Active polypeptide and application thereof in prevention and / or treatment of HPV (human papillomavirus)

Abstract

The invention provides an active polypeptide and application thereof in prevention and / or treatment of HPV (human papillomavirus), and belongs to the technical field of biological medicine. The invention provides a polypeptide with prevention and / or treatment activity, the sequence of the polypeptide is SEQ ID No.1, and the polypeptide can be directly used as a medicine to effectively activate immunity and inhibit and eliminate human papilloma virus; the polypeptide can be used as a medicine for independent injection. The polypeptide can also induce mononuclear cells (PBMC) in vitro, culture CTL cells capable of specifically killing the HPV, and directly kill tumors or stimulate immune response of the body by transfusing the CTL cells back into the body of a patient, so that the HPV is effectively killed, positive HPV in the body of the patient is converted into negative HPV, and the treatment effect is finally achieved. The polypeptide disclosed by the invention can also be used as a medicine for preventing HPV (human papillomavirus), and can effectively excite strong immune response, so that a large number of immune cells are generated in a body to achieve the effects of protection and prevention.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to an active polypeptide and its application in preventing and / or treating HPV. Background Art

[0002] Human papillomavirus (HPV) is a DNA virus. HPV can be divided into low-risk and high-risk types according to its type and pathogenicity. High-risk HPV is closely related to malignant tumors such as penile cancer and common gynecological cervical cancer.

[0003] At present, HPV preventive vaccines in the domestic and foreign markets only play a preventive role. For patients who have already been infected with HPV, especially for patients with long-term persistent HPV infection, the simple injection of HPV preventive vaccine has little therapeutic effect. More importantly, patients with postoperative recurrence of cervical disease are mostly caused by HPV integrated infection, and preventive HPV vaccines with L1 as the target antigen have little therapeutic effect on integrated HPV infection. Therefore, drugs with dual effects of HPV prevention and / or treatment are urgently needed to be developed. In addition, how to further improve the efficacy of preventing and / or treating HPV infection, especially how to simultaneously improve specificity and immune activity, is still a technical problem that needs to be solved by technicians in this field. Summary of the invention

[0004] The present invention provides an active polypeptide and its use in preventing and / or treating HPV. The polypeptide can be used as a drug for preventing and treating HPV, effectively stimulating a strong immune response and generating a large number of immune cells in the body to achieve the effects of protection and prevention.

[0005] The present invention provides a polypeptide having preventive and / or therapeutic activity, wherein the polypeptide has any of the following amino acid sequences: (a) the amino acid sequence shown in SEQ ID No. 1;

[0006] (b) An amino acid sequence having 80% or more homology with the amino acid sequence shown in (a).

[0007] Preferably, the polypeptide can prevent and / or treat diseases associated with human papillomavirus.

[0008] Preferably, the human papillomavirus-related diseases include at least one of the following: cervical cancer, cervical and vaginal intraepithelial neoplasia (CIN I-III, VIN I-III), polyps and warts.

[0009] The present invention also provides the use of the polypeptide in preparing medicines for preventing and / or treating diseases caused by human papillomavirus.

[0010] The present invention also provides a medicine for preventing and / or treating diseases caused by human papillomavirus, wherein the effective ingredient of the medicine comprises the above polypeptide.

[0011] Preferably, the dosage form of the drug includes injection.

[0012] Preferably, the administration method of the drug includes at least one of the following: subcutaneous administration, intradermal administration, intramucosal administration, submucosal administration, intramuscular administration and intraperitoneal administration.

[0013] The present invention also provides CTL cells with the ability to specifically kill cells infected with human papillomavirus, which are obtained by inducing mononuclear cells in vitro based on the above polypeptide.

[0014] The present invention also provides the use of the CTL cells in preparing medicines for preventing and / or treating diseases caused by human papillomavirus.

[0015] The present invention also provides a medicine for preventing and / or treating diseases caused by human papillomavirus, wherein the effective ingredients of the medicine include the above-mentioned CTL cells.

[0016] Beneficial effects: The present invention provides a polypeptide with preventive and / or therapeutic activity, which can be directly used as a drug to effectively activate immunity, inhibit and eliminate human papillomavirus; the polypeptide can be used as a drug for single injection without adding any cytokines such as interleukin 2 (IL-2), interferon α (IFN-α), interferon γ (IFN-γ), granulocyte colony-stimulating factor (GM-CSF), etc.

[0017] After artificial synthesis, the polypeptide of the present invention can also induce mononuclear cells (PBMC) in vitro, cultivate CTL cells with specificity to kill HPV-infected cells, and then be reinfused into the body to stimulate immune response, effectively kill HPV-infected positive cells in the body, and achieve a therapeutic effect.

[0018] The polypeptide of the present invention can also be used as a drug for preventing HPV infection, effectively stimulating a strong immune response and generating a large number of immune cells in the body to achieve the effect of protecting and preventing the progression of infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is the tumor weight (g) of the experimental group and the control group mice in the prevention group model; Note: * indicates that the tumor weight of the experimental group mice in the prevention model is significantly lower than that of the control group mice (P<0.01);

[0020] Figure 2is the tumor weight (g) of the experimental and control mice in the treatment model; Note: * indicates that the tumor weight of the experimental mice in the treatment model is significantly lower than that of the control mice (P<0.01). DETAILED DESCRIPTION

[0021] The present invention provides a polypeptide having preventive and / or therapeutic activity, wherein the polypeptide has any of the following amino acid sequences: (a) the amino acid sequence shown in SEQ ID No. 1;

[0022] (b) An amino acid sequence having 80% or more homology with the amino acid sequence shown in (a).

[0023] The amino acid sequence of the polypeptide of the present invention includes SEQ ID No. 1: KKKKKKKKALQAIELQLTLETIYNSQYSNE, and the polypeptide has the activity of preventing and / or treating diseases associated with human papillomavirus. The diseases associated with human papillomavirus of the present invention preferably include at least one of the following: cervical cancer, cervical and vaginal intraepithelial neoplasia (CIN I-III, VIN I-III), polyps and warts.

[0024] The polypeptide of the present invention can be directly synthesized by chemical synthesis, such as using the Fmoc solid phase synthesis method to artificially synthesize the polypeptide and perform subsequent experimental operations in the examples.

[0025] The present invention also provides the use of the polypeptide in preparing medicines for preventing and / or treating diseases caused by human papillomavirus.

[0026] The polypeptide of the present invention can be directly used as a drug to effectively activate immunity, inhibit and eliminate human papillomavirus. The polypeptide of the present invention can be used as a drug for single injection without adding any cytokines such as interleukin 2 (IL-2), interferon α (IFN-α), interferon γ (IFN-γ), granulocyte colony stimulating factor (GM-CSF), etc.

[0027] The present invention also provides a medicine for preventing and / or treating diseases caused by human papillomavirus, wherein the effective ingredient of the medicine comprises the above polypeptide.

[0028] The drug of the present invention may contain cytokines or may not contain cytokines. In the absence of any cytokines, the polypeptide is used as a drug, and the dosage form of the drug preferably includes an injection, and its administration method preferably includes at least one of the following: subcutaneous administration, intradermal administration, intramucosal administration, submucosal administration, intramuscular administration, and intraperitoneal administration. The dosage of the drug of the present invention preferably depends on the type and severity of the disease, as well as on individual characteristics, including general health status, age, sex, weight, and tolerance to the drug, etc. The dosage can be 1 to 100 mg / kg, or 2 to 20 mg / kg, for example, 2.7 mg / kg, 2.8 mg / kg.

[0029] The solvent of the injection of the present invention is preferably physiological saline. In Example 1 of the present invention, 2 ml of the physiological saline was used to dissolve 1 mg of the polypeptide. In Example 2 of the present invention, it was confirmed by experiments on C57BL / 6 mice that the polypeptide can be used alone as a drug for treatment and prevention, and the administration concentration of the polypeptide when injected as an injection is preferably 25 μg polypeptide / 100 μl injection. Among them, the injection concentration described in Example 1 is the concentration of the in vitro induction test, and the injection concentration described in Example 2 is the concentration of the polypeptide of the present invention as an injection in the in vivo test of C57BL / 6 mice.

[0030] The present invention also provides CTL cells with the ability to specifically kill human papillomavirus, which are obtained by inducing mononuclear cells in vitro using the polypeptide.

[0031] The present invention preferably utilizes the polypeptide to directly load DC cells, efficiently induces the formation of CTL cells that specifically kill HPV, and then directly kills tumors or stimulates the body's immune response to effectively kill HPV-infected cells by reinfusion into the body, thereby achieving a therapeutic effect.

[0032] The present invention also provides the use of the CTL cells in preparing medicines for preventing and / or treating diseases caused by human papillomavirus.

[0033] The medicine of the present invention can effectively stimulate a strong immune response, causing a large number of immune cells to be produced in the body to achieve the effects of protection and prevention.

[0034] The present invention also provides a medicine for preventing and / or treating diseases caused by human papillomavirus, wherein the effective ingredients of the medicine include the above-mentioned CTL cells.

[0035] When the CTL cells are used as the effective ingredient in the present invention, the cell amount is preferably (0.5 to 2)×10 9 / mL, and PBS or saline was used as the solvent.

[0036] In order to further illustrate the present invention, an active polypeptide provided by the present invention and its application in preventing and / or treating HPV are described in detail below in conjunction with examples, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Example 1

[0038] 1. The active polypeptide shown in SEQ ID No. 1 (hereinafter referred to as peptide A) was artificially synthesized using the Fmoc solid phase synthesis method.

[0039] 2. Preparation of CTL cells loaded with peptide A:

[0040] (1) Collect 50 mL of peripheral venous whole blood from the subject using an anticoagulant tube syringe containing sodium heparin, and separate the blood using lymphocyte separation medium by centrifugation at 2000 r / min for 10 min to obtain the subject's PBMC.

[0041] (2) After centrifugation, aspirate the PBMCs as thoroughly as possible.

[0042] (3) Wash twice with 0.9% saline and adjust the cell concentration to 2-2.5×10 6 Pieces / ml.

[0043] (4) The washed and concentration-adjusted PBMCs were inoculated into a culture flask, and the culture flask was placed in a 37° C., 5% CO 2 incubator for 2 hours, and then the adherent cells and the suspended cells were collected.

[0044] (5) Collection of DC cells from the experimental group and the control group: The adherent cells were placed in the culture flasks of the experimental group and the control group, and 15 ml of serum-free lymphocyte culture medium supplemented with rhGM-CSF (1000U / ml) and rhIL-4 (500U / ml) were added respectively, and placed in a 37°C, 5% CO2 incubator to induce DC cells. On the third day, rhGM-CSF (1000U / ml) and rhIL-4 (500U / ml) were added respectively. On the fifth day, the DC cells in the experimental group were loaded with peptide A at a final concentration of 5μg / ml, and the control group was not loaded with any peptide. On the sixth day, TNF-α at a final concentration of 1000U / ml and 1μg / ml PGE2 were added respectively to induce DC cell maturation. On the seventh day, mature adherent DC cells were harvested and prepared for mixed culture.

[0045] (6) Collection of CTL cells: The suspended cells were resuspended in a lymphocyte culture medium containing 5% autologous serum, and the culture flask was coated with CD3 and CD28 monoclonal antibodies at a final concentration of 500 ng / ml 24 hours in advance to adjust the cell concentration to 2×10 6 / ml, add IFN-γ with a final concentration of 1000U / ml, add to the coated culture flask, and then place the culture flask in a 37°C, 5% CO2 incubator for incubation. On the second day, add IL-7 with a final concentration of 10ng / mL, IL-15 with a final concentration of 20ng / ml, and IL-2 with a final concentration of 1000U / ml, and continue incubation in a 37°C, 5% CO2 incubator. On the fourth day, add serum-free lymphocyte culture medium with a final concentration of 5ng / mL IL-7, a final concentration of 10ng / ml IL-15, and a final concentration of 1000U / ml IL-2, and adjust the cell concentration to 2×10 6 On the 6th day, serum-free lymphocyte culture medium containing IL-12 at a final concentration of 5 ng / mL, IL-15 at a final concentration of 10 ng / ml, and IL-2 at a final concentration of 1000 U / ml was added to adjust the cell concentration to 2×10 6 CTL cells were harvested on day 7 for mixed culture.

[0046] (7) The above CTL cells were mixed with the experimental group and control group DC cells at a ratio of 20:1 and cultured in a 37°C, 5% CO2 incubator.

[0047] (8) Serum-free lymphocyte culture medium containing IL-2 at a final concentration of 1000 U / ml was added every 3 days. After 7 days of culture, DC-CTL cells of the experimental group (loaded with peptide A) and the control group (not loaded with peptide) were obtained.

[0048] 3. CD8+T cell detection:

[0049] Detect the frequency of CD8+T cells in DC-CTL cells loaded with peptide A (hereinafter referred to as peptide A-DC-CTL cells), and use DC-CTL cells without peptide loading as a control: collect cells from the test group (peptide A-DC-CTL cells) and the control group (DC-CTL cells without any peptide loading) at the beginning (D0), middle (D13), and late (D16) of the culture, add fluorescently labeled PE-CD8 according to the reagent instructions, and incubate in the dark for 30 minutes. Wash with PBS and resuspend, take 200μl / tube of cells to be tested and test on the machine. Take the average value of 3 tests.

[0050] Table 1 CD8+T cell frequency in CTL cells of each group (%)

[0051] Group D0 D13 D16 Experimental Group 18.9±2.9a 67.8±5.2a 92.8±5.9a Control group 20.3±2.7a 39.1±2.4b 73.3±3.0b

[0052] Note: The same lowercase letters in the same column indicate that P>0.05 under the conditions of the same period, that is, the difference is not significant; different lowercase letters in the same column indicate that P<0.05 under the conditions of the same period, that is, the difference is significant.

[0053] The results are shown in Table 1. In the middle (D13) and late (D16) stages of DC-CTL cell culture, the proportion of CD8+T cells in peptide A-DC-CTL cells (experimental group) was significantly higher than that in DC-CTL cells not loaded with peptide (control group).

[0054] 4. Detection of cytokine IFN-γ and TNF-α secretion:

[0055] The cell supernatants of the DC-CTL cell culture at the initial (D0), mid-term (D13), and late (D16) stages of the experimental group (loaded with peptide A) and the control group (not loaded with any peptide) were collected, and IFN-γ and TNF-α cytokines were detected according to the instructions of the ELISA kit.

[0056] Table 2 IFN-γ cytokine expression levels in cell supernatants of each group (pg / mL)

[0057]

[0058] Note: The same lowercase letters in the same column indicate that P>0.05 under the conditions of the same period, that is, the difference is not significant; different lowercase letters in the same column indicate that P<0.05 under the conditions of the same period, that is, the difference is significant.

[0059] Table 3 TNF-α cytokine expression levels in the cell supernatant of each group (pg / mL)

[0060]

[0061] Note: The same lowercase letters in the same column indicate that P>0.05 under the conditions of the same period, that is, the difference is not significant; different lowercase letters in the same column indicate that P<0.05 under the conditions of the same period, that is, the difference is significant.

[0062] The results are shown in Tables 2 and 3. In the middle (D13) and late (D16) stages of DC-CTL cell culture, the secretion of IFN-γ and TNF-α in the supernatant of peptide A-DC-CTL cells (experimental group) was significantly higher than that in the supernatant of DC-CTL cells (control group) without peptide loading. This result shows that peptide A can promote the secretion of IFN-γ and TNF-α cytokines in CTL cells.

[0063] 5. In vitro killing test of DC-CTL cells in the experimental group and the control group:

[0064] The target cells used in this experiment are SIHA cells, a cervical cancer cell line infected with HPV16.

[0065] Effector cell 1: Peptide A-DC-CTL cells, effector cell 2: DC-CTL cells not loaded with peptide. Specifically divided into 2 groups: the experimental group is effector cell 1 and target cells, and the control group is effector cell 2 and target cells (Table 4). At the same time, complete culture medium is used as blank control group, and each group has 3 replicates.

[0066] Table 4 Grouping of effector cells and target cells in the experimental and control groups

[0067]

[0068] (1) Cultivation of target cells: SIHA cells were cultured in RPMI-1640 medium containing 10-15% fetal bovine serum and placed in an incubator at 37°C and 5% CO2. The cells were subcultured every 3 days. The cell morphology was observed under a microscope, and target cells in the logarithmic growth phase with a cell viability higher than 90% were used for MTT killing experiments with DC-CTL cells in the experimental group and the control group.

[0069] (2) Culture of effector cells: DC-CTL cells were prepared in the same manner as in the test group and the control group in Example 1.

[0070] (3) Select target SIHA cells in the logarithmic growth phase (1×10 5 / mL), and added to the effector cell control well, effector cell test well, and blank well (100 μL / well) of a 96-well plate, respectively, and cultured overnight in a 37°C, 5% CO2 incubator.

[0071] (4) The next day, observe the target cell morphology under a microscope to see whether they are adherent and growing normally. Remove the target cell supernatant and add 100 μL of lymphocyte culture medium to the blank well. Add 100 μL of effector cells to the effector cell test group and the effector cell control group at a ratio of 5:1, 10:1, or 20:1 and incubate in a 37°C, 5% CO2 incubator for 4-8 h.

[0072] (5) The morphology of effector cells and target cells was observed under a 40x inverted microscope, and the inhibition rate of effector cells killing target cells was detected by MTT method: 20 μL of 0.5% MTT reagent was added to all wells and incubated in a 37°C, 5% CO2 incubator for 4 h.

[0073] (6) Centrifuge, remove the supernatant, add 150 μL DMSO, and shake at low speed in a microplate reader for 10 min. After the formazan crystals are completely dissolved, measure the OD value at a wavelength of 490 nm, and calculate the average OD value and killing inhibition rate of each experimental group and control group.

[0074] Killing inhibition rate % = [1-(experimental group OD-effect cell control group OD-blank group OD) / (target cell control group OD-blank group OD)] × 100%

[0075] Statistical methods:

[0076] Statistical analysis was performed using SPSS 12.0 software. Data were expressed as mean ± standard deviation (x ± s). One-way analysis of variance was used for comparison among multiple groups, and t-test was used for comparison of means among groups. The test level was α = 0.05. P < 0.05 indicated significant differences.

[0077] Table 5 In vitro killing rate of CTL cells in different groups with different effector-target ratios

[0078]

[0079] Note: The same lowercase letters in the same column indicate that P>0.05 under the same effect-target ratio, that is, the difference is not significant; different lowercase letters in the same column indicate that P<0.05 under the same effect-target ratio, that is, the difference is significant.

[0080] The results are shown in Table 5. The effect of peptide A-DC-CTL cells (experimental group) in killing SIHA cells was significantly higher than that of DC-CTL cells not loaded with peptide A (control group) in killing SIHA cells, regardless of the effector-target ratio of 5:1, 10:1 or 20:1 (P < 0.05).

[0081] The above data show that peptide A can efficiently induce the formation of specific CTL cells, effectively inhibiting and targeting the killing of HPV16-infected cervical cancer cell lines.

[0082] Example 2

[0083] In this example, 5-8 week old female C57BL / 6 mice were used, and the C57BL / 6 mice were randomly divided into a test group and a control group to establish a prevention and treatment model of C57BL / 6 mice. Specifically, the target cells were TC-1-HPV16 cells (wherein, TC-1-HPV16 cells were C57BL / C (H-2b) mouse lung epithelial cells co-transformed with HPV16E6, E7 and ras genes), and the immune substance injected into the test group mice was the peptide segment A synthesized in Example 1; the control group mice were injected with 100 μl of sterile PBS for control.

[0084] 1. Prevention model:

[0085] Experimental purpose: To prove that the peptide segment A of the present invention can be used as a drug for preventing HPV, effectively stimulating a strong immune response and causing a large number of immune cells to be produced in the body to achieve the effects of protection and prevention;

[0086] The C57BL / 6 mice in the test group were intradermally injected with the peptide A (25 μg / 100 μL) of the present invention on the 1st, 5th, 9th and 13th days, and the C57BL / 6 mice in the control group were intradermally injected with 100 μL of sterile PBS at the same time. Subcutaneous tumor inoculation was performed on the 3rd day after the last immunization. TC-1-HPV16 cells were collected, counted, and the cell concentration was adjusted to 5×10 6 C57BL / 6 mice were subcutaneously inoculated with 100 μL of Tc-1-HPV16 cell suspension (5×10 5 / mouse). The growth of transplanted tumors was observed every 2 days after inoculation. Finally, the mice were killed on the 30th day after inoculation of transplanted tumors, and the tumor tissues were surgically removed and weighed. The spleen cells of the mice were taken for immune cell killing test (Table 6).

[0087] Table 6 Detailed information of the C57BL / 6 mouse prevention model

[0088]

[0089]

[0090] 2. Determination of HPV-specific immune cell activity

[0091] According to the above, single lymphocytes of the spleen of mice in the test group and the control group were obtained, and the counted cells were placed in a 6-well plate, and cultured in an incubator at 37°C, 5% CO2 with RPMI-1640 culture medium containing peptide A or positive stimulant PMA and 10% fetal bovine serum (IL-21000U / ml) for 5 days, respectively, as effector cells of the test group and the control group. And target cells TC-1-HPV16 cells were added at a ratio of 10:1 and 20:1. The specific steps of the in vitro killing rate detection of immune cells were carried out according to the in vitro killing test of DC-CTL cells in the test group and the control group in Example 1.

[0092] Table 7 In vitro killing rate of immune cells with different effector-target ratios in each group

[0093]

[0094] Note: Different lowercase letters in the same column indicate that P < 0.05 under the same effect-target ratio, i.e., significant difference.

[0095] The results are as follows Figure 1 As shown in Table 7, in the prevention model of C57BL / 6 mice, the tumor weight of C57BL / 6 mice in the test group (injected with peptide A) was significantly lower than that of C57BL / 6 mice in the control group ( Figure 1), and the immune cell killing rate of the test group (injection of peptide A) was significantly higher than that of the control group, whether the effect-target ratio was 10:1 or 20:1 (Table 7). The above test results show that the peptide A of the present invention can effectively prevent the growth of HPV16 cervical cancer cells.

[0096] 3. Treatment Model

[0097] Purpose of the experiment: To prove that peptide A can be directly used as a drug to effectively induce HPV-specific CTL cells to immune and kill HPV, and have a therapeutic effect on HPV-induced related diseases.

[0098] After C57BL / 6 mice were allowed to grow adaptively in the experimental environment for one week, TC-1-HPV16 cells were collected, counted, and the cell concentration was adjusted to 5×10 6 On the first day, 100 μl of TC-1-HPV16 cell suspension was subcutaneously inoculated in the front of the right hind limb of mice, i.e., 5×10 5 The growth of the transplanted tumor was observed every 2 days after inoculation. The observation showed that palpable tumors were generated about two weeks after inoculation with TC-1-HPV16 cell suspension. Therefore, mice were immunized two weeks after inoculation with TC-1-HPV16 cell suspension. Specifically, the experimental group was injected with peptide A (25μg / 100μ1) for immunization. The control group mice were controlled with 100μ1 PBS. Among them, peptide A was inoculated once every two days, for a total of 6 times. The mice were killed on the 30th day after inoculation of the transplanted tumor, and the tumor tissue was surgically removed, weighed and photographed. At the same time, mouse spleen cells were taken for CTL killing test (Table 8).

[0099] Table 8 Detailed information of the treatment model of C57BL / 6 mice

[0100] Experimental Group Control group Mouse species 5-8 weeks old female C57BL / 6 mice 5-8 weeks old female C57BL / 6 mice quantity 10 10 Transplantation time Day 1 Day 1 Time of first immunization Day 16 Day 16 2nd immunization time Day 18 Day 18 3 times of immunization Day 20 Day 20 …… …… …… Last immunization time Day 26 Day 26 Amount of drug inoculated Peptide A (25μg / 100μL) 100 μl PBS Vaccination injection method Intradermal injection Intradermal injection Attack tumor cell types TC-1-HPV16 cells TC-1-HPV16 cells Tumor cell mass <![CDATA[5×10 5 ]]> <![CDATA[5×10 5 ]]> Immune cell detection time Day 30 after tumor transplantation Day 30 after tumor transplantation

[0101] 4. HPV-specific immune cell activity

[0102] According to the above, single lymphocytes of the spleen of mice in the test group and the control group were obtained, and the counted cells were placed in a 6-well plate, and cultured in an incubator at 37°C, 5% CO2 with RPMI-1640 culture medium containing peptide A or positive stimulant PMA and 10% fetal bovine serum (IL-21000U / ml) for 5 days, respectively, as effector cells of the test group and the control group. And the target cell TC-1-HPV16 culture well was added at a ratio of 10:1 and 20:1. The specific steps of the in vitro killing rate detection of immune cells were carried out according to the in vitro killing test of DC-CTL cells in the test group and the control group in Example 1.

[0103] Table 9 In vitro killing rate of immune cells with different effector-target ratios in each group

[0104]

[0105]

[0106] Note: Different lowercase letters in the same column indicate that P < 0.05 under the same effect-target ratio, i.e., significant difference.

[0107] The results are as follows Figure 2 As shown in Table 9, in the treatment model of C57BL / 6 mice, the tumor weight of C57BL / 6 mice in the test group (injected with peptide A) was significantly lower than that of C57BL / 6 mice in the control group ( Figure 2 ), and the immune cell killing rate of the test group (injection of peptide A) was higher than that of the control group, whether the effect-target ratio was 10:1 or 20:1 (Table 9). The above test results show that the peptide A of the present invention can effectively treat HPV16 cervical cancer cells.

[0108] All experimental results of the prevention and treatment models of C57BL / 6 mice indicate that the peptide A of the present invention can be directly injected as a drug and can effectively reduce the tumor weight of C57BL / 6 mice ( Figure 1-Figure 2 ), and specifically kill TC-1-HPV16 cells (Table 7, Table 9), to achieve the effect of preventing and treating HPV infection.

[0109] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A polypeptide having preventive and / or therapeutic activity, characterized in that The polypeptide has any of the following amino acid sequences: (a) the amino acid sequence shown in SEQ ID No. 1; (b) An amino acid sequence having 80% or more homology with the amino acid sequence shown in (a).

2. The polypeptide according to claim 1, characterized in that The polypeptides include those for preventing and / or treating diseases associated with human papillomavirus.

3. The polypeptide according to claim 2, characterized in that The diseases associated with human papillomavirus include at least one of the following: cervical cancer, cervical and vaginal intraepithelial neoplasia, polyps and warts.

4. Use of the polypeptide according to any one of claims 1 to 3 in the preparation of a drug for preventing and / or treating diseases caused by human papillomavirus.

5. A drug for preventing and / or treating diseases caused by human papillomavirus, characterized in that: The active ingredient of the drug comprises the polypeptide according to any one of claims 1 to 3.

6. The drug according to claim 5, characterized in that: The dosage form of the drug includes injection.

7. The drug according to claim 6, characterized in that: The administration method of the drug includes at least one of the following: subcutaneous administration, intradermal administration, intramucosal administration, submucosal administration, intramuscular administration and intraperitoneal administration.

8. CTL cells with the ability to specifically kill cells infected with human papillomavirus obtained by inducing mononuclear cells in vitro based on the polypeptide according to any one of claims 1 to 3.

9. Use of the CTL cells according to claim 8 in the preparation of a drug for preventing and / or treating diseases caused by human papillomavirus.

10. A drug for treating diseases caused by human papillomavirus, characterized in that: The active ingredient of the medicine comprises the CTL cell according to claim 8.

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