Application of myeloid cell triggering receptor 2 as a novel coronavirus pneumonia diagnosis or treatment target

A coronavirus and cell technology, applied in disease diagnosis, antiviral agents, respiratory diseases, etc., can solve the problems of high disease mortality rate and poor effect of severe patients, and achieve less trauma, less pain, and easy review Effect

Active Publication Date: 2021-03-19
THE FIFTH AFFILIATED HOSPITAL SUN YAT SEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the above drugs have a certain effect on most patients, the treatment effect

Method used

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  • Application of myeloid cell triggering receptor 2 as a novel coronavirus pneumonia diagnosis or treatment target
  • Application of myeloid cell triggering receptor 2 as a novel coronavirus pneumonia diagnosis or treatment target
  • Application of myeloid cell triggering receptor 2 as a novel coronavirus pneumonia diagnosis or treatment target

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Experimental program
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Embodiment 1

[0040] Select 15 cases of severe (Severe) and 20 cases of non-severe (Non-severe) patients with novel coronavirus pneumonia as the experimental group; select 20 cases of healthy people as the control group.

[0041] Healthy subjects: subjects without clinical symptoms.

[0042] Patients with novel coronavirus pneumonia: SARS-CoV-2 virus nucleic acid test positive, and clinical symptoms manifested as pneumonia.

[0043] Collect 5 mL of peripheral blood from subjects in the experimental group and control group, lyse red blood cells to prepare a single-cell suspension, and adjust the total amount of cells to 5×10 6 For each, add TREM-2 antibody (R&D Company, Cat. No. FAB1278P) and CD4 and CD8 antibodies (both purchased from Biolegend Company) to 100 μL of the system, mix well and protect from light for 30 minutes, and detect by flow cytometry, with FCS / SSC as the gate Analyze the proportion of TREM-2 positive cells in CD4 and CD8 positive cells respectively.

[0044] figure 1 ...

Embodiment 2

[0046] Five cases of critically ill patients with novel coronavirus pneumonia were selected, and the expression of TREM-2 in patients in the early stage (Pre-ICU), severe stage (ICU) and remission stage (Post-ICU) were detected respectively.

[0047] Refer to the "New Coronary Pneumonia Diagnosis and Treatment Program (Trial Seventh Edition)" for the judgment of the above-mentioned severe stage, and take 5 mL of peripheral blood from the patient in 3 periods for the following tests. The red blood cells of the above samples were lysed to prepare a single cell suspension, and the total amount of cells was adjusted to 5×10 6 For each, add TREM-2 antibody (R&D Company, Cat. No. FAB1278P) to 100 μL of the system, mix well and protect from light for 30 minutes, and perform flow cytometry detection. FCS / SSC gates are set to analyze the percentage of TREM-2 positive cells that are CD4 and CD8 positive, respectively. cell ratio.

[0048] figure 2 It is the expression level of TREM-2...

Embodiment 3

[0050] The result is as image 3 As shown, a total of 5 kinds of plasmids were constructed, including Vector blank plasmid, TREM-2 expression plasmid with HA tag (TREM-2-HA), SARS-CoV-2 membrane protein expression plasmid with FLAG tag (M-FLAG), SARS-CoV-2 nucleoprotein expression plasmid with FLAG tag (N-FLAG), angiotensin converting enzyme 2 (ACE2) expression plasmid with FLAG tag (ACE2-FLAG).

[0051] The above 5 kinds of plasmids were combined into 293T cells as shown in the figure, - indicates that the plasmid is not transferred, + indicates that the plasmid is transferred. After 24 hours of transfer, the cells were collected, lysed to obtain proteins, and co-immunoprecipitated to detect the combination of TREM-2 and viral proteins. Input on the left indicates the expression of the above plasmids in the cell lysate detected by immunoblotting. The results show that the above plasmids can be effectively expressed in 293T cells. The black bands are ACE2-FLAG and N-FLAG from...

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Abstract

The invention belongs to the technical field of biology, and discloses an application of a myeloid cell triggering receptor 2 as a novel coronavirus pneumonia diagnosis or treatment target. Through research, TREM-2 is found to be highly expressed in CD4+ and CD8+T cells of peripheral blood of a patient suffering from novel coronavirus pneumonia and is positively correlated with the severity of a disease. The TREM-2 can be used as a marker for novel coronavirus pneumonia diagnosis and prognosis, and a TREM-2-Fc fusion protein (with an action effect of blocking TREM-2 signals) obviously reducesthe levels of cytokines IFN-g, TNF, IL-2 and Granzyme B. TREM-2 expression has significant correlation with severe indexes of aging, lymphocyte count reduction, C-reactive protein and D-dimer increaseproposed by a''novel coronavirus pneumonia diagnosis and treatment scheme''. Intervention of expression of TREM-2 is prompted to be capable of relieving or even be expected to treat novel coronaviruspneumonia.

Description

technical field [0001] The present invention relates to the field of biotechnology, and more specifically, relates to the application of myeloid cell triggering receptor 2 as a target for diagnosis or treatment of novel coronavirus pneumonia. Background technique [0002] Novel coronavirus pneumonia is an acute infectious pneumonia, and its pathogen is a new type of coronavirus not previously found in humans, namely severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2). [0003] According to the existing case data, the main manifestations of novel coronavirus pneumonia are fever, dry cough, fatigue, etc., and a small number of patients are accompanied by upper respiratory tract and digestive tract symptoms such as nasal congestion, runny nose, and diarrhea. Some patients have mild onset symptoms, may have no fever, and mostly recover after 1 week. Most patients have a good prognosis, while a few patients are in critical condition and even die. Severe cases ofte...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577A61K45/00A61K38/17A61K47/68A61P11/00A61P31/14
CPCA61K38/1774A61K45/00A61K47/6811A61P11/00A61P31/14G01N33/577G01N33/6893G01N2333/70503G01N2800/12G01N2800/52
Inventor 黄曦吴永坚王漫妮尹欢
Owner THE FIFTH AFFILIATED HOSPITAL SUN YAT SEN UNIV
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