A method for evaluating the degree of immunodeficiency in an immunodeficiency mouse model

An immunodeficiency mouse, immunodeficiency technology, applied in animal cells, animal husbandry, vertebrate cells, etc., can solve the difficult degree of immunodeficiency, blind construction and use of immunodeficiency mice, unfavorable standards for immunodeficiency mice and promotion issues

Active Publication Date: 2019-05-17
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the continuous emergence of immunodeficient mice, there is still no standard method for assessing the degree of immunodeficiency in mice. It is difficult to objectively evaluate the degree of immunodeficiency in genetically modified mouse strains, which will lead to the blind construction of immunodeficient mice And use, it is also not conducive to the standard unification and promotion of the application of immunodeficiency mice

Method used

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  • A method for evaluating the degree of immunodeficiency in an immunodeficiency mouse model
  • A method for evaluating the degree of immunodeficiency in an immunodeficiency mouse model
  • A method for evaluating the degree of immunodeficiency in an immunodeficiency mouse model

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment one: specific implementation steps:

[0022] Construction of human hematoma cell line with marker gene

[0023] In order to facilitate the promotion and application of the present invention, the human chronic myeloid leukemia cell line K562 is used as the standard (heterogeneous) human blood tumor cell of the immunodeficiency mathematical model, and the human lung adenocarcinoma cell line A549 is used as the standard of the immunodeficiency mathematical model ( Heterogeneous) human solid tumor cells, and marker genes (such as GFP green fluorescent gene, Tdtomato red fluorescent gene, Luciferase luciferase, etc.) were introduced into the K562 cell line. The following describes the GFP green fluorescent gene transfection method, packaging Lenti-GFP virus to infect tumor cell lines, expanding and culturing the infected cells, sorting the successfully infected GFP positive tumor cells, and obtaining GFP positive cells with a purity greater than 98%.

[0024] ...

Embodiment 2

[0034] Utilize the method for evaluating the degree of immunity of mice of the present invention, comprehensively assess the degree of immunodeficiency (TEI) of NSI (NOD-scid IL2rg- / -) mice NSI ),Specific steps are as follows:

[0035] 1.1. HTEI NSI Numerical calculation: Transplant K562-GFP cells of H, M, and L magnitudes into NSI mice, observe the mice every day, and obtain and record the experimental parameters in the following table when the mice become ill:

[0036] Table 1. NSI mouse-human blood tumor transplantation model parameter list

[0037]

[0038] Substitute the above parameters into the formula to get:

[0039]

[0040]

[0041]

[0042] HTEI NSI =average(HTEI( NSI，H) , HTEI (NSI，M) , TEI (NSLL) ) = 0.014

[0043] 1.2. STEI NSI Numerical calculation: transplant A549-GFP cells of H, M, and L order into NSI mice respectively, observe the mice every day, obtain and record the experimental parameters as follows when the mice are ill:

[0044] Ta...

Embodiment 3

[0056] Utilize the method for evaluating the degree of immunity of mice of the present invention, comprehensively assess the degree of immunodeficiency (TEI) of NOD-scid (abbreviation: NS) mice NS ),Specific steps are as follows:

[0057] 1.4. HTEI NS Numerical calculation: Transplant K562-GFP cells of H, M, and L magnitudes into NOD-scid mice, observe the mice every day, obtain and record the experimental parameters when the mice are ill, and record them in the following table:

[0058] Table 3. NOD-scid mouse-human blood tumor transplantation model parameter table

[0059]

[0060] Substitute the above parameters into the formula to get:

[0061]

[0062]

[0063]

[0064] HTEI NS =average(HTEI (NS，H) , HTEI (NS，M) , TEI (NS，L) ) = 0.009

[0065] 1.5. STEI NS Numerical calculation: A549-GFP cells of H, M, and L magnitudes were transplanted into NOD-scid mice, and the mice were observed every day, and the experimental parameters were obtained and recorded ...

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Abstract

The invention relates to the field of biology, and discloses a method for evaluating immunodeficiency degree of an immunodeficient mice model. The method comprises: through transplanting neoplastic hematologic disorder cells and solid tumor cells in different orders of magnitudes into an immunodeficient mice model, establishing a tumor mice model, detecting the proportion of tumor cells GPB in internal and external blood of the tumor mice model, the proportion of tumor cells GBM in bone marrow, and the proportion of tumor cells GSP in spleen, according to proportion parameter indexes, evaluating the immunodeficiency degree of the immunodeficient mice model. The method is advantaged in that the method can quantitatively evaluate defect level of the immunodeficient mice, the method relates to simplify and summarize a mathematic model and a calculation method which are obtained through relations between tumor transplanting efficiency and mice immunodeficiency degree, and through scores of immunodeficiency degree, defect difference degree among the immunodeficient mice can be compared. The method is simple and feasible, and is easy to operate, and the method has good application prospect.

Description

technical field [0001] The invention relates to the field of biology, in particular to a method for evaluating the degree of immunodeficiency in an immunodeficiency mouse model. Background technique [0002] Using immunodeficient mice as hosts, the constructed humanized mouse models and patient-derived xenograft models are important tools for the study of the human immune system and the pathological mechanism of human tumors. In 1963, scientists developed nude mice (Nudemice), which had mutations in the Foxn1 gene and a deterioration or loss of the thymus, which blocked the development of T cells, resulting in a large number of T cells and a defect in the immune system. However, nude mice have B cells and natural killer (NK, natural killer) cells, which are not conducive to maintaining the growth of human cells in mice. In 1983, severe combined immunodeficiency mice (scid, severe combinedimmunodeficiency) appeared, the mouse Prdkc gene defect, resulting in the loss of B cel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A01K67/02
CPCA01K67/02A01K2207/30A01K2227/105A01K2267/0387
Inventor 李鹏叶未肖毅仁蒋治武林思妙姚瑶王素娜李柏衡
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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