CN110590910A - Synthetic peptides capable of enhancing cellular immune response and anti-infection - Google Patents
Synthetic peptides capable of enhancing cellular immune response and anti-infection Download PDFInfo
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- CN110590910A CN110590910A CN201910921479.5A CN201910921479A CN110590910A CN 110590910 A CN110590910 A CN 110590910A CN 201910921479 A CN201910921479 A CN 201910921479A CN 110590910 A CN110590910 A CN 110590910A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
Abstract
The invention relates to a synthetic peptide capable of enhancing cellular immune response and anti-infection effect and application of the synthetic peptide in inducing CD4+Use of T cells to differentiate into Th1 and produce high levels of IFN-gamma. The invention also relates to another synthetic peptide capable of enhancing cellular immune response and anti-infection effect and application of the synthetic peptide in inducing CD4+Use of T cells to differentiate into Th1 and Th17 and produce high levels of IFN-gamma and IL-17A. The invention screens two synthetic peptides which can enhance cellular immune response and anti-infection function and can respectively induce CD4+Differentiation of T cells into Th1 and production of high levels of IFN-gamma and induction of CD4+T cells are differentiated into Th1 and Th17 and produce high-level IFN-gamma and IL-17A, and can be used for improving immunity and anti-infection ability.
Description
Technical Field
The invention belongs to the field of biomedicine, relates to synthetic peptides, and particularly relates to synthetic peptides capable of enhancing cellular immune response and anti-infection effect.
Background
In the anti-infection process, cellular immune response plays an important role, and CD4 is induced by antigen+The T cells are differentiated into different subclasses such as Th1, Th2 and Th17, the Th1 cells mainly secrete IFN-gamma, activate and enhance phagocytosis of macrophages and kill intracellular bacteria, and promote B cells to convert antibody subclasses;th2 cells mainly secrete IL-4, IL-5, IL-13, etc., promote humoral immune response, stimulate IgE, mast cell and eosinophilic granulocyte reaction, and eliminate helminth infection, etc.; th17 cell mainly secretes IL-17 and IL-22, promotes macrophage, monocyte and neutrophil to migrate to infected part, and has effects of resisting extracellular bacteria and fungal infection. Increase antigen-induced CD4+The high level expression of specific IFN-gamma and IL-17A by T cells is the key to vaccine research.
Peptide induced CD4 in vivo+The cellular immune response and the level of IFN-gamma and IL-17 can be used as an immunopotentiator, enhance the immunogenicity of epitope vaccine, improve specific immune response and play a role in preventing bacterial infection. Therefore, it is important to screen synthetic peptides for their ability to enhance cellular immune responses and anti-infectious effects.
Disclosure of Invention
The first purpose of the invention is to provide a synthetic peptide capable of enhancing cellular immune response and anti-infection effect and application thereof.
The second purpose of the invention is to provide another synthetic peptide capable of enhancing cellular immune response and anti-infection effect and application thereof.
The invention is realized by the following technical scheme:
a synthetic peptide capable of enhancing cellular immune response and anti-infection effect, the amino acid sequence of which is shown in SEQ ID No. 1.
The synthetic peptide is used for inducing CD4+Use of T cells to differentiate into Th1 and produce high levels of IFN-gamma.
The amino acid sequence of the synthetic peptide capable of enhancing the cellular immune response and the anti-infection effect is shown as SEQ ID No. 2.
The synthetic peptide is used for inducing CD4+Use of T cells to differentiate into Th1 and Th17 and produce high levels of IFN-gamma and IL-17A.
Adopt above-mentioned technical scheme's positive effect: the invention screens two synthetic peptides which can enhance cellular immune response and anti-infection function and can respectively induce CD4+Differentiation of T cells into Th1 and production of high levels of IFN-gamma and induction of CD4+T cells are differentiated into Th1 and Th17 and produce high-level IFN-gamma and IL-17A, and can be used for improving immunity and anti-infection ability.
Drawings
FIG. 1 is the in vivo induction of CD4 by peptides+(ii) a T cell proliferation outcome;
FIG. 2 is the in vivo induction of CD4 by peptides+Levels of IFN- γ secretion by T cells;
FIG. 3 is the in vivo induction of CD4 by the peptide+T cells secrete IL-17A levels.
Detailed Description
Sources of the biological material in the present invention:
all synthetic peptides were synthesized by Shanghai Chu peptide Biotech, Inc.
The technical solution of the present invention is further described below with reference to specific examples, but the present invention should not be construed as being limited thereto:
example 1
This example illustrates the synthesis of a peptide.
MHC-II molecule restricted peptides were synthesized by on-line software simulation, and 10 derived peptide sequences were synthesized as detailed in Table 1.
TABLE 1 peptide sequences
Example 2
This example illustrates the effect of synthesizing peptides.
1. Immunization of laboratory animals
SPF grade female 6-8 week old BALB/c mice were randomly grouped. After raising for one week, emulsifying the synthetic peptide with Freund's complete adjuvant according to the volume ratio of 1:1, and injecting 100 μ g of peptide into each mouse by leg intramuscular injection; three weeks after the initial immunization, the mice were re-immunized after emulsification of an equal amount of peptide with Freund's incomplete adjuvant. Sterile PBS was also emulsified with adjuvant as a control.
2. Preparation of antigen-presenting cells
Aseptically collecting spleen of non-immunized BALB/c mouse, grinding to obtain cell suspension, adding 1mL erythrocyte lysate, treating at 37 deg.C for 2min, stopping with 10mL RPMI-1640 culture solution, washing cells for three times, adding 10mL RPMI-1640 culture solution (containing 10% fetal calf serum, 100U/mL cyan, and streptomycin), resuspending cells, making into spleen lymphocyte suspension, placing in CO2Incubator (37 ℃, 5% CO)2) Culturing for 24 h; discarding nonadherent cells and cell culture solution, adding 10mL RPMI-1640 culture solution containing 100 μ g mitomycin C, and placing in CO2The adherent cells were blown down and washed 3 times in an incubator to remove mitomycin C, the cells were resuspended using 1mL of RPMI-1640 medium, the count was diluted, and the cell suspension was diluted to 1X 106cells/mL are ready for use.
3、CD4+Preparation of T cells
The spleen lymphocyte suspension was centrifuged, the supernatant was discarded, and the operation was performed according to the instructions of CD4(L3T4) MicroBeads mouse to obtain CD4+T cells. Resuspend cells using 1mL RPMI-1640 medium, dilute count and adjust cell suspension concentration to 5X 106cells/mL are ready for use.
4、CD4+T cell proliferation assay
To a 96-well cell culture plate, 100. mu.L each of the antigen-presenting cells and CD4 were added at an adjusted concentration+T cell suspension and 5. mu.L inducer at a concentration of 1mg/mL, 96 well cell culture plates were placed in CO using equal doses of PBS as negative control2Culturing for 24h in an incubator; then 10. mu.L of CCK-8 solution was added to each well, and the 96 well cell culture plate was placed in CO2Incubating for 4h in an incubator, measuring the absorbance of the sample at the wavelength of 450nm by using a microplate reader, calculating an induction index (SI value), and determining CD4+T cell proliferation. SI ═ OD450 experimental group-OD 450 blank)/(OD 450 non-induced-OD 450 blank group)
5. Cytokine detection
CD4 co-cultured with inducer and antigen-presenting cells for 48h+Transfer of T cell suspension to 2mL EP tubeAnd centrifuging at 2200r/min for 20min, collecting supernatant, and detecting according to the operation process of Dake products, namely Mouse IL-4, IFN-gamma and IL-17A PrecoatedELISA Kit.
6. Statistical analysis
All data were analyzed biometrically using SPSS software for one-way anova, and differences between groups were statistically significant using a t-test, where p is < 0.05, p is < 0.01, and p is < 0.001.
The results are as follows:
1. synthetic peptide induced CD4+T cell proliferation results
Peptide-immunized BALB/c mouse spleen CD4+Specific SI detection of T cells, results that each peptide stimulates CD4+T cells proliferate to different degrees, with peptide 6, peptide 1, peptide 2 and peptide 10 derived stimulating CD4+The proliferation level of T cells is higher, and the SI is more than 1.5; stimulation of CD4 by other derivatized peptides+T cells, SI were all less than 1.5. The results are shown in detail in FIG. 1.
2. Peptide induced CD4 in vivo+Cytokine secretion by T cells
Spleen sorting of peptide immunized mice to obtain CD4+T cells, tested for their induced specificity CD4 following stimulation with each peptide+Levels of IFN-gamma, IL-4 and IL-17 secreted by T cells. As a result, the level of IFN-gamma generated by induction of the peptide 1 and the peptide 2 is obviously increased, and the level of IFN-gamma generated by the peptide 1 is obviously higher than that induced by the peptide 2; the results are shown in detail in FIG. 2. Peptide 2 induced high levels of IL-17, significantly different (p < 0.001) compared to the other peptides; the results are shown in detail in FIG. 3. Peptide 1 induced higher levels of IFN- γ, but decreased IL-17 levels; peptide 2 was able to induce not only higher levels of IFN-. gamma.but also higher levels of IL-17.
In conclusion, the invention screens two synthetic peptides capable of enhancing cellular immune response and anti-infection effect and respectively can induce CD4+Differentiation of T cells into Th1 and production of high levels of IFN-gamma and induction of CD4+T cells are differentiated into Th1 and Th17 and produce high-level IFN-gamma and IL-17A, and can be used for improving immunity and anti-infection ability.
Sequence listing
<110> university of eight agricultural reclamation of Heilongjiang
<120> synthetic peptides capable of enhancing cellular immune response and anti-infectious effects
<130> B012
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Claims (4)
1. A synthetic peptide capable of enhancing cellular immune response and anti-infection effect has an amino acid sequence shown in SEQ ID No. 1.
2. CD4 induction by the synthetic peptide of claim 1+Use of T cells to differentiate into Th1 and produce high levels of IFN-gamma.
3. A synthetic peptide capable of enhancing cellular immune response and anti-infection effect has an amino acid sequence shown in SEQ ID No. 2.
4. The synthetic peptide of claim 2 induced CD4+Use of T cells to differentiate into Th1 and Th17 and produce high levels of IFN-gamma and IL-17A.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114524880A (en) * | 2022-02-10 | 2022-05-24 | 黑龙江八一农垦大学 | RTG fusion protein capable of enhancing immunogenicity and immune protection |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006032472A2 (en) * | 2004-09-22 | 2006-03-30 | Glaxosmithkline Biologicals S.A. | Immunogenic composition for use in vaccination against staphylococcei |
| CN105254719A (en) * | 2015-10-21 | 2016-01-20 | 黑龙江八一农垦大学 | Identification for CD4<+>T cell epitopes of staphylococcus aureus TRAP (target of RNAIII activating protein) and recombinant epitope vaccine adopting CD4<+>T cell epitopes |
| CN109575141A (en) * | 2017-09-29 | 2019-04-05 | 苏州工业园区唯可达生物科技有限公司 | A kind of CD4 helper T lymphocyte epitope fusogenic peptide and its vaccine |
-
2019
- 2019-09-27 CN CN201910921479.5A patent/CN110590910A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006032472A2 (en) * | 2004-09-22 | 2006-03-30 | Glaxosmithkline Biologicals S.A. | Immunogenic composition for use in vaccination against staphylococcei |
| CN105254719A (en) * | 2015-10-21 | 2016-01-20 | 黑龙江八一农垦大学 | Identification for CD4<+>T cell epitopes of staphylococcus aureus TRAP (target of RNAIII activating protein) and recombinant epitope vaccine adopting CD4<+>T cell epitopes |
| CN109575141A (en) * | 2017-09-29 | 2019-04-05 | 苏州工业园区唯可达生物科技有限公司 | A kind of CD4 helper T lymphocyte epitope fusogenic peptide and its vaccine |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114524880A (en) * | 2022-02-10 | 2022-05-24 | 黑龙江八一农垦大学 | RTG fusion protein capable of enhancing immunogenicity and immune protection |
| CN114524880B (en) * | 2022-02-10 | 2023-09-19 | 黑龙江八一农垦大学 | RTG fusion protein capable of enhancing immunogenicity and immunoprotection |
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Inventor after: Tong Chunyu Inventor after: Cui Yudong Inventor after: Li Wanyu Inventor before: Tong Chunlan Inventor before: Cui Yudong Inventor before: Li Wanyu |