Application of major histocompatibility complex II (MHC II) molecules on preparation of drugs for preventing and curing animal immunodepression
An animal immune and molecular technology, applied in the direction of drug combination, peptide preparation method, medical preparations containing active ingredients, etc., can solve the problems of difficult industrialization, complicated operation, high cost, etc., and achieve long action time and small dosage , the effect of short reaction time
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Embodiment 1
[0020] The specific operation method can be carried out as follows:
[0021] a. Process fresh pig spleen;
[0022] Take fresh pig spleen, put it into a sterile beaker immediately, add balanced salt solution to soak for half an hour or immerse in 1% bromogeramine solution, take out the spleen surface with iodine alcohol to disinfect, use dissecting scissors and tweezers to remove spleen fat and capsule, Wash once with balanced salt solution, then cut the spleen into small pieces and transfer them to a plate, and finally cut the spleen into 1mm pieces with ophthalmic scissors 3 For small pieces, wash 2-3 times with balanced salt solution to remove blood cells, pigment substances and cells damaged mechanically during the shredding process;
[0023] b. Digest and disperse tissue pieces;
[0024] Suck off the balanced salt solution washed in the previous step, move the chopped tissue pieces into the Erlenmeyer flask, add 0.25% trypsin solution (PH7.6-7.8) according to 5 times t...
experiment example 1
[0040] Experimental example 1: In vitro virus inhibition experiment
[0041] The specific operation method can be carried out in the following steps:
[0042] a. Preparation of chicken embryo fibroblasts.
[0043] Take chicken embryos aged 9-11 days, prepare chicken embryo fibroblasts in a sterile state, cover a 96-well plate, and prepare for the experiment after they grow into single cells.
[0044] b. Reticuloendotheliosis virus (REV) resuscitation.
[0045] Take and store reticuloendotheliosis virus (REV) in a -86°C refrigerator, and resuscitate according to conventional methods. The TCID50 of the virus is 10 -5 .
[0046] c. Inhibitory effect of different gradients of MHC class II molecules prepared in Example 1 of the present invention on reticuloendothelial hyperplasia virus.
[0047] Add 50 microliters of reticuloendotheliosis virus to each of the blank wells in the 96-well plate except for the first column, then set the first column of the 96-well plate as blank...
Embodiment 2
[0054] a. Using animal peripheral blood as a raw material, a lymphocyte monolayer was prepared according to a conventional method.
[0055] The specific method is: take 30 ml of animal whole blood in a sterile vein, add 0.3 ml of anticoagulant, and let it stand in the separating funnel for about 2-3 hours, collect the plasma above the red blood cell layer, divide it into a sterile centrifuge tube, and use balanced salt The liquid was repeatedly washed 2-3 times, and the centrifugation speed was 800-1400 rpm, and then the white blood cells were cultured with 40 ml of hydrolyzed milk protein containing 30% calf serum. Tighten the bottle stopper and culture in a 37°C incubator or in a spinner. After 2-3 days, the leukocytes will evenly adhere to the wall, and a monolayer of lymphocytes will be formed.
[0056] b. Subculturing Lymphocyte Monolayers
[0057] Before the cells are subcultured, first observe the culture bottle under a microscope to confirm that the cells have grown i...
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