Development of peptide-based vaccine against SARS-CoV

开发针对 SARS-CoV 的肽疫苗

• 批准号: 7413336
• 负责人: GANG LIU
• 金额: $ 33.08万
• 依托单位: INSTITUTE OF MATERIA MEDICA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7413336
• 关键词: Achievement; Amino Acid Sequence; Amino Acids; Animal Model; Animals; Antibodies; Antigens; Antiviral Agents; B-Lymphocyte Epitopes; B-Lymphocytes; Carrier Proteins; Classification; Code; Combinatorial Synthesis; Communicable Diseases; Conjugated Carrier; Coronavirus; Country; Development; Diagnostic Procedure; Enzyme-Linked Immunosorbent Assay; Epidemiology; Epitopes; Evaluation; Genome Stability; Glycoproteins; Human; IL2 gene; Immunization; In Vitro; Individual; Interferon Type II; Interleukin-12; Interleukin-2; Length; Membrane Proteins; Methods; Patients; Peptide Library; Peptide Sequence Determination; Peptides; Peripheral Blood Mononuclear Cell; Proteins; Relative (related person); Serum; Severe Acute Respiratory Syndrome; System; T-Lymphocyte; T-Lymphocyte Epitopes; Technology; Testing; Vaccines; Viral; antibody conjugate; combinatorial; cytokine; design; env Gene Products; genome sequencing; immunogenicity; novel; peptide based vaccine; prevent; prophylactic; protein structure; response; therapeutic vaccine

DESCRIPTION (provided by applicant): Severe acute respiratory syndrome (SARS) caused by coronavirus is an emerging human infectious disease that epidemically spread worldwide. Therefore, the actual diagnostic method, preventing and therapeutic vaccine, and antiviral drugs are urgently demanded. This proposal aims at development of peptide-based vaccine including combinatorial synthesis of overlapped peptide library, identification of B- and T-cell epitopes directly, selection of neutralizing antigens and immunization of animal. 1.Using our coding and "MBGB" combinatorial technology, total 1938 overlapped individual peptides (10 amino acids in length) will be synthesized according to the BJ01 protein sequence including spike glycoprotein, membrane protein, envelope protein, and nucleocapasid protein, which potentially comprise the human immuno-recognizing domains of SARS CoV. 2. B-cell epitopes will be directly determined using ELISA method. The peptides reacted across with at least twelve antibody-positive sera of SARS patients will be determined as being positive. 3. SARS CoV specific T-cell epitopes from above proteins will be identified by peptide's stimulation of peripheral blood mononuclear cells (PBMC) releasing cytokines of recovered SARS patients, such as IFN-gamma, IL2, or IL12 detecting by Elispot method. 4. The elongated peptides covering two or three identified epitopes will be designed, synthesized and tested their antigenicity. Their immunogenicity will be determined when the anti-peptide (peptide-carrier conjugate) antibody from animal sera neutralizes the SARS CoV. 5. Multiple antigens containing both B-cell neutralizing antigens and SARS CoV specific T-cell epitopes will covalently conjugate onto protein carrier which will be able to immunize animals. The protection of SARS CoV challenges and CTL responses will be investigated to evaluate the peptide-based vaccine.

描述（由申请人提供）： 由冠状病毒引起的严重急性呼吸道综合症（SARS）是一种新出现的人类传染病，在全球范围内流行。因此，迫切需要实际的诊断方法、预防和治疗性疫苗以及抗病毒药物。该提案旨在开发基于肽的疫苗，包括重叠肽库的组合合成、直接鉴定B细胞和T细胞表位、中和抗原的选择以及动物的免疫。 1.利用我们的编码和“MBGB”组合技术，根据BJ01蛋白序列，将合成总共1938个重叠的单个肽（长度为10个氨基酸），包括刺突糖蛋白、膜蛋白、包膜蛋白和核衣壳蛋白，这些肽可能包含SARS冠状病毒的人类免疫识别结构域。 2.采用ELISA法直接测定B细胞表位。与至少12份SARS患者抗体阳性血清反应的肽将被确定为阳性。 3.通过肽刺激康复SARS患者的外周血单核细胞（PBMC）释放细胞因子，通过Elispot法检测IFN-γ、IL2或IL12等，鉴定上述蛋白中的SARS CoV特异性T细胞表位。 4. 设计、合成覆盖两个或三个已识别表位的延长肽并测试其抗原性。当动物血清中的抗肽（肽载体缀合物）抗体中和 SARS CoV 时，将确定它们的免疫原性。 5. 含有 B 细胞中和抗原和 SARS CoV 特异性 T 细胞表位的多种抗原将共价结合到蛋白载体上，从而能够对动物进行免疫。将研究对 SARS CoV 攻击和 CTL 反应的保护，以评估基于肽的疫苗。