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Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-induced porcine splenic lymphocyte

A porcine circovirus and lymphocyte technology, which is applied in the field of lymphocyte oxidative stress model, can solve the problems of lack of understanding, poor controllability of modeling effect, and unstable concentration.

Inactive Publication Date: 2016-07-27
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is still a lack of knowledge about whether PCV2 infection can cause changes in the redox state of immune cells, and further research is urgently needed
Currently, the commonly used H 2 o 2 To establish a cellular model of oxidative stress, but H 2 o 2 In the cell culture medium, the metabolism is fast, the concentration is unstable, and the controllability of the modeling effect is poor.

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  • Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-induced porcine splenic lymphocyte
  • Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-induced porcine splenic lymphocyte
  • Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-induced porcine splenic lymphocyte

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Embodiment Construction

[0026] 1. Research ideas

[0027] The invention infects porcine spleen lymphocytes with PCV2, and measures the level of nitric oxide (NO) secreted by cells after virus infection, the level of total reactive oxygen species (ROS) in cells, the content of reduced glutathione (GSH), xanthine oxidase (XOD) activity, myeloperoxidase (MPO) activity, inducible nitric oxide synthase (iNOS) activity, to explore the correlation between PCV2 virus infection amount, infection time and dynamic changes of reactive oxygen species level, and establish the porcine spleen lymphatic In vitro model of cellular oxidative stress.

[0028] 2. Experimental method

[0029] (1) Isolation and culture of porcine spleen lymphocytes: the porcine spleen was washed twice in sterilized PBS, the spleen was ground with a glass syringe core, and the cells were collected by filtering through a 200-mesh filter. Tris-NH 4 After lysing red blood cells with Cl, wash them twice with PBS and once with 10%-FBS-RPMI164...

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Abstract

The invention discloses a method for constructing an oxidative stress model for a porcine circovirus 2 (PCV2) in-vitro-induced porcine splenic lymphocyte. The method comprises the steps of arranging a cell control group and a PCV2 infected group, separately adding an RPMI 1640 nutrient solution and a viral solution of corresponding concentration into each group, carrying out cell adsorption for 2 hours, then, discarding the solutions, adding 1mL of the RPMI 1640 nutrient solution containing 5% of FBS to each group, continuing to carry out culture, separately collecting samples, and applying the samples to the determination on cell activity and cell redox state related indexes. The model has the characteristics of convenience in constructing, relatively low cost, high practicability and the like, a relatively ideal in-vitro model is provided for researching whether PCV2 infection can cause the change of a redox state of an immunocyte or not and an action mechanism of the PCV2 infection, and then, some new thinking may be provided for the treatment on PCV2 infection induced animal diseases.

Description

technical field [0001] The invention belongs to the technical field of lymphocyte oxidative stress models, and in particular relates to a method for constructing a porcine circovirus type 2-induced oxidative stress model of porcine spleen lymphocytes in vitro. Background technique [0002] Porcine circovirus disease caused by porcine circovirus type 2 (PCV2) infection has become a major disease affecting the swine industry worldwide, causing huge economic losses. The main sites of PCV2 virus replication are monocytes-macrophages and antigen-presenting cells of the body. PCV2 virus nucleic acid could be detected, and the main manifestations of PCV2 infection were lymphocyte loss and monocyte infiltration. Therefore, it causes the destruction of the pig's immune system, and the immunity is suppressed. The main site of PCV2 proliferation is B lymphocytes, and B lymphocytes also induce apoptosis of lymphocytes after PCV2 infection, causing immunosuppression. In the process of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/0781C12N7/00
CPCC12N5/0635C12N5/0636C12N5/0646C12N7/00C12N2750/10031
Inventor 胡庭俊尹丹谭红连韦英益郝祝兵杨剑
Owner GUANGXI UNIV