CN109627318A - An epitope polypeptide that blocks chicken PD-1/PD-L1 signaling pathway and its application - Google Patents
An epitope polypeptide that blocks chicken PD-1/PD-L1 signaling pathway and its application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70503—Immunoglobulin superfamily
- C07K14/70521—CD28, CD152
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
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Abstract
The invention discloses a kind of epitope polypeptide for blocking chicken PD-1/PD-L1 signal path and its application, the amino acid sequences of the polypeptide are as follows: SGFYYCGLITFSRSLKVVESSHS.The invention discloses the epitope polypeptides that one can effectively block PD-1/PD-L1 signal path, and inhibit the recovery of situation and reversal to study immunity of organism after blocking PD-1/PD-L1 signal path under chicken immune holddown.The present invention passes through research polypeptide and IBDV infected chicken PBMCs binding ability, and then study the restitution of T lymphocyte Proliferation Ability after polypeptide effect, and the expression conditions of immunological regulation relevant cell factor IL-2, IL-6 and IFN-γ etc., so that immunosupress becomes the potentiality target spot of immunization therapy clinical application caused by the activation of PD-1/PD-L1 signal path, and the blocking inhibitor for polypeptide as a kind of new immunologic test point provides theoretical foundation, to obtain the polypeptide drugs and immunologic adjuvant vaccine based theoretical that improve immunological effect.
Description
Technical field
The present invention relates to block chicken PD-1/PD-L1 signal path epitope polypeptide and its application, belong to molecular pathology with
Immunological technique field.
Background technique
The programmed death factor (programmed death-1, PD-1 or CD279) be a kind of inhibition in T cell surface by
Body, the negative regulator as immune response play key effect in maincenter and periphery immune response.There are two ligands by PD-1
PD-Ls (Ligand of Programmed-1) is respectively PD-L1 and PD-L2, is compared to PD-L1, and PD-L2 expression is more limited
System, but PD-L2 has higher affinity to PD-1.It has been proved that T cell contact expression its ligand PD-L1 of expression PD-1
Or PD-L2 cell when, PD-1/PD-Ls signal path activation, the activation signal of the T cell receptor of inhibition response antigenic stimulus
Function, including proliferation, cytokine secretion and cytotoxicity reduce.It is logical that PD-1/PD-L1 is important immune negative regulator signal
Road plays important negative regulator in organism adaptation is immune and makees as the costimulatory signal of t cell immune response
With.
PD-1 and its ligand, which interact during infection or tumor regression or lower during self tolerance development, exempts from
Epidemic disease response.Chronic antigen stimulation, such as the stimulation occurred during tumor disease or chronic infection, cause T cell to express PD-1
Level increases.Verified tumour occurs and viral persistence infection can cause body T cell inhibitive ability of immunity receptor PD-1
And its expression quantity variation of ligand PD-Ls, and then PD-1/PD-L1 signal path is activated, cause under cytotoxic t cell activity
It adjusts, inhibits the proliferation activity of cell, influence the immune response such as programmed death of each immunocyte, then in central immune and outside
Premunition tolerance and immunosupress signal in week immune response cause the immunosupress and viral persistence infection of body.Resistance
Break can effectively restore after the access or reverse the T cell immune function due to virus infection or the initiation of certain diseases exhaust or
Inhibit, provides new idea and method to improve immune level and inhibitive ability of immunity disease.
It is found in chronic infection system, blocks PD-1/PD-L1 interaction that T cell activity can be enhanced.Chronic lymphatic
Cellularity chorion meningitis virus infecting mouse discovery, by blocking PD-1/PD-L1 also to show the improvement of virus sweep rate
With the recovery of immunity.In addition, by blocking PD-1/PD-L1 that can restore function CD8 tired out during chronic viral infection+T cell.It can restore the T of HIV patient, HCV patient or HBV patient by the monoclonal antibodies block PD-1/PD-L1 of PD-L1
Cells in vitro antigen-specific sexual function.Other than enhancing to the immune response of chronic antigen, also shows and block the way PD-1/PD-L1
Diameter can enhance the reaction to vaccine inoculation, carry out therapeutic vac-cination in the case where being included in chronic infection.By targeting PD-
L1 albumen blocks PD-1/PD-L1 interaction to have been demonstrated that can restore T cells with antigenic specificity in vitro and in vivo is immunized function
Can, including in the case where tumour or chronic infection to the intensified response of vaccine inoculation.Therefore, it is necessary to block PD-L1 and PD-1
The reagent of interaction.
Scientific research institution and drugmaker have carried out the largely list about therapeutic PD-1 and its ligand PD-L1 both at home and abroad at present
Clonal antibody development, some drugs come into clinical investigation phase.A large amount of zooperies and experiment in vitro result of study
Show to close PD-1 and its be combineding with each other for ligand using monoclonal antibody specific, the T cell that immune function is exhausted
Function has obtained effective recovery and reverse, secretion, cytotoxic T cell killing including anti-inflammatory and viral type cytokines
Ability, immune cell propagation activity, ctl response activity, antibody-secreting ability etc., accelerate the removing of internal cause of disease.Therefore, this
So that PD-1 and its ligand-mediated signal path become the potentiality target spot of immunization therapy clinical application.But antibody drug has very
More unfavorable factors, such as high production cost, stability, immunogenicity, and become master in primary drug resistance and secondary resistance
Want obstacle.Therefore, it is introduced into for PD-1/PD-L1 micromolecular inhibitor, micromolecular inhibitor is in dynamics and stability side
Face provides inherent advantages.And the blocking inhibitor for polypeptide as a kind of new immunologic test point provides theoretical foundation.And mesh
Preceding polypeptide is also rarely reported as the relevant information of blocking preparation.
Summary of the invention
In view of the deficiencies of the prior art, the object of the present invention is to provide block the epitope of chicken PD-1/PD-L1 signal path more
Peptide and its application.
To achieve the goals above, the technical scheme adopted by the invention is that:
Block the amino acid sequence of the epitope polypeptide of chicken PD-1/PD-L1 signal path are as follows:
SGFYYCGLITFSRSLKVVESSHS。
Polypeptide is located at the IgV structural domain FG section of PD-1 albumen.
A kind of epitope polypeptide answering in IBDV infection of PBMCs s cell experiment blocking chicken PD-1/PD-L1 signal path
With.
A kind of epitope polypeptide blocking chicken PD-1/PD-L1 signal path answering in the recovery metainfective cell factor of IBDV
With.
It is a kind of block chicken PD-1/PD-L1 signal path epitope polypeptide block PD-1/PD-L1 signal path, improve
Application in PBMCs proliferative capacity.
It is a kind of block chicken PD-1/PD-L1 signal path epitope polypeptide improve cellular immunity in application.
The invention has the advantages that:
Research confirms, after infections chicken cloacal bursa virus IBDV infection, as virus replicates in body, and the expression of PD-1
Amount gradually rises, and PD-1 and the immune negative regulator signal that its ligand PD-L1 interacts and then activation PD-1/PD-L1 is important are logical
Road；The expression of the activation of PD-1/PD-L1 signal path and immunological regulation relevant cell factor IL-2, IL-2 and IFN-γ changes
Between there are certain relationship, the activation of PD-1/PD-L1 signal path causes chicken T lymphocyte immune function and proliferation activity
Decline, cause healthy chicken or subclinical infection chicken immune to inhibit the lasting presence of reaction.The present invention is to chicken PD-1 and PD-L1 phase
The epitope polypeptide of interaction is screened, and is caused to block the activation of the PD-1/PD-L1 signal path due to caused by virus infection
Immunosupress provide theoretical direction.
The invention discloses the epitope polypeptides that one can effectively block PD-1/PD-L1 signal path, and inhibit to chicken immune
Block immunity of organism after PD-1/PD-L1 signal path that the recovery of situation and reversal is inhibited to be studied under state.The present invention
By research polypeptide and IBDV infected chicken PBMCs binding ability, and then study the extensive of T lymphocyte Proliferation Ability after polypeptide effect
The expression conditions of multiple effect and immunological regulation relevant cell factor IL-2, IL-6 and IFN-γ etc., so that PD-1/PD-
Immunosupress becomes the potentiality target spot of immunization therapy clinical application caused by the activation of L1 signal path, and is polypeptide as one
The blocking inhibitor of the new immunologic test point of kind provides theoretical foundation, to obtain the polypeptide drugs of raising immunological effect and being immunized
Adjuvanted vaccines based theoretical.
The screening technique of chicken PD-1 of the invention and PD-Ls interaction epitope polypeptide, for the immune suppression of detection Chickens Infected
The activation of PD-1/PD-L1 signal path in immunosuppressive condition caused by property disease IBDV (Bursal Disease) processed
Provide technology platform, result of study is the mechanism for further disclosing IBDV immunosupress and occurring, and seek can to block or
Immunosuppressive method caused by avian viral infection etc. is reversed to provide theoretical foundation.
Detailed description of the invention
Fig. 1 is the efficient liquid phase figure of polypeptide of the present invention.
Fig. 2 is the mass spectrogram of polypeptide of the present invention.
Fig. 3 is PBMCs microscopic examination result (20 ×) of the invention.
Fig. 4 is PBMCs flow cytometer scatter plot of the invention.
Fig. 5 is polypeptide of the present invention immunofluorescence in conjunction with PBMCs.In figure, A: unrelated polypeptide negative control group；B: polypeptide
1-4 processing group.
Fig. 6 is streaming scatter plot (left side) and curve graph (right side) of the polypeptide of the present invention in conjunction with PBMCs.In figure, upper row: unrelated
Polypeptide negative control group, lower row: polypeptide 1-4 processing group.
Fig. 7 is polypeptide of the present invention to the PBMCs streaming scatter plot (left side) being proliferated and curve graph (right side).In figure, upper row: unrelated
Polypeptide negative control group, lower row: polypeptide 1-4 processing group.
Fig. 8 is polypeptide of the present invention to CD4+、CD8+The activity influence of T lymphocyte proliferation.In figure, A: healthy chicken group；B:
IBDV infected chicken group；C: unrelated polypeptide processing group；D: polypeptide 1-4 processing group.
Fig. 9 is change of the polypeptide of the present invention to the transcriptional level of PBMCs surface cytokine IL-2, IL-6, IFN-γ gene
Change situation.
Specific embodiment
Specific embodiments of the present invention will be described in further detail with reference to embodiments.
Experimental animal and strain in embodiment: 1 age in days healthy chick, raising is to 4 week old in SPF isolator；IBDV,
LD50106/ 0.1mL is separated and is saved for this laboratory.
The synthesis of embodiment 1, polypeptide 1-4
Using RCSB PDB (http://www.rcsb.org/pdb/home/home.do) to the other provenances announced
PD-1 and PD-L1 interaction compound three-dimensional structure carries out retrieval collection, and is reference structures to chicken PD-1 and PD-Ls using it
Interaction epitope is predicted and is analyzed.It is analyzed by sequence analysis, the table on chicken PD-1 complete encoding sequence IgV section
Bit sequence shares three sections, is located on CC``, C```D and FG section, contains possible epitope polypeptide.
By screening, polypeptide 1-4, amino acid sequence are obtained are as follows: SGFYYCGLITFSRSLKVVESSHS (SEQ ID
NO.1), it is located at the IgV structural domain FG section of PD-1 albumen.The synthesis of polypeptide can give professional Peptide systhesis service company,
It can be used conventional method synthesis, such as solid phase polypeptide synthesis, and with FITC labeling polypeptide 1-4.Polypeptide 1-4 is detected using HPLC
Purity, HPLC is about as the result is shown that 11min or so single narrow peak occurs in elution time, shows that Purity is higher
(Fig. 1), mass spectral analysis (MS) analysis is the result shows that amino acid quantity and sequence are correct (Fig. 2).
The combination of polypeptide and PBMCs that embodiment 2, FITC are marked
2.1 peripheral blood mononuclear cells (PBMCs) separation
(1) acquisition IBDV infects venous blood 4mL under chick and healthy chick's wing respectively, and 1mL anti-coagulants (anti-coagulants is added
For 10% (w/v) sodium citrate) to prevent Hemostatic Oral Liquid solidification, the PBS that 5mL contains 2% (v/v) fetal calf serum is added, resists with above-mentioned
Blood coagulation mixing, piping and druming mix, and are uniformly divided into two equal portions；
(2) it is slowly added to 5mL lymphocyte separation medium (note: movement is had to slowly) from the top that step (1) mixes blood,
So that separating liquid and mixing blood form apparent layering；
(3) horizontal rotor 800r/min, room temperature is centrifuged 15min, and (note: having to trim, makes to shake drop among centrifugal process
As low as minimum, centrifuge raising and lowering speed need to be adjusted to it is minimum, to reduce to cellular damage)；
(4) cell can occur significantly to be layered after being centrifuged, and top layer is diluted plasma layer, and centre is transparent separating liquid
Layer, the tunica albuginea layer between blood plasma and separating liquid is buffy coat, and centrifugation bottom of the tube is red blood cell and granulocyte.It is dripped with capillary
Pipe is careful to draw white cell thin in new centrifuge tube, and the PBS that 5mL contains 2% (v/v) fetal calf serum is added, gently hangs
Floating cell, horizontal rotor 800r/min, room temperature are centrifuged 10min, abandon supernatant；
(5) PBS that 5mL contains 2% (v/v) fetal calf serum is added, repeated centrifugation abandons supernatant, 0.5mL PBS is added to obtain the final product
To PBMCs cell suspension.
(6) flow cytometer, trypan blue microscopy PBMCs vigor.Flow cytometer streaming scatter plot (Fig. 3) and trypan blue
Dye (Fig. 4) the result shows that, the cell purity and vigor extracted is all relatively good, can satisfy the demand of subsequent experimental.
2.2 fluorescence microscopes detect polypeptide 1-4 in conjunction with PBMCs
PBMCs obtains IBDV from said extracted and infects chick, is 10 with PBS adjustment PBMCs6/ mL takes 35 μ L to adjust
PBMCs cell suspension afterwards is added 100 μ L, 4% (w/v) paraformaldehyde and above-mentioned cell is fixed on glass slide, alcohol lights
On flame gently calcination to fix cell.200 μ L 0.3% (v/v) Triton X-100 permeabilized cells 20min, PBS washings 3 are added
It is secondary, 2min/ times；37 DEG C closing 40min, PBS washing 3 times in wet box using 5% (w/v) skimmed milk power-PBST solution,
2min/ times；Filter paper gently sucks extra confining liquid, and the FITC labeling polypeptide 1-4 of final concentration of 0.1mg/mL is added, sets simultaneously
Unrelated polypeptide (amino acid sequence LERDEMYGYVNIARNLDGYQ, SEQ ID NO.2) is set as negative control group, 37 in wet box
DEG C be incubated for 60min.PBS is washed 3 times, 5min/ times.Fluorescence microscopy microscopic observation polypeptide 1-4 situation in conjunction with PBMCs.
Fluorescence microscope is the result shows that (Fig. 5), the unrelated polypeptide of FITC label are adjusted to highest condition in fluorescence field intensity
Under do not observe apparent fluorescence yet, nothing to do with polypeptide group is compared, FITC label the visible apparent fluorescence of polypeptide 1-4 processing group
Staining cell carries out gray analysis to it through Image J software, and discovery polypeptide processing group is 3 times of control group gray value or so,
It further confirms that apparent combination occurs for aforementioned polypeptides 1-4 and PBMCs, there is the PD-1 albumen knot preferably with the surface PBMCs
Conjunction ability.
2.3 Flow cytometry polypeptide 1-4 are in conjunction with PBMCs
PBMCs infects chick from IBDV, with PBS adjustment 106The FITC mark of final concentration of 0.1mg/mL is added in/mL
Remember polypeptide 1-4, while unrelated polypeptide group is set as negative control, it is during which more after mixing in being protected from light on ice in conjunction with 2.5h
Secondary suspension cell, PBS wash 5-8 time, 300 mesh nylon screen filtration cell suspensions, flow cytomery polypeptide 1-4 and
PBMCs combination situation, 20000 cells of Flow cytometry.
Flow cytometry results show (Fig. 6) that the polypeptide 1-4 processing group of FITC label is corresponding under 488nm laser
Apparent fluorescence signal is collected into the channel BB515, compared with the unrelated polypeptide group of FITC label is collected into fluorescence signal about
It is higher by two orders of magnitude, it was demonstrated that apparent combination occurs for polypeptide 1-4 and PBMCs, has the PD-1 egg preferably with the surface PBMCs
White binding ability.
PBMCs proliferation variation after embodiment 3, Flow cytometry polypeptide 1-4 are blocked
It (1) is the influence after further research polypeptide 1-4 effect to PBMCs in-vitro multiplication situation after IBDV infection, in
IBDV attacked poison after 7 days, and separation infects the chick of IBDV and is uninfected by the PBMCs of IBDV chick as sample, separation method respectively
With embodiment 2, cell concentration is adjusted to 10 using RPMI-1640 culture medium6A/mL；
(2) carbox fluorescenceindiacetate succinimidyl ester (CFSE) of 10 μm of ol/L of final concentration is added, 37 DEG C are protected from light
It is incubated for 10min, PBS washes away unbonded CFSE, and the fetal calf serum that pre-cooling is added terminates reaction, 1500r/min, room temperature on ice
It is centrifuged 5min, supernatant is abandoned, washes twice, unbonded CFSE is washed away；
(3) the unmarked polypeptide 1-4 of final concentration of 0.1mg/mL is added, is protected from light is incubated for 2.5h on ice, PBS is washed 3 times,
5min/ times, to wash away unbonded polypeptide；
(4) by above-mentioned PBMCs according to every 100 μ L of hole, 20000 every holes go to 12 orifice plates, and each sample does 3 repetitions,
Then the fetal calf serum of 100 μ L inactivation is added, concentration is concanavalin A (ConA) the 5 μ L, concentration 4mmol/L of 10mg/mL
L-Glutamine 20 μ L, 10 μ L (0.22 μm of filter filtration sterilization) chicken serum, is eventually adding final concentration of 100IU/mL penicillin
With 100 μ g/mL streptomysins, 37 DEG C, 5%CO2It is cultivated 3 days in incubator, the variation of flow cytomery CFSE fluorescence signal
Power determines that PBMC proliferation is strong and weak.
Flow cytometer is the result shows that (Fig. 7), and PBMCs proliferative capacity declines to a great extent compared with healthy chicken after IBDV infects 7 days,
Fluorescence signal is higher, and unrelated polypeptide processing has no big influence to the PBMCs ability of cell proliferation of IBDV infection chick.Polypeptide 1-
The PBMCs fluorescence signal peak value of 4 processing groups obviously deviates to the left, and ratio about 67.3% illustrates that polypeptide 1-4 can not only be combined
PD-1 albumen on PBMCs, and PD-1/PD-L1 signal path can also be blocked, Cell-mediated Immunity is improved, further to grind
The immune function for studying carefully polypeptide 1-4 lays the foundation.
CD4 after embodiment 4, Flow cytometry polypeptide blocks+And CD8+T lymphocyte subgroup proliferative effect
Infection IBDV chick after being cultivated in Example 3 be uninfected by the PBMCs of IBDV chick, according to 105A/
ML, every mL cell suspension are separately added into the chicken CD4 that concentration is phycoerythrin (PE) dye marker of 0.5 μ g/ μ L+2 μ L of monoclonal antibody
With the chicken CD8 of Cy5 dye marker+2 μ L of monoclonal antibody, in being protected from light incubation 30min on ice after mixing well, PBS washs cell, 800r/
Min, room temperature are centrifuged 5min, abandon supernatant, and cell finally is resuspended with 1mL 10% (v/v) 1640 complete medium.Select streaming thin
In born of the same parents' instrument under 488nm laser under the channel PE and 640nm laser the channel APC to chicken CD4+T lymphocyte and chicken CD8+T lymph
Cell is collected and identifies.
Flow cytometer is the result shows that (Fig. 8), CD4 in healthy chicken flock+T lymphocyte and CD8+T lymphocyte accounts for outer respectively
Detection in the 7th day finds chicken CD4 after the 23.8% of all blood mononuclear cells and 19.0%, IBDV infected chicken+And CD8+T lymphocyte
It is down to 9.74% and 12.3% respectively, after unrelated polypeptide processing, chicken CD4+And CD8+Without obvious after T lymphocyte and IBDV infection
Difference, respectively 9.42% and 12.4%；Chicken CD4 after polypeptide 1-4 effect+And CD8+T lymphocyte is restored to 15.4% He
14.9%, while CD4+And CD8+Double positive cells ratio is increased to 11.1%, illustrates polypeptide 1-4 to chicken CD4+/CD8+Double positives
The proliferation of T cell has significantly more promotion effect.
Cell factor IL-2, IL-6 and the variation of IFN-γ transcriptional level after embodiment 5, RT-PCR detection polypeptide blocks
For the chicken infected rear caused immunological effect variation of detection IBDV, the 7th day peripheral blood after acquisition IBDV is chicken infected
Liquid, and be uninfected by the same Japanese instar chickling of IBDV for control, separate lymphocyte, after cell count, with contain 10% (v/v) blood
Clear 1640 culture mediums adjustment cell concentration is 106A/mL is added in 96 porocyte culture plates, 100 holes μ L/, and every hole is added
Concentration is the 5 μ L of ConA of 10mg/mL, and L-Glutamine 20 the μ L, 10 μ L that concentration is 4mmol/L (cross and filter out by 0.22 μm of filter
Bacterium) chicken serum, final concentration of 100IU/mL penicillin and 100 μ g/mL streptomysins, it is eventually adding the polypeptide of final concentration 0.1mg/mL
20 μ L of 1-4, does three repetitions, and unrelated polypeptide is arranged as negative control group, and the IBDV infection chick of polypeptide is not added
Healthy chicken flock PBMCs is arranged as control, in 37 DEG C, 5% CO as blank control in PBMCs2It is cultivated in incubator
Cell is collected after 72h, RNA extracts kit (Dalian treasured bioengineering Co., Ltd) is extracted cell total rna, tried using reverse transcription
(the Dalian treasured bioengineering Co., Ltd) reverse transcription of agent box synthesizes cDNA, detects cell using real-time fluorescence quantitative RT-PCR
Factor variations are horizontal, blocking effect of the analysis polypeptide 1-4 to cell factor IL-2, IL-6 and IFN-γ.
The result shows that (Fig. 9), significant change is had occurred in IL-2, IL-6 and IFN-γ transcriptional level after IBDV infection,
Compared with not attacking malicious healthy chicken control group, the 7th day its expression of IL-2 after the virus infection, which drops to, does not attack malicious healthy chicken
1/3 or so (p < 0.05 *) of control group, although IL-6 expression, which is increased, does not attack malicious healthy chicken control group without significant
Difference, the transcriptional level of IFN-γ do not attack malicious healthy chicken control group and improve 6.3 times (p < 0.01 * *), illustrate that chicken is infecting
After IBDV, there is exception in relevant important cytokine IL-2, IL-6 of Organism immunoregulation and IFN-γ expression.Polypeptide
After 1-4 processing, IL-2 transcriptional level does not attack malicious healthy chicken control group and rises 2.3 times, poor between nothing to do with polypeptide processing group
It is different significant；The more unrelated polypeptide processing group of IL-6 transcriptional level is declined, close with malicious healthy chicken control group is not attacked；IFN-γ turns
The more unrelated polypeptide processing group of record level drops to 2.1 times, although being still higher than normal level, downward trend is significant.
Sequence table
<110>Xinxiang University
<120>a kind of epitope polypeptide for blocking chicken PD-1/PD-L1 signal path and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 23
<212> PRT
<213>chicken (Gallus gallus)
<400> 1
Ser Gly Phe Tyr Tyr Cys Gly Leu Ile Thr Phe Ser Arg Ser Leu Lys
1 5 10 15
Val Val Glu Ser Ser His Ser
20
Claims (6)
1. blocking the epitope polypeptide of chicken PD-1/PD-L1 signal path, which is characterized in that the amino acid sequence of the polypeptide are as follows:
SGFYYCGLITFSRSLKVVESSHS。
2. the epitope polypeptide according to claim 1 for blocking chicken PD-1/PD-L1 signal path, which is characterized in that described more
Peptide is located at the IgV structural domain FG section of PD-1 albumen.
3. a kind of epitope polypeptide as described in claim 1 for blocking chicken PD-1/PD-L1 signal path is in IBDV infection of PBMCs s
Application in cell experiment.
4. a kind of epitope polypeptide as described in claim 1 for blocking chicken PD-1/PD-L1 signal path is after restoring IBDV infection
Cell factor application.
5. a kind of epitope polypeptide as described in claim 1 for blocking chicken PD-1/PD-L1 signal path is blocking PD-1/PD-L1
Signal path improves the application in PBMCs proliferative capacity.
6. a kind of epitope polypeptide as described in claim 1 for blocking chicken PD-1/PD-L1 signal path is in improving cellular immunity
Application.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104761633A (en) * | 2015-03-25 | 2015-07-08 | 新乡学院 | Polypeptide inhibiting porcine PD-1/PD-L1 pathway and application thereof |
| US20150299322A1 (en) * | 2012-08-03 | 2015-10-22 | Dana-Farber Cancer Institute, Inc. | Agents That Modulate Immune Cell Activation and Methods of Use Thereof |
| CN108997478A (en) * | 2018-08-06 | 2018-12-14 | 中国药科大学 | A kind of polypeptide and its application with immunologic test point antagonistic activity |
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2019
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150299322A1 (en) * | 2012-08-03 | 2015-10-22 | Dana-Farber Cancer Institute, Inc. | Agents That Modulate Immune Cell Activation and Methods of Use Thereof |
| CN104761633A (en) * | 2015-03-25 | 2015-07-08 | 新乡学院 | Polypeptide inhibiting porcine PD-1/PD-L1 pathway and application thereof |
| CN108997478A (en) * | 2018-08-06 | 2018-12-14 | 中国药科大学 | A kind of polypeptide and its application with immunologic test point antagonistic activity |
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