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Method for researching influence of interferon induced gene IFIT3 on activation and differentiation of B lymphocytes

A technology of interferon induction and B lymphocytes, applied in the field of gene technology, can solve the problems of unclear immunological abnormalities of lupus erythematosus

Pending Publication Date: 2022-03-25
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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Problems solved by technology

At present, the mechanism of immunological abnormalities in systemic lupus erythematosus at home and abroad is still unclear

Method used

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  • Method for researching influence of interferon induced gene IFIT3 on activation and differentiation of B lymphocytes

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Embodiment 1

[0054] The peripheral blood samples were collected in June 2018, and the SLE blood samples came from 4 confirmed volunteers from the Department of Rheumatology and Immunology of the Second Xiangya Hospital of Central South University. All subjects included in our study followed strict inclusion and exclusion criteria. The included SLE patients should follow strict diagnostic criteria, including women of childbearing age under the age of 40, and the diagnosis of SLE complied with the 2009 revision of the American College of Rheumatology classification criteria for SLE.

[0055] A research method for the influence of interferon-inducible gene IFIT3 on the activation and differentiation of B lymphocytes, which comprises the following steps:

[0056] Step 1: Collect peripheral blood from clinically diagnosed SLE patients and healthy controls. Ficoll extracts peripheral blood mononuclear cells (PBMCs). At the same time, immunomagnetic beads are used to sort and purify B cells, and ...

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Abstract

The invention belongs to the technical field of gene science and technology, and particularly relates to a method for researching activation and differentiation of B lymphocytes by an interferon-induced gene IFIT3, which comprises the following steps: step 1, collecting peripheral blood of clinically diagnosed SLE patients and healthy control persons, extracting peripheral blood mononuclear cells (PBMC) by Ficoll, and separating and purifying B cells by adopting immunomagnetic beads; step 2, culturing an in-vitro induced hair growth center B cell (iGCB), and detecting the change condition of an IFIT3 transcript by a fluorescent quantitative PCR (qRTPCR) technology; 3, overexpressing IFIT3 through an iGCB system, inducing and differentiating into plasma cells (iPC), and detecting main regulatory factors IRF4, Pax5 and Bcl6 for differentiating the B cells into the plasma cells through flow analysis; and 4, exploring epigenetic modification of the IFIT3 protein by using technologies such as mass spectrum, co-immunolocalization, Western-blot, molecular cloning and the like, and discussing the influence of the interferon-induced gene IFIT3 on activation and differentiation functions of B lymphocytes, thereby providing a scientific basis for clarifying the pathogenesis of lupus and finding targets for treating lupus in the future.

Description

technical field [0001] The invention relates to the technical field of gene technology, in particular to a research method for the effect of interferon-induced gene IFIT3 on the activation and differentiation of B lymphocytes. Background technique [0002] One of the main functions of B lymphocytes is to produce antibody-mediated humoral immune responses. After BCR gene rearrangement, they can produce antibodies that specifically neutralize antigens. ADCC action and other immune system-related reactions. B lymphocytes can also present soluble antigens and produce cytokines involved in immune regulation. When the immune tolerance of B lymphocytes is abnormal, it can produce abnormal immune response and secrete a large number of autoantibodies and cytokines. A large number of autoantibodies can directly mediate cell destruction or abnormal cell function, thus playing a pathogenic role. Autoantibodies can involve multiple organs and tissues, involving a complex inflammatory r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/50G01N33/68
CPCC12Q1/6883G01N33/6893G01N33/5005C12Q2600/158G01N2800/104
Inventor 黄向阳毛妮彭笑菲陈谊月
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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