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Application of CD200 for preparing systemic lupus erythematosus psychotherapeutic drugs

A CD200-, lupus erythematosus technology, applied in the field of biomedicine, can solve problems such as unclear effect

Inactive Publication Date: 2012-10-03
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite substantial evidence that CD200 / CD200R1 plays an important role in experimental autoimmune diseases, the role of this signaling pathway in human autoimmune diseases such as SLE remains unclear

Method used

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  • Application of CD200 for preparing systemic lupus erythematosus psychotherapeutic drugs
  • Application of CD200 for preparing systemic lupus erythematosus psychotherapeutic drugs
  • Application of CD200 for preparing systemic lupus erythematosus psychotherapeutic drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Flow cytometry detects the expression of CD200 and CD200R1 on the cell surface

[0027] 1) Patients and healthy controls

[0028] A total of 161 patients with SLE who were treated in the Department of Rheumatology of Peking Union Medical College Hospital from 2009 to 2011 were selected (see Table 1). The diagnoses were all in line with the classification criteria revised by the American College of Rheumatology (ACR) in 1987. All patients were female, aged 12 to 55 years (average 29.0±10.2 years). In addition, 95 healthy controls matched by sex and age were selected. This study was approved by the Ethics Committee of Peking Union Medical College Hospital.

[0029] Table 1 Characteristics of SLE patients (n=161)

[0030]

[0031] The expression of CD200 and CD200R1 on the cell surface was detected by flow cytometry. Separate PBMCs from peripheral blood with lymphocyte separation medium, combine with different fluorescently labeled CD4, CD25, CD11c, CD123,...

Embodiment 2

[0043] Example 2 Antagonistic anti-CD200R1 antibody promotes proliferation of SLE CD4+ T cells driven by TCR activation

[0044] Select 5 cases of SLE samples and 5 cases of healthy control samples in Example 1.

[0045] Understanding its effect on CD4 by measuring the effect of soluble CD200Fc and antagonistic anti-CD200R1 antibody on expression of phosphorylated DOK2 molecule + Effects of CD200 / CD200R1 signaling in T cells.

[0046] CD4 + After T cells were pretreated with or without antagonistic anti-CD200R1, they were co-cultured with recombinant human CD200Fc (isotype IgG Fc as a control). After 5 days, the cells were collected, washed twice with cold PBS, and the cell lysate [containing 20mM HEPES (pH7.9 ), 20% glycerol, 1% NP-40, 1mM MgCl 2 , 0.5mM EDTA, 0.1mM EGTA, 1mM DTT, 1mM PMSF and protease inhibitors (BD Biosciences)] lysate, lysate on ice for 1 hour, shake and mix once every 10 minutes, centrifuge at 13,000rpm at 4°C, SDS-PAGE ( Invitrogen) protein electrophor...

Embodiment 3

[0051] Example 3 Increase of apoptotic lymphocytes in SLE with upregulation of CD200 expression

[0052] Select 45 cases of SLE samples and 15 cases of healthy control samples in Example 1.

[0053] PBMC were cultured in PBS to observe spontaneous apoptosis, or PBMC apoptosis was induced by X-ray accelerator (5Gray), cultured in RPMI 1640 complete medium, and incubated in a 37°C carbon dioxide incubator for 48 hours. Annexin V and propidium iodide (propidium iodide, PI) staining were performed to distinguish living cells, apoptotic cells and necrotic cells, and CD200 fluorescent antibody surface staining was performed, and flow cytometry detection was performed. CD200 + live cells, CD200 - live cells, CD200 + Apoptotic cells, CD200 - Apoptotic cells and necrotic cells were sorted by flow cytometry Moflo (Cytomation, USA). Sorting gating was strictly limited to lymphocyte gates defined by forward and lateral angles.

[0054] Experiments have shown that early spontaneous a...

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Abstract

The invention belongs to the field of biomedicine, in particular to application of CD200 or CD200 fusion protein for preparing systemic lupus erythematosus psychotherapeutic drugs. When sufficient exogenous CD200 molecules are given from external bodies, developmental deficits of adjusting T cell of systemic lupus erythematosus patients can be corrected, developmental differentiation of Th17 is reduced, migration chemotaxis functions of the DC is restrained, and the CD200 molecules are indicated to have potential therapeutic value. Preparation of high affinity CD200 recombinant protein is a conventional technique in the field, application is simple, therapeutical effects can be expressed through adjustment of internal body on the adjusting T cell, the Th17 cell and the CD cell in a multi-link mode, and novel psychotherapeutic drugs of systemic lupus erythematosus can be prepared.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of CD200 or CD200 fusion protein in the preparation of medicines for treating systemic lupus erythematosus. Background technique [0002] Systemic lupus erythematosus (SLE) is an autoimmune disease that can affect multiple organs and tissues. Immunoderegulation defects, especially loss of immune tolerance, autoantibody production, immune complex deposition, cytokine imbalance and The uncontrolled inflammatory response is the main immunopathological mechanism. The dysfunction of phagocytosis prevents apoptotic cells from being cleaned up in time, so that a large number of self-antigens are released to induce autoimmunity, which is considered to be an important link in the initiation of the disease, but the internal mechanism is unclear. In addition, CD4 + CD25 high FoxP3 + Regulatory T cells (Tregs), as a key regulatory component to maintain T cell homeos...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P37/02A61P19/04
Inventor 张烜赵丽丹李扬
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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