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Antigen epitope peptide and application thereof of a kind of novel coronavirus T cell

An antigenic epitope and coronavirus technology, applied in the direction of virus antigen components, viral peptides, viruses, etc., can solve the problem of no research reports on the new coronavirus

Active Publication Date: 2021-10-29
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant research report in the field of new coronavirus

Method used

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  • Antigen epitope peptide and application thereof of a kind of novel coronavirus T cell
  • Antigen epitope peptide and application thereof of a kind of novel coronavirus T cell
  • Antigen epitope peptide and application thereof of a kind of novel coronavirus T cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Prediction of New Coronavirus HLA-A2 Restricted Epitopes

[0059] 1. Experimental method

[0060] Using the MHC-I binding tool (http: / / tools.iedb.org / mhci), SARS-CoV-2 Wuhan strain (NC_045512.2), SARS-CoV-GD01 strain (AY278489.2), MERS-CoV strain (KF600612.1) and human coronavirus OC43 strain (KF530099.1) spike (S), membrane (M) and nucleocapsid (N) protein sequences for T cell epitope prediction. The prediction method used was 2.22 (NetMHCpan-EL) recommended by IEDB, and the MHC allele selection was HLA-A*02:01 and HLA-A*02:06 (the most common HLA class I genotypes in the Chinese population) . Variant sequences for each epitope were extracted from the GISAID database (GISAID.org website) for further analysis.

[0061] 2. Experimental results

[0062] A large number of high-efficiency and specific T cell antigen epitope candidates were screened. Its information is shown in Table 1.

[0063] Table 1 Novel coronavirus T cell antigen epitope peptide:

[00...

Embodiment 2

[0066] Example 2 Does Mitomycin Intervention T2A2 Cells Affect the Combination with Antigen Peptides

[0067] 1. Experimental method

[0068] Take logarithmic growth state T2A2 cells (174x CEM.T2( CRL-1992 TM )), seeded into 96-well plate, 10 per well 5 , select different concentrations of mitomycin (MC) (20μg / mL and 10μg / mL) and different action time (20min and 2h) to intervene in T2A2 respectively, set up 6 groups, respectively, group 1: T2A2 blank group, group 2 : T2A2+MC 10μg / mL 2h, group 3: T2A2+MC 20μg / mL 20min, group 4: T2A2+10μM positive control peptide (influenza A M1 polypeptide GILGFVFTL), group 5: T2A2+MC 10μg / mL 2h+10μM positive Control peptide (influenza A M1 polypeptide GILGFVFTL), group 6: T2A2+MC 20 μg / mL 20 min+10 μM positive control peptide (influenza A M1 polypeptide GILGFVFTL). Each group has 3 replicate wells with a final volume of 200 μL. After incubation at 37°C for 4 hours, centrifuge and wash twice, label with FITC anti-human HLA-A2 (β2m) antibo...

Embodiment 3

[0071] Example 3 Mitomycin inhibits T2A2 cell proliferation

[0072] 1. Experimental method

[0073] Take T2A2 cells in the logarithmic growth state and plant them into 96-well plates, 10 per well 5 , take mitomycin (MC 20μg / mL), the action time (20min) to intervene in T2A2, set up 3 groups, respectively T2A2 blank group, label T2A2 with CFSE (5μM) (day 0), and label T2A2 with CFSE (5μM) (Day 7), each group had three replicate wells, and the final volume was 200 μL. Placed at 37°C, 5% CO 2 The cells were cultured in the incubator, centrifuged and washed twice after the corresponding time, and detected by flow cytometry. The experiment was carried out 3 times.

[0074] 2. Experimental results

[0075] After mitomycin MC intervened in T2A2, it could inhibit the cell proliferation ( figure 1 C and panel D). figure 1 C shows that CFSE labeling was successful (the fluorescence peak of the middle group shifted to the right on day 0). After the 7th day, CFSE still showed a si...

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Abstract

The invention discloses an antigenic epitope peptide of novel coronavirus T cells and application thereof. The amino acid sequence of the antigenic epitope peptide is shown in any one of SEQ ID NO: 1-13. The pMHC complex monomer was prepared by using it, and the pMHC complex multimer was further prepared, which can be used for the detection of antigen-specific T cells in the peripheral blood of patients who recovered from novel coronavirus infection, and for the activation of T cells in vitro Experimentally, these new coronavirus T cell antigen epitope peptides can be applied to new coronavirus-related immune detection and vaccine development, which is worthy of in-depth research and vigorous promotion.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a novel coronavirus T cell antigen epitope peptide and its application. Background technique [0002] The new coronavirus is the pathogen of a new type of infectious pneumonia that occurred at the end of 2019. At present, the source of the virus is still unclear. Gene sequencing shows that it is related to bat SARS-like coronavirus, severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East Respiratory Syndrome MERS-CoV is highly homologous. There is no specific drug report yet. The immune response mechanism of the human immune system to the new coronavirus, including antigen recognition, immune clearance mechanism, and immune memory protection mechanism, is not clear. [0003] According to the research on SARS and other coronaviruses with high homology of the new coronavirus, T cell immune response plays an important role in the body's antiviral defense and immu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/165C07K19/00G01N33/569A61K39/215A61P31/14
CPCC07K14/005C07K14/70539G01N33/56983A61K39/12A61P31/14C12N2770/20022C07K2319/00G01N2333/165C12N2770/20034
Inventor 陈国兵肖潺潺邓洁萍邱聪龄王鹏程罗钧洪高利娟
Owner JINAN UNIVERSITY
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