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Application of IL-12p40(-/-)IL-2R alpha(-/-) mouse model

1. The technology of il-12p40 and mouse model, applied in the field of medical biology, can solve the problems of lacking animal models of autoimmune myelofibrosis

Inactive Publication Date: 2017-10-10
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, animal models of autoimmune myelofibrosis are lacking

Method used

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  • Application of IL-12p40(-/-)IL-2R alpha(-/-) mouse model
  • Application of IL-12p40(-/-)IL-2R alpha(-/-) mouse model
  • Application of IL-12p40(-/-)IL-2R alpha(-/-) mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Establishment of IL-12p40(- / -)IL-2Rα(- / -) mice

[0034] IL-2Rα (- / -) mice (B6.129S4-Il2ratm1Dw) [Willerford, D.M., et al. (1995). Immunity 3 (4): 521-530] of C57BL / 6J background and IL-12p40 (- / -) mouse (B6.129S1-Il12btm1Jm) [Magram, J., et al. (1996). Immunity 4 (5): 471-481] was purchased from Jackson Laboratory, USA (The Jackson Laboratory, Maine, USA, http : / / www.jax.org / ), reared in a special pathogen-free (SPF) environment.

[0035] We crossed IL-2Rα(+ / -) mice with IL-12p40(- / -) mice to obtain IL-12p40(+ / -)IL-2Rα(+ / -) mice; 12p40(- / -) mice were crossed to obtain IL-12p40(- / -)IL-2Rα(+ / -) mice. The IL-12p40(- / -)IL-2Rα(- / -) mice and IL-12p40(- / -)IL-2Rα(+ / -) mice used in the experiment were all produced by IL-12p40(- / -) IL-2Rα(+ / -) mice were bred. Since the mutant genes of IL-12p40 and IL-2Rα both contain a neo gene, the method for identifying the mouse IL-12p40 gene is to use genotyping to identify the wild-type gene of IL-12p40 and identify the mous...

Embodiment 2

[0036] Embodiment 2. routine blood test

[0037] The peripheral blood of the mice was collected into an anticoagulant tube, 150 μL of blood was drawn, and blood routine data were collected by an automatic hematology analyzer.

[0038] see results figure 1 , Compared with IL-12p40(- / -)IL-2Rα(+ / -) mice, the peripheral red blood cell count (RBC) of IL-12p40(- / -)IL-2Rα(- / -) mice, Hemoglobin content (HGB), hematocrit (HCT) and white blood cell count (WBC) all decreased significantly, but platelet count (PLT) did not change significantly.

Embodiment 3

[0039] Example 3. Histopathological examination

[0040] The liver, spleen and bone marrow tissues of the mice were fixed in 4% neutral formaldehyde for 1-2 days. The bone marrow tissue was put into decalcification solution (5% hydrochloric acid+5% acetic acid) for 24 hours. Tissues were dehydrated and embedded in paraffin, and sectioned into 4 μm slices. H&E staining was performed after dewaxing the thin sections. Bone marrow tissue showed fibrosis by silver staining of reticular fibers.

[0041] see results figure 2 , image 3 ,From figure 2 Spleen of IL-12p40(- / -)IL-2Rα(- / -) mice compared with IL-12p40(- / -)IL-2Rα(+ / -) mice can be seen in HE staining Structural changes occurred, the distinction between red and white pulp was not obvious, and a large number of megakaryocytes appeared. Hematopoietic islands exist in the liver of IL-12p40(- / -)IL-2Rα(- / -) mice. These results demonstrate extramedullary hematopoiesis in the spleen and liver of IL-12p40(- / -)IL-2Rα(- / -) mi...

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Abstract

The invention disclosesan application of an IL-12p40 (- / -) IL-2R alpha (- / -) mouse model and belongs to the technical field of medical biology. Compared with IL-12p40 (- / -) IL-2R alpha (+ / -) mouse as a control mouse, an IL-12p40(- / -)IL-2R alpha(- / -) mouse is abnormal in a hematopoietic system in the following aspects: quantities of red blood cells and leukocytes in peripheral blood are reduced; CD4<+>T cells and CD8<+>T cells in bone marrow are abundantly infiltrated, proportion and quantity of LSK cells (Lin<->c-Kit<+>Sca-1<+> cells)are increased, hematopoietic capacity of the marrow declines, and reticular fibers appear in the marrow; splenomegaly occurs, and the spleen and the liver show extramedullary hematopoiesis. Therefore, the IL-12p40(- / -)IL-2R alpha(- / -) mouse can be takenas an animal model to be used for screening autoimmune myelofibrosis treatment drugs.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and in particular relates to the application of an IL-12p40(- / -)IL-2Rα(- / -) mouse model in screening drugs for treating autoimmune myelofibrosis. Background technique [0002] Myelofibrosis (Myelofibrosis) is a hematopoietic system disease, mainly in the elderly, the main feature is the hyperplasia of reticular fibrous tissue in the bone marrow, usually manifested as abnormal increase of hematopoietic stem cells, myeloid cell proliferation, anemia, splenomegaly, spleen and Liver extramedullary hematopoiesis, decreased survival rate, etc. In the vast majority of cases, patients with myelofibrosis are accompanied by gene mutations such as JAK2, CALR, and MPL, and myeloid cell tumors occur. The pathogenesis of myelofibrosis is considered to be mainly the activation of fibroblasts and the production of a large amount of collagen caused by gene mutations leading to hematopoietic stem cell differe...

Claims

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Application Information

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IPC IPC(8): A61K49/00
CPCA61K49/0008
Inventor 廉哲雄姚远李亮
Owner SOUTH CHINA UNIV OF TECH
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