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IL-6-IL-27 compound and application thereof in preparation of antiviral drugs

An antiviral drug, IL-27 technology, applied in the field of biomedical detection, to achieve the effect of inhibition of replication level

Active Publication Date: 2021-11-02
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are few drugs capable of resisting multiple viral infections in the prior art, so it is necessary to develop a new drug capable of resisting multiple viral infections

Method used

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  • IL-6-IL-27 compound and application thereof in preparation of antiviral drugs
  • IL-6-IL-27 compound and application thereof in preparation of antiviral drugs
  • IL-6-IL-27 compound and application thereof in preparation of antiviral drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The construction of embodiment 1 expression vector

[0060] All expression plasmids in this patent are constructed on the pcDNA3.1(+) vector, and cloned using the homologous recombination cloning kit of Qingdao Biotechnology Co., Ltd. Total cellular RNA was extracted from SeV-infected human THP1 cells, and the cDNAs of human IL-6, EBi3 and IL-27A were amplified and cloned directly into the pcDNA3.1(+) vector. IL-6 was tagged with C-terminal 3×HA, EBi3 was tagged with C-terminal Myc, and IL-27A was tagged with C-terminal V5. In the IL-27 expression plasmid, the coding sequences of EBi3 and IL27A were linked with the sequence GSGSGGSGGSGSGKL coding the peptide chain, and a C-terminal V5 tag was added. The IL-6 fluorescent protein consists of IL6, GFP and the C-terminal ER retention signal KDEL, named IL6-GFP-KDEL. The IL-27 fluorescent protein consists of IL27, DsRed and a C-terminal KDEL signal, named IL27-DsRed-KDEL.

[0061] In addition, plasmids overexpressing MyD8...

Embodiment 2

[0086] Embodiment 2, endogenous interaction

[0087] 1. Serum samples

[0088] The serum samples of 10 patients with influenza virus infection and the throat swab samples of 45 patients with influenza virus infection used in this patent are all from Hubei Provincial Center for Disease Control and Prevention. Collection of clinical samples was performed in accordance with the principles of the Declaration of Helsinki and was approved by the Institutional Review Board of Wuhan University in accordance with the Guidelines for the Protection of Human Subjects. All study participants provided written informed consent for sample collection and subsequent analysis.

[0089] 2. Processing of serum samples

[0090] Serum samples from 10 IAV-infected patients were obtained from Hubei Provincial Center for Disease Control and Prevention. Serum samples were mixed and divided into three groups, and 1ml samples were taken from each group for CoIP experiments. The rest of the samples wer...

Embodiment 3

[0096] Example 3 Determination of interactions by co-immunoprecipitation

[0097] The IL-6-HA, IL-27-V5, IL-27A-V5 and EBi3-Myc expression plasmids constructed in Example 1 were co-transformed in different combinations. After 30 hours of co-transformation, the cells were harvested and an appropriate amount of cells were added for lysis Buffer solution (containing protease inhibitors), lysed on ice for 30 minutes, and the cell lysate was centrifuged at the maximum speed for 15 minutes at 4°C, and the supernatant was taken to obtain the total cell protein. Among them, the KD of IL-6-HA, IL-27-V5, IL-27A-V5 and EBi3-Myc proteins are 25KD, 28KD, 54KD and 34KD respectively;

[0098] Take a small amount of lysate for Western blot analysis (input analysis);

[0099] Add 1 μg of the corresponding antibody to the remaining lysate (the remaining lysate is divided into 4 parts, one part is incubated with 1ug anti-HA antibody, one part is incubated with 1ug IgG, one part is incubated wit...

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Abstract

The invention provides an IL-6-IL-27 compound and an application thereof in preparation of an antiviral drug. The IL-6-IL-27 compound is a compound formed by interaction of IL-6 and IL-27. The invention proves that the infection of various viruses, including influenza virus (IAV), vesicular stomatitis virus (VSV), enterovirus 71 (EV71), I-type herpes simplex virus (HSV-I) and Sendai virus (SeV), can be inhibited by using the IL-6-IL-27 compound for treatment. Cells pretreated by the IL-6-IL-27 compound are infected by the virus, and compared with a control group, the replication level of the virus is inhibited to a great extent. Therefore, the IL-6-IL-27 compound has the activity of resisting virus infection. The discovery of the new biological function can be used for researching the potential function of the IL-6-IL-27 compound in the aspect of inhibiting virus infection.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to IL-6-IL-27 complex and its application in the preparation of antiviral drugs. Background technique [0002] Viral infection can cause a variety of diseases in the human body. Influenza virus infection is a major factor causing influenza. It is highly contagious and spreads quickly, mainly through droplets in the air, contact between people or contact with contaminated items. Typical clinical symptoms are: sudden onset of high fever, general pain, significant fatigue and mild respiratory symptoms. The autumn and winter seasons of each year are its high incidence period. Enterovirus 71 mainly causes hand, foot and mouth disease, and can also cause a variety of neurological diseases such as aseptic meningitis, brainstem encephalitis and polio-like paralysis. Patients manifest as low-grade fever, runny nose, loss of appetite, mouth pain, vomiting, and diarrhea. Small...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/85A61K38/20A61P31/14A61P31/16A61P31/22
CPCC07K19/00C07K14/5412C07K14/54C12N15/85A61P31/14A61P31/16A61P31/22C12N2800/107A61K38/00A61K2300/00Y02A50/30
Inventor 朱应左麒程智逵佘应龙
Owner WUHAN UNIV
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