Modification method and application of in-vitro immunocyte preparation

A technology of immune cells and preparations, applied in the field of cell biology, can solve problems such as failure to improve function, difficulty in obtaining NSCs, and body damage.

Pending Publication Date: 2017-06-13
SHANXI DATONG UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

With the deepening of NSCs research, more and more theoretical and technical issues need to be further improved and standardized: ①The source selection of NSCs: NSCs are located deep in the nerve tissue, and obtaining autologous NSCs will cause certain damage to the body; if Harvesting NSCs from fetuses, even from aborted fetuses, is ethically problematic
②Number of NSCs: Studies have found that NSCs have a significant dose-dependent effect on the impr...

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  • Modification method and application of in-vitro immunocyte preparation
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  • Modification method and application of in-vitro immunocyte preparation

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Embodiment Construction

[0031] 1. Animals and materials

[0032] 100 female C57BL / 6 mice aged 8-10 weeks were purchased from Beijing Weitong Lihua Company. Myelin oligodendrocyte glycoprotein 35-55 polypeptide (MOG 35-55 ) was prepared by Xi'an Lianmei Biotechnology Co., Ltd.; pertussis toxin (PTX) was purchased from ALEXIS company; complete Freund's adjuvant (CFA), saponin (saponin), rabbit anti-mouse CD4 antibody was purchased from Sigma company; Mycobacterium tuberculosis (TB), PE-labeled IFN-γ, IL-10, IL-17, TGF-β, CD16 / 32, IL-12, CD40, CD206, CD14, Arginase-1 (Arg-1) antibody purchase From BD Company; rabbit anti-mouse nitric oxide synthase (iNOS) antibody was purchased from EnZo Company; fetal bovine serum (BSA) was purchased from Gibco Company; IL-10, IFN-γ, IL-17, IL-1β, TNF -α, IL-6 assay ELISA kits were purchased from Peprotech, USA; CD11b MicroBeads, T cell MicroBeads, and LS separation columns were purchased from Miltenyi, Germany; Fasudil was purchased from Tianjin Hongri Pharmaceutica...

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Abstract

The invention provides a modification method and application of an in-vitro immunocyte preparation. The method comprises the following steps: preparing an EAE (Experimental Autoimmune Encephalomyelitis) model by utilizing a MOG-35-55 peptide fragment in an immunizing manner; inducing encephalitogenic immunocyte: after immunizing, co-culturing spleen T cells and spleen macrophages, and Fasudil for 72 hours at the 9 day, so as to obtain the encephalitogenic immunocyte: Fasudil modified T cells and Fasudil modified macrophages. A treatment effect on an EAE mouse by Fasudil modified immunocyte is observed and a result proves that the Fasudil in-vitro modified immunocyte can be used for effectively alleviating clinical symptoms of the EAE mouse; the treatment effect of the Fasudil modified macrophages is better than that of the Fasudil modified T cells. Furthermore, the Fasudil in-vitro modified immunocyte also has the advantages of small side effect, low treatment cost and the like, and has good clinical application prospect on the treatment of neurodegenerative diseases.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a modification method and application of an in vitro immune cell preparation. Background technique [0002] Multiple sclerosis (MS) is an immune inflammatory demyelinating / degenerative disease of the central nervous system (CNS) whose etiology and pathogenesis have not been fully elucidated. Dysfunction, even paralysis, places a heavy burden on families and society. [0003] So far, there is no effective cure for MS, mainly to inhibit the progression of inflammatory demyelinating lesions, prevent the deterioration of the acute phase of the disease and relapse in the remission phase, and symptomatic and supportive therapy to relieve the pain caused by neurological dysfunction. At present, the treatment of MS patients in China is even more difficult, including drugs approved by the FAD for the treatment of the disease, such as β-interferon (IFN-β), glatimir acetate (GA), natalizumab (N...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/0786A61K35/17A61K45/06A61K35/15A61P37/02A61P25/28
CPCA61K35/15A61K35/17A61K45/06C12N5/0636C12N5/0645
Inventor 刘春云郭尚德尉杰忠李艳花肖保国马存根
Owner SHANXI DATONG UNIV
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