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A novel long non-coding RNA, namely lnc-Dpf3, and a sequence, immunological effects and use thereof

A non-coding and sequence technology, applied in the fields of biotechnology and medicine, can solve the problems of unclear mechanism of long-chain non-coding RNA immune function regulation, no reports, etc., to block the progress of autoimmune diseases and inhibit inflammation damage effect

Inactive Publication Date: 2019-05-03
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, no specific long non-coding RNAs involved in the regulation of dendritic cell chemotaxis and / or chemotactic dendritic cell state and / or function have been reported so far, so it is unclear how long non-coding RNAs affect Mechanism of immune function regulation by chemotactic dendritic cells

Method used

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  • A novel long non-coding RNA, namely lnc-Dpf3, and a sequence, immunological effects and use thereof
  • A novel long non-coding RNA, namely lnc-Dpf3, and a sequence, immunological effects and use thereof
  • A novel long non-coding RNA, namely lnc-Dpf3, and a sequence, immunological effects and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1. Culture process of dendritic cells

[0083] Mouse bone marrow cells plus 100ng / mL mouse GM-CSF and 10ng / mL mouse IL-4 cytokine (R&D Systems, Minneapolis, MN) were cultured in a cell culture incubator at 37°C [RPMI-1640 (PAA) cell culture solution containing 10% (v / v) FCS (PAA)] for six days, added LPS (100ng / ml, sigma) to stimulate for 24 hours, induced mature dendritic cells, and then anti-CD11c magnetic beads (Miltenyi Biotech) sorting. Purified dendritic cells obtained by sorting.

Embodiment 2

[0084] Embodiment 2: lnc-Dpf3 quantitative real-time PCR (qRT-PCR) detection

[0085] According to literature records, CCL19 and CCL21 can act on CCR7 on the surface of dendritic cells to induce chemotactic migration (Ohl, L., Mohaupt, M., Czeloth, N., Hintzen, G., Kiafard, Z., Zwirner, J., Blankenstein, T., Henning, G., and R. (2004). CCR7 governs skin dendritic cell migration under inflammatory and steady-state conditions. Immunity 21, 279-288.).

[0086] The dendritic cells ( figure 1 a) Dendritic cells stimulated by CCL19 (50ng / ml) and CCL21 (50ng / ml) (R&D) at various time points ( figure 1 b), using TRIzol (Invitrogen Company) to extract the RNA sample therein. qRT-PCR uses SYBR RT-PCR kit (Takara Company, SYBR Green Realtime PCR Master Mix Code: QPK-201), and operates on LightCycler (Roche Company) real-time quantitative PCR instrument.

[0087] The quantitative primers for qRT-PCR detection of lnc-Dpf3 are:

[0088] 5'-GTGCCACCTACTGTACACCT-3' (upstream, SEQ ID N...

Embodiment 3

[0100] Example 3: Flow cytometry detection of lnc-Dpf3 expression in chemotactic dendritic cells of draining lymph nodes after FITC skin stimulation

[0101] Apply 100 microliters of 5mg / ml FITC (Invitrogen, dissolved in 50:50 (v / v) acetone-dibutyl phthalate (Sigma Aldrich)) to the right back skin of the mouse without hair, at different time points The right inguinal lymph node was taken, FITC+ cells were chemotactic dendritic cells, and FITC- cells were non-chemotactic dendritic cells. Flow cytometry was used to detect chemotactic dendritic cells at different time points after FITC was applied. The proportion of cells and the expression level of lnc-Dpf3 in chemotactic dendritic cells.

[0102] The cell flow cytometry antibody labeling was carried out according to its standard operating procedures. FACSLSRII flow cytometer was used for flow cytometry, and the software was FACSDiva (BD Biosciences). For specific steps, please refer to the papers previously published by our l...

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Abstract

The invention relates to the technical field of biology, and particularly relates to a novel long non-coding RNA, and a sequence, immunological effects and use thereof. The invention relates to applications of the long non-coding RNA (namely lnc-Dpf3) shown as SEQ ID NO:1 or complementary or homologous RNA sequences thereof or expression products thereof or inhibitors or synergists thereof in regulation of dendritic cell chemotaxis capability and / or states and / or functions of chemotactic dendritic cells, or applications in preparation of functional medicines or kits regulating dendritic cell chemotaxis capability and / or states and / or functions of chemotactic dendritic cells. The long non-coding RNA can be applied for regulating dendritic cell chemotaxis capability and / or states and / or functions of chemotactic dendritic cells, and / or further applied for regulating body immune response and steady state balance, and preventing and treating allergic diseases such as contact dermatitis, andautoimmune diseases such as colitis, and applied for tumor immunotherapy and / or prognosis evaluation. The long non-coding RNA has a wide application prospect.

Description

technical field [0001] The present invention relates to the fields of biotechnology and medicine, specifically, a long non-coding RNA——lnc-Dpf3, which plays a role in regulating dendritic cell chemotaxis ability and / or chemotaxis dendritic cell state and / or or functions in the application. Background technique [0002] Mammalian genomes can transcribe a variety of long noncoding RNAs (long noncoding RNAs, lncRNAs), but the sequences and functions of only a few lncRNAs have been clarified. In recent years, the role of lncRNA in the physiological process of life and disease process has attracted continuous attention, but there are not many studies on the relationship between lncRNA and the immune system, especially the reports of lncRNA in dendritic cells are almost blank. [0003] Dendritic cells are a key bridge connecting innate immunity and adaptive immunity, and play a key regulatory role in activating the body's anti-pathogen immune response and maintaining autoimmune t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P37/04A61K31/713G01N33/53
Inventor 刘娟曹雪涛
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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