An immunological composition and its use in the preparation of vaccines

Abstract

The application provides an immune composition and its use in the preparation of varicella-zoster vaccine, and a varicella-zoster vaccine prepared by the same, the immune composition comprising a subunit component and a recombinant adenovirus vector component. The subunit component provided by the application is a recombinant protein derived from varicella-zoster virus, and the recombinant adenovirus vector component is formed by genetic engineering recombination of an antigen gene sequence derived from varicella-zoster virus and an adenovirus vector. The immune composition provided by the application can effectively stimulate humoral immunity, cellular immunity and mucosal immunity by inhalation administration, intramuscular injection administration and microneedle administration, and is used for preventing or improving infectious diseases caused by varicella-zoster virus, and has a more optimal prevention effect and a wide range.

Description

Technical Field

[0001] This invention relates to the field of vaccine technology, and to an immune composition and its use in the preparation of a varicella-zoster vaccine, as well as the varicella-zoster vaccine prepared therefrom. Background Technology

[0002] Vaccines have gained widespread public acceptance as the most economical and effective means of preventing infectious diseases. To date, vaccine delivery methods and types have undergone numerous upgrades and evolutions. In terms of delivery technology, there has been a gradual shift from traditional injection to non-invasive delivery, with the successful commercialization of inhaled vaccine products in recent years sparking a research boom. Regarding vaccine types, currently widely used vaccines include inactivated vaccines, live attenuated vaccines, protein subunit vaccines, and nucleic acid vaccines, each with its own advantages and limitations.

[0003] After the initial infection with the varicella-zoster virus (VZV) causes chickenpox, it remains dormant in the sensory ganglia for a long time. When the body's immunity declines, the virus can be reactivated, causing shingles and serious complications such as postherpetic neuralgia (PHN). Vaccination against shingles is the most effective way to prevent the disease. Currently, the varicella-zoster vaccines approved for marketing worldwide mainly include two technical routes: live attenuated vaccines and recombinant subunit vaccines. Representative products include GlaxoSmithKline's (GSK) Shingrix (recombinant subunit vaccine) and BEGC Biotech's Ganwei (live attenuated vaccine).

[0004] Despite significant progress in existing varicella-zoster vaccines, several limitations remain. Currently dominant recombinant subunit vaccines require adjuvants, such as AS01B adjuvant in Shingrix. The recombinant varicella-zoster vaccine disclosed in patent CN113683704B is also composed of varicella-zoster virus r-gE fusion protein and an adjuvant. However, the addition of adjuvants can induce strong adverse reactions, significantly impacting people's willingness to be vaccinated. Furthermore, some existing vaccine adjuvant systems themselves have technical bottlenecks. Patent application number 200410033878.1 uses aluminum hydroxide combined with CpG as an adjuvant, where aluminum hydroxide acts as a carrier for CpG. The combined effect stimulates immunity, but the resulting cellular immunity is weak (see the background technology of patent CN113683704B).

[0005] This invention overcomes the above limitations and innovatively develops a mixed vaccine with low antigen content and no need for adjuvants. Furthermore, by combining it with a preferred route of administration, it achieves significant humoral, cellular, and mucosal immune effects, providing an innovative solution for the prevention and control of varicella-zoster virus. Summary of the Invention

[0006] This invention provides an immune composition and its use in the preparation of a varicella-zoster vaccine, as well as the varicella-zoster vaccine prepared therefrom. The immune composition of this invention comprises: (A) a recombinant protein derived from the varicella-zoster virus; and (B) a recombinant adenovirus vector encoding an antigen derived from varicella-zoster virus. The immune composition of this invention is suitable for the preparation of a varicella-zoster vaccine and is suitable for the preparation of inhaled, injected, or microneedle vaccine formulations.

[0007] This invention provides an immune composition and a vaccine prepared therefrom, specifically, which can be used to prepare a varicella-zoster vaccine. The varicella-zoster vaccine has a low antigen content, is adjuvant-free, and can effectively stimulate humoral immunity, cellular immunity, and mucosal immunity. It can be used to prevent or improve infectious diseases caused by varicella-zoster virus, with better preventive effect and wider range.

[0008] This invention provides an immune composition comprising: (A) Recombinant protein derived from varicella-zoster virus; and, (B) A recombinant adenovirus vector encoding an antigen derived from varicella-zoster virus.

[0009] The applicable dose range of the recombinant protein derived from varicella-zoster virus contained in the immune composition is 0.1~2.0 μg / μl; preferably, the applicable dose range of the recombinant protein derived from varicella-zoster virus is 0.2~0.5 μg / μl; more preferably, the applicable dose of the recombinant protein derived from varicella-zoster virus is 0.5 μg / μl.

[0010] In some specific embodiments, the applicable content of recombinant protein derived from varicella-zoster virus in the immune composition is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, in units of ug / μl.

[0011] The applicable dose range of the recombinant adenovirus vector contained in the immune composition is 2 × 10⁻⁶. 7 ~8×10 8 VP / μl; preferably, the applicable dose range of the recombinant adenovirus vector is 1×10⁻⁶. 8 ~4×10 8 VP / μl; More preferably, the applicable dose of the recombinant adenovirus vector is 2 x 10⁻⁶. 8 VP / μl.

[0012] In some specific embodiments, the applicable content of the recombinant adenovirus vector contained in the immune composition is 2 x 10⁻⁶. 7 2.5x10 7 3x10 7 3.5x10 7 4x10 7 4.5x10 6 5x10 7 5.5x10 7 6x10 7 6.5x10 7 7x10 7 7.5x10 7 8x10 7 8.5x10 7 9x10 7 9.5x10 7 1x10 8 1.5x10 8 2x10 8 2.5x10 8 3x10 8 3.5x10 8 4x10 8 4.5x10 8 5x10 8 5.5x10 8 6x10 8 6.5x10 8 7 x 10 8 7.5x10 8 8x10 8 Unit: VP / μl.

[0013] The recombinant adenovirus vector is non-replicating and has a deletion in at least one of the early region 1 (E1 region) and early region 3 (E3 region) of the adenovirus.

[0014] The adenovirus described in this invention is any one of the seven subgroups of human adenovirus or simian adenovirus A, B, C, D, E, F, and G. Any one of the seven subgroups (A, B, C, D, E, F, G) of simian adenoviruses, such as sAd1, sAd10, sAd11, sAd12, sAd13, sAd14, sAd15, sAd16, sAd18, sAd19, sAd2, sAd20, sAd23, sAd25, sAd26, sAd27, sAd28, sAd29, sAd3, sAd30, sAd31, sAd32, sAd33, sAd34, sAd35, sAd37, sAd38, sAd39, sAd4, sAd40, sAd43, sAd44, sAd45, sAd46, sAd47, sAd48, sAd49, sAd6, sAd9, AdY25, AdC8, AdC22, AdC30, AdC37, AdC11. AdC3, AdC17, AdC19, AdC31, AdC20, AdC24, AdC16, AdC26, AdC82,AdC5, AdC7, AdC44, AdC38, AdC43, AdC63, AdC147, AdC73, AdC6, AdC55, AdC83,AdC143, AdC144, AdC145, AdC10, AdC28, AdC9, AdC6 Any one of the seven subgroups A, B, C, D, E, F, and G of Rhesus adenovirus, such as RhAd51, RhAd52, RhAd53, RhAd54, RhAd55, RhAd56, RhAd57, RhAd58, RhAd59, RhAd60, RhAd61, RhAd62, RhAd63, RhAd64, RhAd65, RhAd66, RhAd67, CyAd1 Any one of the seven subgroups A, B, C, D, E, F, and G of Gorilla adenovirus, such as GoAd B20, GoAd B22, GoAd B21, GoAd B10, GoAd B11, GoAd B12, GoAd B13, GoAd B14, GoAd B19, GoAd E1, GoAd C10, GoAd C11, GoAd C12, GoAd C13, GoAd C14, GoAd C15, GoAd C16, GoAd C17, GoAd C18, GoAd1, GoAd 2; Any one of the seven subgroups of baboon adenovirus, A, B, C, D, E, F, and G, such as BaAd 3; Human adenoviruses include any one of the seven subgroups A, B, C, D, E, F, and G, such as: Ad1, Ad10, Ad11, Ad12, Ad14, Ad15, Ad17, Ad18, Ad19, Ad2, Ad20, Ad21, Ad22, Ad23, Ad25, Ad26, Ad27, Ad28, Ad29, Ad3, Ad30, Ad31, Ad32, Ad33, Ad34, Ad35, Ad36, Ad38, Ad39, Ad4, Ad40, Ad41, and Ad42. Any one of Ad43, Ad44, Ad45, Ad46, Ad47, Ad48, Ad49, Ad5, Ad50, Ad51, Ad52, Ad53, Ad54, Ad55, Ad56, Ad57, Ad6, Ad60, Ad61, Ad62, Ad63, Ad64, Ad65, Ad66, Ad68, Ad7, Ad89, Ad81, Ad86, Ad8E, AdD8, AdD9, AdD37, AdD10, AdD13, AdD110, and AdD109.

[0015] Preferably, the human adenovirus vector is AdHu5 or AdHu26.

[0016] Preferably, the simian adenovirus vector is of the ChAdOx1 type.

[0017] In some specific embodiments, the recombinant protein derived from varicella-zoster virus and the antigen derived from varicella-zoster virus are varicella-zoster virus glycoprotein (gE).

[0018] In some specific embodiments, the sequence of the varicella-zoster virus glycoprotein (gE) is SEQ ID NO:1 or SEQ ID NO:2.

[0019] In some specific embodiments, the immune composition is administered via inhalation, microneedle, or intramuscular injection.

[0020] Preferably, the inhalation method includes, but is not limited to, dry powder inhalation or nebulized inhalation.

[0021] The present invention also provides a varicella-zoster vaccine prepared from any of the above-described immune compositions, wherein the varicella-zoster vaccine does not contain an adjuvant.

[0022] In some specific embodiments, the varicella-zoster vaccine may be formulated as an inhaled formulation, an injectable formulation, or a microneedle formulation.

[0023] In some specific embodiments, the inhaled formulation includes nasal inhalation or oral inhalation.

[0024] Preferably, the inhaled drug delivery formulation includes a liquid inhaled formulation or a dry powder inhaled formulation.

[0025] In some specific embodiments, the applicable dose range of the recombinant protein derived from varicella-zoster virus contained in the vaccine formulation of the present invention is 0.1~2.0 μg / μl; preferably, the applicable dose range of the recombinant protein derived from varicella-zoster virus is 0.2~0.5 μg / μl; more preferably, the applicable dose of the recombinant protein derived from varicella-zoster virus is 0.5 μg / μl.

[0026] In some specific embodiments, the applicable content of recombinant protein derived from varicella-zoster virus in the vaccine is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, in units of ug / μl.

[0027] The applicable dose range of the recombinant adenovirus vector described in this invention is 2 x 10⁻⁶. 7 ~8x10 8 VP / μl; preferably, the applicable dose range of the recombinant adenovirus vector is 1x10. 8 ~4x10 8 VP / μl; More preferably, the applicable dose of the recombinant adenovirus vector is 2 x 10⁻⁶. 8 VP / μl.

[0028] In some specific embodiments, the appropriate content of the recombinant adenovirus vector contained in the vaccine formulation is 2 x 10⁻⁶. 7 2.5x10 7 3x10 7 3.5x10 7 4x10 7 4.5x10 6 5x10 7 5.5x10 7 6x10 7 6.5x10 7 7x10 7 7.5x10 7 8x10 7 8.5x10 7 9x10 7 9.5x10 7 1x10 81.5x10 8 2x10 8 2.5x10 8 3x10 8 3.5x10 8 4x10 8 4.5x10 8 5x10 8 5.5x10 8 6x10 8 6.5x10 8 7 x 10 8 7.5x10 8 8x10 8 Unit: VP / μl.

[0029] In some specific embodiments, the recombinant protein derived from varicella-zoster virus and the antigen derived from varicella-zoster virus are glycoproteins (gE) derived from varicella-zoster virus.

[0030] The present invention also provides the use of any of the above-described immune compositions and vaccines in the prevention and / or treatment of varicella-zoster.

[0031] It should be understood that the present invention is not limited to the specific methods and experimental conditions described, as these methods and conditions can vary. It should also be understood that the terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0032] While any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the invention, preferred methods and materials are described below. All publications mentioned in this specification are incorporated herein by reference in their entirety.

[0033] The terms used in this specification have meanings that are generally accepted and known to those skilled in the art; however, for the sake of convenience and completeness, specific terms and their meanings are given below and throughout the specification.

[0034] When used in this specification and the appended claims, terms may be interpreted in both narrow and broad senses unless the context clearly specifies otherwise. Thus, for example, the term "method" includes one or more methods and / or steps, such as those described herein and / or methods and / or steps that will become apparent to a person skilled in the art upon reading this specification, etc.

[0035] The term "comprising" means both "containing" and "consisting of". For example, a composition that "contains" X can consist of only X or may contain other substances, such as X+Y.

[0036] The term "antigen" generally refers to a biomolecule, typically a protein, peptide, polysaccharide, lipid, or conjugate, containing at least one epitope to which an antibody that recognizes it can selectively bind; or, in some cases, an immunogenic substance capable of stimulating antibody production or a T-cell response, or both, in an animal, including compositions injected into or ingested by an animal. An immune response can be induced on the whole molecule or on one or more different parts of the molecule (e.g., epitopes or haptens). The term can be used for a single molecule or a group of homogeneous or heterogeneous antigen molecules. Antigens are recognized by antibodies, T-cell receptors, or other elements of specific humoral and / or cellular immunity. The term "antigen" includes all relevant antigenic epitopes. Epitopes of a particular antigen can be identified using a variety of methods well known in the art. For example, conformational epitopes can be identified by determining the spatial conformation of amino acids, for example, by methods such as X-ray crystallography and 2D nuclear magnetic resonance. Furthermore, for the purposes of this invention, the term "antigen" can also be used for proteins having modifications such as deletions, additions, and substitutions (generally conserved in nature, but they may not be conserved), in their native sequence, provided that the protein retains the ability to elicit an immune response. Such modifications may be intentional, such as as a result of site-directed mutagenesis, or due to the use of specific synthetic or genetic engineering methods, or they may be accidental, such as due to mutations in the host that produces the antigen. Furthermore, antigens can be produced, obtained, or isolated from microorganisms such as bacteria, or they can be from the entire microorganism. Similarly, oligonucleotides or polynucleotides expressing antigens, such as in nucleic acid immunization protocols, are also included in this definition. Synthetic antigens are also included, such as multiepitopes, flanking epitopes, and other recombinant or synthetic antigens.

[0037] The term "immune composition" refers to a composition containing at least one antigen that induces an immune response in an animal.

[0038] As used herein, the term “prevention” (including its variants, such as “treatment” or “being treated”) means any one or more of the following: (i) prevention of infection or reinfection, as in the case of routine vaccination; (ii) reduction of severity or elimination of symptoms; and (iii) substantial or complete elimination of a specific pathogen or disease. Therefore, treatment can be preventive (before infection) or therapeutic (after infection). According to the invention, preventive or therapeutic treatment can be used. According to one specific embodiment of the invention, compositions and methods for treating host animals against infection caused by varicella-zoster virus are provided, including through preventive and / or therapeutic immunization. The methods of the invention can be used to preventively and / or therapeutically induce immunity in patients. The methods of the invention can also be applied to patients for biomedical research purposes.

[0039] The term "patient" refers to a mammal, bird, fish, reptile, or any other animal. The term "patient" also includes humans. The term "patient" also includes pets. Non-limiting examples of pets include: dogs, cats, pigs, rabbits, rats, mice, gerbils, hamsters, guinea pigs, ferrets, birds, snakes, lizards, fish, turtles, and frogs. The term "patient" also includes pets. Non-limiting examples of domesticated animals include: alpacas, bison, camels, cattle, deer, pigs, horses, llamas, mules, donkeys, sheep, goats, rabbits, reindeer, yaks, chickens, geese, and turkeys.

[0040] The term "applicable dose" refers to the human dose. When calculated per unit body weight (mg / kg), the immunization dose for mice is approximately 12-13 times that for humans. This is based on the scientific principle of body surface area conversion: the Km coefficient for mice is approximately 3, while for humans it is approximately 37, resulting in a conversion factor of approximately 12.3. However, in terms of the total dose and the actual setup of vaccine experiments, mice typically use 1 / 10 to 1 / 5 of the human dose as the starting gradient for immunogenicity evaluation (References: "The Influence of Body Weight Factor on Human Equivalent Dose Conversion Calculation in Human Tolerance Trials", "Discussion on the Dose-Effect Relationship of Toad Venom Based on Human Equivalent Dose").

[0041] Treatment can be administered using a single-dose or multiple-dose regimen. Multiple-dose regimens can be used for primary immunization and / or booster immunization. In multiple-dose regimens, different doses can be administered via the same or different routes.

[0042] Immunogenicity, or an effective amount, of the immunogenic composition can be determined in a dose-response study, in which patients are immunized with incremental increments of the immunogenic composition and the immune response is analyzed to determine the optimal dose. The initial dose in the study can be determined based on immunization data from animal models. The dose may vary depending on individual circumstances. This amount can be determined in routine trials using methods known to those skilled in the art.

[0043] An immunologically effective amount of the immunogenic composition is administered to the patient at an appropriate dose to initiate an immune response. The dosage may vary depending on individual circumstances, such as age and weight. This amount can be determined in routine testing using methods known to those skilled in the art.

[0044] The present invention has the following beneficial effects: 1) The present invention provides a method for preparing and applying a vaccine for the prevention and / or treatment of varicella-zoster virus, wherein the vaccine comprises recombinant protein and recombinant adenovirus vector components derived from varicella-zoster virus, and does not include adjuvants, thereby avoiding the side effects caused by adjuvants and having higher safety. 2) The varicella-zoster vaccine provided by the present invention includes a recombinant protein derived from the varicella-zoster virus, namely varicella-zoster glycoprotein (gE), and a recombinant adenovirus vector, which is formed by genetic engineering recombination of the gE gene sequence and the adenovirus vector. This can greatly reduce the antigen content and achieve significant humoral and cellular immune effects. 3) The immune composition provided by the present invention can be administered via inhalation, microneedle administration, or injection to induce immunity. It can not only effectively stimulate humoral and cellular immunity, but also generate mucosal immunity, and can be used to effectively prevent or improve infectious diseases caused by varicella-zoster virus. Attached Figure Description

[0045] Figure 1 Serum IgG test results for recombinant adenovirus vector at different dosages; Figure 2 Recombinant adenovirus vectors at different dosages, CD4 + Results of cytokine detection in T cells; Figure 3 : Results of serum IgG detection of gE protein at different drug doses; Figure 4 gE protein and CD4 at different dosages + Results of cytokine detection in T cells; Figure 5 Serum IgG test results for different administration methods; Figure 6 Different administration methods, CD4 + Results of cytokine detection in T cells. Detailed Implementation

[0046] The following examples illustrate some embodiments of the present invention. However, it should be understood that these examples are for illustrative purposes only and are not intended to be definitive in defining the terminology and scope of the invention. It should be understood that conditions outside or below the specified range may also be used, although less convenient, given typical reaction conditions (e.g., temperature, reaction time, etc.). Unless otherwise stated, all parts and percentages given in this specification are by weight, and all temperatures are in degrees Celsius.

[0047] Furthermore, the following embodiments utilize standard methods well-known and conventional to those skilled in the art, unless otherwise stated. As described above, the following embodiments are provided for illustrative purposes and should not be construed as limiting the scope of the invention in any way.

[0048] Example 1: Preparation of Varicella-Zoster Vaccine The varicella-zoster vaccine is composed of a subunit component, gE protein (sequence shown in SEQ ID NO: 1 or SEQ ID NO: 2), and a recombinant adenovirus vector component, CVE, mixed in a specific ratio. The gE protein was purchased from Novizan, and the preparation process of the recombinant adenovirus vector CVE is as follows: (1) Recombinant shuttle plasmid and adenovirus backbone vector recombination construction The target antigen gene was codon-optimized, and a strong CMV promoter and a PolyA terminator were ligated to construct an antigen expression cassette. The expression cassette was then inserted into the multiple cloning site of the chimpanzee adenovirus backbone plasmid with an E1 region deletion via enzyme digestion-ligation or bacterial homologous recombination. The cassette was transformed into competent *E. coli*, plated on antibiotic-resistant plates, and amplified by single colony picking. The plasmid was extracted, and identified by PCR, double enzyme digestion, and sequencing to verify the complete correctness of the recombinant plasmid sequence.

[0049] (2) Large-scale preparation and linearization of recombinant adenovirus plasmids Positive strains were cultured in a shaker to expand the volume of endotoxin-free plasmids, ensuring plasmid purity and concentration. Specific restriction endonucleases were used to digest the linearized recombinant adenovirus whole-genome plasmid, removing the prokaryotic backbone sequence. The digestion products were purified and recovered to obtain high-purity linearized viral genomic DNA for cell transfection.

[0050] (3) HEK293 cell culture and primary virus packaging HEK293 cells were cultured in complete medium under adherent / suspension conditions at 37°C and 5% CO2. When cell confluence reached 70%–80%, the medium was replaced with fresh medium. Linearized recombinant adenovirus DNA was transfected into HEK293 cells using transfection reagents. Cells were cultured continuously for 7–10 days, with daily microscopic observation of cell status until typical CPE cytopathic effects appeared: cells became rounded, shrunken, clustered, and partially detached. All cells and the culture supernatant were collected, and the cells were subjected to 3–5 freeze-thaw cycles to fully lyse the cells and release intracellular adenovirus particles. Cell debris was removed by low-speed centrifugation, and the supernatant was the P0 generation recombinant chimpanzee adenovirus seed culture.

[0051] (4) Virus seed bank amplification and establishment Fresh HEK293 cells were infected with P0 generation virus at low MOI, and P1 and P2 generation working seeds were obtained through stepwise amplification. The seeds were aliquoted and cryopreserved to establish a master seed bank and a working seed bank. Samples were taken to test viral titer, sterility, exogenous microorganisms, and gene stability.

[0052] (5) Large-scale cell expansion and culture Cell scaling-up: Cell culture flasks → cell factory → large-scale bioreactor, serum-free suspension culture on a large scale. Cell density and viability are monitored. Once the cell density reaches the target during the logarithmic growth phase, recombinant adenovirus working seed is inoculated at the appropriate MOI. Virus adsorption culture is performed for 1–2 hours, fresh culture medium is added, and culture continues for 48–72 hours. After widespread cytopathic effects occur, the cell culture medium and cell pellet are collected uniformly.

[0053] (6) Sample pretreatment and crude purification The entire material is centrifuged at low temperature to remove large cell debris and impurities. Benzobonase nuclease is then added for isothermal digestion to degrade host cell genomic DNA and free RNA, reducing subsequent impurity residues. Hypotonic lysis combined with mechanical disruption completely releases intracellularly encapsulated adenovirus particles. Deep filtration provides preliminary clarification, removing fine suspended matter and reducing chromatographic load.

[0054] (7) Chromatographic fine purification Anion exchange chromatography: Utilizing the specific binding of adenovirus surface charges, gradient elution removes host proteins, residual nucleic acids, and broken virus particles. Molecular sieve / gel filtration chromatography: Based on differences in molecular particle size, it separates intact viruses, empty capsids, and aggregates, further improving vaccine purity. The entire process is conducted in a low-temperature, aseptic environment to maintain the stability of the virus particle structure.

[0055] (8) Ultrafiltration concentration and buffer replacement The purified virus solution was concentrated using a tangential flow ultrafiltration (TFF) system. Simultaneously, it was replaced with vaccine-specific buffer solution to adjust pH, osmotic pressure, and ion concentration. Samples were taken to detect the virus particle count (VP) and activity titer, and adjustments were made to the target formulation concentration. Finally, a 0.22 μm sterile membrane was used for terminal sterilization filtration to obtain a sterile and qualified recombinant adenovirus vector CVE stock solution.

[0056] Example 2: Study on Immunization Dosage of Recombinant Adenovirus Vector (CVE) Fifty-four healthy, 6-8 week old SPF-grade female C57BL / 6 mice were randomly divided into 9 groups (n=6 per group). Groups 1-5 were immunized intradermally (ID) at day 0 and 4 weeks with the recombinant adenovirus vector varicella-zoster vaccine (CVE+gE) prepared in Example 1. Group 6 was immunized intradermally (ID) at day 0 and 4 weeks with replication-deficient chimpanzee adenovirus vector varicella-zoster vaccine (gE+ChAdOx1). Group 7 was immunized intradermally (ID) at day 0 and 4 weeks with varicella-zoster vaccine (gE). Group 8 was immunized intradermally (ID) at day 0 and 4 weeks with recombinant adenovirus vector (CVE). Group 9 was immunized intradermally (ID) with physiological saline at day 0 and 4 weeks. Each group was administered the vaccine twice. Each dose of vaccine has a volume of 0.5 ml, and for each specific group, the content of gE protein and recombinant adenovirus vector in each dose is 2.5 μg and 1 × 10⁻⁶, respectively. 9 VP. See Table 1 for details. Blood and spleen were collected two weeks (6 weeks) after the second immunization. Serum antigen-specific IgG antibody titers were detected by indirect ELISA, and spleen CD4 was detected by ICS. + The percentages of TNF-α, IFN-γ, and IL-2 cytokines in T cells were statistically analyzed using Graphpad Prism 8.0 software. The results showed that, regarding humoral immunity, two weeks after secondary immunization, the serum antigen-specific IgG antibody titer in group 7 mice was significantly lower than that in other groups, while groups 1-4 had higher levels of humoral immunity. Figure 1 Regarding cellular immunity: CD4 in group 7 + The levels of TNF-α, IFN-γ, and IL-2 cytokines in T cells were significantly lower than in other groups. Cellular immunity levels were higher in groups 1-4, and CD4 levels were higher in groups 1-4. + There were no significant differences in IFN-γ and IL2 in T cells, but CD4 in group 3 was significantly different. + TNF-α is optimal in T cells ( Figure 2 According to experimental data, the optimal dosage range for CVE is 0.5 × 10⁻⁶. 9 VP~4×10 9 VP, the optimal dose is 1×10 9 VP.

[0057] Table 1 Example 3: Study on gE protein immunotherapy dosage Forty-two healthy, 6-8 week old SPF-grade female C57 mice were randomly divided into seven groups (n=6 per group). Groups 1-5 were immunized intradermally (ID) with the recombinant adenovirus vector varicella-zoster vaccine (CVE+gE) prepared in Example 1 at day 0 and 4 weeks, respectively. Group 6 was immunized intradermally (ID) with recombinant adenovirus vector (CVE) at day 0 and 4 weeks, respectively. Group 7 was immunized intradermally (ID) with physiological saline at day 0 and 4 weeks, respectively. Each group received two doses. Two weeks (6 weeks) after the second immunization, blood and spleens were collected. Serum antigen-specific IgG antibody titers were detected by indirect ELISA, and spleen CD4 counts were detected by ICS. + The percentages of TNF-α, IFN-γ, and IL-2 cytokines in T cells were statistically analyzed using Graphpad Prism 8.0 software. The results showed that, regarding humoral immunity: there was no significant difference in humoral immunity levels after secondary immunization among groups 1-5, but group 5 showed a decreasing trend, and the humoral immunity level in group 6 was significantly lower than that in groups 1-4. Figure 3 Regarding cellular immunity: CD4 in groups 1-4 + There was no significant difference in TNF-α expression within T cells. Groups 3 and 4 showed the highest IFN-γ expression levels, while groups 1-3 showed the highest CD4+ expression levels. + There was no significant difference in IL2 levels within T cells, but it was better than in group 4. Figure 4 According to experimental data, the optimal dose of gE protein is 2.5 μg.

[0058] Table 2 Example 4: Study on different administration methods Eighteen healthy, 6-8 week old SPF-grade female C57 mice were randomly divided into three groups of six each. Based on the immunization dosage results from Examples 2 and 3, and considering the combined effects of gE protein and recombinant adenovirus vector dosages on safety, the optimal content of varicella-zoster virus glycoprotein (gE) was selected as 2.5 μg, and the optimal content of recombinant adenovirus vector was selected as 1 x 10⁻⁶ μg. 9 VP was prepared according to the preparation method in Example 1. Group 1 was immunized by inhalation, Group 2 by intradermal injection, and Group 3 by intramuscular injection. Specific immunization routes and dosages for each group are detailed in Table 3. Two weeks (6 weeks) after the second immunization, blood and spleens were collected. Serum antigen-specific IgG antibody titers were detected by indirect ELISA, and spleen CD4 counts were detected by ICS. +The percentages of TNF-α, IFN-γ, and IL-2 cytokines in T cells were statistically analyzed using Graphpad Prism 8.0 software. The results showed that, compared with the negative control group, mice treated with different routes of administration all exhibited significant humoral and cellular immune responses, with no significant differences between the different treatment groups. Figure 5 and Figure 6 ).

[0059] Table 3 The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0060] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. An immune composition, characterized in that, The immune composition comprises: (A) Recombinant protein derived from varicella-zoster virus; and, (B) A recombinant adenovirus vector encoding an antigen derived from varicella-zoster virus.

2. The immune composition according to claim 1, characterized in that, The dosage range of the recombinant protein derived from varicella-zoster virus is 0.1~2.0 μg / μl; the dosage range of the recombinant adenovirus vector is 2 x 10⁻⁶. 7 ~8x10 8 VP / μl.

3. The immune composition according to claim 2, characterized in that, The dosage range of the recombinant protein derived from varicella-zoster virus is 0.2~0.5 μg / μl; the applicable dosage range of the recombinant adenovirus vector is 1x10. 8 ~4x10 8 VP / μl.

4. The immune composition according to any one of claims 1-3, characterized in that, The recombinant adenovirus vector is non-replicating and has a deletion in at least one of the early region 1 (E1 region) and early region 3 (E3 region) of the adenovirus.

5. The immune composition according to claim 4, characterized in that, The recombinant adenovirus vector is derived from a human adenovirus vector or a simian adenovirus vector; preferably, the human adenovirus is AdHu5 or Ad26; and the simian adenovirus vector is ChAdOx1.

6. The immune composition according to claim 1, characterized in that, The recombinant protein derived from varicella-zoster virus and the antigen derived from varicella-zoster virus are varicella-zoster virus glycoprotein (gE).

7. The immune composition according to claim 6, characterized in that, The sequence of the varicella-zoster virus glycoprotein (gE) is SEQ ID NO:1 or SEQ ID NO:

2.

8. The immune composition according to any one of claims 1-7, characterized in that, The immune composition can be administered via inhalation, microneedle, or intramuscular injection.

9. A varicella-zoster vaccine prepared from the immune composition according to any one of claims 1-8.

10. The varicella-zoster vaccine according to claim 9, characterized in that, The varicella-zoster vaccine does not contain adjuvants.

11. The varicella-zoster vaccine according to claim 9 or 10, characterized in that, The varicella-zoster vaccine is available in formulations including inhaled, injectable, or microneedle formulations.

12. The varicella-zoster vaccine according to claim 11, characterized in that, The inhalation formulation includes nasal inhalation or oral inhalation; preferably, the inhalation formulation includes a liquid inhalation formulation or a dry powder inhalation formulation.

13. Use of the immune composition of any one of claims 1-8 or the vaccine of any one of claims 9-12 in the prevention and / or treatment of varicella-zoster.

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