriculture Animals Antibodies Antiviral Agents Attention Binding Binding Sites Biological Biological Assay Cells Chlamydia trachomatis Clinical Collaborations Complex Data Data Set Development Dose Drug Kinetics Epithelial Epithelium Evaluation Exhibits Fermentation Gel Genetic Goals HIV HIV-1 Hemophilus Histology Histopathology Homologous Gene Human Human Herpesvirus 2 Human Papillomavirus Immune Immune response In Vitro Infection Inflammation Inflammatory Response Intravaginal Administration Investigation Lectin Leukocyte Chemotaxis Leukocytes Mannose Measurement Measures Modeling Molecular Mus Mutate Oryctolagus cuniculus Peptides Pharmaceutical Preparations Pharmacology and Toxicology Phenotype Placebo Control Plant Leaves Play Predisposition Prevention Production Proteins Recombinant Proteins Recombinants Recruitment Activity Research Resistance Risk Factors Role Safety Series Sexual Transmission Sexually Transmitted Diseases Simplexvirus Site-Directed Mutagenesis Solutions Structure Sum Surface System Testing Tissues Tobacco Toxic effect Toxicology Vaccines Vagina Vertebrates Virus base chemokine cost cytokine genital herpes hydroxyethylcellulose immunogenic immunogenicity in vivo inhibitor/antagonist insight irritation lymphocyte proliferation microbicide molecular marker mutant pathogen practical application prevent public health relevance rectal microbicide research study resistance mechanism response sugar transmission process vaginal microbicide

项目摘要

DESCRIPTION (provided by applicant): For practical application, microbicides must be available in massive amounts, and must be produced cost- effectively. These requirements might eliminate recombinant proteins from consideration as costs of production in cell-based fermentation systems are prohibitively high. We developed a practical solution by expressing and purifying recombinant forms of the algal lectin Griffithsin (GRFT), the most potent HIV-1 entry inhibitor yet described, from tobacco leaf tissues. GRFT has no known homologs in vertebrates, and hence has the potential to be immunogenic and thus possibly to induce an inflammatory response incompatible with its use as a microbicide. This is a theoretical concern that applies to most, if not all, peptide-based microbicides, and it is of vital importance to the field that this phenomenon be investigated more thoroughly. We will generate a mutant of GRFT that lacks lectin activity to allow us to answer the question whether lectin activity plays a role in induction of mucosal inflammatory responses. We will use a comprehensive mouse vaginal administration model, developed by our consultant and collaborator Dr. Betsy Herold, to evaluate the mucosal toxicity of hydroxyethylcellulose-formulated GRFT. Toxicology endpoints will include histology, measurement of molecular markers of inflammation, and phenotypes of leukocyte infiltrate in vaginal tissues. Since immunogenicity of protein microbicides is a concern, we will also measure immune responses to the vaginal microbicide in animals in which we have intentionally raised an anti-GRFT immune response. We will use Dr. Herold's elegant mouse genital herpes simplex virus (HSV) infectivity assay that functions as a surrogate measurement of microbicide safety. Susceptibility to challenge with HSV-2 in animals treated with GRFT will inform us whether the complex of biological responses to treatment with GRFT can increase susceptibility to infection with HSV-2, and hence to possible enhanced susceptibility to HIV-1 infection in humans. We will also study the mechanism of resistance to GRFT in a mutant HIV-1 Clade C virus that we isolated in a selection experiment. Collectively, the data that we generate in the course of this research will provide a comprehensive assessment of the utility of GRFT as a component of a vaginal microbicide to prevent HIV-1 transmission. PUBLIC HEALTH RELEVANCE: In the absence of an effective vaccine against HIV, there is an urgent need for alternative strategies to prevent sexual transmission of HIV, such as vaginally-applied microbicidal gels. We aim to answer the question whether proteins called lectins, which bind the sugar structures found on the surface of the AIDS virus, generate an inflammatory response when administered in the vagina of mice. These studies will provide important information that will help determine whether lectins have acceptable toxicity profiles to justify further development as microbicides.

描述（由申请人提供）：为了实际应用，杀菌剂必须大量供应，并且必须经济有效地生产。这些要求可能会排除重组蛋白的考虑，因为基于细胞的发酵系统的生产成本过高。我们通过从烟叶组织中表达和纯化藻类凝集素 Griffithsin (GRFT) 的重组形式开发了一种实用的解决方案，GRFT 是迄今为止描述的最有效的 HIV-1 进入抑制剂。 GRFT 在脊椎动物中没有已知的同源物，因此具有免疫原性的潜力，因此可能诱导与其作为杀微生物剂的用途不相容的炎症反应。这是一个理论上的问题，适用于大多数（如果不是全部）基于肽的杀微生物剂，并且更彻底地研究这种现象对于该领域至关重要。我们将产生缺乏凝集素活性的 GRFT 突变体，以便我们回答凝集素活性是否在诱导粘膜炎症反应中发挥作用的问题。我们将使用由我们的顾问和合作者 Betsy Herold 博士开发的综合小鼠阴道给药模型来评估羟乙基纤维素配制的 GRFT 的粘膜毒性。毒理学终点将包括组织学、炎症分子标记物的测量以及阴道组织中白细胞浸润的表型。由于蛋白质杀微生物剂的免疫原性是一个问题，我们还将测量我们有意提高抗 GRFT 免疫反应的动物对阴道杀微生物剂的免疫反应。我们将使用 Herold 博士的优雅的小鼠生殖器单纯疱疹病毒 (HSV) 感染性测定，该测定可作为杀菌剂安全性的替代测量。接受 GRFT 治疗的动物对 HSV-2 攻击的敏感性将告诉我们，对 GRFT 治疗的复杂生物反应是否会增加对 HSV-2 感染的易感性，从而可能增加人类对 HIV-1 感染的易感性。我们还将研究我们在选择实验中分离出的突变型 HIV-1 C 型病毒的 GRFT 抗性机制。总的来说，我们在本研究过程中生成的数据将对 GRFT 作为阴道杀微生物剂成分预防 HIV-1 传播的效用提供全面评估。公共卫生相关性：在缺乏有效的艾滋病毒疫苗的情况下，迫切需要采取替代策略来预防艾滋病毒的性传播，例如阴道使用的杀菌凝胶。我们的目的是回答这样一个问题：凝集素蛋白与艾滋病病毒表面的糖结构结合，在给小鼠阴道注射时是否会产生炎症反应。这些研究将提供重要信息，有助于确定凝集素是否具有可接受的毒性特征，以证明进一步开发为杀菌剂的合理性。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Studies in a Murine Model Confirm the Safety of Griffithsin and Advocate Its Further Development as a Microbicide Targeting HIV-1 and Other Enveloped Viruses.

DOI：
10.3390/v8110311
发表时间：
2016-11-17
期刊：
Viruses
影响因子：
0
作者：
Kouokam JC;Lasnik AB;Palmer KE
通讯作者：
Palmer KE

Characterization of the hypersensitive response-like cell death phenomenon induced by targeting antiviral lectin griffithsin to the secretory pathway.

DOI：
10.1111/pbi.12917
发表时间：
2018-10
期刊：
Plant biotechnology journal
影响因子：
13.8
作者：
Kim BM;Lotter-Stark HCT;Rybicki EP;Chikwamba RK;Palmer KE
通讯作者：
Palmer KE

Preformulation Characterization of Griffithsin, a Biopharmaceutical Candidate for HIV Prevention.

DOI：
10.1208/s12249-021-01931-0
发表时间：
2021-02-24
期刊：
AAPS PharmSciTech
影响因子：
3.3
作者：
Kramzer LF;Hamorsky KT;Graebing PW;Wang L;Fuqua JL;Matoba N;Lasnik AB;Moncla BJ;Zhang J;Palmer KE;Rohan LC
通讯作者：
Rohan LC

Bulk production of the antiviral lectin griffithsin.