Medicine for treating immunological rejection in corneal transplantation

A technology of transplant immunity and rejection, applied in the field of immunosuppression in corneal transplantation, can solve the problems of no research, no research on immune rejection inhibition, prolonging the survival time of corneal grafts, etc.

Active Publication Date: 2021-10-29
BEIJING TONGREN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, prior to this application, no studies have considered CD4 + LAP + Tregs cells are used to suppress immune rejection of corneal transplantation, and there is no research on the CD4 + LAP + The regulatory mechanism of Tregs cells regulating immune rejection was studied; this application not only verified the CD4 + LAP + The role of Tregs cells in maintaining corneal transplantation immune tolerance and transplantation immune rejection has also been systematically studied, and it has been found that by regulating ROCK protein (mainly ROCK2 protein), it can effectively inhibit corneal transplantation rejection and prolong corneal transplantation. Corneal graft survival time

Method used

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  • Medicine for treating immunological rejection in corneal transplantation
  • Medicine for treating immunological rejection in corneal transplantation
  • Medicine for treating immunological rejection in corneal transplantation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: CD4 + LAP + Preparation of Tregs cells

[0026] Spleen of Balb / c mice was aseptically isolated, milled and filtered through a 200-mesh filter to prepare cell suspension. mouse CD4 + LAP + Tregs cell isolation kit to isolate CD4 + LAP + For Tregs cells, the purity of CD4+LAP+Tregs cells was analyzed by flow cytometry and the isolated CD4 + LAP + Tregs cell count. Adjust cell concentration to 1 x 10 6 / ml, add CD3 / 28 antibody beads (according to the ratio of cells and beads: 2:1) and add IL-2 with a concentration of 1000ng / ml for culture. On day 5-7 of culture, perform anti-CD4 FITC, anti-LAP PerCP-Cy5.5 staining, and use flow cytometry to sort out CD4 + LAP + Tregs cells were purified and back-tested after sorting, and the expression of LAP in the in vitro culture and expansion group could be (85.76±5.01)%.

Embodiment 2

[0027] Example 2: Construction of mouse corneal transplant rejection model

[0028] BALB / c (H-2d) mice were used as recipients, C57BL / 6 (H-2b) mice were used as donors, and both donor and recipient mice were male, 8-10 weeks old. Prepare the mouse corneal transplantation model according to the following operations, and verify the CD4 + LAP + The role of Tregs cells:

[0029] (1) Mice were anesthetized: 4% chlorine hydrate was used for intraperitoneal injection. After anesthesia, foreign objects in the oral cavity of the mice were removed and a horizontal position was maintained. During the anesthesia process, pay attention to the mouse insulation.

[0030] (2) Corneal transplantation in mice: 11-0 simple interrupted suture was used for the operation method. Preparation of implants: One hour before operation in C57 mice, the pupils were dilated with compound tropicamide, the implants were drilled with a 2.5mm drill, and the microtweezers were cut along the imprint, taking ...

Embodiment 3

[0041] Example 3: CD4 + LAP + Tregs cells in the treatment of corneal transplantation rejection in mice

[0042] After the mouse corneal transplantation operation of embodiment 3 finishes, inject 10 microliters concentration and be 5x10 to every group of mice 5 / ml CD4 prepared in Example 1 + LAP + Tregs cells; and CD4 not injected + LAP + Allogeneic keratoplasty mice with Tregs were used as controls. The results showed that injection of CD4 + LAP + In mice with Tregs cells, the survival time of the implants was significantly prolonged, compared with the control group, the average extension was 15±1.32 days.

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Abstract

The invention relates to application of a CD4 + LAP + Tregs cell or an ROCK2 protein inhibitor capable of regulating expression and functions of the CD4 + LAP + Tregs cell in treatment of corneal transplantation immunological rejection. A CD4 + LAP + Tregs cellular pathway is used for treating corneal transplantation immunological rejection, and compared with an existing NKT cellular pathway, the CD4 + LAP + Tregs cellular pathway is more targeted, safer and more effective. Especially, the ROCK2 protein inhibitor is used for treatment, other side effects of injected cells on organs in the direct cell treatment process can be avoided, and good clinical application value is achieved.

Description

technical field [0001] The invention relates to the related field of immunosuppression in corneal transplantation, in particular to a method of using regulatory T cells or ROCK protein inhibitors to induce corneal transplantation immune tolerance and inhibit the occurrence of corneal transplantation immune rejection. Background technique [0002] Corneal blindness is a type of eye disease in which the loss of visual function is caused by the loss of corneal function. It is usually caused by diseases or trauma to the eye, resulting in varying degrees of damage to the cornea, turbidity, rupture or infection. Usually, the main causes of corneal blindness are: infectious keratopathy, non-infectious keratitis, corneal degeneration and corneal dystrophy, keratoconus, ocular trauma, eyelid injury and dry eye disease. Because of corneal lesion or corneal trauma, only the cornea loses its transparency and becomes cloudy, but the internal structure of the eyeball is normal, so if the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/17A61K45/00A61K31/517A61P37/06
CPCA61K35/17A61K45/00A61K31/517A61P37/06
Inventor 接英李上臧云晓潘志强杨珂田磊
Owner BEIJING TONGREN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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