Application of peritoneal cells to induction of differentiation of T cells into regulatory T cells

A technology of cell induction and cell differentiation, applied in the field of medicine, can solve the problem of no T cell report

Active Publication Date: 2019-08-30
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there is no report on the production of regulatory T cells induced by peritoneal cells

Method used

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  • Application of peritoneal cells to induction of differentiation of T cells into regulatory T cells
  • Application of peritoneal cells to induction of differentiation of T cells into regulatory T cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Study on the composition and characteristics of peritoneal cells

[0043] 1. Acquisition of peritoneal cells

[0044] 1. Mice aged 6-7 weeks were killed by cervical dislocation, soaked in 75% alcohol for 1-2 minutes, moved into ultra-clean bench, fixed on dissecting board in supine position, and disinfected abdominal skin with alcohol.

[0045] 2. Use a 10ml syringe fitted with a 25g needle to draw 8mL of pre-cooled RPMI-1640 medium into the abdominal cavity, rub the abdomen lightly, then cut a small opening in the neck with ophthalmic scissors, cut the abdominal skin along the midline of the abdomen through the small opening, and expose the abdominal muscles.

[0046] 3. Under sterile conditions, draw the cell suspension with a 10ml syringe and transfer it into a centrifuge tube.

[0047] 4. After counting, centrifuge at 1200rpm for 5min, resuspend the cells in RPMI-1640 culture medium and dilute to the target concentration.

[0048] 2. The composition and...

Embodiment 2

[0052] Example 2 The role of peritoneal cells in inducing the differentiation of CD4+ T cells into regulatory T cells

[0053] Use 1ml 2×10 6 / ml according to the peritoneal cells obtained in Example 1 and 1ml 2×10 6 Spleen T cells / ml were co-cultured in an incubator with 5% carbon dioxide at 37 degrees Celsius for 6 or 12 hours, CD3+CD4+CD25+Foxp3+ regulatory T cells and CD3+CD4+IL in CD4+ T cells were detected by flow cytometry Percentage of -17+Th17 cells. The experiment was repeated 3 times. See the experimental results figure 2 .

[0054] figure 2 It was shown that CD3+CD4+CD25+Foxp3+regulatory T cells were significantly increased and CD3+CD4+IL-17+Th17 cells were significantly decreased when peritoneal cells were co-cultured with spleen T cells for 6 or 12 hours. The above experimental results show that the co-culture of peritoneal cells and T cells can effectively promote the differentiation of CD4+ T cells into regulatory T cells, thereby promoting the producti...

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Abstract

The invention relates to an application of peritoneal cells to induction of differentiation of T cells into regulatory T cells, belongs to the field of medicines, aims to solve the problems that obtaining enough regulatory T cells from tissue for treatment is difficult, and few obtaining methods exist, and provides an application of peritoneal cells to induction of differentiation of T cells intoregulatory T cells. The method comprises the following steps of firstly, detecting the expression situation of various markers on surfaces of the peritoneal cells, confirming whether the peritoneal cells are a mixture of various suppressor cells of B cells, macrophages, the T cells and the like, and confirming that the peritoneal cells have immunosuppressive action; and then performing co-cultureon the peritoneal cells and the T cells, wherein the results confirm that CD3+CD4+CD25+Foxp3+ regulatory T cells are obviously increased, and the peritoneal cells can induce the differentiation of T cells into regulatory T cells, and can promote generation of the regulatory T cells. The invention provides a new method for preparing the regulatory T cells.

Description

technical field [0001] The invention relates to the use of peritoneal cells to induce T cells to differentiate into regulatory T cells, and belongs to the field of medicine. Background technique [0002] Regulatory T cells (Treg) have a good immunosuppressive effect in the treatment of inflammatory and autoimmune diseases, but it is very difficult to obtain enough regulatory T cells from tissues for treatment. Therefore, it is necessary to study a method to induce the generation of regulatory T cells. [0003] Peritoneal cells (PCs) are an important cell population in the peritoneal cavity. Recent studies have found that B1a cells are mainly distributed in the abdominal cavity and thorax, and are positive for CD5 and CD11b. B1a cells can secrete IL-10 and play an important role in mucosal immunity. In addition, peritoneal cells express CD206mRNA strongly positive, and CD206 is a characteristic marker of M2 macrophages, M2 macrophages can reduce inflammatory cytokines and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0637C12N2506/11C12N2506/115
Inventor 刘婷江咏梅
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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