Application of CD11c<+>B cells to respects of diagnosis and treatment of sepsis

A technology for sepsis and B cells, which is applied in the field of CD11c+B cells in the diagnosis and treatment of sepsis, and achieves the effects of rapid detection, low price, and simple and quick operation

Inactive Publication Date: 2020-03-27
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

[0004] In recent years, studies have found that a group of age-associated B cells (Age associated B cells) highly express CD11c, also known as CD11c+ B cells. So far, there is no international research on the relationship between CD11c+ B cells and sepsis. Related reports

Method used

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  • Application of CD11c&lt;+&gt;B cells to respects of diagnosis and treatment of sepsis
  • Application of CD11c&lt;+&gt;B cells to respects of diagnosis and treatment of sepsis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1CD11c + Increased proportion of B cells in sepsis patients

[0025] The peripheral blood of the subjects was collected, including 40 cases of healthy subjects and 50 cases of sepsis patients.

[0026] Specifically include the following steps:

[0027] (1) Use lymphocyte separation medium to separate PBMC, take 1×10 5 ~1×10 7 1 cell, add 1ml of reaction buffer, and centrifuge at 1500 rpm for 5 minutes;

[0028] (2) Specific antibody staining reaction: 100μl reaction system contains: PBMC is 1x10 5 ~1x10 7 For each cell, 100 μl of reaction buffer PBS, 3 μl of CD11c antibody or isotype control antibody IgG, 2 μl of CD19 antibody, and incubate the above reaction tube on ice (0-4°C) for 30 minutes in the dark. Add 1ml of reaction buffer, centrifuge at 1500 rpm for 5 minutes, repeat 3 times;

[0029] (3) Resuspend the cells in 500ul of reaction buffer, and detect with a flow cytometer;

[0030] (4) Judgment of results: Use the isotype control antibody as a ne...

Embodiment 2

[0032] Example 2CD11c + B cells promote mortality in mice

[0033] 1. Sorting CD11c+ B cells by flow cytometry, and injecting 5×10 cells into the tail vein 6 CD11c+ B cells in the mice, 48 days later, the mice were intraperitoneally infected with LPS, and then the survival rate of the mice was observed and recorded. Then based on the same LPS concentration, the model was established again, and the mice in each group were sacrificed 12 hours later, and the lungs were taken for pathological section observation. Preparation of pathological slices: cut tissues were soaked in 4% paraformaldehyde, and fixed at room temperature for 24 hours. Pour off the formaldehyde fixative, add pH7.4 BSA, replace it every 8 hours, store at 4°C, and change the solution for a total of 3 days. Open the oven and heat up to 62°C to dissolve the paraffin. Alcohol gradient dehydration (30%, 50%, 70%, 80%, 90%, 95%, 100% twice), replaced every 30min. 50% alcohol + 50% xylene for 30 minutes, 100% xyle...

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Abstract

The invention belongs to the technical field of cellular immunity treatment. Firstly, specificity B cell subgroups and CD11c<+>B cells are applied to diagnosis of sepsis. Through cell surface molecular detection, quick detection of the sepsis can be realized. A detection method and a detection reagent kit have the advantages of being high in sensitivity, good in specificity, simple and quick to operate and the like, and besides, are low in cost and suitable for promotion and application.

Description

technical field [0001] The present invention relates to the technical field of cellular immunotherapy, more specifically, to CD11c + Application of B cells in the diagnosis and treatment of sepsis. Background technique [0002] Sepsis refers to the systemic inflammatory response syndrome (SIRS) caused by infection. It is clinically confirmed that there are bacteria or highly suspicious infection foci, and it is the main cause of death in critically ill patients. According to its severity, it can be divided into sepsis, severe sepsis and septic shock. Its pathogenesis is mainly due to the pathological damage of the body caused by the "inflammatory storm" produced by immune cells. Although various countries have devoted a lot of manpower, material and financial resources to the research of sepsis, its mortality rate is still high. One of the main reasons is the lack of effective diagnostic indicators and early effective drug treatment. Although the latest sepsis diagnostic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P31/04G01N15/14
CPCA61K45/00A61P31/04G01N15/14G01N2015/0065G01N2015/1006
Inventor 黄曦刘娇吴永坚
Owner THE FIFTH AFFILIATED HOSPITAL SUN YAT SEN UNIV
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