Product for inhibiting and/or preventing influenza virus secondary bacterial infection

A technology for influenza virus infection and prevention of influenza, which is applied in the field of biomedicine and can solve problems affecting the prevention and control of severe influenza and death

Inactive Publication Date: 2016-06-01
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Seriously affected the prevention and con

Method used

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  • Product for inhibiting and/or preventing influenza virus secondary bacterial infection
  • Product for inhibiting and/or preventing influenza virus secondary bacterial infection
  • Product for inhibiting and/or preventing influenza virus secondary bacterial infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1, IAV can promote the activity of human lung epithelial cells A549 secreting TGF-β

[0066] 1. Preparation of test samples

[0067] 1. Cultivate A549 cells, inoculate 48-well plate, 0.5-1×10 5 / 0.5ml per well. Place at 37°C, 5% CO 2 Cultivate overnight in an incubator;

[0068] 2. Cells grow to 90-100% confluence, washed 1-2 times with PBS;

[0069] 3. Add 270 μl / well serum-free DMEM medium;

[0070] 4. Add 30 μl / well H1N1 influenza virus PR8 virus solution (0.5-1×10 4 TCID50) or PBS control (30 μl / well PBS), after mixing. Place at 37°C, 5% CO 2 Cultivate in the incubator for 45min;

[0071] 5. Wash the cells 1-2 times with PBS to remove the virus liquid;

[0072] 6. Add 300 μl of DMEM complete culture solution, and collect the supernatant at different time points of culture, the time points are 2h, 4h, 6h;

[0073] 7. After the collected cell supernatant was centrifuged at 1000 g at 4°C for 10 minutes to remove cell debris, it was stored at -20°C or ...

Embodiment 2

[0083] Example 2. IAV promotes the expression of integrin α5 and fibronectin on the surface of A549 cells, which can be inhibited by TGF-β signaling pathway inhibitor SB431542 or SIS3

[0084] 1. Preparation of test samples

[0085] 1. Culture A549 cells, press 1-2×10 5 / 1ml per well to inoculate a 24-well plate at 37°C 5% CO 2 Incubate overnight in an incubator;

[0086] 2. When the cells are 80-90% confluent, wash them twice with sterile PBS and add 0.5-1×10 4 The H1N1 influenza virus PR8 / 500μl of TCID50 per hole, the culture medium is serum-free DMEM (+4% (4g / 100mL) BSA); 50mM, dissolved in DMSO); the control group only added solvent (0.5μl DMSO). Place at 37°C 5% CO 2 Incubate for 18 hours in the incubator. At the same time, a control group treated as follows was set up in the experiment: no virus PR8 infection and only solvent (0.5 μl DMSO) was added, which was recorded as DMSO / PBS group.

[0087] 2. Flow Cytometry (FACS)

[0088] 1. 0.25% (0.25g / 100ml) trypsin 10...

Embodiment 3

[0097] Example 3, IAV can promote the adhesion of type A streptococcus, pneumococcus and staphylococcus aureus to A549 cells, and this promoting effect can be inhibited by TGF-β signaling pathway inhibitor SB431542 or SIS3

[0098] 1. Infection of cells with PR8 virus

[0099] Same as Step 1 of Example 2.

[0100] 2. Bacterial adhesion test

[0101] Bacteria to be tested: Streptococcus type A (S.pyogenes) M1 type, Streptococcus pneumoniae (S.pneumococcus) T19F, Staphylococcus aureus (S.aureus) RN6390 (FnBPA+B+).

[0102] 1. Bacterial culture preparation: inoculate the bacteria into the corresponding culture medium (suspend with 10ml PBS for cultured colony, stand-by), place at 37°C 5% CO 2 Cultivate in the incubator for 17-18h. Centrifuge at 3000g for 10min, remove the culture medium (or PBS), and then add 10ml of PBS to wash once. Add an appropriate amount of serum-free DMEM (+4% (4g / 100mL) BSA), and adjust the amount of bacteria to 0.5-1×10 7 CFU / ml.

[0103] Among them,...

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PUM

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Abstract

The invention discloses a product for inhibiting and/or preventing influenza virus secondary bacterial infection. The invention provides an application of a TGF-beta signaling pathway inhibitor, namely an application of a TGF-beta signaling pathway inhibitor in preparation of a product for inhibiting and/or preventing influenza virus secondary bacterial infection. The TGF-beta signaling pathway inhibitor is at least one of the following inhibitors: an anti-TGF-beta monoclonal antibody, a soluble TGF-beta receptor, a TGF-beta antisense oligonucleotide, an inhibitor for inhibiting TGF-beta expression or inhibiting activity, a small-molecule inhibitor of a TGF-beta receptor and a small-molecule inhibitor of a TGF-beta signaling pathway downstream factor. By research results of a key role of TGF-beta in A-type influenza virus secondary bacterial infection, an impact factor for inhibiting TGF-beta signaling pathway or regulating TGF-beta is brought forward, and the effect of preventing settling and infection of bacteria at respiratory tract due to influenza virus infection is achieved. The product of the invention has the advantage of early-stage, high-efficiency and wide-spectrum blocking of bacterial adhesion.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a product for inhibiting and / or preventing secondary bacterial infection of influenza virus. Background technique [0002] The infection caused by influenza A virus (influenza Avirus, IAV) is one of the diseases with the highest morbidity and mortality in the world, causing about 5 million people to be infected and 250,000 to 500,000 people to die every year. Secondary bacterial pneumonia is the leading cause of death in influenza patients. In 2008, the US National Institutes of Health retested autopsy lung tissue sections from 58 patients who died of 1918 influenza, and confirmed that the main lesion was severe bacterial lung infection. Scientists retrieved all autopsy studies on 1918 influenza deaths published between 1919 and 1929, with a total of more than 8,000 cases from 15 countries, and found that pleural fluid of 80% of patients who died of influenza was positive for bacterial c...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K38/18A61K48/00A61P31/16A61P31/04A61P11/00
Inventor 王北难李宁王小爽范鑫
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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