Extracellular vesicle and application thereof in prevention and treatment of HPV (human papillomavirus) infection

Abstract

The invention belongs to the technical field of biological medicine, and particularly discloses an extracellular vesicle and application thereof in prevention and treatment of HPV infection. The preparation method comprises the following steps: carrying out co-fermentation on composite probiotics, collecting a fermentation supernatant, and carrying out ultrafiltration to obtain a composite probiotic fermentation active substance; adding into a mesenchymal stem cell complete culture medium to prepare a treatment culture medium; culturing the mesenchymal stem cells by using a treatment culture medium, collecting a culture solution, centrifuging, and collecting supernate; and sequentially carrying out differential centrifugation, filtering with a 0.22 mu m filtering membrane, and purifying to obtain the extracellular vesicles. The extracellular vesicle loaded antiviral component derived from mesenchymal stem cells has excellent biocompatibility, low immunogenicity and targeting property, can effectively neutralize HPV virus and inhibit proliferation of infected cells, and provides a novel stem cell derived treatment strategy for prevention and treatment of HPV.

Description

Extracellular vesicles and their application in preventing and treating HPV infection Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an extracellular vesicle and its application in the prevention and treatment of HPV infection. Background Technology

[0002] Human papillomavirus (HPV) infection is a major causative factor for various malignant tumors, including cervical cancer, anal cancer, and oropharyngeal cancer, especially persistent infection with high-risk HPV types. Globally, the prevention and control of HPV-related diseases is a significant public health challenge. Currently, the main prevention and control measures for HPV infection include preventative vaccines and therapeutic surgeries or physical therapies targeting the lesions. However, preventative vaccines are ineffective against existing infections, and their accessibility and coverage remain limited; while therapeutic methods are mostly invasive procedures, potentially leading to risks such as bleeding, infection, and cervical insufficiency, and cannot fundamentally eliminate the virus, resulting in the possibility of recurrence. Therefore, developing a novel biotherapy strategy that can safely and effectively inhibit or eliminate HPV infection has significant clinical implications and application value. In recent years, extracellular vesicles derived from mesenchymal stem cells have shown great promise in regenerative medicine and disease treatment due to their biological functions, low immunogenicity, good biocompatibility, and targeted delivery potential. Extracellular vesicles carry bioactive substances such as proteins, nucleic acids, and lipids, which can mediate intercellular communication and regulate the physiological and pathological processes of recipient cells. Existing studies have explored its anti-tumor, anti-inflammatory, and immunomodulatory effects, but how to specifically enhance its efficacy against specific pathogens (such as HPV) remains a challenge for current research. Summary of the Invention

[0003] This invention aims to provide an extracellular vesicle derived from mesenchymal stem cells and its application in the prevention and treatment of HPV infection. The vesicle is obtained by inducing mesenchymal stem cells to secrete through the fermentation of probiotic active substances. It can target and interfere with the survival and proliferation of HPV-infected cells and has significant antiviral capabilities.

[0004] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: a method for preparing extracellular vesicles, comprising the following steps: S1, co-fermenting compound probiotics, collecting the fermentation supernatant, ultrafiltration, and obtaining compound probiotic fermentation active substances; S2, adding the compound probiotic fermentation active substances obtained in S1 to mesenchymal stem cell complete culture medium to prepare a treatment culture medium; S3, culturing mesenchymal stem cells in the treatment culture medium prepared in S2, collecting the culture medium, centrifuging, and collecting the supernatant; S4, purifying the supernatant collected in S3 by differential centrifugation and filtration through a 0.22 μm filter membrane to obtain extracellular vesicles.

[0005] Preferably, in S1, the compound probiotics consist of Lactobacillus crispatus, Lactobacillus rhamnosus, and Lactobacillus plantarum.

[0006] Preferably, in S1, the compound probiotics are composed of Lactobacillus crispatus ATCC33820, Lactobacillus rhamnosus ATCC 7469, and Lactobacillus plantarum ATCC 14917 in an inoculation ratio of 1-1.5:1.5-2:1.

[0007] Preferably, in S2, the compound probiotic fermentation active substance prepared in S1 is added to the mesenchymal stem cell complete culture medium at a volume ratio of 5-20% to prepare a treatment culture medium.

[0008] Preferably, in S3, the cultivation conditions are as follows: cultivation at 37-42℃ and 5-10% CO2 for 24-48 hours.

[0009] Preferably, in S4, the specific process parameters for differential centrifugation are as follows: the culture medium is centrifuged at 4°C in the following order: 300-500×g for 8-12 min, and the precipitate is discarded; the supernatant is centrifuged at 2000-3000×g for 15-25 min, and the precipitate is discarded; the supernatant is centrifuged at 10000-12000×g for 30-40 min, and the precipitate is discarded; the final supernatant is collected.

[0010] The present invention also provides extracellular vesicles prepared by the aforementioned preparation method.

[0011] The present invention also provides the use of the extracellular vesicles in the preparation of medicaments for the prevention and / or treatment of HPV infection.

[0012] The present invention also provides a pharmaceutical composition for the prevention and / or treatment of HPV infection, comprising the extracellular vesicles and a pharmaceutically acceptable carrier.

[0013] Preferably, the pharmaceutically acceptable carrier is one or more of the following: solvent, dispersant, suspending agent, surfactant, isotonic agent, thickener, preservative, solid binder, or lubricant.

[0014] Compared with existing technologies, this invention has the following advantages and technical effects: This invention utilizes co-fermentation of active substances from compound probiotics to specifically induce mesenchymal stem cells, causing them to secrete extracellular vesicles loaded with antiviral components. These stem cell-derived vesicles possess excellent biocompatibility, low immunogenicity, and targeting ability, effectively neutralizing HPV viruses and inhibiting the proliferation of infected cells, providing a novel stem cell-derived therapeutic strategy for HPV prevention and treatment. These vesicles can directly interact with HPV viral particles, exhibiting an inhibition rate of up to 78.5% against HPV16 pseudovirus infection. Furthermore, these vesicles can effectively inhibit the proliferation of SiHa cells, strongly induce apoptosis, and arrest the cell cycle at the G0 / G1 phase. These extracellular vesicles can target and interfere with the survival and proliferation of HPV-infected cells, providing a promising non-surgical, minimally invasive therapeutic candidate for treating persistent HPV infection and related lesions.

[0015] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0016] Figure 1 shows the statistical changes in TNF-α concentration in macrophages under different treatment groups; Figure 2 shows the statistical changes in IL-6 concentration in macrophages under different treatment groups; Figure 3 shows the statistical changes in IL-10 concentration in macrophages under different treatment groups; Figure 4 shows the statistical changes in IL-12p70 concentration in macrophages under different treatment groups; Figure 5 shows the statistical changes in the expression ratio of CD80 (M1 type marker) on the surface of macrophages in different treatment groups; Figure 6 shows the statistical changes in the expression ratio of CD206 (M2 type marker) on the surface of macrophages in different treatment groups; Figure 7 shows the statistical changes in the viral infection inhibition rate in different treatment groups. Detailed Implementation

[0017] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0018] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0019] Source of test materials: Unless otherwise specified, all other test materials and instruments in this invention are conventional test materials in the field and can be purchased through commercial channels.

[0020] Example 1: An extracellular vesicle, prepared by the following steps: S1, Lactobacillus crispatus ATCC 33820, Lactaseibacillus rhamnosus ATCC 7469, and Lactiplantibacillus plantarum ATCC 14917 was inoculated into MRS liquid medium and anaerobically activated to the logarithmic growth phase at 37℃. The three probiotics were inoculated at a volume ratio of 1:1.8:1 and co-inoculated into MRS fermentation medium. Fermentation was carried out under anaerobic conditions at 37℃ for 48 hours. After fermentation, the fermentation broth was centrifuged at 4℃ and 8000×g for 15 minutes. The supernatant was collected and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 10 kDa. The filtrate was replaced with sterile PBS buffer to obtain concentrated compound probiotic fermentation active material, which was stored at -80℃ for later use. S2: The compound probiotic fermentation active material prepared in S1 was added to DMEM complete medium at a volume ratio of 10%, mixed thoroughly, and filtered through a 0.22 μm filter membrane for sterilization to prepare the treatment medium. S3: Human umbilical cord-derived mesenchymal stem cells were routinely cultured in DMEM complete medium containing 10% fetal bovine serum. The well-grown third-generation mesenchymal stem cells were digested, counted, and then cultured at 5×10⁻⁶ cells / mL. 4 The cells were seeded at a density of cells / well in 6-well plates. After the cells adhered, the original culture medium was discarded, and 2 mL of the above-treated culture medium was added to each well. The plates were then incubated at 37°C and 5% CO2 for 36 h. The culture medium was collected, centrifuged at 200×g for 10 min, and the supernatant was collected. S4. The supernatant collected in S3 was subjected to differential centrifugation at 4°C: centrifuged at 400×g for 10 min, and the supernatant was collected; centrifuged at 2500×g for 20 min, and the supernatant was collected; centrifuged at 11000×g for 35 min, and the final supernatant was collected. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and centrifuged at 100,000×g for 70 min at 4°C. The supernatant was discarded, and the precipitate was resuspended in an appropriate amount of sterile PBS to obtain purified extracellular vesicles (EVs) derived from mesenchymal stem cells.

[0021] Example 2: An extracellular vesicle was prepared using the following steps: S1. *Lactobacillus crispatus* ATCC 33820, *Lactobacillus rhamnosus* ATCC 7469, and *Lactobacillus plantarum* ATCC 14917 were inoculated into MRS liquid medium and anaerobically activated to the logarithmic growth phase at 37°C. The three probiotics were then co-inoculated into MRS fermentation medium at a volume ratio of 1.5:1.5:1 and co-fermented at 39°C under anaerobic conditions for 36 hours. After fermentation, the fermentation broth was centrifuged at 4°C and 8000×g for 15 minutes. The supernatant was collected and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 10 kDa. The filtrate was then replaced with sterile PBS buffer. The concentrated compound probiotic fermentation active substance was obtained and stored at -80℃ for later use; S2, the compound probiotic fermentation active substance prepared in S1 was added to DMEM complete medium at a volume ratio of 10%, mixed evenly, and filtered through a 0.22μm filter membrane for sterilization to prepare the treatment medium; S3, mesenchymal stem cells derived from human umbilical cord were routinely cultured in DMEM complete medium containing 10% fetal bovine serum. The third-generation mesenchymal stem cells with good growth were digested, counted, and then cultured at 8×10⁻⁶. 4 The cells were seeded at a density of cells / well in 6-well plates. After the cells adhered, the original culture medium was discarded, and 2 mL of the above-treated culture medium was added to each well. The plates were then incubated at 39°C and 8% CO2 for 48 h. The culture medium was collected, centrifuged at 200×g for 10 min, and the supernatant was collected. S4. The supernatant collected in S3 was subjected to differential centrifugation at 4°C: centrifuged at 300×g for 12 min, and the supernatant was collected; centrifuged at 2000×g for 25 min, and the supernatant was collected; centrifuged at 10000×g for 40 min, and the final supernatant was collected. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and centrifuged at 100,000×g for 70 min at 4°C. The supernatant was discarded, and the precipitate was resuspended in an appropriate amount of sterile PBS to obtain purified extracellular vesicles derived from mesenchymal stem cells.

[0022] Example 3: An extracellular vesicle, prepared by the following steps: S1, Lactobacillus crispatus ATCC 33820, Lactaseibacillus rhamnosus ATCC 7469, and Lactiplantibacillus plantarum ATCC 14917 was inoculated into MRS liquid medium and anaerobically activated to the logarithmic growth phase at 37℃. The three probiotics were inoculated at a volume ratio of 1.5:2:1 and co-inoculated into MRS fermentation medium. Co-fermentation was carried out at 39℃ under anaerobic conditions for 36 hours. After fermentation, the fermentation broth was centrifuged at 4℃ and 8000×g for 15 minutes. The supernatant was collected and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 10kDa. The filtrate was replaced with sterile PBS buffer to obtain concentrated compound probiotic fermentation active material, which was stored at -80℃ for later use. S2: The compound probiotic fermentation active material prepared in S1 was added to DMEM complete medium at a volume ratio of 20%, mixed thoroughly, and filtered through a 0.22μm filter membrane for sterilization to prepare the treatment medium. S3: Human umbilical cord-derived mesenchymal stem cells were routinely cultured in DMEM complete medium containing 10% fetal bovine serum. The well-grown third-generation mesenchymal stem cells were digested, counted, and then cultured at 8×10⁻⁶. 4 The cells were seeded at a density of cells / well in 6-well plates. After the cells adhered, the original culture medium was discarded, and 2 mL of the above-treated culture medium was added to each well. The plates were then incubated at 42°C and 10% CO2 for 24 h. The culture medium was collected, centrifuged at 200×g for 10 min, and the supernatant was collected. In step S4, the supernatant collected in step S3 was subjected to differential centrifugation at 4°C: centrifuged at 500×g for 8 min, and the supernatant was collected; centrifuged at 3000×g for 15 min, and the supernatant was collected; centrifuged at 12000×g for 30 min, and the final supernatant was collected. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and centrifuged at 100,000×g for 70 min at 4°C. The supernatant was discarded, and the precipitate was resuspended in an appropriate amount of sterile PBS to obtain purified extracellular vesicles derived from mesenchymal stem cells.

[0023] Comparative Example 1: An extracellular vesicle, prepared by the following steps: S1, Lactiplantibacillus plantarum ATCC 14917 was inoculated into MRS liquid medium and anaerobically activated to the logarithmic growth phase at 37℃. It was then inoculated into MRS fermentation medium and fermented anaerobically at 37℃ for 48 hours. After fermentation, the fermentation broth was centrifuged at 4℃ and 8000×g for 15 minutes, and the supernatant was collected. The supernatant was concentrated by ultrafiltration using a 10kDa molecular weight cutoff membrane. The filtrate was replaced with sterile PBS buffer to obtain concentrated probiotic fermentation active material, which was stored at -80℃ for later use. S2: The probiotic fermentation active material prepared in S1 was added to DMEM complete medium at a volume ratio of 10%, mixed thoroughly, and filtered through a 0.22μm filter membrane for sterilization to prepare the treatment medium. S3: Human umbilical cord-derived mesenchymal stem cells were routinely cultured in DMEM complete medium containing 10% fetal bovine serum. The well-grown third-generation mesenchymal stem cells were digested, counted, and then cultured at 5×10⁻⁶. 4 The cells were seeded at a density of cells / well in 6-well plates. After the cells adhered, the original culture medium was discarded, and 2 mL of the above-treated culture medium was added to each well. The plates were then incubated at 37°C and 5% CO2 for 36 h. The culture medium was collected, centrifuged at 200×g for 10 min, and the supernatant was collected. S4. The supernatant collected in S3 was subjected to differential centrifugation at 4°C: centrifuged at 400×g for 10 min, and the supernatant was collected; centrifuged at 2500×g for 20 min, and the supernatant was collected; centrifuged at 11000×g for 35 min, and the final supernatant was collected. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and centrifuged at 100,000×g for 70 min at 4°C. The supernatant was discarded, and the precipitate was resuspended in an appropriate amount of sterile PBS to obtain extracellular vesicles.

[0024] Comparative Example 2: An extracellular vesicle was prepared using the following steps: S1. Human umbilical cord-derived mesenchymal stem cells were routinely cultured in DMEM complete medium containing 10% fetal bovine serum. The well-grown third-generation mesenchymal stem cells were digested, counted, and then cultured at 5 × 10⁻⁶ cells / mL. 4 The cells were seeded at a density of cells / well in 6-well plates. After the cells adhered, the original culture medium was discarded, and 2 mL of serum-free culture medium was added to each well. The plates were then incubated at 37°C and 5% CO2 for 36 h. The culture medium was collected, centrifuged at 200×g for 10 min, and the supernatant was collected. S2. The supernatant collected in S1 was subjected to differential centrifugation at 4°C: 400×g for 10 min, and the supernatant was collected; 2500×g for 20 min, and the supernatant was collected; 11000×g for 35 min, and the final supernatant was collected. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and centrifuged at 100,000×g for 70 min at 4°C. The supernatant was discarded, and the precipitate was resuspended in an appropriate amount of sterile PBS to obtain extracellular vesicles.

[0025] The effects of the extracellular vesicles provided in Examples 1-3 and Comparative Examples 1-2 were verified through the following experiments.

[0026] 1. The effect of extracellular vesicles on macrophage polarization: The specific experimental protocol is as follows: Human mononuclear cell line THP-1 was used as a model. First, it was stimulated with 100 nM phorbol ester (PMA) for 48 h to induce differentiation into resting-state (M0) macrophages. Morphological observation confirmed their adhesion and extension. Subsequently, the M0 macrophages were divided into five groups for intervention: Group A: THP-1 cells without PMA induction; Group B: PMA-induced M0 macrophages + fresh culture medium; Group C: M0 + LPS 100 ng / mL; Group D: M0 + LPS 100 ng / mL + extracellular vesicles from Example 1 20 μg / mL; Group E: M0 + LPS 100 ng / mL + extracellular vesicles (EVs) from Comparative Example 1 20 μg / mL; Group F: M0 + LPS 100 ng / mL + extracellular vesicles (EVs) from Comparative Example 2 20 μg / mL. All treatments lasted for 24 h.

[0027] After treatment, the cell culture supernatant was collected, and the levels of key cytokines were detected by enzyme-linked immunosorbent assay (ELISA). The results are shown in Figures 1-4. Simultaneously, cells were collected for flow cytometry analysis to detect the expression of surface markers CD80 (M1 type) and CD206 (M2 type). The results are shown in Figures 5-6.

[0028] As shown in Figures 1-4, compared with the LPS group, the EVs provided in Example 1 significantly inhibited the secretion of pro-inflammatory factors TNF-α, IL-6, and IL-12p70, while greatly increasing the secretion of the anti-inflammatory factor IL-10. The EVs provided in Comparative Examples 1 and 2 showed only slight regulatory effects, far lower than the regulatory effect of Example 1.

[0029] As shown in Figures 5 and 6, the EV treatment in the experimental group significantly reduced the proportion of cells expressing CD80 from 82.5% in the LPS group to 35.2%, while increasing the proportion of cells expressing CD206 from 12.6% to 58.7%, indicating that it effectively reversed the macrophage phenotype from the pro-inflammatory M1 phenotype to the anti-inflammatory / repair M2 phenotype.

[0030] In vitro anti-HPV activity verification: HPV pseudovirus neutralization experiment, the specific experimental protocol is as follows: susceptible 293FT cells were infected with HPV16 pseudovirus (PsV) carrying luciferase reporter gene.

[0031] The experiment was divided into the following groups: Virus control group (CON group): HPV PsV only; Group D: HPV PsV + EVs prepared in Example 1, 20 μg / mL; Group E: HPV PsV + EVs prepared in Comparative Example 1, 20 μg / mL; Group F: HPV PsV + EVs prepared in Comparative Example 2, 20 μg / mL; Blank group (cells only).

[0032] After pre-incubating each EV sample with HPV16 PsV at 37°C for 1 hour, 293FT cells were infected. Cells were lysed 48 hours after infection, and luciferase activity (RLU value) was measured. The viral infection inhibition rate was calculated using the following formula: The experimental results are shown in Figure 7.

[0033] As shown in Figure 7, compared with the virus control group, the EVs of Example 1 significantly inhibited viral infection, with an inhibition rate of 78.5%. In contrast, the inhibition rates of EVs of Comparative Example 1 and Comparative Example 2 were only 22.3% and 15.8%, respectively. This result demonstrates that the extracellular vesicles prepared by the method of this invention have highly efficient HPV virus neutralization capabilities and can directly block the initial infection of host cells by the virus.

[0034] HPV-positive cell model experiment: The HPV16-positive cervical cancer cell line SiHa was used. The experimental groups were as follows: blank control group (CON group): routine culture; group D: added with Example 1 EVs, 20 μg / mL; group E: added with Comparative Example 1 EVs, 20 μg / mL; group F: added with Comparative Example 2 EVs, 20 μg / mL.

[0035] After 72 hours of treatment, the cells in each group were analyzed as follows: cell proliferation inhibition was detected by CCK-8 assay; cell apoptosis was detected by Annexin V-FITC / PI double staining flow cytometry; and cell cycle distribution was analyzed by PI single staining flow cytometry. The results are shown in Table 1.

[0036] Table 1. Effects of extracellular vesicles on HPV16-positive SiHa cells in each group

[0037] As shown in Table 1, regarding cell proliferation, the EVs treatment in Example 1 inhibited SiHa cell proliferation by 61.9%, significantly outperforming the two comparative studies. Secondly, regarding apoptosis, flow cytometry analysis revealed that the EVs in Example 1 significantly increased the late apoptosis rate of SiHa cells from 5.9% in the control group to 33.4%. Finally, regarding the cell cycle, the EVs treatment in Example 1 resulted in significant cell cycle arrest in the G0 / G1 phase for SiHa cells, with a substantial reduction in the proportion of cells in the S phase.

[0038] In summary, the extracellular vesicles prepared by co-fermentation of compound probiotics to induce mesenchymal stem cells have dual anti-HPV biological functions: on the one hand, they can directly neutralize HPV virus particles and effectively block their infection of host cells, demonstrating preventive potential; on the other hand, for infected HPV-positive cells, they can effectively inhibit cell proliferation, induce apoptosis, and cause cell cycle arrest, demonstrating therapeutic potential.

[0039] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for preparing extracellular vesicles, characterized in that, The process includes the following steps: S1, co-fermenting the compound probiotics, collecting the fermentation supernatant, ultrafiltration, and obtaining the compound probiotic fermentation active substance; S2, adding the compound probiotic fermentation active substance obtained in S1 to the complete culture medium for mesenchymal stem cells to prepare a treatment culture medium; S3, culturing the mesenchymal stem cells in the treatment culture medium prepared in S2, collecting the culture medium, centrifuging, and collecting the supernatant; S4, purifying the supernatant collected in S3 by differential centrifugation and filtration through a 0.22 μm filter membrane to obtain extracellular vesicles.

2. The preparation method according to claim 1, characterized in that, In S1, the compound probiotics consist of Lactobacillus crispatus, Lactaseibacillus rhamnosus, and Lactiplantibacillus plantarum.

3. The preparation method according to claim 2, characterized in that, In S1, the compound probiotics are composed of Lactobacillus crispatus ATCC 33820, Lactaseibacillus rhamnosus ATCC 7469, and Lactiplantibacillus plantarum ATCC 14917 in an inoculation ratio of 1-1.5:1.5-2:

1.

4. The preparation method according to claim 1, characterized in that, In S2, the compound probiotic fermentation active substance prepared in S1 is added to the mesenchymal stem cell complete culture medium at a volume ratio of 5-20% to prepare a treatment culture medium.

5. The preparation method according to claim 1, characterized in that, In S3, the specific cultivation conditions are: cultivation at 37-42℃ and 5-10% CO2 for 24-48 hours.

6. The preparation method according to claim 1, characterized in that, In S4, the specific process parameters for differential centrifugation are as follows: the culture medium is centrifuged at 4°C in the following order: 300-500×g for 8-12 min, and the precipitate is discarded; the supernatant is centrifuged at 2000-3000×g for 15-25 min, and the precipitate is discarded; the supernatant is centrifuged at 10000-12000×g for 30-40 min, and the precipitate is discarded; the final supernatant is collected.

7. Extracellular vesicles prepared by the preparation method according to any one of claims 1-6.

8. The use of the extracellular vesicles as described in claim 7 in the preparation of medicaments for the prevention and / or treatment of HPV infection.

9. A pharmaceutical composition for the prevention and / or treatment of HPV infection, characterized in that, It includes the extracellular vesicles of claim 7, and also includes a pharmaceutically acceptable carrier.

10. The pharmaceutical composition according to claim 9, characterized in that, The pharmaceutically acceptable carrier is one or more of the following: solvent, dispersant, suspending agent, surfactant, isotonic agent, thickener, preservative, solid binder, or lubricant.

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