DC vaccine and DC-CTL method for sensitizing with tumor antigen polypeptides mediated by cell-penetrating peptide and transfecting with CD40L

A technology of DC-CTL, tumor antigen peptide, applied in the direction of anti-tumor drugs, cancer antigen components, vertebrate antigen components, etc., to achieve the effect of easy transportation and storage, complete removal of tumor cells, and no carcinogenicity

Pending Publication Date: 2021-09-28
浙江圣希澳医学科技有限公司
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Problems solved by technology

[0015] Membrane-penetrating peptide-mediated tumor antigen polypeptide sensitization and transfection of CD40L DC vaccine and

Method used

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  • DC vaccine and DC-CTL method for sensitizing with tumor antigen polypeptides mediated by cell-penetrating peptide and transfecting with CD40L

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Embodiment Construction

[0118] The DC-CTL method for sensitizing and transfecting CD40L with a tumor antigen polypeptide mediated by penetrating peptides is characterized in that: the specific method includes:

[0119] S1: DC and CTL were cultured in vitro; the surface morphology and internal structure of DC were observed by inverted microscope, scanning electron microscope and transmission electron microscope; the expression of CD1a, CD83, CD80, CD86 and HLA-DR on the surface of DC was detected by flow cytometry ; use the thiazolium blue (MTT) method to detect the proliferation-promoting effect of DC cells on T cells in the mixed lymphocyte test (MLR); use the LDH method (lactate dehydrogenase killing test) to detect the specific killing activity of CTL on tumor cells; The in vitro culture of DC and CTL includes standard procedures for in vitro induction of DC culture, standard operating procedures for in vitro CTL high-efficiency expansion and culture technology, identification of dendritic cells, f...

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PUM

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Abstract

The invention belongs to the field of biological medicine and the technical field of cell therapy, and particularly relates to a DC vaccine and a DC-CTL method for sensitizing with tumor antigen polypeptides mediated by cell-penetrating peptide and transfecting with CD40L. The method comprises the following steps: culturing DC and CTL in vitro; jointly applying a supermotif method for antigen epitope prediction, a polynomial scheme and a quantitative motif method to predict the HLA-A2 restrictive tumor antigen peptide and verify the feasibility of antigen epitope prediction; constructing an h-CD40L expression system; transfecting the CD40L expression vector into the DC cells; and carrying out in-vitro and in-vivo immune function test on the DC cells expressing CD40L after sensitization of the CTL with the tumor antigen peptides mediated by the cell-penetrating peptide. On the basis of synthesis and screening of tumor antigen peptides with killing effects on various tumors and a DC induction culture technology, a tumor antigen peptide library is continuously improved, DC cell culture is optimized, and the transfection of the CD40L expression vector into the DC cells, the DC vaccine and DC-CTL method of DC cells transfected with CD40L expression vector and the test of in vitro immune function of DC cells expressing CD40L after sensitization with the tumor antigen peptides mediated by the cell-penetrating peptide are dicussed.

Description

technical field [0001] The invention belongs to the field of biomedicine and the field of cell therapy technology, in particular to a DC vaccine and a DC-CTL method for sensitizing and transfecting CD40L mediated by a membrane-penetrating peptide. Background technique [0002] On the basis of the successful establishment of in vitro DC cell induction culture technology and the prediction of HLA-A2 restricted tumor antigen peptides and the establishment of a tumor antigen peptide library by synthesis and screening, in-depth research was carried out at different times to promote DC maturation, different concentrations of antigen peptide loading DC, and different methods. The effects of antigen-loaded peptides and CD40L transfected DCs on the morphology, phenotype and function of DCs were investigated to optimize the DC maturation promotion scheme, improve the loading efficiency of DCs, the efficiency of DC antigen presentation and the specific anti-tumor response mechanism medi...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/0784C12N5/10A61K39/00A61P35/00G01N21/84G01N23/04G01N23/20G01N15/00G01N15/14
CPCC12N5/0638C12N5/0639A61K39/0011A61P35/00G01N21/84G01N23/04G01N23/20G01N15/00G01N15/14C12N2510/00C12N2502/1121C12N2502/1114C12N2502/99A61K2039/5154A61K2039/5156A61K2039/5158A61K2039/585G01N2015/0065
Inventor 徐健李陶张扬
Owner 浙江圣希澳医学科技有限公司
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