Application of parabacteroides dielsii and / or parabacteroides gold in preparation of immunopotentiator of vaccine

Abstract

The invention relates to the technical field of immunization. The invention discloses parabacteroides dieldrii and parabacteroides goldsteinii, an application of the parabacteroides dieldrii and / or the parabacteroides goldsteinii in preparation of an immunopotentiator of a vaccine, and the immunopotentiator of the vaccine, and particularly discloses the parabacteroides dieldrii and the parabacteroides goldsteinii, the application of the parabacteroides dieldrii and / or the parabacteroides goldsteinii in preparation of the immunopotentiator of the vaccine. The parabacteroides dieldrii and / or the parabacteroides gold can be used as an immunopotentiator, can activate immunity, enhance antibody titer and shorten antibody generation reaction time, and can remarkably promote proliferation of B cells, plasma cells and memory B cell subgroups in a hair growth center.

Description

Technical Field

[0001] This application belongs to the technical field of immunization, specifically relating to the use of *Pseudomonas diffusa* and / or *Pseudomonas quinquefolius* in the preparation of an immune enhancer for a vaccine, and an immune enhancer for a vaccine. Background Technology

[0002] Gut microbiota are an important component of the human gut microbiota, playing a vital regulatory role in metabolism, cognition, and immunity. *Parabacteroides* genus ( Parabacteroides *Pseudomonas dilataniae* is a core member of the human gut microbiota, with a detection rate exceeding 90% in human gut sample databases. Parabacteroides distasonis , P. distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii , P. goldsteinii These are two species of bacteria in the genus *Pseudomonas*, both of which are Gram-negative. Studies have shown that they have protective effects against diseases such as obesity, non-alcoholic fatty liver disease, and diabetes.

[0003] The immune response is a series of biological activities initiated by the body's immune system after recognizing an antigenic stimulus, including the activation, proliferation, differentiation, and production of effector molecules by immune cells. The level of the immune response after vaccination directly affects the effectiveness of immunization. Current vaccine systems still suffer from limitations due to insufficient immunogenicity, particularly traditional vaccine platforms such as inactivated vaccines. Insufficient antigenic epitope presentation and limited immune activation lead to low immune response efficiency. This problem is particularly prominent in vulnerable groups such as the elderly and immunocompromised individuals, manifesting as insufficient antibody titers, incomplete immune memory formation, and decreased protective efficacy, severely restricting the preventive effect and application value of vaccines. Therefore, developing immune enhancers for vaccines that safely and efficiently promote the vaccine immune response can effectively protect vulnerable populations and block disease progression. Summary of the Invention

[0004] To address the problems of the prior art, this application provides *Pseudomonas difficile* and / or *Pseudomonas quinquefolius*, their use in the preparation of an immune enhancer for a vaccine, and an immune enhancer for a vaccine.

[0005] Specifically, this application relates to the following aspects: 1. A type of *Pseudomonas dilataniae* ( Parabacteroides distasonis It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 35213.

[0006] 2. A type of *Pseudomonas jini* ( Parabacteroides goldsteinii It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 35214.

[0007] 3. The *Pseudomonas dilataniae* described in item 1 ( Parabacteroides distasonis ) and / or the *Pseudomonas jini* described in item 2 ( Parabacteroides goldsteinii Its use in the preparation of immune enhancers for vaccines.

[0008] 4. The use according to item 3, wherein the *Pseudomonas dilataniae* ( Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii ( ) Enhance antibody titer after vaccination and / or shorten vaccine response time; and / or The application of the aforementioned *Pseudomonas dilatatus* ( Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii This increases the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccine recipients.

[0009] 5. The use according to item 3 or 4, wherein the *Pseudomonas dignitaria* ( Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii It can be administered before, after, or at the same time as vaccination.

[0010] 6. The use according to any one of items 3-5, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine, or a peptide vaccine; Preferably, the vaccine is selected from any one or more of the following groups: rabies vaccine, influenza vaccine, pneumonia vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand-foot-and-mouth disease vaccine, and HPV vaccine.

[0011] 7. An immune enhancer for a vaccine, comprising *Pseudomonas difficile* as described in item 1 (… Parabacteroides distasonis ) and / or the *Pseudomonas jini* described in item 2 ( Parabacteroides goldsteinii ).

[0012] 8. The immune enhancer according to claim 7, wherein the immune enhancer enhances the titer of antibodies after vaccination and / or shortens the vaccine response time; and / or The administration of the immune enhancer increases the proportion of germinal center B cells, plasma cells, and / or memory B cells in the vaccinated subject; and / or The immune enhancer is administered before, after, or simultaneously with vaccination.

[0013] 9. The immune enhancer according to item 7 or 8, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine, or a peptide vaccine; Preferably, the vaccine is selected from any one or more of the following groups: rabies vaccine, influenza vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand-foot-and-mouth disease vaccine, and HPV vaccine.

[0014] 10. The immune enhancer according to any one of claims 7-9, wherein the immune enhancer further comprises a pharmaceutically acceptable carrier.

[0015] Beneficial effects: This application provides two strains, one of which is *Pseudomonas difficile* with accession number CGMCC No. 35213. Parabacteroides distasonis ) and *Pseudomonas kiuri* with accession number CGMCC No. 35214 ( Parabacteroides goldsteinii These two strains can act as immune enhancers, activating immunity, increasing antibody titers, and shortening antibody production time. Specifically, administering these strains to mice followed by rabies vaccine significantly increased the titer of rabies virus-specific antibodies. Further studies revealed that these two strains significantly promoted the proliferation of germinal center B cells, plasma cells, and memory B cell subsets, increasing their proportion. Attached Figure Description

[0016] Figure 1 The bacterial extraction process is shown.

[0017] Figure 2 Experimental protocol demonstrating the effectiveness of antibiotic treatment in mice followed by vaccination.

[0018] Figure 3 This shows the colonization of the bacteria in mice after intervention.

[0019] Figure 4 The results show the antibody production in mice after bacterial intervention.

[0020] Figure 5 Flow cytometry analysis results of mouse spleen cell populations after bacterial intervention. Detailed Implementation

[0021] The present application is further illustrated below with reference to embodiments. It should be understood that the embodiments are only used to further illustrate and explain the present application and are not intended to limit the present application.

[0022] Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. While similar or identical methods and materials may be applied in experimental or practical applications, materials and methods are described herein. In case of conflict, the definitions included herein shall prevail. Furthermore, materials, methods, and examples are for illustrative purposes only and are not intended to be limiting. The present application is further described below with reference to specific embodiments, but is not intended to limit the scope of the application.

[0023] As used throughout the specification and claims, the terms "comprising" or "including" are open-ended and should be interpreted as "comprising but not limited to". The subsequent descriptions in the specification are preferred embodiments for carrying out this application; however, these descriptions are for the purpose of understanding the general principles of the specification and are not intended to limit the scope of this application. The scope of protection of this application shall be determined by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0024] It should be understood that the embodiments of this application described herein include embodiments that are "composed of" and / or "substantially composed of". References to values ​​or parameters of "about" herein include (and describe) variations of that value or parameter itself. For example, a reference to "about X" includes a description of "X".

[0025] As used herein, references to “not” values ​​or parameters generally refer to and describe “except” values ​​or parameters. For example, “The method is not used to treat type X cancer” means that the method is used to treat cancers other than type X.

[0026] As used in this article, the term “approximately XY” has the same meaning as “approximately X to approximately Y”.

[0027] As used herein and in the appended claims, the singular forms “a / an” and “the” include the plural objects unless the context clearly indicates otherwise. It should also be noted that claims may be drafted to exclude any optional elements. Therefore, this statement is intended as a preliminary basis for the use of exclusive terms such as “only” or “merely” in conjunction with the description of the elements of the claim, or for the use of the limitation of “no”.

[0028] As used herein, the term "and / or" in words such as "A and / or B" is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, as used herein, the term "and / or" in words such as "A, B and / or C" is intended to include each of the following embodiments: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0029] As used herein, the term "vaccine" refers to any preparation of an antigen or immunizing substance that is suitable for stimulating active immunity in animals or humans.

[0030] As used herein, the term "antigen" means any substance that, when introduced into an immune-active human or animal, stimulates a humoral and / or cell-mediated immune response. The antigen may be a pure substance, a mixture of substances, or a particulate matter (including cells, cell fragments, or cell-derived fragments) or a live (usually attenuated) organism or virus. Examples of suitable antigens include, but are not limited to, proteins, glycoproteins, lipoproteins, peptides, carbohydrates / polysaccharides, lipopolysaccharides, toxins, viruses, bacteria, fungi, and parasites. Antigens may be natural (naturally expressed or produced), synthetic, or derived from recombinant DNA methodologies familiar to those skilled in the art.

[0031] As used in this article, the term "immunostimulant" refers to a substance that can enhance the immune function of an animal. It is mainly used to enhance the body's anti-tumor and anti-infection capabilities, correct immune deficiencies, activate one or more immune-active cells, enhance the body's specific and non-specific immune functions, restore low immune function to normal, or have an adjuvant effect, enhance the immunogenicity of antigens used in combination with it, and accelerate the induction of immune response; or replace immune-active components lacking in the body, producing an immune substitution effect; or have a bidirectional regulatory effect on the body's immune function, bringing excessively high or low immune function towards normal.

[0032] As used herein, the term "subject" refers to a mammal, which can be a human (i.e., a male or female of any age, such as a pediatric subject (e.g., an infant, child, or adolescent) or an adult subject (e.g., a young adult, middle-aged adult, or elderly person)) or a non-human animal. In some specific embodiments, the subject is a human.

[0033] As used herein, the term “administration” means the implantation, absorption, ingestion, injection, inhalation, or other introduction of a drug or reagent into or onto a subject.

[0034] As used herein, the terms “therapeutic effective amount,” “therapeutic effective dose,” “effective amount,” and “effective dose” refer to the amount or dose of a compound or composition that, when administered to a subject, is capable of treating, preventing, or improving the subject’s condition, disease, or disorder. In other words, the amount is “therapeuticly effective” when administered to a subject. The actual amount will vary depending on a variety of factors, including but not limited to the specific condition, disease, or disorder being treated, prevented, or improved; the severity of the condition; the patient’s weight, height, age, and health; and the route of administration.

[0035] As used herein, the term “treatment” means, to a certain extent, the eradication, reduction, improvement, or reversal of the signs or symptoms of a health condition, disease, or disorder, and includes, but is not limited to, the complete cure of the condition, disease, or disorder. Treatment can be curative, improving, or partially improving a disorder. “Treatment” can also include improving or enhancing a condition or characteristic, for example, bringing a particular system in the body to a state of high health or homeostasis.

[0036] As used herein, the term "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form suitable for contact with human or animal tissues without causing excessive toxicity, irritation, allergic reactions, other problems, or complications, and with a reasonable benefit / risk ratio. In some embodiments, pharmaceutically acceptable compounds, materials, compositions, and / or dosage forms refer to those compounds, materials, compositions, and / or dosage forms approved by regulatory agencies (such as the U.S. Food and Drug Administration, the China National Medical Products Administration, and the European Medicines Agency) or listed in recognized pharmacopoeias (such as the United States Pharmacopeia, the Chinese Pharmacopoeia, and the European Pharmacopoeia) for use in animals, particularly humans.

[0037] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or delivery vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, used to carry or transport the strain of this application from one location, body fluid, tissue, organ (internal or external), or body part to another location, body fluid, tissue, organ, or body part. A pharmaceutically acceptable carrier may be a delivery vehicle, diluent, excipient, or other material that can be used in contact with animal tissue without excessive toxicity or adverse reactions. Exemplary pharmaceutically acceptable carriers include, but are not limited to, sugars, starch, cellulose, malt, astragalus gum, gelatin, Ringer's solution, alginate, isotonic saline, buffers, etc. In this application, the pharmaceutically acceptable carrier is added in amounts commonly used by those skilled in the art.

[0038] Germinal centers are microanatomical structures that form within secondary lymphoid organs after exposure to antigens. They are crucial sites for B cell proliferation, high-frequency mutation, and antibody affinity maturation. Within the germinal center, germinal center B cells undergo mutation and selection, with the assistance of T cells, to undergo fate selection. B cells with high affinity for antigens tend to differentiate into plasma cells, while others differentiate into memory B cells. Through this process, germinal centers are the foundation for the generation of high-affinity antibodies and the formation of long-term immune memory.

[0039] In the phrase "increased proportion of germinal center B cells, plasma cells, and / or memory B cells," "cell proportion" means that this cell population accounts for a significant portion of CD19. + The ratio of the total number of B cells. For example, the proportion of B cells in the germinal center is the percentage of the total number of B cells in the germinal center to CD19. + The ratio of the total number of B cells; for example, the proportion of plasma cells as a percentage of the total CD19 cell count. + The ratio of the total number of B cells; for example, the proportion of memory B cells is the percentage of total memory B cells in CD19. + The ratio of the total number of B cells. In this application, the cell content of memory B cells in the subject can be detected by flow cytometry after obtaining peripheral blood mononuclear cells to evaluate the effect; wherein, the peripheral blood mononuclear cells of the subject can be obtained by conventional methods known to those skilled in the art.

[0040] This application provides a *Pseudomonas dilatatus* (… Parabacteroides distasonis The accession number is CGMCC No. 35213, which was deposited on July 11, 2025 at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postcode: 100101.

[0041] This application also provides a strain of *Pseudomonas jinnsis* (… Parabacteroides goldsteinii The accession number is CGMCC No. 35214, which was deposited on July 11, 2025 at the China General Microbiological Culture Collection Center (CGMCC), No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postcode: 100101.

[0042] This application provides the aforementioned *Pseudomonas dilataniae* (… Parabacteroides distasonis Its use in the preparation of immune enhancers for vaccines.

[0043] This application provides the aforementioned *Pseudomonas jini* (… Parabacteroides goldsteinii Its use in the preparation of immune enhancers for vaccines.

[0044] This application provides the aforementioned *Pseudomonas dilataniae* (… Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii Its use as an immune enhancer in the preparation of vaccines.

[0045] In some implementations, the *Pseudomonas dilataniae* ( Parabacteroides distasonis ) shorten vaccine response time; in some implementations, the *Pseudomonas dignitaria* ( Parabacteroides distasonis ) enhances the titer of antibodies after vaccination; in some implementations, the *Pseudomonas dignitaria* ( Parabacteroides distasonis This shortens vaccine response time and enhances antibody titers after vaccination. For example, the anti-rabies virus IgG titer in mice vaccinated with rabies vaccine was approximately 2.5 times that of the control group on day 14, approximately 1.5 times that of the control group on day 20, and approximately 1.7 times that of the control group on day 27. This demonstrates that the *Pseudomonas dilataniae* (…) Parabacteroides distasonis It can significantly enhance the titer of antibodies after rabies vaccination and significantly shorten the vaccine response time.

[0046] In some implementations, the *Pseudomonas kiuri* ( Parabacteroides goldsteinii ) shorten vaccine response time; in some implementations, the *Pseudomonas kiuri* ( Parabacteroides goldsteinii ) enhances the titer of antibodies after vaccination; in some implementations, the *Pseudomonas aeruginosa* ( Parabacteroides goldsteinii This shortens vaccine response time and enhances antibody titers after vaccination. For example, the anti-rabies virus IgG titer in mice vaccinated with rabies vaccine was approximately 2.5 times that of the control group on day 14, approximately 1.7 times that of the control group on day 20, and even approximately 3 times that of the control group on day 27. This demonstrates that the *Pseudomonas jini* (… Parabacteroides goldsteinii It can significantly enhance the titer of antibodies after rabies vaccination and significantly shorten the vaccine response time.

[0047] In some embodiments, the *Pseudomonas dignitaria* is applied in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii This is to enhance the titer of antibodies after vaccination and / or shorten the vaccine response time.

[0048] In some implementations, the *Pseudomonas dilataniae* (…) is applied. Parabacteroides distasonis This results in an increase in the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccinated subjects. In some embodiments, the *Pseudomonas ginseng* (…) is administered…Parabacteroides goldsteinii This results in an increase in the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccinated subjects. In some embodiments, the *Pseudomonas dignitaria* is administered in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii This increases the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccinated subjects. For example, compared to subjects who received a rabies vaccine but did not receive any *Pseudomonas*, those vaccinated with *Pseudomonas* dichotoma (*Pseudomonas* dichotoma) showed a higher proportion of germinal center B cells, plasma cells, and / or memory B cells. Parabacteroides distasonis Furthermore, in subjects vaccinated against rabies, the proportion of germinal center B cells increased from 3.51% to 8.16% (or even higher, such as around 8.5%), an increase of at least 132%; the proportion of plasma cells increased from 0.93% to 1.84% (or even higher, such as around 2.0%), an increase of at least 98%; and the proportion of memory B cells increased from 44.9% to 63.9% (or even higher, such as around 65%), an increase of at least 42%. For example, compared with subjects who received rabies vaccine but did not receive any Parabacteroides, subjects who received Parabacteroides goldsteinii and rabies vaccine showed an increase in the proportion of germinal center B cells from 3.51% to 8.39% (or even higher, such as around 8.5%), an increase of at least 139%; an increase in the proportion of plasma cells from 0.93% to 2.03% (or even higher, such as around 2.2%), an increase of at least 118%; and an increase in the proportion of memory B cells from 44.9% to 60.5% (or even higher, such as around 63%), an increase of at least 35%.

[0049] In some implementations, the *Pseudomonas dilataniae* ( Parabacteroides distasonis ) is administered before vaccination. In some embodiments, the *Pseudomonas kiuri* ( Parabacteroides goldsteinii ) is administered prior to vaccination. In some embodiments, the *Pseudomonas difficile* (*P. difficile*) is administered in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii ) is administered before vaccination. In some embodiments, the *Pseudomonas dilataniae* ( Parabacteroides distasonis It is administered after vaccination. In some embodiments, the *Pseudomonas kiuri* (…) is used. Parabacteroides goldsteinii It is administered after vaccination. In some embodiments, the *Pseudomonas difficile* (*P. difficile*) is administered in any combination of proportions.Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii It is administered after vaccination. In some embodiments, the *Pseudomonas difficile* (…) is administered… Parabacteroides distasonis ) and vaccination are carried out simultaneously. In some embodiments, the *Pseudomonas kiuri* (…) is administered. Parabacteroides goldsteinii ) and vaccination are carried out simultaneously. In some embodiments, the *Pseudomonas difficile* (*P. difficile*) is administered in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii It is administered simultaneously with vaccination.

[0050] In this application, for example, refer to Figure 5 It can be seen that administering *Pseudomonas dilatatus* (a type of bacteria) alone to the subjects... Parabacteroides distasonis ) or Parabacterium kiuri ( Parabacteroides goldsteinii Following rabies vaccination, both methods can increase the proportion of germinal center B cells, plasma cells, and memory B cells in the subject. Germinal centers are the foundation for generating high-affinity antibodies and forming long-term immune memory. Plasma cells (also known as effector B cells) are responsible for the efficient synthesis and secretion of antibodies, while memory B cells are responsible for driving a faster and stronger antibody immune response when the body is reinfected with the same pathogen. Therefore, *Pseudomonas difficile* (… Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii When administered alone or in combination, both primarily function to increase the proportion of core immune cells, thereby enhancing the overall immune level of the subject. Therefore, those skilled in the art should understand that regardless of whether *Pseudomonas difficile* (or *Pseudomonas difficile*) is administered to the subject... Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii When receiving any vaccine, *Pseudomonas dilataniae* (…) Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii All of these can produce an effect that enhances the immune level of the subject: for example, an increase in the proportion of germinal center B cells and / or plasma cells and / or memory B cells. Therefore, in this application, there are no other limitations on the vaccine; the administration of the *Pseudomonas difficile* (of this application) to the subject... Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii Following administration of any one or more vaccines in the conventional sense, the *Pseudomonas difficile* (of this application) Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteiniiAll of these can activate the aforementioned immune response, thereby enhancing the subject's immune level. In some embodiments, the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine, or a peptide vaccine. In some embodiments, the vaccine is not limited and is exemplary selected from any one or more of the following group: rabies vaccine, influenza vaccine, pneumococcal vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand-foot-and-mouth disease vaccine, and HPV vaccine.

[0051] This application provides an immune enhancer for a vaccine. In some embodiments, the immune enhancer includes the aforementioned *Pseudomonas difficile* (…). Parabacteroides distasonis In some embodiments, the immune enhancer includes the aforementioned *Pseudomonas kiwifruit* (…). Parabacteroides goldsteinii In some embodiments, the immune enhancer includes the aforementioned *Pseudomonas difficile*. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii In some embodiments, the immune enhancer is *Pseudomonas difficile* (as described above). Parabacteroides distasonis In some embodiments, the immune enhancer is *Pseudomonas kiuri* (as described above). Parabacteroides goldsteinii In some embodiments, the immune enhancer is *Pseudomonas difficile* (as described above). Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii (In any proportion)

[0052] In some implementations, the *Pseudomonas dilataniae* ( Parabacteroides distasonis ) shorten vaccine response time; in some implementations, the *Pseudomonas dignitaria* ( Parabacteroides distasonis ) enhances the titer of antibodies after vaccination; in some implementations, the *Pseudomonas dignitaria* ( Parabacteroides distasonis This shortens vaccine response time and enhances antibody titers after vaccination. For example, the anti-rabies virus IgG titer in mice vaccinated with rabies vaccine was approximately 2.5 times that of the control group on day 14, approximately 1.5 times that of the control group on day 20, and approximately 1.7 times that of the control group on day 27. This demonstrates that the *Pseudomonas dilatatus* (…) Parabacteroides distasonis It can significantly enhance the titer of antibodies after rabies vaccination and significantly shorten the vaccine response time.

[0053] In some implementations, the *Pseudomonas kiuri* ( Parabacteroides goldsteinii ) shorten vaccine response time; in some implementations, the *Pseudomonas kiuri* ( Parabacteroides goldsteinii ) enhances the titer of antibodies after vaccination; in some implementations, the *Pseudomonas aeruginosa* ( Parabacteroides goldsteiniiThis shortens vaccine response time and enhances antibody titers after vaccination. For example, the anti-rabies virus IgG titer in mice vaccinated with rabies vaccine was approximately 2.5 times that of the control group on day 14, approximately 1.7 times that of the control group on day 20, and approximately 3 times that of the control group on day 27. This demonstrates that the *Pseudomonas aeruginosa* (…) Parabacteroides goldsteinii It can significantly enhance the titer of antibodies after rabies vaccination and significantly shorten the vaccine response time.

[0054] In some embodiments, the *Pseudomonas dignitaria* is applied in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii This is to enhance the titer of antibodies after vaccination and / or shorten the vaccine response time.

[0055] In some implementations, the *Pseudomonas dilataniae* (…) is applied. Parabacteroides distasonis This results in an increase in the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccinated subjects. In some embodiments, the *Pseudomonas ginseng* (…) is administered… Parabacteroides goldsteinii This results in an increase in the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccinated subjects. In some embodiments, the *Pseudomonas dignitaria* is administered in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii This increases the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccine recipients.

[0056] In some implementations, the *Pseudomonas dilataniae* ( Parabacteroides distasonis ) is administered before vaccination. In some embodiments, the *Pseudomonas kiuri* ( Parabacteroides goldsteinii ) is administered prior to vaccination. In some embodiments, the *Pseudomonas difficile* (*P. difficile*) is administered in any combination of proportions. Parabacteroides distasonis ) and Parabacterium kiuri ( Parabacteroides goldsteinii ) Administer before vaccination.

[0057] In some embodiments, the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine, or a peptide vaccine. In some embodiments, the vaccine is exemplary selected from any one or more of the following groups: rabies vaccine, influenza vaccine, pneumococcal vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand-foot-and-mouth disease vaccine, and HPV vaccine.

[0058] Example Specific embodiments of the present application will now be described in more detail with reference to the accompanying drawings. While specific embodiments of the present application are shown in the drawings, it should be understood that the present application can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided to enable a more thorough understanding of the present application and to fully convey the scope of the present application to those skilled in the art.

[0059] Example 1 The two strains of *Plasmobacterium* provided in this application: one is *Plasmobacterium digiri* (… Parabacteroides distasonis The strain, *Pseudomonas jini*, was deposited on July 11, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postcode: 100101, with accession number CGMCC No. 35213; in this application, it is referred to as *Pseudomonas jini* 35213 (live). Parabacteroides goldsteinii The gene was deposited on July 11, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postcode: 100101, with accession number CGMCC No. 35214; and is referred to in this application as Parabacterium kiuri 35214 (live).

[0060]

[0061]

[0062] Strain screening method: Refer to Figure 1 Healthy adult subjects were given arachidonic acid at a dose of 500 mg daily for 17 days. Afterward, fecal samples were collected for screening and bacterial strain extraction. The specific implementation plan was as follows: 1. Dissolve 1g of feces in 9mL of sterile physiological saline and mix well to obtain 10g of the solution. -1 ; Take 1 mL of the pre-mixed sample and mix it into 9 mL of bacterial physiological saline, as 10 mL of this solution. -2 Dilute 10 according to this series -1 Up to 10 -6 .

[0063] 2. Select 10 -4 10 -5 and 10 -6 For each gradient, take 100 μL of each and spread it onto DSMZ_Medium330 solid medium, with 3 plates for each gradient. 3. Take one uncoated solid culture medium and nine coated culture media and incubate them at 37°C for anaerobic incubation for 48 hours; 4. According to 10 -4 10 -5 and 10 -6 To assess the growth of bacteria on petri dishes, select petri dishes with colonies of 300-500. Using a sterile inoculation needle, pick 20-30 single colonies based on the colony morphology of Bacteroides and inoculate them into 2 mL centrifuge tubes containing 1.5 mL of sterile DSMZ_Medium330 liquid medium. Incubate anaerobically for 48 h. 5. Take 10 μL of the bacterial culture from each tube and streak it onto DSMZ_Medium330 solid medium for purification. After streaking, incubate anaerobically for 48 h. 6. From each streaked agar plate, pick one single colony and inoculate it into a 2 mL centrifuge tube containing 1.5 mL of sterile DSMZ_Medium330 liquid medium. Incubate anaerobically for 48 h. 7. Perform 16S rDNA sequencing on the preserved strains and compare the sequencing results with the NCBI database to obtain strain species information.

[0064] Example 2 This application also provides the use of the above-mentioned *Pseudomonas difficile* 35213 and *Pseudomonas kiuri* 35214 as immune enhancers.

[0065] In a pseudo-germless mouse single-bacterial transplantation experiment, rabies vaccine injection showed a significant increase in rabies virus-specific antibody levels and a significant shortening of antibody production time.

[0066] Specifically, the implementation includes the following steps: Reference Figure 2 Six-week-old male C57BL / 6J mice were used for antibiotic clearance via gavage for two weeks (days -15 to -1). The antibiotics included vancomycin (2 mg / day / mouse), neomycin (4 mg / day / mouse), metronidazole (4 mg / day / mouse), and penicillin (4 mg / mouse / day). The date of antibiotic discontinuation was recorded as day 0.

[0067] One day after stopping antibiotic gavage (Day 1), administer 10... 8 cfu / ml *Pseudomonas difficile* 35213, 10 8 Mice were administered 200 μl of cfu / ml Parabacterium ginseng 35214 bacterial suspension by gavage once daily for 14 days.

[0068] The experimental mice were grouped as shown in Table 1 below.

[0069] Table 1

[0070] The first dose of rabies vaccine was administered on day 7 following continuous oral gavage with PBS, at a dose of 10 mg / L. 6 Focus-forming unit (FFU).

[0071] Bacterial / PBS was administered via gavage until day 14. Fecal samples were collected, and 16S RNA was used to detect bacterial colonization after transplantation. Additionally, 100 μl of tail vein blood was collected to detect rabies virus-specific antibody levels. (Refer to...) Figure 3 and Figure 4 The study found that the antibody seroconversion time point of the *Pseudomonas dilatatus* 35213 group and the *Pseudomonas kiuri* 25314 group was significantly earlier than that of the control group, and the titer level of rabies virus-specific antibodies in the two groups was significantly higher than that in the control group, with statistically significant differences.

[0072] After gavage with probiotics / PBS until day 14, the diet was discontinued, and the animals were fed a normal diet until day 20. Blood was collected from the tail vein to test for rabies virus-specific antibody levels. (Refer to...) Figure 4 On day 20, the antibody titer levels in the *Pseudomonas diffusa* 35213 group and the *Pseudomonas quinquefolia* 25314 group were significantly higher than those in the control group, and the differences were statistically significant.

[0073] On day 21 of intervention, a second rabies vaccine was administered, and the animal was kept in a no-intervention environment. On day 27, the animal was euthanized and blood was collected for antibody level testing; spleen lymphocytes were also collected for flow cytometry analysis. (Refer to...) Figure 4The antibody level results showed that the antibody titers in the *Parabacterium difficile* 35213 group and the *Parabacterium kiwifruit* 25314 group were significantly higher than those in the control group, with statistically significant differences. Furthermore, the antibody titer in the *Parabacterium kiwifruit* 25314 group was significantly higher than that in the *Parabacterium difficile* 35213 group (P<0.05), indicating a better effect in promoting vaccine efficacy. (Reference) Figure 5 Flow cytometry results showed that in the *Pseudomonas dilataniae* 35213 group and the *Pseudomonas quinquefolius* 25314 group, germinal center B cells, which are key to enhancing acquired immunity and generating long-term, highly effective immunity, significantly proliferated. Subsequently, plasma cells, which are directly responsible for antibody secretion, significantly increased. At the same time, the memory B cell subset, which is key to long-term antibody immune memory, also significantly increased, which was statistically significant.

[0074] The description in this disclosure is provided for illustrative and descriptive purposes only and is not intended to be exhaustive or to limit the disclosure to its forms. Many modifications and variations will be apparent to those skilled in the art. The embodiments were chosen and described in order to better illustrate the principles and practical application of this disclosure and to enable those skilled in the art to understand this disclosure and to design various embodiments with various modifications suitable for a particular purpose.

Claims

1. A type of *Pseudomonas dilataniae* ( Parabacteroides distasonis It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 35213.

2. A type of *Pseudomonas jini* ( Parabacteroides goldsteinii It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 35214.

3. The *Pseudomonas dilataniae* as described in claim 1 ( Parabacteroides distasonis ) and / or the *Pseudomonas jini* as described in claim 2 ( Parabacteroides goldsteinii Its use in the preparation of immune enhancers for vaccines.

4. The use according to claim 3, wherein the *Pseudomonas dignitaria* ( Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii ( ) Enhance antibody titer after vaccination and / or shorten vaccine response time; and / or The application of the aforementioned *Pseudomonas dilatatus* ( Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii This increases the proportion of germinal center B cells, plasma cells, and / or memory B cells in vaccine recipients.

5. The use according to claim 3 or 4, wherein the *Pseudomonas dignitaria* ( Parabacteroides distasonis ) and / or Parabacterium kiuri ( Parabacteroides goldsteinii It can be administered before, after, or at the same time as vaccination.

6. The use according to any one of claims 3-5, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine, or a polypeptide vaccine; Preferably, the vaccine is selected from any one or more of the following groups: rabies vaccine, influenza vaccine, pneumonia vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand-foot-and-mouth disease vaccine, and HPV vaccine.

7. An immune enhancer for a vaccine, comprising *Pseudomonas difficile* as described in claim 1. Parabacteroides distasonis ) and / or the *Pseudomonas jini* as described in claim 2 ( Parabacteroides goldsteinii ).

8. The immune enhancer according to claim 7, wherein the immune enhancer enhances the titer of antibodies after vaccination and / or shortens the vaccine response time; and / or The administration of the immune enhancer increases the proportion of germinal center B cells, plasma cells, and / or memory B cells in the vaccinated subject; and / or The immune enhancer is administered before, after, or simultaneously with vaccination.

9. The immune enhancer according to claim 7 or 8, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine, or a polypeptide vaccine; Preferably, the vaccine is selected from any one or more of the following groups: rabies vaccine, influenza vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand-foot-and-mouth disease vaccine, and HPV vaccine.

10. The immune enhancer according to any one of claims 7-9, wherein the immune enhancer further comprises a pharmaceutically acceptable carrier.

Images