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Polypeptide, and polypeptide composition and application thereof in tumor immunity

A polypeptide composition and fusion polypeptide technology, applied in the field of medical immunology, can solve the problems of induced tolerance, low quantity, low immunogenicity, etc.

Pending Publication Date: 2020-08-11
SHANGHAI CELL THERAPY GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the clinical limitations of using short peptide-loaded DC in DC-based tumor vaccines, the number of effector CTLs and CD8+ memory T cells produced is not high, and the short peptide is easy to degrade. The immunogenicity is low, multiple administrations are required, and tolerance may be induced, and its immune response is transient and / or low-level, and the in vivo and in vitro effects are not good. A polypeptide and a combination of polypeptides are provided. Drugs and their application in tumor immunity

Method used

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  • Polypeptide, and polypeptide composition and application thereof in tumor immunity
  • Polypeptide, and polypeptide composition and application thereof in tumor immunity
  • Polypeptide, and polypeptide composition and application thereof in tumor immunity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0160] The separation of embodiment 1PBMC cell and dendritic cell (DC)

[0161] Isolation of PBMC cells was performed as follows:

[0162] 1. Take 200-400 μL from the blood of the donor subject and measure the concentration of white blood cells with a white blood cell analyzer.

[0163] 2. Preheat Ficoll in a 20°C water bath for more than 20 minutes.

[0164] 3. According to the measured white blood cell concentration, dilute each blood sample with PBS to a white blood cell concentration of 5-9×10 9 / L, the dilution factor is calculated according to the situation.

[0165] 4. Ficoll separation

[0166] Calculate the amount of Ficoll according to the ratio of volume to blood sample: Ficoll=4:3. Add the diluted blood sample to Ficoll slowly along the wall of the centrifuge tube at a uniform speed. After completion, gently hold the centrifuge tube and put it into the centrifuge. Adjust the speed of the centrifuge to 800g for 25 minutes, adjust the speed up to 1g / s, and the sp...

Embodiment 2D

[0182] The detection of embodiment 2DC aggregation effect

[0183] 1) Spread the DCs isolated in Example 1 on a 24-well plate, and add 1 mL of cells to each well at a density of 1×10 6 / mL of DC suspension;

[0184] 2) Divide the wells covered with DC into 7 groups, namely DC control, DC+polypeptide composition 1, DC+MPC1, DC+HB 100-108 - MPC1, DC+polypeptide composition 2, DC+MPC2 and DC+HB 100-108 -MPC2, 3 parallel auxiliary wells in each group, respectively add corresponding polypeptide compositions or polypeptide constructs (MPC) to each experimental group, wherein the final concentrations of polypeptide compositions 1 and 2 are both 60 μg / mL (polypeptide composition The concentration of each polypeptide in the product is 20μg / mL), all polypeptide constructs (MPC1, HB 100-108 -MPC1, MPC2, HB 100-108 The final concentration of -MPC2) is 40 μg / mL, 37 ℃ 5% CO 2 Incubate for 6 hours;

[0185] 3) The cells were washed with PBS, and each group selected one of the three aux...

Embodiment 3

[0188] Fluorescence microscope detection of the DC absorption level of the polypeptide construct of embodiment 3

[0189] 1) Spread the DCs isolated in Example 1 on a 24-well plate, and add 1 mL of cells to each well at a density of 1×10 6 / mL of DC suspension;

[0190]2) Add 10 μg, 20 μg, 40 μg and 80 μg of FITC-labeled different polypeptide constructs (MPC1 and HB 100-108 -MPC1), 37°C 5% CO 2 Incubate for 3 hours and 6 hours respectively, and observe the absorption level of the polypeptide construct by DCs under a fluorescence microscope.

[0191] The result is as Figure 4-Figure 7 shown. Figure 4-Figure 7 DC responses to different concentrations of MPC1 and HB are shown, respectively 100-108 - MPC1 uptake levels after incubation for 3h, 6h. Figure 4 , 6 are the absorption levels of DCs for different concentrations of MPC1 after incubation for 3h and 6h, respectively. Figure 4 , 6 It was shown that the uptake of MPC1 by DCs after incubation for 6h was significan...

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Abstract

The invention provides a polypeptide composition which comprises a peptide fragment derived from one or more of eEF2, HSP105, MCL-1, MUC1, PSF1 and Survivin, and a separated fusion polypeptide comprising peptide fragments which are covalently linked and derived from two or more of eEF2, HSP105, MCL-1, MUC1, PSF1 and Survivin. The polypeptide and the polypeptide composition provided by the invention are high in immunogenicity, can effectively induce DC maturation, can significantly stimulate proliferation of T cells and secretion of IFN-gamma after being incubated with DC and thus can improve lethality of the T cells, is prevented from inducing DC apoptosis and prevented from inducing up-regulation of the expression of an immunosuppressive receptor of DC, and have extremely excellent immunecell treatment potential and potential anti-tumor effect.

Description

technical field [0001] The invention relates to the field of medical immunology, in particular to a polypeptide, a composition containing the polypeptide and its application in tumor vaccines. Background technique [0002] Dendritic cells (DC) are considered to be a unique type of immune cells because of their ability to initiate and regulate innate immunity and acquired immunity. DC itself plays a very critical role in the immune monitoring of tumors. Under normal circumstances, DCs are usually maintained in an immature and unactivated state until they are exposed to optimal immune stimuli, such as inflammatory cytokines, microbial factors or endogenous alarmins (Nie, Y. et al. al. (2016) Alarmins and antitumor immunity. Clin. Ther. 38, 1042–1053). Once activated, DC will rapidly mature and process the antigen, and present the processed antigen to T cells through the Major Histocompatibility Complex (MHC) molecules on its surface. Although all DCs are specialized antigen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N5/0784C12N5/0786A61K39/00A61P35/00
CPCC07K14/82C07K14/47C12N5/0645C12N5/0639A61K39/0011A61P35/00C07K2319/00
Inventor 莫南德·阿德维希陈楚蒙金华君刘祥箴钱其军
Owner SHANGHAI CELL THERAPY GRP CO LTD
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