Application of inhibitor of immune checkpoint in preparation of drug for treating diseases related to glaucoma and other ocular immune damage mechanisms
A technology of immune checkpoint and immune damage, applied in sensory diseases, drug combination, immunoglobulin, etc., can solve the problems of weakening immune response, destroying and strengthening immune homeostasis
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Embodiment 1
[0173] 1. Test object
[0174] Anti-CD28 antibody, anti-CD86 antibody, anti-CD80 antibody and IgG, and the above antibodies were made into blocking antibodies respectively.
[0175] 2. Establish a mouse model of glaucoma:
[0176] Mice were anesthetized by intraperitoneal injection of a mixture of ketamine (120 mg / kg) and xylazine (12 mg / kg). A short-term increase in intraocular pressure was induced by injecting polystyrene microparticles (Invitrogen, Oregon, USA) with a diameter of 15 μm into the anterior chamber of the right eye of adult C57BL / 6J mice. The particle injection concentration is 5.0×10 6 / ml. First, a 30G needle (BD, USA) was used to make a small puncture port in the center of the right cornea of the mouse, and then a glass micro-syringe was used to inject 3 to 4 microliters of particles into the anterior chamber. The normal control group injected the same volume of PBS into the anterior chamber in the same way.
[0177] According to the above method, a total of 20...
Embodiment 2
[0185] 1. Test object
[0186] Anti-CD28 antibody, anti-CD86 antibody, anti-CD80 antibody and IgG, and the above antibodies were made into blocking antibodies respectively.
[0187] 2. Animal model:
[0188] Purchase spontaneously high intraocular pressure transgenic mice DBA / 2J (Sibano Biotechnology Co., Ltd.), the spontaneous optic nerve damage of the mice began to appear at 6 months, DBA / 2J mice were used as the test group and randomly divided into 4 groups, 5 mice in each group, and the detection data of 3 months old DBA / 2J mice in each group were used as controls.
[0189] DBA / 2J mice were injected with anti-CD28 antibody, anti-CD86 antibody, anti-CD80 antibody or IgG into the vitreous cavity of mice in each test group once a week from 6 months of age, each administration concentration was 0.1μg / ml, The dosage is 2μL / head. Half of the mice in each group were sacrificed when they were 8 months old in the test group, and the other half were sacrificed when they were 12 months old...
Embodiment 3
[0198] 1. Test object
[0199] Anti-CD28 antibody, IgG, and the above antibodies were made into blocking antibodies.
[0200] 2. Establish a mouse model of glaucoma:
[0201] According to the method shown in Example 1, a mouse model of glaucoma was constructed.
[0202] A total of 15 glaucoma mouse models were constructed according to the above method, and they were randomly divided into 3 groups. After the above-mentioned establishment of the glaucoma mouse model, the vitreous cavity of each group of model mice was injected with anti-CD28 antibody or IgG at a concentration of 1 μg / ml and a dose of 2 μl / mouse.
[0203] 3. Detection
[0204] (1) One week after the injection, the peripheral blood of each group of glaucoma model mice was taken, and CD4 was detected by flow cytometry according to the method shown in Example 2. + IFNγ + / IL-4 + / IL-17 + cell;
[0205] (2) Use CD45RO and CD45RA to label mouse peripheral blood CD4 + FoxP3 + T cells and detect CD4 + FoxP3 + The content of T cell...
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