CN113817028A

CN113817028A - Polypeptide formulations

Info

Publication number: CN113817028A
Application number: CN202010999771.1A
Authority: CN
Inventors: 杨金波; 申載敏; 刘珊; 冉祥; 宋巧玲; 郑英柱; 姚丹; 杨萌琳; 赵晨阳
Original assignee: Qingdao Marine Biomedical Research Institute Co Ltd
Current assignee: Qingdao Marine Biomedical Research Institute Co Ltd
Priority date: 2020-09-22
Filing date: 2020-09-22
Publication date: 2021-12-21

Abstract

On the basis of AI-Deep-Learning technology research, the invention aims at activating MHC-I immune system, researches on structural protein of CoViD-19 virus, predicts a polypeptide sequence with the length of 9-14 amino acids, can obviously activate the immune system of an organism, and lays a foundation for designing safe and effective polypeptide vaccine.

Description

Polypeptide formulations

Technical Field

The present invention relates to biological products, in particular to polypeptide products.

Background

Activated T lymphocytes play an important role in anti-tumor, anti-infection, and autoimmune diseases. Whereas activation of T cells requires two distinct signals. The first signal comes from the interaction of TCR (T cell receptor) and antigen peptide-MHC (major histocompatibility complex) complex, triggering a series of antigen-specific T cell responses, including the activation of intracellular calcineurin, MAPK and NF- κ B two signaling pathways, resulting in downstream molecular events that initiate the expression of a variety of genes, including IL-2, IL-2L receptor (CD25), CD69, CD154, etc. The second signal comes from the co-stimulatory signal generated by the binding of a B7 family molecule on APC (antigen presenting cell) to its ligand CD28 family molecule on T cell, such as B7-1, B7-2 binding to CD28, CTLA-4, this pathway is called the classical B7 pathway. The absence of the second signal renders the T cell anergic, a suitable co-stimulatory signal may reduce the need for the first signal upon T cell activation, and an inhibitory co-stimulatory signal may attenuate the immune response or cause immune tolerance.

The polypeptide vaccine is prepared by synthesizing protective polypeptide or epitope of pathogenic microorganism by chemical synthesis or genetic engineering, connecting the protective polypeptide or epitope to macromolecular carrier, and adding adjuvant. Compared with the conventional vaccine and the gene recombinant vaccine, the polypeptide/epitope vaccine does not contain the virus genome information which poses potential threat to animals, does not integrate or recombine virus genes and host cell genes, does not pose potential threat to animals or human beings, and is a safe vaccine. The polypeptide vaccine has the advantages of strong specificity, high safety, easy storage and the like, and can be optimized by utilizing the linear series connection of multiple antigen epitopes, the MAP space mode, the lipopeptide mode and the epitope gene recombination mode, so that the immune effect of the vaccine is enhanced. The allergen polypeptides from cats immunize the body, successfully induce the production of regulatory T cells, form immune tolerance, and have entered clinical phase II trials. The epitope derived from myelin is used for inducing regulatory CD8+ T cells, and can inhibit the autoimmune encephalomyelitis of mice. In animal experiments, the polypeptide vaccine targeting angiotensin II has a certain curative effect when applied to the prevention and treatment of hypertension and heart failure, and the formation of atherosclerosis can be reduced by using polypeptide from apolipoprotein B100 to induce specific CTL response.

The monomeric MHC/antigen peptide complex and TCR have low affinity and cannot be used directly for the detection of epitope-specific CTLs (cytotoxic T lymphocytes). Using the binding of the thiol group of cysteine to biotin, 1 piece of fluorescently labeled streptavidin was coupled to 4 pieces of biotin-labeled MHC/antigenic peptide complexes to form MHC/antigenic peptide-tetramers. Tetramers greatly increase MHC/antigen peptide stability and its affinity for TCR, which, after binding to TCR, can be detected by flow cytometry. Research shows that a plurality of HTNV (Hantaan virus) -GP (glycoprotein) CTL epitopes with HLA limitation are obtained by flow cytometry analysis detection and identification through a tetramer staining technology.

CoViD-19 is RNA-Virus, and the total genome has a base Sequence length of 29,903, and it is assumed that 10 putative proteins (CDS) produced by CoViD-19 are present.

The protein created by the CoViD-19 is a substance (mainly protein, glycoprotein, etc.) constituting virus particles, and therefore, theoretically, can be detected by the immune system of a healthy human body, and has a high probability of natural cure. However, since CoViD-19 is a relatively novel variant, it takes a long time for our immune system to recognize and respond to CoViD-19, which can lead to fatal results if the immune system develops into a severe infection or disease state before it is started. As we have experienced many times in our mankind, if a "vaccine" of CoViD-19 could be produced quickly or easily, it is likely to prevent in advance the current world-wide crisis faced by the infectivity/fatality/fear of CoViD-19.

Although the immune system of humans is not fully understood, it is well known that proteins produced by "foreign infectious agents (pathogens or viruses)" may produce "immune defense mechanisms" due in part to the clear recognition by our immune system.

Among immune system systems, the MHC-I/MHC-II signal pathway is a relatively detailed immune system, and Artificial Intelligence (AI) which has recently been attracting attention makes the accumulated data in contact with the human body relatively abundant. Our researchers have been studying AI-Deep-Learning technology, which can learn and predict the important "Peptide immune antigen-Peptide ImmunoAntigen" for activating immune system by MHC-I, and finally regulating various diseases by immunomodulation, based on the understanding and research that our researchers study MHC-I activated immune system by AI-Deep-Learning technology means, and predict polypeptides that can be recognized by our immune system for protein structure of CoViD-19 virus, which is of great significance for diagnosis and treatment of virus.

Disclosure of Invention

CoViD-19 can produce 10 proteins in total, and the researchers aim at activating the MHC-I immune system by means of AI-Deep-Learning technology, and most of the predicted peptide sequence with the highest possibility is composed of 9-14 amino acids and 37 in total aiming at the protein structure of the CoViD-19 virus (see Table 1).

Accordingly:

in a first aspect, the invention provides a polypeptide having immune activation, the amino acid sequence of which encodes one or more of SEQ ID NOs: 1-37 (see Table 1) or a derivative polypeptide having at least 95% sequence identity with the amino acid sequence thereof.

Further wherein the amino acid sequence of the polypeptide is encoded by SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or a derivative polypeptide having at least 95% sequence identity to its amino acid sequence.

In a second aspect, the invention provides a viral therapeutic composition comprising one or more polypeptides having an amino acid sequence encoding SEQ ID NOs 1-37 (see table 1) or derivatives thereof having at least 95% sequence identity to the amino acid sequence thereof. The derivative polypeptide can have the same or similar immune activation effect with the corresponding polypeptide in SEQ ID NO 1-37, and preferably the amino acid sequence of the derivative polypeptide has the consistency of 98 percent or 99 percent.

Further, the virus therapeutic composition preferably comprises SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or one or more derivative polypeptides having at least 95% sequence identity to the amino acid sequence thereof. The derivative polypeptide is capable of hybridizing to the corresponding SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33, preferably the amino acid sequences of the derived polypeptides are 98% identical or have a similar immune activation, such as 99%.

In a second aspect, the invention provides a virus detection composition comprising one or more polypeptides having an amino acid sequence encoding a polypeptide sequence of SEQ ID NOs 1-37 (see Table 1) or a derivative polypeptide having at least 95% sequence identity to its amino acid sequence. The derivative polypeptide can have the same or similar immune activation effect with the corresponding polypeptide in SEQ ID NO 1-37, and preferably the amino acid sequence of the derivative polypeptide has the consistency of 98 percent or 99 percent.

Further, the virus detection composition preferably comprises SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or one or more derivative polypeptides having at least 95% sequence identity to the amino acid sequence thereof. The derivative polypeptide is capable of hybridizing to the corresponding SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33, preferably the amino acid sequences of the derived polypeptides are 98% identical or have a similar immune activation, such as 99%.

In a third aspect, the invention provides vectors (e.g. plasmids and recombinant viral vectors) and host cells comprising vectors which express one or more polypeptides having an amino acid sequence encoding SEQ ID NO:1 to 37 (see table 1) or a polypeptide derived therefrom, preferably a polypeptide having an amino acid sequence encoding SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or one or more derivative polypeptides having at least 95% sequence identity to the amino acid sequence thereof. The derivative polypeptide can be compared with a corresponding sequence SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33, preferably the amino acid sequences of the derived polypeptides are 98% identical or have a similar immune activation, such as 99%.

In a fourth aspect the invention provides a method of performing one or more of the following on a subject:

a, up-regulation of cytokines;

inducing proliferation of T cells;

c: promoting antigen-specific T cell immunity;

d: promote CD4+ and/or CD8+ T cell activation;

the method comprises administering to the subject any one of the viral therapeutic compositions described herein, preferably a viral therapeutic composition comprising SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or one or more derivative polypeptides having at least 95% sequence identity to the amino acid sequence thereof. The derivative polypeptide is capable of hybridizing to the corresponding SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33, preferably the amino acid sequences of the derived polypeptides are 98% identical or have a similar immune activation, such as 99%.

Further, the present invention provides a method for treating or preventing an immune-related disorder comprising administering to a subject in need thereof an effective amount of any one of the viral therapeutic compositions of the present invention, said immune-related disorder comprising viral infections including coronavirus infection, influenza virus infection, hepatitis virus infection, and the like.

Further, this treatment is used in combination with another method for treating the viral infection, including other validated effective methods known in the art, such as invasion/fusion inhibitors: apavir, maraviroc, and the like; integrase inhibitors: letergevir; maturation inhibitors Bevirimat; neuraminidase inhibitors: oseltamivir, zanamivir, peramivir, and the like.

The virus of the present invention further includes any viral peptide, polypeptide protein, and any viral microorganism capable of eliciting an immune response, such as hepatitis a, hepatitis b, hepatitis c, herpes simplex, HIV (human immunodeficiency virus), HPV, influenza virus, respiratory syncytial virus, and the like.

The pharmaceutical compositions of the present invention for administration to a subject further include transferring, delivering, introducing or transporting the drug or agent thereof to the subject in any manner, including orally, topically, intravenously, intramuscularly, intranasally, subcutaneously, and the like. The present invention also contemplates the use of a device or term to administer the medicament. Further, the medicament comprises a pharmaceutically acceptable excipient, such as any carrier, diluent, adjuvant or others.

Drawings

FIG. 1: detecting formation of peptide-HLA-I complex

FIG. 2: percent binding of tetramer to CD8+ T cells at 2 hours

FIG. 3: percentage binding of tetramer to CD8+ T cells at 4 hours

FIG. 4: conditions in which tetramer activated CD8+ T cells at 2 hours

FIG. 5: conditions in which tetramer activated CD8+ T cells at 4 hours

Detailed Description

The invention is further illustrated by the following examples.

Example 1: peptide-HLA-I complex formation

The experiments were performed according to the Monomer/Tetramer production protocol of the easYmer HLA-A02: 01 MHC Tetramers Kit from immunaware manufacturer.

1. An appropriate amount of the peptide to be tested (see Table 1) was weighed and dissolved in dimethyl sulfoxide to 1 mM.

2. Positive control peptide (positive control peptide supplied by Kit Immunaware easyme HLA-A02: 01 MHC Tetramers Kit), and sample peptide to be tested was diluted to 25. mu.M with double distilled water.

3. Each reaction tube was prepared on ice according to the following table (see Table 2), and was repeatedly blown and beaten by a pipette to mix well, thereby avoiding the generation of bubbles.

4. And incubating for 48 hours at 18 ℃ after sealing by a sealing tape for standby.

No	size	sequence	No	size	sequence
						1#	1,380	GVAPGTAVLRQW	19#	1,380	RTVYDDGARRVW
2#	1,265	VAPGTAVLRQW	20#	1,265	TVYDDGARRVW
						3#	1,150	VAIKITEHSW	21#	1,035	VSFLAHIQW
4#	1,150	GRVDGQVDLF	22#	1,150	RRVVFNGVSF
						5#	1,265	GRVDGQVDLFR	23#	1,265	YLFDESGEFKL
6#	1,150	KRFKESPFEL	24#	1,035	LYDKLVSSF
						7#	1,035	KRVDWTIEY	25#	1,150	TTDPSFLGRY
8#	1,610	VTDVTQLYLGGMSY	26#	1,150	TEIDPKLDNY
						9#	1,035	IYNDKVAGF	27#	1,265	TEIDPKLDNYY
10#	1,150	VENPDILRVY	28#	1,265	AEAELAKNVSL
						11#	1,035	KLFDRYFKY	29#	1,380	IALKGGKIVNNW
12#	1,150	KSAGFPFNKW	30#	1,035	LLLDRLNQL
						13#	1,035	VENPHLMGM	31#	1,495	ITVEELKKLLEQW
14#	1,150	QEYADVFHLY	32#	1,150	EELKKLLEQW
						15#	1,150	LTNDNTSRYW	33#	1,035	NRFLYIIKL
16#	1,495	ARFPKSDGTGTIY	34#	1,150	KRFDNPVLPF
						17#	1,265	ISMDNSPNLAW	35#	1,150	TEKSNIIRGW
18#	1,035	FLLPSLATV	36#	1,495	RSYLTPGDSSSGW
									37#	1,495	YRFNGIGVTQNVL

Table 1: polypeptide sequence

Table 2: conditions for preparation of peptide-HLA-I complex

	Positive control	Negative control	Sample peptides
				Double distilled water	3μl	4μl	3μl
Peptide (25. mu.M)	1μl		1μl
				6 Xbuffer	1μl	1μl	1μl
easymer	lμl	lμl	lμl

Example 2: flow cytometry-based detection of peptide-HLA-I Complex formation

1. Mu.l of the peptide-HLA-I complex solution was added to 46. mu.l of the dilution buffer and mixed well.

2. Mu.l of peptide-HLA-I complex diluent was added to 50. mu.l of dilution buffer and mixed well.

3. Take 40. mu.l of step 2 dilution to a new EP tube.

4. Streptavidin-coated magnetic beads (6-8 μm) were diluted 45-fold with dilution buffer, 20 μ l each tube was added, and mixed well.

5. Incubate at 37 degrees for 1 hour on a shaker.

6. Add 160. mu.l FACS solution to each tube.

7. Centrifuge at 700g for 3min and discard the supernatant.

8. 200 μ l FACS solution was resuspended, centrifuged at 700g for 3min and the supernatant discarded, the procedure repeated and washed twice more.

9. PE-labeled anti-human monoclonal antibody BBM.1 was diluted 200-fold with FACS solution.

10. Add 50. mu.l antibody dilution to each tube and resuspend, incubate at 4 ℃ in dark for 30 min.

11. Add 150. mu.l FACS solution, centrifuge at 700g for 3min and discard the supernatant.

12. 200 μ l FACS solution was resuspended, centrifuged at 700g for 3min and the supernatant discarded, the procedure repeated and washed twice more.

13. 200 μ l FACS solution was resuspended and analyzed by flow cytometry.

The results show (see fig. 1): the fluorescence intensity of the 18#, 23#, and 30# polypeptides is similar to that of the positive control peptide, indicating that the 18#, 23#, and 30# polypeptides can successfully bind with HLA-I.

Example 3: tetramer correlation:

the experiment was performed according to the Monomer/Tetramer production protocol of easYmer HLA-A02: 01 MHC Tetramers Kit from immunaware manufacturer.

And (3) preparing a tetramer.

1. mu.L of the prepared 500nM polypeptide 18#, 23#, 30# and HLA-I monomer complex was placed in a 0.2mL centrifuge tube and 0.48. mu.L of Streptavidin-BV421 (BD; Cat # 563259; 0.1mg/mL) was added.

2. After mixing, incubation was carried out at 4 ℃ for 1 hour in the dark.

Isolation of human Peripheral Blood Mononuclear Cells (PBMC)

1. Human venous anticoagulated whole blood (EDTA) was taken and diluted with an equal volume of physiological saline.

2. And adding a proper amount of lymphocyte separation liquid into a 15mL centrifuge tube, flatly spreading the diluted blood above the liquid level of the separation liquid, and keeping the interface between the two liquid levels clear.

3. Centrifuge horizontally at room temperature for 600g, 35 min.

4. After centrifugation, the tube can be seen to be divided into three layers, the upper layer is blood plasma and normal saline, the lower layer is mainly red blood cells and granulocytes, and the middle layer is lymphocyte separation fluid. At the interface between the upper and middle layers, there is a white cloud narrow band mainly composed of mononuclear cells, including lymphocytes and monocytes, and platelets.

5. And (4) inserting a pipette into the cloud layer, and sucking the mononuclear cells. Put into another 15mL centrifuge tube, add more than 5 times volume of physiological saline, centrifuge 250g, 10min, wash cells 1 times.

6. 10ml PBS or cell washing solution is added, 10. mu.L is counted, centrifuged 250g for 10min, and the cells are washed 1 time.

Co-incubation of tetramers with PBMC

1. New 1.5mL centrifuge tubes were removed by adding 2X 10^6 PBMC per tube, adding 200. mu.L FACS buffer, centrifuging 700g for 3min, and discarding the supernatant.

2. The tetramer was diluted to 300nmol with FACS buffer, resuspended PBMC in 40. mu.L of diluted tetramer per tube, and incubated at 37 ℃ for 2-4 hours in the dark.

3. The cells were washed once with pre-cooled FACS buffer.

4. Flow-through antibodies (CD3-percp-cy5.5, CD8-APC, CD4-PE-cy7, CD69-PE, from BioLegend) were added and stained for 30min at 4 ℃.

5. The cells were washed once with FACS buffer and tested on the machine.

The results show that after the tetramer is incubated with the PBMC for 2 hours, the flow results detect the positive control peptide tetramer and the T cells with double positive of CD8+, but not the T cells with double positive of 18#, 23#, 30# polypeptide tetramer and CD8+, indicating that after the tetramer is incubated with the PBMC for 2 hours, the positive control peptide tetramer is combined with the CD8+ T cells, and the 18#, 23# and 30# polypeptide tetramer is not combined with the CD8+ T cells (see fig. 2);

after the tetramer is incubated with the PBMC for 4 hours, the flow results detect T cells with positive control peptide, 23# polypeptide tetramer and CD8+ double positive, but no T cells with 18#, 30# tetramer and CD8+ double positive, indicating that 18#, 30# polypeptide tetramer is not bound to CD8+ T cells after the tetramer is incubated with the PBMC for 4 hours, and the positive control peptide tetramer, 23# polypeptide tetramer is bound to CD8+ T cells (see fig. 3).

The results show that after the tetramer is incubated with the PBMC for 2 hours, the flow results detect that the positive control peptide tetramer has CD69+ positive T cells in the cells combined with the CD8+ T cells, but no 18#, 23#, 30# polypeptide tetramer has CD69+ positive T cells in the T cells combined with the CD8+ T cells, which indicates that the positive control peptide tetramer can significantly activate the CD8+ T cells after the tetramer is incubated with the PBMC for 2 hours, and the 18#, 23#, 30# polypeptide tetramer cannot activate the CD8+ T cells. (see FIG. 4)

The results show that after the tetramer is incubated with PBMC for 4 hours, the flow results detect that T cells positive for CD69+ in the T cells bound to CD8+ T cells by the polypeptide tetramer 23#, but not detect that T cells positive for CD69+ in the T cells bound to CD8+ T cells by the polypeptide tetramer 18#, 30# and indicate that the positive control peptide, polypeptide tetramer 18#, 30# and PBMC can not activate CD8+ T cells after being incubated with the tetramer for 4 hours, and the polypeptide tetramer 23# can significantly activate CD8+ T cells. (see FIG. 5).

Sequence listing

<110> Qingdao Marine biological medicine research institute

<120> polypeptide preparation

<160>37

<170> Patent In Version 2.1

<210> 1

<211>12

<212> PRT

<213> "Artificial sequence"

<400> GVAPGTAVLRQW

<210> 2

<211>11

<212>PRT

<213> "Artificial sequence"

<400> VAPGTAVLRQW

<210> 3

<211>10

<212>PRT

<213> "Artificial sequence"

<400> VAIKITEHSW

<210> 4

<211>10

<212>PRT

<213> "Artificial sequence"

<400> GRVDGQVDLF

<210> 5

<211>11

<212>PRT

<213> "Artificial sequence"

<400> GRVDGQVDLFR

<210>6

<211>10

<212>PRT

<213> "Artificial sequence"

<400> KRFKESPFEL

<210>7

<211>9

<212>PRT

<213> "Artificial sequence"

<400> KRVDWTIEY

<210>8

<211>14

<212>PRT

<213> "Artificial sequence"

<400> VTDVTQLYLGGMSY

<210>9

<211>9

<212>PRT

<213> "Artificial sequence"

<400> IYNDKVAGF

<210>10

<211>10

<212>PRT

<213> "Artificial sequence"

<400> VENPDILRVY

<210>11

<211>9

<212>PRT

<213> "Artificial sequence"

<400> KLFDRYFKY

<210>12

<211>10

<212>PRT

<213> "Artificial sequence"

<400> KSAGFPFNKW

<210>13

<211>9

<212>PRT

<213> "Artificial sequence"

<400> VENPHLMGM

<210>14

<211>10

<212>PRT

<213> "Artificial sequence"

<400> QEYADVFHLY

<210>15

<211>10

<212>PRT

<213> "Artificial sequence"

<400> LTNDNTSRYW

<210>16

<211>13

<212>PRT

<213> "Artificial sequence"

<400> ARFPKSDGTGTIY

<210>17

<211>11

<212>PRT

<213> "Artificial sequence"

<400> ISMDNSPNLAW

<210>18

<211>9

<212>PRT

<213> "Artificial sequence"

<400> FLLPSLATV

<210>19

<211>12

<212>PRT

<213> "Artificial sequence"

<400> RTVYDDGARRVW

<210>20

<211>11

<212>PRT

<213> "Artificial sequence"

<400> TVYDDGARRVW

<210>21

<211>9

<212>PRT

<213> "Artificial sequence"

<400> VSFLAHIQW

<210>22

<211>10

<212>PRT

<213> "Artificial sequence"

<400> RRVVFNGVSF

<210>23

<211>11

<212>PRT

<213> "Artificial sequence"

<400> YLFDESGEFKL

<210>24

<211>9

<212>PRT

<213> "Artificial sequence"

<400> LYDKLVSSF

<210>25

<211>10

<212>PRT

<213> "Artificial sequence"

<400> TTDPSFLGRY

<210>26

<211>10

<212>PRT

<213> "Artificial sequence"

<400> TEIDPKLDNY

<210>27

<211>11

<212>PRT

<213> "Artificial sequence"

<400> TEIDPKLDNYY

<210>28

<211>11

<212>PRT

<213> "Artificial sequence"

<400> AEAELAKNVSL

<210>29

<211>12

<212>PRT

<213> "Artificial sequence"

<400> IALKGGKIVNNW

<210>30

<211>9

<212>PRT

<213> "Artificial sequence"

<400> LLLDRLNQL

<210>31

<211>13

<212>PRT

<213> "Artificial sequence"

<400> ITVEELKKLLEQW

<210>32

<211>10

<212>PRT

<213> "Artificial sequence"

<400> EELKKLLEQW

<210>33

<211>9

<212>PRT

<213> "Artificial sequence"

<400> NRFLYIIKL

<210>34

<211>10

<212>PRT

<213> "Artificial sequence"

<400> KRFDNPVLPF

<210>35

<211>10

<212>PRT

<213> "Artificial sequence"

<400> TEKSNIIRGW

<210>36

<211>13

<212>PRT

<213> "Artificial sequence"

<400> RSYLTPGDSSSGW

<210>37

<211>13

<212>PRT

<213> "Artificial sequence"

<400> YRFNGIGVTQNVL

Claims

1. A polypeptide having immune activating effect, wherein the amino acid sequence of the polypeptide is encoded by one or more of SEQ ID NO 1-37 or derivative polypeptides having at least 95% sequence identity with the amino acid sequence thereof.

2. The polypeptide of claim 1, wherein the amino acid sequence of the polypeptide encodes the amino acid sequence set forth in SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or a derivative polypeptide having at least 95% sequence identity to its amino acid sequence.

3. A viral therapeutic composition comprising one or more of the amino acid sequences encoded as SEQ ID NOs 1-37 or a derivative polypeptide having at least 95% sequence identity to the amino acid sequence thereof.

4. The composition of claim 3, comprising an amino acid sequence encoded as SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or a derivative polypeptide having at least 95% sequence identity to its amino acid sequence.

5. A virus detection composition comprising one or more of the amino acid sequences encoded as SEQ ID NOs 1-37 or a derivative polypeptide having at least 95% sequence identity to the amino acid sequence thereof.

6. The composition of claim 5, comprising an amino acid sequence encoded as SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or a derivative polypeptide having at least 95% sequence identity to its amino acid sequence.

7. A host cell comprising an expression vector capable of expressing one or more of the amino acid sequences encoded as SEQ ID NOs 1-37 or derivative polypeptides having at least 95% sequence identity to the amino acid sequences thereof.

8. Use of a polypeptide, which refers to one or more of the amino acid sequences encoded as SEQ ID NOs 1-37 or a derivative polypeptide having at least 95% sequence identity to the amino acid sequence thereof, for the preparation of a viral therapeutic composition that promotes activation of CD4+ and/or CD8+ T cells.

9. Use of a polypeptide, which refers to one or more of the amino acid sequences encoded as SEQ ID NOs 1-37 or derivative polypeptides having at least 95% sequence identity to their amino acid sequences, for the preparation of a pharmaceutical composition for the treatment of a viral infection.

10. The use of claim 9, wherein the polypeptide is a polypeptide whose amino acid sequence encodes the amino acid sequence of SEQ ID NO: 8. SEQ ID NO: 11. SEQ ID NO: 17. SEQ ID NO: 18. SEQ ID NO: 23. SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 33 or a derivative polypeptide having at least 95% sequence identity to its amino acid sequence.