Mouse macrophage human complement membrane attack complex in vitro model as well as construction and application

Abstract

The invention belongs to the technical field of immune systems, and particularly relates to a mouse macrophage human complement membrane attack complex in vitro model as well as a construction method and application. The mouse macrophage human complement membrane attack complex in vitro model disclosed by the invention is built and formed through the following steps: (1), injecting a thioglycollate culture medium of which 1mL is sterilized and the mass percentage is 3 percent into a mouse abdominal cavity, extracting ascitic fluid after two days, performing centrifugation, removing supernatant and red cells, adding DF 12 culture medium to re-prepare cell suspension, and dropwise adding the cell suspension on a hole plate or a slide to obtain primary macrophage; and (2), adding C5b-6, C7, C8 and C9 into a culture solution of the primary macrophage according to different demands, and performing incubation to form the mouse macrophage human complement in vitro model. The amount of formed MAC(Membrane Attack Complex) can be controlled according to different amount of the human complement components C5b-6, C7, C8 and C9 required to be added in experiments, so that the effects of performing dissolved damage and killing cells or regulating and controlling the cells are achieved.

Description

technical field [0001] The invention belongs to the technical field of the immune system, and in particular relates to an in vitro model of a mouse macrophage human complement membrane attack complex and a construction method and application thereof. Background technique [0002] The complement system is an important part of the human immune system, which participates in the body's specific and non-specific immune mechanisms, manifested as antimicrobial defense responses, immune regulation, and mediated immune pathological damage responses. The complement system is a protein response system composed of more than 30 kinds of serum proteins with cascade enzymatic reactions and self-regulatory mechanisms. Under the stimulation of activators, complement proteins can be activated through the classical or alternative pathways. In the common terminal pathway of the three complement activation pathways, the complement component C5b can be stably combined with C6 to form C5b6, which ...

AI Technical Summary

Problems solved by technology

Therefore, there are the following problems: 1. The complement response may be activated by the classical pathway, the alternative pathway, and the MBL pathway. Their activators are antigen-antibody immune complexes, bacterial fungal viruses, etc., and mannose on the surface of pathogenic microorganisms, etc. Wait

2. The complement provided by the serum is the complement that has not been sheared. When the serum is extracted from a normal animal or human body, it may not be able to form a membrane attack complex because there is no activator but an active complement component.

3. The complement content in the serum of each individual is different, and quantitative analysis is required after each extraction, but the quantitative analysis can only know the total amount of complement, and cannot clarify the content of each complement, so the repeatability is not good

Without knowing the content of each complement, the conditions required for the experiment cannot be controlled

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