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Construction method of rhesus monkey immunosuppression model and application of rhesus monkey immunosuppression model in CAR-T cell implantation

A technology of immunosuppression and construction methods, which is applied in the direction of pharmaceutical formulas, organic active ingredients, preparations for in vivo tests, etc., to achieve significant effects, reduce risks, and have small side effects

Pending Publication Date: 2022-04-12
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention solves the problem that reinfused CAR-T cells can survive in rhesus monkeys through the establishment of a rhesus monkey immunosuppressive model

Method used

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  • Construction method of rhesus monkey immunosuppression model and application of rhesus monkey immunosuppression model in CAR-T cell implantation
  • Construction method of rhesus monkey immunosuppression model and application of rhesus monkey immunosuppression model in CAR-T cell implantation
  • Construction method of rhesus monkey immunosuppression model and application of rhesus monkey immunosuppression model in CAR-T cell implantation

Examples

Experimental program
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Effect test

Embodiment 1

[0051] Example 1: Construction of a new rhesus monkey model of immunosuppression

[0052] 1. Experimental animal information

[0053] Source: Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences.

[0054] Experimental animal production license number: SCXK (Dian) K2017-0003 (Issuing agency: Kunming Science and Technology Bureau).

[0055] Domestication and breeding license number of national key protected wild animals: Diankun Taming Fan 201602 (Issuing agency: Kunming Forestry Bureau).

[0056] Experimental animal use license: SYXK (Dian) K2017-0008; issuing unit: Kunming Science and Technology Bureau.

[0057] Animal sex, age, weight range information:

[0058] Table 1 List of Animals

[0059]

[0060] 2. Environmental conditions and feeding management

[0061] 1) Breeding environment: ordinary environment (monkey ordinary animal laboratory of our institution).

[0062] Environmental Condition Index

[0063] Temperature: 16~26°C (daily...

Embodiment 2

[0150] Example 2: In order to further verify whether the immunosuppressive model established in this experiment can engraft CAR-T cells, three rhesus monkeys were selected in this experiment, and the same doses of cyclophosphamide and fludala phosphate were used as in experiments 2 to 4 The monkeys were pretreated by the sideline injection, and the animals were given immunosuppressants on day D-7, and the immunosuppressants were given continuously for three days, and the drugs were stopped for four days.

[0151] In this experiment, before immunosuppressive pretreatment, 20-30ml of peripheral blood was collected from monkeys, and PBMCs were sorted, and then monkey-derived CAR-T was prepared. The monkeys were subjected to immunosuppressive pretreatment according to the aforementioned method, and whether the pretreatment was successful was detected by hematology and FACS. After the detection indicators all met the criteria for successful immunosuppression, one rhesus monkey was ...

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Abstract

The invention discloses a construction method of a rhesus monkey immunosuppression model and application of the rhesus monkey immunosuppression model in CAR-T cell implantation. 25mg / (kg / d) cyclophosphamide is combined with 2.5 mg / (kg / d) fludarabine to pre-treat rhesus monkeys, a disease model consistent with the change trend of various immune indexes under the human immunosuppression condition is established, and the reduction ranges of the white blood cell number (WBC) and the lymphocyte absolute number (LYM) in hematology exceed 50%; a flow cytometry detection result shows that CD3 is reduced by more than 40%, and the immunosuppression animal model is successfully built; the model is used for implanting CAR-T cells so as to be used for researching CAR-T immunotherapy. According to the method, the rhesus monkey immunosuppression animal model can be quickly and accurately established, sufficient immunosuppression can be ensured during CAR-T cell implantation of animals, so that CAR-T cells are alive in rhesus monkeys, and a more ideal and reliable non-human primate experimental animal model and a perfect evaluation system are provided for preclinical research of CAR-T immunotherapy.

Description

technical field [0001] The invention belongs to the field of constructing animal models, and in particular relates to a method for constructing a rhesus monkey immunosuppressive model and its application in CAR-T cell transplantation. Background technique [0002] Today's tumor treatment methods are increasing day by day. In addition to conventional surgery, radiotherapy and chemotherapy, tumor immunotherapy, as the "fourth pillar" of cancer treatment, has attracted widespread attention. Medicine brings new hope. Among them, chimeric antigen receptor engineered T (CAR-T) cell therapy is a cell therapy method that has developed very rapidly in recent years, that is, by modifying T cells in vitro so that they can target tumor antigens and generate corresponding immune responses. Great success has been achieved in multiple clinical studies. Chimeric Antigen Receptor T Cell Therapy (CAR-T) refers to the isolation of T lymphocytes from the patient, after in vitro culture, trans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K31/675A61K31/7076
Inventor 李冰赵献军
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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