Method and preparation for preventing mycoplasma hyorhinis from infecting cells

A technology of Mycoplasma hyorrhinosus and cells, which is applied in the directions of medical preparations, antibodies, and antibacterial drugs containing active ingredients, can solve the problems of lack of specificity in preventing Mycoplasma hyorrhinosus infecting cells, and the molecular mechanism of Mycoplasma hyorrhinosus infecting cells is not clear. The effect of preventing the infection of cells by Mycoplasma hyorhinosa

Active Publication Date: 2014-11-12
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the molecular mechanism of Mycoplasma hyorhina infecting cells is still unclear,

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method and preparation for preventing mycoplasma hyorhinis from infecting cells
  • Method and preparation for preventing mycoplasma hyorhinis from infecting cells
  • Method and preparation for preventing mycoplasma hyorhinis from infecting cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1, Mycoplasma hyorhinois infection of mammalian cells depends on its surface membrane protein p37

[0061] In this example, it was confirmed by common cell ELISA experiments reported in literature that both Mycoplasma hyorhinosus and p37 protein can bind to cells respectively.

[0062] Cell ELISA experiments first inoculated gastric cancer cells MGC803 and AGS (these cells were from the tumor cell bank of the Chinese Academy of Medical Sciences) in 96-well plates and cultured them for 48 hours, and then added 1ⅹ10 3 CCU, 1ⅹ10 4 CCU, 1ⅹ10 5 CCU / mL of M. hyorhini to infect cells. After the infection lasted for 24 hours, wash with PBS three times, fix with 0.4% glutaraldehyde at room temperature for 10 minutes, and block with 5% milk powder / PBST at room temperature for 2 hours, and then add anti-Mycoplasma hyorhini antibody, enzyme-labeled secondary antibody and substrate solution in sequence, Finally, measure OD with ELISA READER 492nm . Higher readings indic...

Embodiment 2

[0072] Example 2, N-terminal polypeptide of p37 blocks mycoplasma infected cells

[0073] The amino-terminal sequence of p37 (the 2nd to 23rd amino acid sequence of the p37 protein sequence, denoted as p37-2-23: LKKLKNFILFSSIFSPIAFAIS, SEQ ID No.1) is unique to Mycoplasma hyorhina, while in other types of Mycoplasma Does not exist (Dudler R, Schmidhauser C, Parish RW, et al. A mycoplasma high-affinity transport system and the in vitro invasiveness of mouse sarcoma cells. (1988) The EMBO Journal 7, 3963-3970).

[0074] In order to explore the role played by the amino terminal of p37 in mediating the infection of cells by Mycoplasma hyorhina, in this example, GST fusion protein was used to detect its binding to cells by ELISA. It was found that the amino-terminal polypeptide fusion protein (GST-p37-2-23) expressed by prokaryotic recombinant expression could bind to gastric cancer cell lines in a time- and concentration-gradient dependent manner, while the p37 protein lacking the...

Embodiment 3

[0079] Example 3, p37 protein and host ANXA2 protein interact through the amino terminal respectively

[0080] In order to find host cell surface receptor molecules that interact with p37, this example first enriches the p37-interacting proteins in the cell lysate by GST pull-down technology, and then finds them out by SDS-PAGE and Coomassie brilliant blue staining. difference band ( Figure 3A Indicated by the arrow in the middle), the band was identified by MALDI-TOF mass spectrometry, and the polypeptide fragment was found to be the Annexin A2 sequence ( Figure 3A ), indicating that the interacting protein of p37 is Annexin A2 (hereinafter referred to as ANXA2).

[0081] In order to verify that there is an interaction between p37 and Annexin A2, in this example, p37 and ANXA2 antibodies (the antibody was purchased from Novus Biotechnology) were also used to pass the conventional two-way co-immunoprecipitation experiment, and the results showed that immunoprecipitation wit...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a new method and preparation for preventing mycoplasma hyorhinis from infecting cells. Particularly, the invention provides an application of an antagonist for mycoplasma hyorhinis membrane proteins P37 or host cell membrane proteins Annexin A2 in preparation of preparations for preventing mycoplasma hyorhinis from infecting cells or promoting cell migration. According to the invention, the fact that cells are infected with mycoplasma hyorhinis through the interaction between 22 amino acid segments at the amino terminal of a membrane protein P37 on the surface of the mycoplasma hyorhinis and 25 amino acid segments at the amino terminal of a cell membrane protein Annexin A2 is mainly found, so that mycoplasma hyorhinis can be prevented from infecting cells by using anti P37 protein antibodies or antibodies of amino terminal polypeptides of mycoplasma hyorhinis P37, amino terminal polypeptides of Annexin A2, or anti host cell membrane proteins Annexin A2.

Description

technical field [0001] The present invention relates to a method and a preparation for preventing mycoplasma hyorhina from infecting cells, in particular to the discovery that mycoplasma hyorhina infects cells through the interaction between the amino-terminal 22 amino acid fragments of the membrane protein P37 on its surface and the cell membrane protein Annexin A2, and provides a solution against The application of the antagonist of mycoplasma hyorhina membrane protein P37 or host cell membrane protein Annexin A2 in the preparation of preparations for preventing mycoplasma hyorhina from infecting cells or promoting cell migration, and the method for preventing mycoplasma hyorhina from infecting cells or promoting cell migration. Background technique [0002] Mycoplasma belongs to the mollusc class of prokaryotes. It is the smallest microorganism that exists in nature and can replicate independently. Its size is about 0.1m. (Lo SC. Mycoplasmas and AIDS. In: Maniloff J, McEl...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K39/40A61K48/00A61K45/00A61P31/04
CPCA61K39/40A61K45/00A61K48/00
Inventor 寿成超段红英陈玲杨华孟麟
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap