Preparation and application of IL-15 isoform protein of mice

A mouse, protein technology, applied in applications, peptide/protein components, animal/human proteins, etc.

Active Publication Date: 2010-09-15
KUSN BEIRUIKANG BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This isoform was found in mouse small intestinal epithelial cells when the immune system was in a normal state, but there was no report on the production of IL-15 isoform protein when the immune system was in a stimulated state

Method used

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  • Preparation and application of IL-15 isoform protein of mice
  • Preparation and application of IL-15 isoform protein of mice
  • Preparation and application of IL-15 isoform protein of mice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1, Preparation of purified functional unlabeled mouse IL-15 isoform protein

[0058] 1. Extraction of total cellular RNA

[0059] Take the spleen of C57BL / 6 mice, prepare spleen cell suspension, stimulate mouse spleen cells with 0.1 μg / ml LPS for 48 hours, obtain the stimulated C57BL / 6 mouse spleen cell suspension, add 1 ml of lysis buffer Trizol, and pipet repeatedly. Mix well, dissolve the cells in Trizol, incubate the samples at room temperature for 5 min to lyse the cells; add 0.2 ml of chloroform, mix vigorously for 15 s, incubate the samples at room temperature for 3-5 min, centrifuge at 12000g for 15 min at 4°C; transfer the upper colorless aqueous phase to EP tube, add 0.5ml isopropanol, mix well, ice bath for 10min; centrifuge at 12000g for 10min at 4℃, carefully remove the supernatant; add 1ml 75% ethanol, invert and mix, centrifuge at 4℃, 7500g for 5min, wash twice ; Discard the supernatant completely, dry the RNA slightly, dissolve it in 20 μl RNase...

Embodiment 2

[0142] In this example, the concentration of LPS used in step 1 was 10 μg / ml, and the action time was 24 hours. The rest of the reagents and procedures are exactly the same.

Embodiment 3

[0144] In this example, the concentration of LPS used in step 1 was 20 μg / ml, and the action time was 24 hours; the remaining reagents and operation steps were exactly the same.

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PUM

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Abstract

The invention relates to a preparation method of monoclonal protein of mice, in particular to preparation and application of IL-15 isoform protein of mice. In the preparation method, the IL-15 isoform protein of the mice is prepared in the excitation state of an immunity system, the antagonistic effect of the IL-15 isoform protein on IL-15 protein of the normal mice is detected through in-vitro experiments, and the application of the isoform protein in clinical treatment such as autoimmune diseases and the like is provided. The study proves that pulpar cells which are not stimulated by LPS cannot detect the isoform, but the isoform can be detected in macrophage and B cells which are stimulated by the LPS and are from primary culture or cell strains, and the isoform is possible to have important negative feedback regulate and control effect under the condition that the immune system is activated. In the preparation method, the isoform protein is discovered under the condition that the immune system is activated for the first time, and is subjected to correlational studies of the functions thereof, and thus a more direct evidence is provided for the effect of the isoform protein in the regulation and control of organism immunity.

Description

technical field [0001] The invention relates to the preparation of a mouse monoclonal antibody protein, in particular to the preparation and application of a mouse IL-15 subtype protein. Background technique [0002] IL-15mRNA (interleukin 15, IL-15) is widely expressed in various tissues, including placenta, skeletal muscle, kidney, heart, lung, brain, intestinal epithelium, and is also expressed in myeloid hematopoietic stem cells such as macrophages ,Dendritic Cells. IL-15 is structurally homologous to interleukin 2 (IL-2). Therefore, IL-15 has some similar functions to IL-2, such as stimulating T cell activation and proliferation, enhancing NK cell killing activity and promoting B cells to produce immunoglobulins. Recent studies have found that IL-15 plays an important role in the differentiation, proliferation and activation of NK cells, NKT cells and intestinal epithelial cells. IL-15 and IL-17 are very important for the homeostasis regulation of CD8 memory T cells. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/24C12N15/70C07K14/54A61K38/20A61P37/02A61P31/16A61P11/00A61P19/02A61P19/04A61P29/00A61P1/00A61P17/06A61P1/16A61P31/14A61P11/06A61P21/00A61P39/00A61P9/00
Inventor 刘海燕胡博
Owner KUSN BEIRUIKANG BIOLOGICAL TECH
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