WO2023080501A1 - Method of providing information for predicting immune response to sars-cov-2 vaccine through gut microbiota and functional biomarker profiles, and method of providing subject-customized vaccine information - Google Patents

Abstract

The present invention relates to a method of providing information for predicting immune response to SARS-CoV-2 vaccination and a method of providing customized vaccine information, and more specifically, the present invention verifies that the taxonomic composition of a gut microbiota, metabolites produced thereby, and functional profiles thereof are significantly correlated with the immune response after vaccination, and thus can be used to predict the immune response to SARS-CoV-2 vaccination or provide subject-customized vaccine information. According to the present invention, by identifying the taxonomic composition and functional biomarkers of a subject's gut microbiota prior to vaccination, the subject's humoral immune response level after vaccination can be predicted and information can be provided for vaccine administration that would induce an effective immune response and fewer side effects in the subject.

Description

Information provision method for predicting immune response of SARS-COV-2 vaccine through intestinal microbiome and functional biomarker profile and subject-specific vaccine information provision method

The present invention relates to a method for providing information for predicting an immune response upon vaccination with SARS-CoV-2 and a method for providing customized vaccine information, and more specifically, to a taxonomic composition of the intestinal microflora and metabolites produced by microorganisms. And functional biomarkers including them are confirmed to have a significant correlation with the immune response after vaccination, and intestinal microbial biomarkers and functional markers showing good or bad immune responses are identified to determine the effect of SARS-CoV-2 after vaccination. It is to predict the immune response or to provide customized vaccine information for the subject.

Coronavirus disease 2019 (COVID-19) is a novel respiratory infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It spread from Wuhan, China in December 2019 and is now causing a pandemic and causing a serious health crisis worldwide. As of March 3, 2022, more than 442 million confirmed cases of COVID-19 and more than 5.9 million deaths have been recorded worldwide.

Global efforts to overcome the COVID-19 pandemic have led to the rapid development of various vaccines against SARS-CoV-2. Vaccination is known to be the best means of overcoming the pandemic, but the immunogenicity of vaccines can vary from person to person. The post-vaccination immune response depends on intrinsic factors (e.g. age, sex, genetics, comorbidity and pre-existing immunity), extrinsic factors (e.g. nutrition, environment, behavioral factors) and vaccine (e.g. vaccine type, adjuvant, route of administration). may be affected by a variety of factors, including those related to

Recent studies have shown that the human gut microbiome is an important determinant of the basic immune status and immune response to vaccines. The gut microbiome consists of more than 100 trillion bacteria of more than 150 species, which play important roles in the development, direction, and priming of the immune system, influencing both innate and acquired immunity. It is known that local microbiome-immune system interactions occur in the small and large intestine mainly through B cells, T cells and antigen-presenting cells, which can also respond to a significant number of biomolecules produced by the microbiome (de Jong SE, Olin A, Pulendran B. The Impact of the Microbiome on Immunity to Vaccination in Humans. Cell Host Microbe 2020; 28: 169-179.).

Several studies have reported that the microbiome can play an important role in regulating the immune response to vaccines, including oral vaccines such as cholera and rotavirus vaccines, as well as parenteral vaccines such as hepatitis B, tetanus, and influenza vaccines. Oh JZ, Ravindran R, Chassaing B, et al. TLR5-mediated sensing of gut microbiota is necessary for antibody responses to seasonal influenza vaccination. Immunity 2014; 41: 478-492. and Harris VC, Armah G, Fuentes S, et al. Significant Correlation Between the Infant Gut Microbiome and Rotavirus Vaccine Response in Rural Ghana. J Infect Dis 2017; 215: 34-41.). In a recent human microbiome intervention study using a trivalent influenza vaccine, it was reported that a decrease in microbial diversity increases inflammation and reduces vaccine response (Hagan T, Cortese M, Rouphael N, et al. Antibiotics-Driven Gut Microbiome Perturbation Alters Immunity to Vaccines in Humans. Cell 2019; 178: 1313-1328.e13.).

In the case of a COVID-19 vaccine that can exclude the effect of pre-existing immunity through previous exposure to the same antigen or vaccination, it is important to evaluate the effect of the gut microbiome on vaccine immunogenicity.

Enhancing the effectiveness of existing vaccines is as important as developing effective new vaccines to the success of global efforts against the COVID-19 pandemic. Current vaccinations are non-individual and uniform, and do not consider individual variability in immunological responses. Accordingly, the present inventors have completed the invention of a method for providing information capable of increasing vaccine efficacy through the correlation between the above-described intestinal microbial community, their metabolites, and functional biomarkers and immune responses.

The technical problem to be achieved by the present invention is to provide an information providing method for predicting an immune response through the composition of the intestinal microflora of a subject before vaccination at the time of SARS-CoV-2 vaccination.

Another technical problem to be achieved by the present invention is to provide subject-specific vaccination information for determining a vaccine that can most efficiently induce an immune response in a subject through the composition of the subject's intestinal microflora.

Another technical problem to be achieved by the present invention is to provide probiotics for immunity enhancement and/or health functional food for immunity enhancement.

Another technical problem to be achieved by the present invention is to provide an information providing method for predicting an immune response through the intestinal metabolome and functional biomarkers of a subject before vaccination at the time of SARS-CoV-2 vaccination.

Another technical problem to be achieved by the present invention is to provide subject-specific vaccination information for determining a vaccine capable of inducing the most efficient immune response in a subject through the composition of the subject's intestinal metabolites and functional biomarkers.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to solve the above problems, the present invention is Parasutterella genus, Eubacterium_g23 PAC001034_s, Blautia_uc, Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, La Genus chnospiraceae PAC001043_g and Lachnospiraceae PAC001043_g Provided is a method for providing vaccination information tailored to a subject, comprising detecting any one or more microorganisms from the group consisting of PAC001449_s.

According to one side, the Eubacterium_g23 PAC001034_s species has a 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 1, the Eubacterium_g5 LT907848_s species has a 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 2, the Roseburia cecicola species has a 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 3, the Romboutsia timonensis species has a 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 4, and the Clostridium PAC001136_s species has SEQ ID NO: 5 nucleotide and 16s rRNA having 98% or more homology, and the Lachnospiraceae PAC001043_g PAC001449_s species may have 16s rRNA having 98% or more homology with nucleotide of SEQ ID NO: 6, respectively.

According to one side, the vaccine may be a vaccine against corona virus.

According to one side, after the above step, if at least one of the groups consisting of Parasutterella genus, Eubacterium_g23 PAC001034_s, and Blautia_uc is detected in the biological sample isolated from the subject, the step of determining the subject as having a good immune response to the viral vector vaccine can include more.

According to one side, the viral vector vaccine may be an adenovirus vector vaccine.

According to one side, after the above step, in the biological sample isolated from the subject, Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_g genus and Lachnospiraceae PAC001043_ At least one of the groups consisting of g PAC001449_s When this is detected, a step of determining a subject having a good immune response to the messenger ribonucleic acid (mRNA) vaccine may be further included.

According to another embodiment of the present invention, intestinal microorganisms of the genus Parasutterella, Eubacterium_g23 PAC001034_s, Blautia_uc, Ruminococcaceae PAC000661_g, Eubacterium_g5 LT907848_s, the genus Romboutsia, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae The group consisting of the genus PAC001043_g and Lachnospiraceae PAC001043_g PAC001449_s A composition for providing vaccination information tailored to a subject, including an agent for detecting any one or more of the above, is provided.

According to another embodiment of the present invention, a kit for providing vaccination information tailored to a subject, including the composition, is provided.

According to another embodiment of the present invention, genus Parasutterella, Eubacterium_g23 PAC001034_s, Blautia_uc, genus Ruminococcaceae PAC000661_g, genus Eubacterium_g5 LT907848_s, genus Romboutsia, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC00 1043_g, consisting of Lachnospiraceae PAC001043_g PAC001449_s and combinations thereof Probiotics for immunity enhancement and/or health functional food for immunity enhancement comprising at least one selected from the group are provided.

The present invention relates to endoglucanase (KEGG ortholog K01179), fumarate hydratase class I (KEGG ortholog K01676), two-component regulatory system LytTR family sensor kinase (two -component regulatory system, LytTR family, sensor kinase, KEGG ortholog K02478), anthranilate synthase component I orthologs (KEGG ortholog K01657), butyryl-CoA dehydrogenase , KEGG ortholog K00248), two-component system, OmpR family, response regulator CpxR (two-component system, OmpR family, response regulator CpxR, KEGG ortholog K07662), peptide/nickel transport system permease protein protein, KEGG ortholog K02033), putative ABC transport system ATP-binding protein (KEGG ortholog K02003), aminocarboxymuconate-semialdehyde decarboxylase (KEGG ortholog K03392 ), iron complex transport system ATP-binding protein (KEGG ortholog K02013), aromatic-L-amino-acid/L-tryptophan decarboxylase, KEGG ortholog K01593), glycine cleavage system transcriptional repressor (KEGG ortholog K03567), cobalt/nickel transport system ATP-binding protein (KEGG ortholog K02006), RNA 2', 3'-cyclic 3'-phosphodiesterase (RNA 2', 3'-cyclic 3'-phosphodiesterase, KEGG ortholog K01975), propionyl-CoA carboxylase beta chain (propionyl-CoA carboxylase beta chain, KEGG ortholog K01966), putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase small subunit (KEGG ortholog K00534), Betaine/carnitine transporter BCCT family (BCCT family, KEGG ortholog K03451), HlyD family secretion protein (KEGG ortholog K02022), thiosulfate/3-mercaptopyruvate sulfurtransferase thiosulfate/3-mercaptopyruvate sulfurtransferase (KEGG ortholog K01011), serine palmitoyltransferase (KEGG ortholog K00654), CRP/FNR family transcriptional regulator anaerobic regulatory protein , KEGG ortholog K01420), podocalyxin-like (KEGG ortholog K06817), ribosomal protein L11 methyltransferase (KEGG ortholog K02687), pyrroline-5-carboxylate reductase (pyrroline-5-carboxylate reductase, KEGG ortholog K00286), 23S rRNA (guanosine 2251-2'-O)-methyltransferase (23S rRNA (guanosine2251-2'-O)-methyltransferase, KEGG ortholog K03218), anaerobic Ribonucleoside-triphosphate reductase activating protein (anerobic ribonucleoside-triphosphate reductase activating protein, KEGG ortholog K04068), epoxyqueuosine reductase (KEGG ortholog K09765), putative sigma-54 regulatory protein (putative sigma -54 modulation protein, KEGG ortholog K05808), 2-iminobutanoate/2-iminopropanoate deaminase (2-iminobutanoate/2-iminopropanoate deaminase, KEGG ortholog K09022), rod shape protein RodA (rod shape Detecting at least one functional biomarker from the group consisting of determining protein RodA, KEGG ortholog K05837), competency protein ComEA (competence protein ComEA, KEGG ortholog K02237), and acrylaminoacyl-peptidase (KEGG ortholog K01303) Including, it provides a method for providing subject-specific vaccination information.

According to one side, the vaccine may be a vaccine against corona virus.

According to one side, after the above step, in a biological sample isolated from the subject, consisting of endoglucanase, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinase and anthranilate synthase component I ortholog. When any one or more functional biomarkers in the group are detected, a step of determining a subject having a good immune response to the viral vector vaccine may be further included.

According to one side, after the above steps, in the biological sample isolated from the subject, butyryl-CoA dehydrogenase, the two-component control system OmpR family response regulator CpxR, the peptide / nickel transport system fermease protein and putative ABC transport When one or more functional biomarkers from the group consisting of the system ATP-binding protein (KEGG ortholog K02003) are detected, a step of determining the subject as having a poor immune response to the viral vector vaccine may be further included.

According to one side, the viral vector vaccine may be an adenovirus vector vaccine.

According to one side, after the above step, in the biological sample isolated from the subject, aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid / L-tryptophan dicarboxyl lase, glycine cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'-phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, If at least one functional biomarker from the group consisting of serine palmitoyltransferase, CRP/FNR family transcriptional regulator anaerobic regulatory protein and pocalyxin-analog is detected, a good immune response to the messenger ribonucleic acid (mRNA) vaccine A step of determining the object as an object may be further included.

According to one side, after the above step, in the biological sample isolated from the subject, ribosomal protein L11 methyltransferase, pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O)- Methyltransferase, anaerobic ribonucleoside-triphosphate reductase activator protein, epoxyquiosine reductase, putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase , If at least one functional biomarker from the group consisting of rod-shaped determinant protein RodA, competence protein ComEA, and acrylaminoacyl-peptidase is detected, a step of determining a subject with a poor immune response to the messenger ribonucleic acid (mRNA) vaccine is further included. can include

According to another embodiment of the present invention, endoglucanase, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinase, anthranilate synthase component I ortholog, butyryl-CoA dehydrogenase , two-component regulatory system OmpR family response regulator CpxR, peptide/nickel transport system fermease protein, putative ABC transport system ATP binding protein (KEGG ortholog K02003), aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid/L-tryptophan decarboxylase, glycine cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'- Phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, serine palmitoyltransferase, CRP/FNR family transcriptional regulator anaerobic regulatory protein, pocalyxin-analog, ribosomal protein L11 methyltransferase, Pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O)-methyltransferase, anaerobic ribonucleoside-triphosphate reductase active protein, epoxyquiosine reductase , functional biologics of any one or more of the group consisting of putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase, rod-shaped crystal protein RodA, competence protein ComEA, and acrylaminoacyl-peptidase. A composition for providing vaccination information tailored to a subject, including an agent for detecting a marker, is provided.

According to another embodiment of the present invention, a kit for providing vaccination information tailored to a subject including the composition is provided.

According to the information providing method, the composition of the intestinal microflora taxa of the subject before inoculation, the composition of the intestinal metabolites and functional biomarkers can be identified, and the degree of humoral immune response of the subject after inoculation can be predicted.

In addition, through this, it is possible to provide information for administering a vaccine that effectively causes an immune response to a subject and has few side effects.

The effects of the present invention are not limited to the above effects, and should be understood to include all effects that can be inferred from the detailed description of the present invention or the configuration of the invention described in the claims.

1 is a schematic diagram of sample collection and survey of the present invention.

Figure 2 shows the change in alpha diversity (Shannon), beta diversity and ternary plot after administration of two different vaccine platforms.

3 shows changes in ACE and CHAO alpha diversity indices after vaccination.

4 shows changes in Jackknife and NP Shannon alpha diversity indices after vaccination.

5 shows changes in Simpson's and Phylogenetic Diversity alpha diversity indices after vaccination.

6 is a three-way plot (a: phylum, b: strong) according to the classification stage, showing changes in the microbiota for the ChAdOx1 vaccination group and the BNT162b2 vaccination group.

7 is a three-way plot (c: neck, d: family) according to the classification stage, showing the changes in the microbiota for the ChAdOx1 vaccination group and the BNT162b2 vaccination group.

8 is a three-way plot (e: species) according to the classification stage, showing the changes in the microbiota for the ChAdOx1 vaccination group and the BNT162b2 vaccination group.

Figure 9 shows baseline differences in microbiome species richness (ACE) and distance between sets with respect to immunogenicity of two different vaccine platforms.

10 shows changes in CHAO and Jackknife alpha diversity indices by dividing a good immune response group and a poor immune response group after inoculation.

11 shows changes in NP Shannon and Shannon alpha diversity indices after inoculation by dividing a good immune response group and a poor immune response group.

Figure 12 shows the change of the Simpson's and Phylogenetic Diversity alpha diversity indices by dividing the excellent immune response group and the poor immune response group after inoculation.

13 shows the results of linear discriminant analysis effect size (LEfSe) analysis for identifying taxonomic biomarkers for taxa with p < 0.05.

Figure 14 shows the results of linear discriminant analysis effect size (LEfSe) analysis for identifying functional biomarkers for metabolites with p < 0.05.

The present inventors found that the gut microbiome affects the immune response after SARS-CoV-2 vaccination, and that adenoviral vectors and vaccine components induce changes in the gut microbiome to affect the immune response after repeated vaccination. It was confirmed that the present invention was completed, and specific details for carrying out the present invention are as follows.

In order to solve the above problems, the present invention is Parasutterella genus, Eubacterium_g23 PAC001034_s, Blautia_uc, Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, La Genus chnospiraceae PAC001043_g and Lachnospiraceae PAC001043_g Provided is a method for providing vaccination information tailored to a subject, comprising the step of detecting any one or more microorganisms from the group consisting of PAC001449_s. The biological sample may preferably be feces of a subject, but is not limited thereto.

Preferably, the Eubacterium_g23 PAC001034_s species has a 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 1, and the Eubacterium_g5 LT907848_s species has a 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 2, the Roseburia cecicola The species has 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 3, the Romboutsia timonensis species has 16s rRNA having 98% or more homology with nucleotide of SEQ ID NO: 4, and the Clostridium PAC001136_s species has SEQ ID NO: 5 16s rRNA having 98% or more homology with the nucleotide, the Lachnospiraceae PAC001043_g PAC001449_s species may have 16s rRNA having 98% or more homology with the nucleotide of SEQ ID NO: 6, respectively.

The 16s rRNA is an rRNA constituting the 30S subunit of the prokaryotic ribosome, and most of the base sequences are significantly conserved, while high base sequence diversity is shown in some sections. In particular, since there is little diversity among homologous species, but there is diversity between different species, prokaryotes can be usefully identified by comparing 16S rRNA sequences.

Preferably, the genus Parasutterella, the genus Blautia, the genus Ruminococcaceae PAC000661_g, the genus Romboutsia, and the genus Lachnospiraceae PAC001043_g may include all microorganisms of subclasses, and the 16s rRNA of each microbial species included therein is per Ezbiocloud, NCBI genbank, etc. It can be confirmed through methods known in the art.

Detection of the microorganisms may be achieved through a known detection method known in the art, preferably through a detection agent. The detection agent is a primer or probe capable of specifically detecting organic biomolecules such as proteins, nucleic acids, lipids, glycolipids, glycoproteins, or sugars (monosaccharide, disaccharide, oligosaccharide, etc.) that are specifically present in the microorganism. , antisense oligonucleotides, LNA (Locked Nucleic Acids), aptamers or antibodies, and preferably fusion primers targeting a specific region of the 16S rRNA gene.

As used herein, the term "provision of subject-specific vaccination information" means that, based on the microbial taxa and functional marker information, it is possible to predict whether a subject will have an excellent or poor immune response to a specific vaccine and show the best immune response to the subject. It refers to the provision of vaccination information that is available. The subject may be a mammal including a human.

The excellent immune response group and the poor immune response group can be classified by the anti-SARS-CoV-2-S IgG titer in the blood at 3 weeks after the completion of the second vaccine vaccination, preferably anti-SARS-CoV-2 If the -S IgG titer is 2500 U/mL or more, it can be classified as an excellent immune response group, and if it is 1000 U/mL or less, it can be classified as a poor immune response group. According to the present invention, by identifying the correlation between humoral immunity levels and the composition of the intestinal microbiota, an intestinal microbial profile that can serve as a biomarker of the immune response was secured, and accordingly, information capable of predicting the immune response of the subject Provides a method of delivery.

According to one side, the vaccine may be a vaccine against corona virus. Preferably, the coronavirus vaccine may be either a viral vector vaccine or an mRNA vaccine, and more preferably, the viral vector vaccine may be ChAdOx1 and the mRNA vaccine may be BNT162b2.

According to one side, after the above step, if at least one of the groups consisting of Parasutterella genus, Eubacterium_g23 PAC001034_s, and Blautia_uc is detected in the biological sample isolated from the subject, the step of determining the subject as having a good immune response to the viral vector vaccine can include more. Preferably, the viral vector vaccine may be an adenovirus vector vaccine. As verified in Example 3 of the present invention, when the genus Parasutterella, Eubacterium_g23 PAC001034_s, and Blautia_uc are present in the intestinal microflora before inoculation (V1), an excellent immune response is obtained 3 weeks after inoculation (V3). It was confirmed as a biomarker of excellent immune response. A subject with a good immune response refers to a subject with an excellent humoral immune response after vaccination, and preferably may be a subject whose blood concentration of anti-SARS-CoV-2 S IgG is 2500 U/mL or more.

According to one side, after the above step, in the biological sample isolated from the subject, Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_g genus and Lachnospiraceae PAC001043_ At least one of the groups consisting of g PAC001449_s When this is detected, a step of determining a subject having a good immune response to the messenger ribonucleic acid (mRNA) vaccine may be further included. As verified in Example 3 of the present invention, the Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_g genus and Lachnospiraceae PAC001043_g PAC0 Microbial taxa of 01449_s at the time point before inoculation (V1) When present in the intestinal microflora, it showed an excellent immune response at the time point (V3) 3 weeks after completion of inoculation, and was identified as a biomarker of excellent immune response.

According to another embodiment of the present invention, intestinal microorganisms of the genus Parasutterella, Eubacterium_g23 PAC001034_s, Blautia_uc, Ruminococcaceae PAC000661_g, Eubacterium_g5 LT907848_s, the genus Romboutsia, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae A group consisting of the genus PAC001043_g and Lachnospiraceae PAC001043_g PAC001449_s A composition for providing vaccination information tailored to a subject, including an agent for detecting any one or more of the above, is provided. Description of the detection agent is as described above.

According to another embodiment of the present invention, a kit for providing vaccination information tailored to a subject, including the composition, is provided.

The kit may include a detection agent as an essential component. As a non-limiting example, in the case of an RT-PCR kit, in addition to a fusion primer complementary to the specific nucleotide of each microorganism, a test tube or other suitable container, reaction buffer, deoxynucleotides (dNTPs), Taq-polymerase and reverse transcriptase, DNase, RNase inhibitors, DEPC-water, sterile water, and the like. In addition, the kit may include various tools and reagents known in the art that facilitate the measurement of microbial levels, such as suitable carriers, labeling substances capable of generating a detectable signal, stabilizers, and the like.

According to the present invention, it was confirmed that vaccination also affects the species diversity of intestinal microorganisms. The species diversity may preferably be alpha diversity, more preferably any one or more selected from the group consisting of ACE, Chao1, Jackknife, Shannon, NPShannon, Simpson and Phylogenetic diversity, most preferably ACE alpha diversity can be As verified in Example 2 below, there was a significant correlation between the species diversity of the intestinal microflora before inoculation and the excellent immune response after inoculation. In the group having a high ACE alpha diversity index before vaccination, an excellent immune response with an anti-S IgG titer of 2500 or more was observed after completion of vaccination.

The present inventors found that the following group 1 and group 2 gut microbial taxa had a significant correlation with the immune response after vaccination, respectively.

Group 1: Genus Parasutterella, Eubacterium_g23 PAC001034_s, Blautia_uc, Genus Ruminococcaceae PAC000661_g, Genus Eubacterium_g5 LT907848_s, Genus Romboutsia, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_ The group consisting of the genus g and Lachnospiraceae PAC001043_g PAC001449_s; and

Group 2: the group consisting of Anaerotignum PAC001031_s, Bifidobacterium animalis, Bacteroides dorei and Megasphaera indica;

As shown in Example 3 below, the microorganisms belonging to the first group were a relatively dominant taxa of the intestinal microflora before inoculation of the group having an excellent immune response after inoculation. Preferably, Parasutterella genus, Eubacterium_g23 PAC001034_s and Blautia_uc were relatively dominant in the ChAdOx1 inoculated group with excellent immune response, and in the BNT162b2 inoculated group with excellent immune response, Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia ti monensis, Clostridium The genera PAC001136_s, Lachnospiraceae PAC001043_g and Lachnospiraceae PAC001043_g PAC001449_s belonged to the relatively dominant taxa.

As shown in Example 4 below, the microorganisms belonging to the second group were relatively dominant taxa of the intestinal microflora before inoculation of the group with poor immune response after inoculation. Preferably, Anaerotignum PAC001031_s and Bifidobacterium animalis were relatively dominant in the poor immune response group of the ChAdOx1 inoculated group, and Bacteroides dorei and Megasphaera indica belonged to relatively dominant taxa in the poor immune response group of the BNT162b2 inoculated group.

Specifically, the present invention relates to genus Parasutterella, Eubacterium_g23 PAC001034_s, Blautia_uc, genus Ruminococcaceae PAC000661_g, genus Eubacterium_g5 LT907848_s, genus Romboutsia, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043 Genus _g, selected from the group consisting of Lachnospiraceae PAC001043_g PAC001449_s and combinations thereof Provides probiotics for enhancing immunity comprising at least one.

In addition, the present invention is Parasutterella genus, Eubacterium_g23 PAC001034_s, Blautia_uc, Ruminococcaceae PAC000661_g genus, Eubacterium_g5 LT907848_s, Romboutsia genus, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_g At least one selected from the group consisting of the genus Lachnospiraceae PAC001043_g PAC001449_s and combinations thereof It provides a health functional food for enhancing immunity comprising a.

In the present invention, "probiotics" refers to microorganisms having antibacterial and enzymatic activities that help balance intestinal microorganisms and products produced by the microorganisms. In addition, probiotics are defined as live bacteria in the form of single or complex strains that are supplied to humans or animals in the form of dry cells or fermentation products to improve the intestinal flora.

In the present invention, "immunity enhancement" means strengthening and/or improving the immunity of an organism. It includes those that enhance immunity against diseases or have good immune responses to vaccines.

The health functional food of the present invention can be used in various ways such as pharmaceuticals, foods, and beverages for improving the intestinal environment. Functional foods of the present invention include, for example, various foods, candy, chocolate, beverages, gum, tea, vitamin complexes, health supplements, etc., and can be used in the form of powders, granules, tablets, capsules or beverages.

The health functional food of the present invention may be added to food or beverages for the purpose of improving the intestinal environment. At this time, the amount of the extract in the food or beverage is generally 0.01 to 50% by weight, preferably 0.1 to 20% by weight of the total food weight of the health functional food composition of the present invention, and the health drink composition is 100 ml Based on 0.02 to 10 g, it may be added at a rate of preferably 0.3 to 1 g.

The health beverage composition of the present invention has no particular limitations on the liquid component except for containing the strain as an essential component in the indicated ratio, and may contain various flavors or natural carbohydrates as additional components like conventional beverages. Examples of the natural carbohydrates described above are monosaccharides such as glucose and fructose, disaccharides such as maltose, sucrose, and lactose, oligosaccharides, polysaccharides such as dextrins and cyclodextrins, and sugar alcohols such as xylitol, sorbitol, and erythritol. . As flavoring agents other than those described above, flavoring agents (stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic sweeteners (saccharin, aspartame, etc.) can advantageously be used. The proportion of is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, It may contain organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected from the range of 0 to about 50 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention relates to endoglucanase (KEGG ortholog K01179), fumarate hydratase class I (KEGG ortholog K01676), two-component regulatory system LytTR family sensor kinase (two -component regulatory system, LytTR family, sensor kinase, KEGG ortholog K02478), anthranilate synthase component I orthologs (KEGG ortholog K01657), butyryl-CoA dehydrogenase , KEGG ortholog K00248), two-component system, OmpR family, response regulator CpxR (two-component system, OmpR family, response regulator CpxR, KEGG ortholog K07662), peptide/nickel transport system permease protein protein, KEGG ortholog K02033), putative ABC transport system ATP-binding protein (KEGG ortholog K02003), aminocarboxymuconate-semialdehyde decarboxylase (KEGG ortholog K03392 ), iron complex transport system ATP-binding protein (KEGG ortholog K02013), aromatic-L-amino-acid/L-tryptophan decarboxylase, KEGG ortholog K01593), glycine cleavage system transcriptional repressor (KEGG ortholog K03567), cobalt/nickel transport system ATP-binding protein (KEGG ortholog K02006), RNA 2', 3'-cyclic 3'-phosphodiesterase (RNA 2', 3'-cyclic 3'-phosphodiesterase, KEGG ortholog K01975), propionyl-CoA carboxylase beta chain (propionyl-CoA carboxylase beta chain, KEGG ortholog K01966), putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase small subunit (KEGG ortholog K00534), Betaine/carnitine transporter BCCT family (BCCT family, KEGG ortholog K03451), HlyD family secretion protein (KEGG ortholog K02022), thiosulfate/3-mercaptopyruvate sulfurtransferase thiosulfate/3-mercaptopyruvate sulfurtransferase (KEGG ortholog K01011), serine palmitoyltransferase (KEGG ortholog K00654), CRP/FNR family transcriptional regulator anaerobic regulatory protein , KEGG ortholog K01420), podocalyxin-like (KEGG ortholog K06817), ribosomal protein L11 methyltransferase (KEGG ortholog K02687), pyrroline-5-carboxylate reductase (pyrroline-5-carboxylate reductase, KEGG ortholog K00286), 23S rRNA (guanosine 2251-2'-O)-methyltransferase (23S rRNA (guanosine2251-2'-O)-methyltransferase, KEGG ortholog K03218), anaerobic Ribonucleoside-triphosphate reductase activating protein (anerobic ribonucleoside-triphosphate reductase activating protein, KEGG ortholog K04068), epoxyqueuosine reductase (KEGG ortholog K09765), putative sigma-54 regulatory protein (putative sigma -54 modulation protein, KEGG ortholog K05808), 2-iminobutanoate/2-iminopropanoate deaminase (2-iminobutanoate/2-iminopropanoate deaminase, KEGG ortholog K09022), rod shape protein RodA (rod shape Detecting at least one functional biomarker from the group consisting of determining protein RodA, KEGG ortholog K05837), competency protein ComEA (competence protein ComEA, KEGG ortholog K02237), and acrylaminoacyl-peptidase (KEGG ortholog K01303) Including, it provides a method for providing subject-specific vaccination information. The biological sample may preferably be feces of a subject, but is not limited thereto.

The KEGG ortholog used to define the functional biomarker refers to a group of homologous genes in the genome provided by the Kyoto Encyclopedia of Genes and Genomes.

As used herein, the term "functional biomarker" is a biomarker selected to predict the immune response of a subject based on the function in the metabolic pathway of a metabolite, preferably the metabolic pathway and It may be a relevant metabolomic biomarker.

Detection of the functional biomarker may be performed using a method known in the art. When the component to be detected is an amino acid, it may be a primer, probe, aptamer, or antibody capable of complementary binding, and in the case of a protein, LC- It can be done through MS (liquid chromatography-mass spectrometry) technique or ELISA (enzyme-linked immunosorbent assay), and in other cases, it can be detected through reagents or chemicals that can quantify them.

According to one side, after the above step, in a biological sample isolated from the subject, consisting of endoglucanase, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinase and anthranilate synthase component I ortholog. When any one or more functional biomarkers in the group are detected, a step of determining a subject having a good immune response to the viral vector vaccine may be further included.

According to one side, after the above steps, in the biological sample isolated from the subject, butyryl-CoA dehydrogenase, the two-component control system OmpR family response regulator CpxR, the peptide / nickel transport system fermease protein and putative ABC transport When one or more functional biomarkers from the group consisting of the system ATP-binding protein (KEGG ortholog K02003) are detected, a step of determining the subject as having a poor immune response to the viral vector vaccine may be further included.

As verified in Example 5 of the present invention, functional biomarkers of the endoglucanase, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinase, and anthranilate synthase component I ortholog were inoculated. If detected at all time points (V1), an excellent immune response was shown at 3 weeks after completion of inoculation (V3), which was confirmed as a biomarker of excellent immune response, butyrill-CoA dehydrogenase, two-component control system OmpR family 3 weeks after inoculation (V3 ) showed a poor immune response, and was identified as a biomarker of poor immune response. A subject with a good immune response refers to a subject with an excellent humoral immune response at 3 weeks after completion of vaccination, preferably a subject whose blood concentration of anti-SARS-CoV-2 S IgG is 2500 U/mL or more can be The subject with a poor immune response refers to a subject with a poor humoral immune response at 3 weeks after completion of vaccination, preferably a subject whose blood anti-SARS-CoV-2 S IgG concentration is 1000 U/mL or less can be

According to one side, the viral vector vaccine may be an adenovirus vector vaccine.

According to one side, after the above step, in the biological sample isolated from the subject, aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid / L-tryptophan dicarboxyl lase, glycine cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'-phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, If at least one functional biomarker from the group consisting of serine palmitoyltransferase, CRP/FNR family transcriptional regulator anaerobic regulatory protein and pocalyxin-analog is detected, a good immune response to the messenger ribonucleic acid (mRNA) vaccine A step of determining the object as an object may be further included.

According to one side, after the above step, in the biological sample isolated from the subject, ribosomal protein L11 methyltransferase, pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O)- Methyltransferase, anaerobic ribonucleoside-triphosphate reductase activator protein, epoxyquiosine reductase, putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase , If at least one functional biomarker from the group consisting of rod-shaped determinant protein RodA, competence protein ComEA, and acrylaminoacyl-peptidase is detected, a step of determining a subject with a poor immune response to the messenger ribonucleic acid (mRNA) vaccine is further included. can include

As verified in Example 5 of the present invention, the aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid / L-tryptophan decarboxylase, glycine Cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'-phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP -binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, serine palmitoyltransfer When functional biomarkers of lase, CRP/FNR family transcriptional regulator anaerobic regulatory protein, and pocalyxin-analog were detected before inoculation (V1), an excellent immune response was shown at 3 weeks after inoculation (V3) , It was identified as a biomarker of an excellent immune response, and the ribosomal protein L11 methyltransferase, pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O) -methyltransferase, Anaerobic ribonucleoside-triphosphate reductase active protein, epoxyquiosine reductase, putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase, rod-shaped crystal protein When functional biomarkers of RodA, competency protein ComEA, and acrylaminoacyl-peptidase were detected before inoculation (V1), a poor immune response was shown at 3 weeks after inoculation (V3), which is considered a biomarker of poor immune response. Confirmed. A subject with a good immune response refers to a subject with an excellent humoral immune response at 3 weeks after completion of vaccination, preferably a subject whose blood concentration of anti-SARS-CoV-2 S IgG is 2500 U/mL or more can be The subject with a poor immune response refers to a subject with a poor humoral immune response at 3 weeks after completion of vaccination, preferably a subject whose blood anti-SARS-CoV-2 S IgG concentration is 1000 U/mL or less can be

According to another embodiment of the present invention, endoglucanase, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinase, anthranilate synthase component I ortholog, butyryl-CoA dehydrogenase , two-component regulatory system OmpR family response regulator CpxR, peptide/nickel transport system fermease protein, putative ABC transport system ATP binding protein (KEGG ortholog K02003), aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid/L-tryptophan decarboxylase, glycine cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'- Phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, serine palmitoyltransferase, CRP/FNR family transcriptional regulator anaerobic regulatory protein, pocalyxin-analog, ribosomal protein L11 methyltransferase, Pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O)-methyltransferase, anaerobic ribonucleoside-triphosphate reductase active protein, epoxyquiosine reductase , functional biologics of any one or more of the group consisting of putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase, rod-shaped crystal protein RodA, competence protein ComEA, and acrylaminoacyl-peptidase. A composition for providing vaccination information tailored to a subject, including an agent for detecting a marker, is provided. The detection agent may be suitable for the detection method of the functional biomarker described above.

According to another embodiment of the present invention, a kit for providing vaccination information tailored to a subject including the composition is provided.

The kit may include a detection agent as an essential component. As a non-limiting example, in the case of an ELISA kit, a capture antibody, a detection antibody, a buffer, a test tube or other appropriate container, a reaction buffer, sterile water, and the like may be included. In addition, the kit may include various tools and reagents known in the art that facilitate measurement of functional biomarkers, such as suitable carriers, labeling substances capable of generating detectable signals, stabilizers, and the like.

The present inventors found that the functional biomarker populations of group 1 and group 2 below each had a significant correlation with the immune response after vaccination.

Group 1: endoglucanases, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinases, anthranilate synthase component I orthologs, aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP-binding protein, aromatic-L-amino acid/L-tryptophan decarboxylase, glycine cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3' -phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted proteins, thiosulfate/3-mercaptopyruvate sulfurtransferase, serine palmitoyltransferase, CRP/FNR family transcriptional regulator anaerobic regulatory proteins and pocalyxin-analogs; and

Group 2: butyryl-CoA dehydrogenase, two-component regulatory system OmpR family response regulator CpxR, peptide/nickel transport system permease protein, putative ABC transport system ATP binding protein (KEGG ortholog K02003), ribosomal protein L11 Methyltransferase, pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O)-methyltransferase, anaerobic ribonucleoside-triphosphate reductase active protein, epoxy quiosine reductase, putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase, rod-shaped determinant protein RodA, competence protein ComEA and acrylaminoacyl;

As shown in Example 5 below, functional markers belonging to the first group were relatively dominant functional biomarkers of the intestinal microflora before inoculation of the group in which the immune response was excellent after completion of inoculation. Preferably, endoglucanase, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinase, and anthranilate synthase component I orthologs were relatively dominant in the superior immunoreactive group of the ChAdOx1 inoculum, and BNT162b2 Aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid/L-tryptophan decarboxylase, glycine cleavage system transcriptional repressor, Cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'-phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, serine palmitoyltransferase, CRP/FNR The family of transcriptional regulators anaerobic regulatory proteins and pocalyxin-analogues belonged to the relatively predominant functional biomarkers.

As shown in Example 5 below, the microorganisms belonging to the second group were relatively dominant functional biomarkers before inoculation of the group with poor immune response after inoculation. Preferably, butyryl-CoA dehydrogenase, two-component control system OmpR family response regulator CpxR, peptide / nickel transport system fermease protein and putative ABC transport system ATP binding protein ( KEGG ortholog K02003) was relatively dominant, and ribosomal protein L11 methyltransferase, pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O) -Methyltransferase, anaerobic ribonucleoside-triphosphate reductase activator protein, epoxyquiosine reductase, putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate diamina enzyme, rod-shaped crystal protein RodA, competence protein ComEA, and acrylaminoacyl were among the relatively predominant functional biomarkers.

Hereinafter, embodiments will be described in detail with reference to the accompanying drawings. However, since various changes can be made to the embodiments, the scope of the patent application is not limited or limited by these embodiments. It should be understood that all changes, equivalents or substitutes to the embodiments are included within the scope of rights.

Terms used in the examples are used only for the purpose of explanation and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "include" or "have" are intended to designate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, but one or more other features It should be understood that the presence or addition of numbers, steps, operations, components, parts, or combinations thereof is not precluded.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiment belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in the present application, they should not be interpreted in an ideal or excessively formal meaning. don't

In addition, in the description with reference to the accompanying drawings, the same reference numerals are given to the same components regardless of reference numerals, and overlapping descriptions thereof will be omitted. In describing the embodiment, if it is determined that a detailed description of a related known technology may unnecessarily obscure the gist of the embodiment, the detailed description will be omitted.

Example 1: Experimental method

1-1: Experiment outline

A cohort study was conducted with adult COVID-19 vaccinated subjects, including 27 BNT162b2 and 26 ChAdOx1 vaccinated subjects. Samples were excluded from participants who had a history of drugs that may have affected the gut microbiome, such as antibiotics, laxatives, and motility drugs within 1 month, or who had a history of or tested positive for SARS-CoV-2 by nasopharyngeal PCR test. Blood and stool samples were collected at each time point before the first inoculation (V1), before the second inoculation (V2) and 3 weeks after the second inoculation (V3). We performed 16S rRNA gene sequencing of fecal samples and analyzed the gut microbiome profile in relation to the humoral immune response to SARS-CoV-2 spike protein after vaccination. The study was approved by the Institutional Review Board (2021GR0097) of Korea University Guro Hospital, and all participants gave informed consent to participate in the study. All procedures were performed in accordance with the ethical standards of institutional and national research committees and the 1964 Declaration of Helsinki and its subsequent amendments or similar ethical standards.

1-2: Sample collection and processing

Stool samples were collected using Fecal Swab DNA Preservation & Transport Kits (Noble Bio, Hwaseong, Korea) with nucleic acid preservation medium. Fecal samples from fecal smear transport medium were stored at -80 °C. Blood samples were collected in serum separation tubes by venipuncture, centrifuged at 2500 rpm for 10 min at -4 °C, and the serum-containing supernatant was pipetted into clean plastic screw cap vials and stored at -80 °C.

1-3: Immunoassay for quantitative measurement of antibodies to SARS-CoV-2 spike protein

Anti-S antibody titers were measured using the anti-SARS-CoV-2 S assay kit (Roche, Switzerland) according to the protocol of Elecsys®. A titer below the lower limit of quantification was set at a value of 0.4. Based on data on immune correlation of protection and association between anti-S IgG and neutralizing antibody titers, participants with an IgG titer of 2500 or greater in V3 were labeled as good immune responders, and participants with titers less than 1000 were labeled as having poor immune responses. labeled as a group. In addition, participants with an IgG titer of 120 or more in V2 were labeled as the V2 excellent immune response group, and participants with a V2 titer of less than 40 were labeled as the V2 poor immune response group.

1-4: Microbial analysis

Total DNA from the microbiota was extracted using the FastDNA® SPIN Kit for Soil (MP Biomedicals, Southern California, USA) according to the manufacturer's instructions. PCR amplification was performed using fusion primers targeting the V3-V4 region of the 16S rRNA gene using the extracted DNA as a template. Primers of SEQ ID NOs: 7 and 8 in Table 1 were used for amplification. The highlighted part below is the target site.

Primer name Sequence (5'->3') SEQ ID NO: 7 341F AGATGTGTATAAGAGACAG CCTACGGGNGGCWGCAG SEQ ID NO: 8 805R AGATGTGTATAAGAGACAG GACTACHVGGGTATCTAATCC

The primer of SEQ ID NO: 7 was fused to the 3' end of 5'-AATGATACGGCGACCACCGAGATCTACAC-XXXXXXXXTCGTCGGCAGCGTC-3' composed of P5 graft binding, i5 index, and NexTera consensus. The primer of SEQ ID NO: 8 was fused to the 3' end of 5'-CAAGCAGAAGACGGCATACGAGAT-XXXXXXXXGTCTCGTGGGCTCGG-3' composed of P7 graft binding, i7 index, and NexTera consensus. The "X" represents the Illumina NexTera barcode region. PCR amplification was performed under the following conditions: initial denaturation at 95 °C for 3 min, followed by 25 cycles of denaturation at 95 °C for 30 sec, primer binding at 55 °C for 30 sec, elongation at 72 °C for 30 sec. , and a final elongation step at 72 °C for 5 min. PCR products were confirmed by electrophoresis using a 1% agarose gel and then visualized using the Gel Doc system (Bio-Rad, Hercules, CA, USA). Amplified products were purified using CleanPCR kit (CleanNA). Equal concentrations of purified products were pooled and short fragments (off-target products) were removed using the CleanPCR kit (CleanNA). Quality and product size were evaluated with a Bioanalyzer 2100 system (Agilent, Palo Alto, Calif., USA) using a DNA 7500 chip. Mixed amplicons were pooled and sequenced at ChunLab, Inc. (Seoul, Korea) using the Illumina MiSeq Sequencing System (Illumina, San Diego, CA, USA) according to the manufacturer's instructions.

1-5: DNA analysis pipeline

Raw readings were first quality checked and low quality (<Q25) readings were filtered out using Trimmomatic (ver. 0.32). After QC processing, paired-end sequence data was merged with default parameters using the fastq_mergepairs command of VSEARCH ver.2.13.4. Next, the primers were trimmed using the Myers-Miller alignment algorithm at a similarity cutoff of 0.8 (Myers EW, Miller W. Optimal alignments in linear space. Comput Appl Biosci 1988; 4: 11-7.). Non-specific amplicons, i.e., those that do not encode 16S rRNA, were detected using the nhmmer algorithm of the HMMER software package version 3.2.1 with hmm profile (Wheeler TJ, Eddy SR. nhmmer: DNA homology search with profile HMMs. Bioinformatics 2013; 29: 2487-2489.). Unique reads were extracted and redundant reads were clustered into unique reads using VSEARCH's deep-full-length command (Rognes T, Flouri T, Nichols B, et al. VSEARCH: a versatile open source tool for metagenomics. PeerJ 2016; 4: e2584.). EzBioCloud 16S rRNA database (Yoon SH, Ha SM, Kwon S, et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol 2017; 67: 1613-1617.) It was used for taxonomic assignment using VSEARCH's usearch_global command, after which a more accurate pairwise alignment was performed. Chimeric reads were filtered to obtain reads of <97% similarity by reference-based chimeric reads detection using the CHIME algorithm and EzBioCloud's non-chimeric 16S rRNA database. After chimeric lyse filtering, lyses that were not identified at the species level (<97% similarity) in the EzBioCloud database were compiled and de novo clustering was performed using the cluster_fast command to generate additional operational taxonomic units (OTUs). Next, OTUs with single reads (singletons) were excluded from further analysis. Secondary analysis including diversity calculation and biomarker discovery was performed using ChunLab, Inc.'s own program.

The following alpha diversity indices were estimated: ACE (Chao A, Lee S-M. Estimating the Number of Classes via Sample Coverage. J Am Stat Assoc1992; 87: 210-217.), Chao1 (Chao A. Estimating the population size for capture -recapture data with unequal catchability. Biometrics 1987; 43: 783-791.), Jackknife (Burnham KP, Overton WS. Robust Estimation of Population Size When Capture Probabilities Vary Among Animals. Ecology 1979; 60: 927-936.), Shannon (Magurran AE. Measuring biological diversity: John Wiley & Sons, 2013.), NPShannon (Chao A, Shen T-J. Nonparametric estimation of Shannon's index of diversity when there are unseen species in sample. Environ Ecol Stat 2003; 10: 429-443 .), Simpson and Phylogenetic diversity (Faith DP. Conservation evaluation and phylogenetic diversity. Biol Conserv 1992; 61: 1-10.), several algorithms (Jensen-Shannon (Lin J. Divergence measures based on the Shannon entropy. IEEE Trans Inf Theory 1991; 37: 145-151.), Bray-Curtis (Beals EW. Bray-Curtis ordination: an effective strategy for analysis of multivariate ecological data. Adv Ecol Res 1984; 14: 1-55.), Generalized UniFrac (Chen J, Bittinger K, Charlson ES, et al. Associating microbiome composition with environmental covariates using generalized UniFrac distances. Bioinformatics 2012; 28: 2106-2113.) and Fast UniFrac (Hamady M , Lozupone C, Knight R. Fast UniFrac: facilitating high-throughput phylogenetic analyzes of microbial communities including analysis of pyrosequencing and PhyloChip data. ISME J 2010; 4: 17-27.) to calculate beta diversity distances. The biomarker uses a statistical comparison algorithm (linear discriminant analysis effect size, LEfSe] (Segata N, Izard J, Waldron L, et al. Metagenomic biomarker discovery and explanation. Genome Biol 2011; 12: R60.) Logarithmic scoring of the above linear discriminant analysis is presented as the LDA score All microbiome count data were normalized to the number of 1000 reads before further use All assays mentioned above were performed in ChunLab , Inc.'s bioinformatics cloud platform, EzBioCloud 16S-based MTP.

1-6: Statistical analysis

All continuous variables are expressed as median ± interquartile range (IQR, range between first and third quartile) deviations, and differences between different groups were compared using non-parametric tests. Categorical variables were expressed as numbers (percentages). The Mann-Whitney U test was used to compare ChAdOx1 and BNT162b2 vaccinated groups, and the Wilcoxon signed-rank test was used to compare identical vaccinated groups pairwise. Statistical analysis was performed using R Statistical software version 4.1.2 (R Foundation for Statistical Computing, Vienna, Austria).

Example 2. Blood test

A total of 159 blood samples were collected from 53 fully vaccinated participants before the first dose (V1), before the second dose (V2) and 3 weeks after the second dose (V3) (Fig. 1). All 159 blood samples (81 from BNT162b2 inoculated subjects and 78 from ChAdOx1 inoculated subjects) were scored for anti-SARS-CoV-2 S IgG.

The baseline characteristics of the participants according to the type of vaccine inoculated and the antibody response are shown in Tables 2 to 4 below. Table 2 below shows blood test results of all of the ChAdOx1 and BNT162b2 inoculation groups, and Tables 3 and 4 below show details of dividing the ChAdOx1 and BNT162b2 inoculation groups into excellent and poor immune response groups, respectively.

ChAdOx1 BNT162b2 V1 V2 V3 V1 V2 V3 age 37.0±24.0 31.0±8.5 Female ratio (%) 20 (50.0) 20 (50.0) BMI 23.4±5.7 20.9±4.9 Anti-SARS-CoV-2 S IgG (U/mL) 0.4 73.4±84.5 1373±3320.5 0.4 70.3±162.7 1981±1423.0 MCHC (g/dL) 32.8±0.8 33.5±0.6 33.5±0.8 33.0±0.9 33.1±0.9 33.1±1.0 Albumin (g/dL) 4.50±0.40 4.50±0.20 4.45±0.32 4.40±0.30 4.60±0.30 4.50±0.30

excellent immune response group poor immune response V1 V2 V3 V1 V2 V3 age 40.0±22.0 34.5±16.0. Female ratio (%) 7 (50.0) 7 (50.0) BMI 24.1±6.5 20.9±4.4 Anti-SARS-CoV-2 S IgG (U/mL) 0.4 69.4±100.6 4278.0±2663.0 0.4 58.9±84.2 522.0±496.0 MCHC (g/dL) 32.6±0.9 33.3±0.7 33.5±0.8 32.9±0.6 33.8±0.4 33.9±0.7 Albumin (g/dL) 4.40±0.40 4.50±0.30 4.40±0.45 4.60±0.35 4.50±0.17 4.45±0.20

excellent immune response group poor immune response V1 V2 V3 V1 V2 V3 age 35.0±18.0 35.0±11.0 Female ratio (%) 6 (60.0) 4 (40.0) BMI 23.5±7.1 20.6±10.1 Anti-SARS-CoV-2 S IgG (U/mL) 0.4 181.0±120.9 3183.0±3222.5 0.4 12.9±37.6 808.0±351.3 MCHC (g/dL) 33.0±0.6 32.9±0.65 33.0±1.1 33.0±1.3 33.5±1.4 33.5±1.1 Albumin (g/dL) 4.40±0.25 4.50±0.50 4.50±0.40 4.25±0.58 4.80±0.38 4.55±0.55

The geometric mean titer (GMT) of anti-SARS-CoV-2 S IgG at baseline (V1) was 0.4 U/mL in all participants. Regardless of vaccine type, higher antibody titers were observed in participants who received 2 doses than those who received 1 dose. It was confirmed that there was no significant difference in the GMT of anti-SARS-CoV-2 S IgG titers between the ChAdOx1 vaccinated group and the BNT162b2 vaccinated group in V1, V2 and V3.

In the ChAdOx1 vaccinated group, compared to V1, mean corpuscular hemoglobin concentration (MCHC) was increased in V2 or V3. In the BNT162b2 vaccinated group, albumin levels were increased in both V2 and V3 compared to V1.

Example 3. Changes in the biodiversity of the intestinal microflora after vaccination

Stool samples were collected at three consecutive time points (V1, V2 and V3) from 53 participants to investigate changes in the gut microbiota in response to vaccination (Fig. 1). Of the 159 stool samples collected, 30 were excluded from analysis due to poor quality and 129 stool samples were used for sequencing (63 from ChAdOx1 inoculum and 66 from BNT162b2 inoculum). The humoral immune response was evaluated using corresponding blood samples.

It was confirmed that the average community abundance and microbial diversity (alpha diversity) gradually decreased in the group vaccinated with ChAdOx1 vaccine. However, there was no significant difference in these parameters in the BNT162b2 vaccinated group (Fig. 2A). Using the Wilcoxon rank sum test, it was confirmed that Shannon diversity decreased in V2 and V3 compared to V1 in the ChAdOx1 inoculum (FIG. 4d). Similar to alpha diversity, principal coordinates analysis (PCoA) of Bray-Curtis distance (beta diversity) using PERMANOVA in ChAdOx1 inoculum (Fig. 2B) showed no significant differences in V2 (p = 0.028) or V3 (p = 0.028) compared to V1. = 0.001) of metagenomes, it was confirmed that there was a distinct inter-set distance.

Changes in the microbiome of V1, V2, and V3 are plotted in three-way compartments to show the distribution of bacterial taxonomic groups and relative abundances in each vaccination group (relative abundance >1%) when compared to those observed after vaccination with BNT162b2. Significant changes were confirmed after vaccination with ChAdOx1. The ternary divisions of genus and other taxonomic levels (phylum, class, order, family and species) are shown in Figure 2C and Figures 6-8, respectively.

Example 4. Analysis of correlation between humoral immunogenicity and intestinal microflora after vaccination

To evaluate the role of the human gut microbiota in the humoral immune response to vaccines, participants were divided into two groups, good and poor immune responders, according to anti-SARS-CoV-2 S IgG titers at V3. Comparing the two groups, greater baseline alpha diversity in V1 was observed in the good immunoreactors (FIG. 9A and FIGS. 10-12). Specific figures are Median ACE Index, 195.61; IQR, 82.46 and 141.78; IQR, 44.58, p < 0.005.

Consistent with this, in the case of the ChAdOx1 inoculated group, higher bacterial species richness was shown in the good immune responder group than the poor immune responder group using the Wilcoxon rank sum test. Specific figures are Median ACE Index, 188.78; IQR, 64.23 and median, 138.52; IQR, 55.36, p = 0.016. However, although a similar trend was observed, there was no statistically significant difference in baseline alpha diversity between good and poor responders among BNT162b2 inoculated subjects. Beta diversity results evaluated by PCoA of Bray-Curtis distance showed no significant difference in distance between sets (Fig. 9B).

The relative abundance of bacterial taxa was assessed to determine microbiological factors (taxonomic biomarkers in V1 based on antibody titers in V3) that influence antibody production to the COVID-19 vaccine. LEfSe was used to differentiate the composition of the gut microbiome based on the immune response in both the ChAdOx1 and BNT162b2 groups (FIG. 13A, 13B). As can be seen in the figure, the pre-vaccination intestinal microbiome of the excellent immune response group in the ChAdOx1 inoculation group was detected the Parasutterella genus, Eubacterium_g23 PAC001034_s and Blautia_uc, and the poor immune response group PAC001031_s and Bifidobacterium animalis group were detected. In BNT162b2 inoculated subjects, the gut microbiome of the superior immune response group was Ruminococcaceae PAC000661_g, Eubacterium_g5 LT907848_s, Romboutsia, Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_g, and Lachnospiraceae PAC00104. 3_g PAC001449_s genus was detected, and Bacteroides dorei and Megasphaera were detected in the poor immune response group. indica was predominantly detected.

In addition, biomarkers of an excellent immune response after vaccination were analyzed for the time point before vaccination, and biomarkers were determined based on an LDA score of 2.5 or higher. showed up

ChAdOx1 (taxonomic markers V1 : V3) taxon name LDA score
(log 10) p-value genus Parasutterella 3.55382 0.02402 Eubacterium_g23 PAC001034_s 2.95184 0.02381 Blautia_uc 2.72821 0.04829

BNT162b2 (taxonomic markers V1 : V3) taxon name LDA score
(log 10) p-value Genus Ruminococcaceae PAC000661_g 3.72866 0.01823 Eubacterium_g5 LT907848_s 3.61881 0.02987 genus Romboutsia 3.58706 0.04067 Roseburia cecicola 3.56697 0.04119 Romboutsia timonensis 3.55002 0.04067 Clostridium PAC001136_s 3.39063 0.01823 Genus Lachnospiraceae PAC001043_g 3.30157 0.04119 Lachnospiraceae PAC001043_g PAC001449_s 3.20569 0.01823

Example 5. Correlation analysis between humoral immunogenicity and functional profile after vaccination

Based on the categorical difference in V1 of each participant, the functional profile predicting the humoral immune response after vaccination between the intestinal microflora of the good and poor immune responders was identified and shown in FIG. 14, LDA Functional markers were selected based on the effect size of 1.5 or more, and specific LDA score values and p-values are shown in Table 7 for ChAdOx1 and Table 8 for BNT162b2.

Functional markers in ChAdOx1 (V1: V3) marker KEGG orthologs LDA score
(log 10) p-value endoglucanase K01179 2.207036 0.020 anthranilate synthase component I K01657 1.603838 0.039 fumarate hydratase, class I K01676 1.666197 0.039 two-component system, LytTR family, sensor kinase K02478 1.658674 0.020 butyryl-CoA dehydrogenase K00248 -1.525516 0.014 Two-component system, OmpR family, response regulator CpxR K07662 -1.813081 0.039 peptide/nickel transport system permease protein K02033 -2.062776 0.039 putative ABC transport system ATP-binding protein K02003 -2.393052 0.020

Here, a marker having a positive LDA score is a marker that was dominant in the good immune response group, and a marker having a negative LDA score is a marker that was dominant in the poor immune response group. LEfSe, or LDA effect size, is the absolute value of the LDA score.

In the excellent immune response group after ChAdOx1 inoculation, endoglucanase (LEfSe, 2.21, p=0.02, KEGG ortholog K01179), fumarate hydratase class I (LEfSe, 1.67, p=0.04, KEGG ortholog K01676), two-component regulatory system (LytTR family, sensor kinase, LEfSe, 1.66, p = 0.02, KEGG ortholog K02478) and anthranilate synthase component I ortholog (anthranilate synthase component I orthologs, LEfSe, 1.60, p=0.04, KEGG ortholog K01657) were detected, and butyryl-CoA dehydrogenase (LEfSe, 1.53, p=0.014, KEGG ortholog K00248) was detected in the poor immune response group. , two-component system, OmpR family, response regulator CpxR (two-component system, OmpR family, response regulator CpxR, LEfSe, 1.81, p = 0.039, KEGG ortholog K07662), peptide / nickel transport system permease protein (peptide / nickel transport system permease protein, LEfSe, 2.062, p=0.039, KEGG ortholog K02033) and putative ABC transport system ATP-binding protein (LEfSe, 2.39, p=0.020, KEGG ortholog K02003). detected (FIG. 14A and Table 6). In the Kyoto Encyclopedia of Genes and Genomes (KEGG) module PTS system using the MinPath algorithm, galactitol-specific II component (LefSe, 2.50, p=0.014), KEGG cutin, suberine and wax biosynthesis (LefSe, 2.36, p=0.039) were confirmed in significant amounts. When using the PICRUSt algorithm, KEGG module multidrug resistance and efflux pump EmrAB were abundantly detected in the good responders.

Functional markers in BNT162b2 (V1: V3) marker KEGG Ortholog LDA score
(log 10) p-value aminocarboxymuconate-semialdehyde decarboxylase K03392 2.397185 0.019016 iron complex transport system ATP-binding protein K02013 2.180459 0.019016 aromatic-L-amino-acid/L-tryptophan decarboxylase K01593 2.00677 0.033006 glycine cleavage system transcriptional repressor K03567 1.957881 0.010515 cobalt/nickel transport system ATP-binding protein K02006 1.957086 0.033006 RNA 2',3'-cyclic 3'-phosphodiesterase K01975 1.922388 0.033006 propionyl-CoA carboxylase beta chain K01966 1.910368 0.033006 putative ABC transport system ATP-binding protein K02021 1.88189 0.019016 ferredoxin hydrogenase small subunit K00534 1.872734 0.033006 betaine/carnitine transporter, BCCT family K03451 1.838406 0.033006 HlyD family secretion proteins K02022 1.838199 0.019016 thiosulfate/3-mercaptopyruvate sulfurtransferase K01011 1.807469 0.033006 serine palmitoyltransferase K00654 1.77698 0.019016 CRP/FNR family transcriptional regulator, anaerobic regulatory protein K01420 1.77055 0.010515 podocalyxin-like K06817 1.767455 0.033006 ribosomal protein L11 methyltransferase K02687 -1.739812 0.033006 pyrroline-5-carboxylate reductase K00286 -1.749211 0.019016 23S rRNA (guanosine2251-2'-O)-methyltransferase K03218 -1.790968 0.019016 anaerobic ribonucleoside-triphosphate reductase activating protein K04068 -1.803531 0.019016 epoxyqueuosine reductase K09765 -1.807832 0.033006 putative sigma-54 modulation protein K05808 -1.817599 0.033006 2-iminobutanoate/2-iminopropanoate deaminase K09022 -1.825438 0.033006 rod shape determining protein RodA K05837 -1.826586 0.033006 competency ComEA K02237 -1.842621 0.033006 acylaminoacyl-peptidase K01303 -1.87819 0.033006

Here, a marker having a positive LDA score is a marker that was dominant in the good immune response group, and a marker having a negative LDA score is a marker that was dominant in the poor immune response group. LEfSe, or LDA effect size, is the absolute value of the LDA score.

KEGG orthologs abundantly observed in the BNT162b2 inoculated group are shown in FIG. 14B and Table 8, and functional markers were selected based on the LDA effect size of 1.6. In the excellent immune response group after inoculation with BNT162b2, aminocarboxymuconate-semialdehyde decarboxylase (LEfSe, 2.397185, p=0.019016, KEGG ortholog K03392), iron complex transport system ATP binding protein (iron complex transport) system ATP-binding protein, LEfSe, 2.180459, p=0.019016, KEGG ortholog K02013), aromatic-L-amino-acid/L-tryptophan decarboxylase, LEfSe, 2.006770 , p=0.033, KEGG ortholog K01593), glycine cleavage system transcriptional repressor (LEfSe, 1.957881, p=0.010515, KEGG ortholog K03567), cobalt/nickel transport system ATP-binding protein (cobalt/nickel transport system ATP-binding protein, LEfSe, 1.957086, p=0.033006, KEGG ortholog K02006), RNA 2', 3'-cyclic 3'-phosphodiesterase (RNA 2', 3'-cyclic 3'-phosphodiesterase, LEfSe, 1.922388, p=0.033006, KEGG ortholog K01975), propionyl-CoA carboxylase beta chain (LEfSe, 1.910368, p=0.033006, KEGG ortholog K01966), putative ABC transport system ATP- putative ABC transport system ATP-binding protein (LEfSE, 1.88189, p=0.019016, KEGG ortholog K02021), ferredoxin hydrogenase small subunit (LEfSe, 1.872734, p=0.033006, KEGG ortholog K00534) , betaine/carnitine transporter, BCCT family, LEfSe, 1.838406, p=0.033006, KEGG ortholog K03451), HlyD family secretion protein (LEfSe, 1.838199, p=0.019016, KEGG ortholog K02022), thiosulfate/3-mercaptopyruvate sulfurtransferase (LEfSe, 1.807469, p=0.033006, KEGG ortholog K01011), serine palmitoyltransferase (LEfSe , 1.77698, p=0.019016, KEGG ortholog K00654), CRP/FNR family transcriptional regulator, anaerobic regulatory protein, LEfSe, 1.77055, p=0.010515, KEGG ortholog K01420) and pocalyxin -A analogue (podocalyxin-like, LEfSe, 1.767455, p=0.033006, KEGG ortholog K06817) was detected, and ribosomal protein L11 methyltransferase (LEfSe, 1.739812, p=0.033006, KEGG ortholog K02687), pyrroline-5-carboxylate reductase (LEfSe, 1.749211, p=0.019016, KEGG ortholog K00286), 23S rRNA (guanosine 2251-2'-O)-methyl Transferase (23S rRNA (guanosine2251-2'-O)-methyltransferase, LEfSe, 1.790968, p=0.019016, KEGG ortholog K03218), anaerobic ribonucleoside-triphosphate reductase activating protein protein, LEfSe, 1.803531, p=0.019016, KEGG ortholog K04068), epoxyqueuosine reductase (LEfSe, 1.807832, p=0.033006, KEGG ortholog K09765), putative sigma-54 modulation protein protein, LEfSe, 1.817599, p=0.033006, KEGG ortholog K05808), 2-iminobutanoate/2-iminopropanoate deaminase (LEfSe, 1.825438, p=0.033006, KEGG ortholog K09022), rod shape determining protein RodA (rod shape determining protein RodA, LEfSe, 1.826586, p = 0.033006, KEGG ortholog K05837), competency protein ComEA (competence protein ComEA, LEfSe, 1.842621, p = 0.033006, KEGG ortholog K02237) and acrylaminoacyl-peptidase (LEfSe, 1.87819, p=0.033006, KEGG ortholog K01303) were abundantly detected (FIG. 14B and Table 8). When using the MinPath algorithm, the ethylmalonyl pathway module (LEfSe, 2.69, p = 0.011) was enriched in the good immune responder group, whereas the VicK-VicR (cell wall metabolism) two-component regulatory system was found in the poor immune responder group. module (VicK-VicR (cell-wall metabolism) two-component regulatory system module, LefSe, p = 0.033) and DNA replication pathway (DNA replication pathway, LEfSe, 2.92, p = 0.019) were detected in abundance. In the PICRUSt algorithm, ketone body biosynthesis, acetyl-CoA -> acetoacetate/3-hydroxybutyrate/acetone module (LEfSe, 2.50, p = 0.019) and tryptophan metabolism in the superior immunoreactive group The tryptophan metabolism pathway (LEfSe, 2.40, p = 0.011) was detected, and in the poor immune response group, gluconeogenesis, oxaloacetate => fructose-6P, LEfSe, 2.64, p = 0.033) and metabolic pathways (LEfSe, 3.18, p = 0.019) were detected.

As described above, although the embodiments have been described with limited drawings, those skilled in the art can apply various technical modifications and variations based on the above. For example, the described techniques may be performed in an order different from the method described, and/or components of the described system, structure, device, circuit, etc. may be combined or combined in a different form than the method described, or other components may be used. Or even if it is replaced or substituted by equivalents, appropriate results can be achieved.

Therefore, other implementations, other embodiments, and equivalents of the claims are within the scope of the following claims.

Claims (20)

In biological samples isolated from the subject, genus Parasutterella , Eubacterium_g23 PAC001034_s, Blautia_uc , genus Ruminococcaceae PAC000661_g , genus Eubacterium_g5 LT907848_s, genus Romboutsia , Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC0 01043_g and detecting one or more microorganisms from the group consisting of Lachnospiraceae PAC001043_g PAC001449_s . The method of claim 1, wherein the Eubacterium_g23 PAC001034_s species has 16s rRNA represented by SEQ ID NO: 1, the Eubacterium_g5 LT907848_s species has 16s rRNA represented by SEQ ID NO: 2, and the Roseburia cecicola species has 16s rRNA represented by SEQ ID NO: 3 , The Romboutsia timonensis species has 16s rRNA represented by SEQ ID NO: 4, the Clostridium PAC001136_s species has 16s rRNA represented by SEQ ID NO: 5, and the Lachnospiraceae PAC001043_g PAC001449_s species has 16s rRNA represented by SEQ ID NO: 6, respectively. , A method for providing subject-specific vaccination information. According to claim 1, Wherein the vaccine is a vaccine against coronavirus. According to claim 1, After the step, if at least one of the groups consisting of Parasutterella genus, Eubacterium_g23 PAC001034_s , and Blautia_uc is detected in the biological sample isolated from the subject, determining that the subject has a good immune response to the viral vector vaccine method. The method of claim 4, wherein the viral vector vaccine is an adenovirus vector vaccine. According to claim 1, After the above step, in the biological sample isolated from the subject, Ruminococcaceae PAC000661_g genus , Eubacterium_g5 LT907848_s, Romboutsia genus , Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC001043_g genus And if any one or more of the group consisting of Lachnospiraceae PAC001043_g PAC001449_ s is detected, further comprising the step of determining that the subject has a good immune response to the messenger ribonucleic acid (mRNA) vaccine. Intestinal microorganisms Genus Parasutterella , Eubacterium_g23 PAC001034_s, Blautia_uc , Genus Ruminococcaceae PAC000661_g , Genus Eubacterium_g5 LT907848_s, Genus Romboutsia , Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Lachnospiraceae PAC00104 3_g in And Lachnospiraceae PAC001043_g PAC001449_ s of the group consisting of an agent for detecting any one or more, a composition for providing subject-specific vaccination information. The method of claim 7, wherein the Eubacterium_g23 PAC001034_s species has 16s rRNA represented by SEQ ID NO: 1, the Eubacterium_g5 LT907848_s species has 16s rRNA represented by SEQ ID NO: 2, and the Roseburia cecicola species has 16s rRNA represented by SEQ ID NO: 3 , The Romboutsia timonensis species has 16s rRNA represented by SEQ ID NO: 4, the Clostridium PAC001136_s species has 16s rRNA represented by SEQ ID NO: 5, and the Lachnospiraceae PAC001043_g PAC001449_s species has 16s rRNA represented by SEQ ID NO: 6, respectively. , A composition for providing subject-specific vaccination information. A kit for providing vaccination information tailored to a subject, comprising the composition of claim 7. Genus Parasutterella , Eubacterium_g23 PAC001034_s, Blautia_uc , Genus Ruminococcaceae PAC000661_g , Genus Eubacterium_g5 LT907848_s, Genus Romboutsia , Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Genus Lachnospiraceae PAC001043_g , Lachnospiraceae PAC001043_g PAC001449_ s and combinations comprising at least one selected from the group consisting of Probiotics for immunity enhancement. Genus Parasutterella , Eubacterium_g23 PAC001034_s, Blautia_uc , Genus Ruminococcaceae PAC000661_g , Genus Eubacterium_g5 LT907848_s, Genus Romboutsia , Roseburia cecicola, Romboutsia timonensis, Clostridium PAC001136_s, Genus Lachnospiraceae PAC001043_g , Lachnospiraceae PAC001043_g PAC001449_ s and combinations comprising at least one selected from the group consisting of A food composition for enhancing immunity. In a biological sample isolated from the subject, endoglucanase (KEGG ortholog K01179), fumarate hydratase class I (KEGG ortholog K01676), two-component regulatory system LytTR family sensor kinase system, LytTR family, sensor kinase, KEGG ortholog K02478), anthranilate synthase component I orthologs (KEGG ortholog K01657), butyryl-CoA dehydrogenase (KEGG ortholog K00248), two-component system, OmpR family, response regulator CpxR (two-component system, OmpR family, response regulator CpxR, KEGG ortholog K07662), peptide/nickel transport system permease protein (KEGG ortholog K02033), putative ABC transport system ATP-binding protein (KEGG ortholog K02003), aminocarboxymuconate-semialdehyde decarboxylase (KEGG ortholog K03392), iron iron complex transport system ATP-binding protein (KEGG ortholog K02013), aromatic-L-amino-acid/L-tryptophan decarboxylase (KEGG ortholog K01593), glycine cleavage system transcriptional repressor (KEGG ortholog K03567), cobalt/nickel transport system ATP-binding protein (KEGG ortholog K02006), RNA 2' , 3'-cyclic 3'-phosphodiesterase (RNA 2', 3'-cyclic 3'-phosphodiesterase, KEGG ortholog K01975), propionyl-CoA carboxylase beta chain , KEGG ortholog K01966), putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase small subunit (KEGG ortholog K00534), betaine/ Betaine/carnitine transporter BCCT family (BCCT family, KEGG ortholog K03451), HlyD family secretion protein (KEGG ortholog K02022), thiosulfate/3-mercaptopyruvate sulfurtransferase /3-mercaptopyruvate sulfurtransferase, KEGG ortholog K01011), serine palmitoyltransferase (KEGG ortholog K00654), CRP/FNR family transcriptional regulator, anaerobic regulatory protein, KEGG ortholog K01420), podocalyxin-like (KEGG ortholog K06817), ribosomal protein L11 methyltransferase (KEGG ortholog K02687), pyrroline-5-carboxylate reductase (pyrroline-5-carboxylate reductase) 5-carboxylate reductase, KEGG ortholog K00286), 23S rRNA (guanosine2251-2'-O)-methyltransferase (23S rRNA (guanosine2251-2'-O)-methyltransferase, KEGG ortholog K03218), anaerobic ribonucleo Side-triphosphate reductase activating protein (anaerobic ribonucleoside-triphosphate reductase activating protein, KEGG ortholog K04068), epoxyqueuosine reductase (KEGG ortholog K09765), putative sigma-54 modulation protein protein, KEGG ortholog K05808), 2-iminobutanoate/2-iminopropanoate deaminase (2-iminobutanoate/2-iminopropanoate deaminase, KEGG ortholog K09022), rod shape determining protein RodA , KEGG ortholog K05837), competency protein ComEA (competence protein ComEA, KEGG ortholog K02237) and acrylaminoacyl-peptidase (acylaminoacyl-peptidase, KEGG ortholog K01303). , A method for providing subject-specific vaccination information. According to claim 12, Wherein the vaccine is a vaccine against coronavirus. According to claim 12, After the above step, in the biological sample isolated from the subject, at least one of the group consisting of endoglucanases, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinases, and anthranilate synthase component I orthologs. If a functional biomarker is detected, the method further comprising determining that the subject has a good immune response to the viral vector vaccine. According to claim 12, After the above steps, in the biological sample isolated from the subject, butyryl-CoA dehydrogenase, two-component control system OmpR family response regulator CpxR, peptide / nickel transport system permease protein and putative ABC transport system ATP binding protein ( KEGG ortholog K02003), if any one or more functional biomarkers from the group consisting of are detected, the method further comprising the step of determining the subject as having a poor immune response to the viral vector vaccine. The method according to claim 14 or 15, wherein the viral vector vaccine is an adenovirus vector vaccine. According to claim 12, After the above steps, aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L-amino acid / L-tryptophan decarboxylase, glycine cleavage in the biological sample isolated from the subject system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'-phosphodiesterase, propionyl-CoA carboxylase beta chain, putative ABC transport system ATP- binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercaptopyruvate sulfurtransferase, serine palmitoyltransferase When at least one functional biomarker from the group consisting of Aze, CRP/FNR family transcriptional regulator anaerobic regulatory protein and pocalyxin-analog is detected, determining a subject with a good immune response to a messenger ribonucleic acid (mRNA) vaccine How to further include. According to claim 12, After the above step, in the biological sample isolated from the subject, ribosomal protein L11 methyltransferase, pyrroline-5-carboxylate reductase, 23S rRNA (guanosine 2251-2'-O)-methyltransferase, Anaerobic ribonucleoside-triphosphate reductase active protein, epoxyquiosine reductase, putative sigma-54 regulatory protein, 2-iminobutanoate/2-iminopropanoate deaminase, rod-shaped crystal protein When at least one functional biomarker from the group consisting of RodA, the competence protein ComEA, and acrylaminoacyl-peptidase is detected, the method further comprising determining that the subject has a poor immune response to the messenger ribonucleic acid (mRNA) vaccine. . Endoglucanases, fumarate hydratase class I, two-component regulatory system LytTR family sensor kinases, anthranilate synthase component I orthologs, butyryl-CoA dehydrogenase, two-component regulatory system OmpR family response regulators CpxR, peptide/nickel transport system permease protein, putative ABC transport system ATP binding protein (KEGG ortholog K02003), aminocarboxymuconate-semialdehyde decarboxylase, iron complex transport system ATP binding protein, aromatic-L- Amino acid/L-tryptophan decarboxylase, glycine cleavage system transcriptional repressor, cobalt/nickel transport system ATP-binding protein, RNA 2', 3'-cyclic 3'-phosphodiesterase, propionyl-CoA Carboxylase beta chain, putative ABC transport system ATP-binding protein (KEGG ortholog K02021), ferredoxin hydrogenase subunit, betaine/cartinine transporter BCCT family, HlyD family secreted protein, thiosulfate/3-mercap Topyruvate sulfurtransferase, serine palmitoyltransferase, CRP/FNR family transcriptional regulator anaerobic regulatory protein, pocalyxin-analog, ribosomal protein L11 methyltransferase, pyrroline-5-carboxylate reducta enzyme, 23S rRNA (guanosine 2251-2'-O)-methyltransferase, anaerobic ribonucleoside-triphosphate reductase activating protein, epoxyquiosine reductase, putative sigma-54 regulatory protein, 2- A subject comprising an agent that detects a functional biomarker of any one or more of the group consisting of iminobutanoate/2-iminopropanoate deaminase, rod-shaped crystal protein RodA, competence protein ComEA, and acrylaminoacyl-peptidase. A composition for providing customized vaccination information. A kit for providing vaccination information tailored to a subject, comprising the composition of claim 19.

Images