CN114450023A - Vaccine composition against severe fever with thrombocytopenia syndrome virus - Google Patents

Abstract

The present invention provides a vaccine composition against SFTS virus using Gn or Gc protein or NP protein or a mixture thereof, based on the result of an experiment that a recombinant protein, NP protein (full-length sequence protein), or a mixture thereof, in which the extracellular domain (extracellular domain) of Gn or Gc protein of fever with thrombocytopenia syndrome (STFS) is fused to the Fc region of a human antibody, provides protective immunity in a first-type interferon receptor knock-out (KO) mouse.

Description

Vaccine composition against severe fever with thrombocytopenia syndrome virus

Technical Field

The invention relates to a vaccine composition for severe fever with thrombocytopenia syndrome virus.

Background

Febrile with thrombocytopenia syndrome (SFTS) is an infectious disease caused by infection with the virus of febrile with thrombocytopenia syndrome (SFTSV). Since the first report in china in 2009, there have been relevant case reports in asian countries such as china, korea, japan, and vietnam. STFSV comprises as a genome three segmented ribonucleic acids (RNAs) encoding an RNA polymerase (RdRp), two envelope glycoproteins (Gn, Gc), a Nucleocapsid (NP), and a nonstructural protein (NS). When the virus is infected, fever, gastrointestinal complications, leukopenia and thrombocytopenia are accompanied, and the fatality rate reaches 5-20 percent (Lancet infection Dis 2018,18,1127, 1137, doi:10.1016/S1473-3099(18) 30293-7). There is no therapeutic agent so far and vaccine development is underway.

STFS vaccine development is currently in the early research stage, and recently reports have been made on the fact that Vaccines or deoxyribonucleic acid (DNA) Vaccines using recombinant viral vectors can provide protective immunity in animal models of infection (NPJ Vaccines 2019,4,5, doi:10.1038/s 41541-018-0096-y; Nat Commun 2019,10,3836, doi:10.1038/s 41467-019-11815-4). It has been confirmed that vaccines using NS can induce the formation of antibodies, but do not provide protective immunity in a mouse infection model (Viral Immunol 2015,28,113-122, doi: 10.1089/vim.2014.0100). It has been reported that when micromanipulation is used to immunize against DNA encoding NP as well as NS genes, specific T cell responses of the antigen can be induced, but there is currently no way to determine whether protective immunity can be provided in animal infection models (Colloids Surf B Biointerfaces 2017,159,54-61, doi:10.1016/j. colsurffb.2017.07.059). Recently, there is a report that formation of neutralizing antibodies can be induced when a recombinant virus (rVSV) expressing Gn and Gc proteins is immunized once, and perfect protective immunity can be induced in a knock-out (KO) mouse infected with STFSV of a lethal dose (NPJ Vaccines 2019,4,5, doi:10.1038/s 41541-018-. Furthermore, it has been reported that when DNAs expressing five genes of SFTSV, respectively, are simultaneously immunized, when two DNAs encoding Gn and Gc are simultaneously immunized, and when three DNAs encoding NP, NS, and RdRp are simultaneously immunized, it is possible to provide immunoprotection against lethal infection of aged weasel (over four years old) susceptible to SFTSV (Nat Commun 2019,10,3836, doi:10.1038/s 41467-019-11815-4). However, no report has been made so far on what degree of protective immunity can be provided when each of the structural proteins (Gn, Gc, NP) is immunized, and no research results have been published on the efficacy of subunit protein vaccines using the proteins.

The invention discloses vaccine efficacy of Gn protein, Gc protein and NP protein.

Detailed Description

Technical problem

The purpose of the present invention is to provide a vaccine composition against SFTS virus, which uses Gn protein, Gc protein and NP protein.

Other and specific objects of the present invention will be described in the following.

Means for solving the problems

As shown in the following examples, the present inventors comparatively verified whether recombinant units (Gn-Fc and Gc-Fc) fusing extracellular domains (extracellular domains) of Gn or Gc proteins to the Fc portion of human antibodies and NP proteins (full-length sequence proteins) could immunize first-type interferon receptor KO mice alone and provide immunoprotection. As a result, all mice immunized with Gc-Fc protein died, but the survival time was statistically increased compared to the control group (group immunized with Fc alone), and it was confirmed that 50% of the mice immunized with Gn-Fc protein survived and 60% of the mice immunized with NP protein survived, respectively, thereby providing significant protective immunity effects. In addition, when the first type interferon receptor KO mouse was immunized after the Gn-Fc and Gc-Fc were mixed with NP and whether the protective immunity was elevated or not was confirmed, the survival rate was improved in the case of the mixed immunity of Gc-Fc and NP by 80%, in the case of the mixed immunity of Gn-Fc and NP by 70%, and in the case of the mixed immunity of Gn-Fc and Gc-Fc and NP by 60%, thereby confirming that the survival rate was improved in the case of the mixed immunity as a whole as compared with the case of the single immunity.

The present invention is based on the above-described experimental results, and the vaccine composition of the present invention relates to a vaccine composition against SFTS virus, which contains, as an active ingredient, the extracellular domain of Gc protein, the extracellular domain of Gn protein, the full-length NP protein, or a mixture of two or more thereof.

In the present invention, particularly, the extracellular domain of the Gc protein or the extracellular domain of the Gn protein may be fused to the Fc region of a human antibody in order to enhance the in vivo half-life, and in the case of fusion with Fc, a hinge (hinge) sequence having 2 to 15 amino acids may be arranged in order to prevent the fusion proteins from interfering with the formation of a three-dimensional structure therebetween. In addition, in order to produce the protein as described above by using a host cell by means of gene recombination, isolation and purification can be more easily achieved by disposing a signal peptide (signal peptide) sequence at the N-terminal as described in the following examples. Since the sequence as described above is removed when the corresponding protein is secreted to the outside of the cell, it is not present in the isolated and purified protein.

In the present specification, "vaccine" refers to prevention of infection or re-infection of a corresponding pathogen, reduction or elimination of symptoms caused by the corresponding pathogen, or substantial or complete elimination of the corresponding pathogen or a disease caused by the corresponding pathogen by inducing an immune response to the corresponding pathogen in a host, i.e., a human. Thus, the vaccine composition of the present invention may be administered to a human for prophylactic purposes prior to infection with the corresponding pathogen, or for therapeutic purposes after infection with the corresponding pathogen. Wherein the "immune response" refers to a humoral immune response, a cellular immune response, or both.

In addition, in the present specification, the "active ingredient" refers to an ingredient that can induce an immune response alone or together with a carrier that cannot induce an immune response by itself. Thus, the active ingredient itself need not be immunogenic (capable of inducing an immune response). In the present invention, the active ingredient is preferably, but not necessarily, purified. The term "purified" as used herein means a state in which cell components, culture medium components thereof, and the like (which are contaminants) in the original organism (i.e., the microorganism transformed with the microorganism of the present invention) in which the corresponding target substance is present are substantially reduced or removed. The state where the contaminants are substantially reduced or removed means that the purity is 50% or more, preferably 90% or more, and more preferably 99% or more. The purity can be evaluated by a method known in the related art such as chromatography column, gel electrophoresis, and immunological analysis, and the purification method can be a method known in the related art to be described later.

The vaccine composition of the present invention can be manufactured in any suitable and pharmaceutically acceptable formulation. For example, the composition may be prepared in the form of a solution or suspension for extemporaneous administration, a concentrated stock solution suitable for dilution prior to administration, or a reconstitutable form such as a lyophilized, freeze-dried or frozen preparation.

The vaccine compositions of the present invention may include a pharmaceutically acceptable carrier when formulated. As for the pharmaceutically acceptable carrier that can be used in formulating the vaccine composition, there are listed and stipulated in the pharmacopoeias of korean nationality or other countries, particularly in the pharmacopoeias of the united states, japan and europe, and specific information can be referred to these pharmacopoeias.

The carrier, as mentioned above, may generally include, for example, diluents, excipients, stabilizers, preservatives and the like. Suitable diluents include, for example, non-aqueous solvents such as propylene glycol, polyethylene glycol, and vegetable oils (e.g., olive oil and peanut oil), or aqueous solvents such as saline (preferably 0.8% saline) and water containing a buffer medium (preferably 0.05M phosphate buffer), as suitable excipients, such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, anhydrous skim milk, glycerol, propylene, glycol, water and ethanol, suitable stabilizers include, for example, carbohydrates such as sorbitol, mannitol, starch, chitosan, glutamic acid, and glucose, and proteins such as animal, vegetable, and microbial proteins (e.g., milk powder, serum albumin, and casein). Examples of suitable preservatives include thimerosal, gentamicin, neomycin, nystatin, amphotericin B, tetracycline, penicillin, streptomycin, and polymyxin B.

The vaccine composition of the present invention may further comprise an antigen adjuvant (adjuvant). An antigen adjuvant may be composed of more than one substance that can enhance an immune response to an antigen. Adjuvants can function as tissue stocks for slow release of antigen and/or as activators of the lymphatic system for non-specifically potentiating immune responses (Hood et al, Immunology, Second ed.,1984, benjamin/Cummings: Menlo Park, California, p.384). The antigen adjuvant may be, for example, freund's complete adjuvant, freund's incomplete adjuvant, saponin, gel-like aluminum adjuvant, surface active substance (such as lysolecithin, polyether glycol, polyanion, peptide, oil and hydrocarbon emulsion, etc.), vegetable oil (such as cotton seed oil, peanut oil or corn oil, etc.), vitamin E acetate, etc.

Aluminum adjuvants are most widely used as antigen adjuvants applicable to human bodies, and the Aluminum adjuvants described above are used in gel form of Aluminum salts such as Aluminum phosphate (Aluminum phosphate), Potassium Aluminum sulfate (Potassium sulfate), and Aluminum hydroxide (Aluminum hydroxide). It has been known that an aluminum adjuvant as described above can potentiate an immune response by eliciting a Th2-type immune response (Sokolovska A et al, vaccine.2007Jun 625(23): 4575-85; O' Hagan DT AND Rappuoli R., Pharm Res.2004-type immune response)Sep; 21(9):1519-30.). The Alhydrogel from Invivogen used in the examples of the present invention is a gel-like suspension (colloidal) of aluminum hydroxide (aluminum hydroxide). The aluminum adjuvant as described above can be produced AND used as it is or purchased as a product distributed on the market according to a production method known in the related art (R.Bomford. immunological additives AND vaccines. NATO ASI Series 1989; 179: 35-41; Vogel FR AND Powell MF. pharm Biotechnol.1995; 6: 141-. As products distributed on the market, examples of the products include Aluminum hydroxide Gel product from Sigma and Aluminum hydroxide Gel product from BRENNTAG, including 2% Aluminum hydroxide Gel product from Invivogen used in the examples described below^TMProducts, etc. are examples.

The vaccine composition of the present invention can be produced in any unit volume. The unit volume refers to the amount of the active ingredient and the pharmaceutically acceptable carrier contained in a single product packaged for the purpose of administering to a human being more than once, and appropriate amounts of the active ingredient and the carrier as described above are amounts that can function as a vaccine upon more than one vaccination with the vaccine composition of the present invention, and the amounts as described above can be determined by non-clinical or clinical means within the ordinary abilities of the relevant practitioner.

The vaccine composition of the present invention can be preferably administered by parenteral means such as rectal, transdermal, intravenous injection, intra-arterial injection, intramuscular injection, intradermal injection, subcutaneous injection, intraperitoneal injection, and intraventricular injection.

The vaccine composition of the present invention may be administered by a modified release system. For example, as the modified release system as described above, there may be exemplified such a liposome, an implantable osmotic pump, a transdermal patch, and the like. Preferably, the active ingredient is delivered by liposome means. For the delivery of active ingredients by means of Liposomes, reference may be made, for example, to the literature [ Langer, Science 249: 1527-. For the mode of delivery of other active ingredients, reference may be made to the literature [ Langer, supra; sefton, CRC Crit.Ref.biomed.Eng.14:201(1987), document [ Buchwald et al, Surgery 88:507(1980) ], document [ Saudek et al, N.Engl.J.Med.321:574(1989) ], document [ Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres, Boca Raton, Florida (1974) ], document [ Controlled Drug bioavailability.drug Product Design and Performance, Smolen and Ball, Wiley, New York (1984) ], document [ Ranger and Peppas, J.Macromol.Sci.Rev.Chem.351.19823: Hough et al [ 19825: 9 J.J.Macromol.19825 ], Neugn.J.1989 et al, and New York (1989). The documents mentioned above are considered to be part of the present description.

The dosage of the vaccine composition of the present invention can be determined by the characteristics of the medical staff, for example, age, body weight, sex, symptoms, complications, and patients with other diseases.

In addition, the time interval and the number of administrations may be determined depending on the dosage form used or the half-life of the active ingredient in blood, etc.

As described above, the dose amount and the time interval and the number of times of administration should be decided by the judgment of medical staff and by the individual, but generally as an appropriate dose amount, it should be decided within a range of about 0.1 to 10mg per 1kg of the body weight of the individual on a one-day basis, the dose interval should be decided within a range of 3 to 10 days, and the number of times of administration should be decided within a range of 1 to 5 times.

Each protein used as an active ingredient in the vaccine composition of the present invention can be produced by a DNA recombination technique known in the related art using a gene encoding the protein. The method as described above, comprising: (i) producing an expression vector comprising a gene encoding each of the proteins; (ii) transforming said expression vector in a host cell; (iii) culturing the host cell after the transformation; and, (iv) isolating and extracting said individual proteins from said cultureAnd (4) a pure step. The present specification discloses genes encoding the respective proteins, and as to methods for producing the respective proteins by a DNA recombination technique using the genes, specifically, as to expression vector constitution or recombinant DNA technique including a signal peptide sequence of a regulatory sequence such as a promoter or a terminator, a selection marker, and the like, a method of transformation of a trait, constitution of a culture medium, a method of culture after transformation of a trait, and the like, a considerable number of documents have been accumulated in the related industries, and as to each related matter, reference can be made to these documents. Specifically, reference may be made to, for example, the literature (nucleic acid Res.14:5399-5467,1986), the literature (Tet.Lett.27:5575-5578,1986), the literature (nucleic acid Res.4:2557,1977), the literature (Lett.,28:2449,1978), the literature (Sambrook et al, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (2001)), the literature (F M Autobel et al, Current Protocols in Molecular Biology, John Wiley amp; Sons, Inc. (1994)), the literature (Marston, F (1987) DNA Cloning technologies), the literature (Cohen, S.N.et al, Proc.Natl.Acac.Sci.9: 9, USA (1987) DNA Cloning technologies, WO 3: WO 3, Cell 4727, Cell 94, WO 3, Cell 580, Cell 4727, Cell J. (1988), the literature (Cell J.90, W.90, WO 3, WO 52, Cell 4727, WO 3, WO 52, Cell 94, K. No. 5, K. A, 1:841(1982)), in literature (Wong, T.K.et al., Gene,10:87(1980)), in literature (Gopal, mol.cell biol.,5: 1188-containing 1190(1985)), in literature (Yang et al., Proc.Natl.Acad.Sci.,87: 9568-containing 9572(1990))

(Maniatis et al, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1982)), literature (Hitzeman et al, J.biol.chem.,255,12073-12080(1980)), literature (Luchansky et al. mol.Microbiol.2,637-646(1988)), literature (Sambrook et al, Microbiol.2,637-646(1988)), and literature (Sambrook)

Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory Press, US (1989)), literature (Ausubel et al, Current Protocols in Molecular Biology, Jon Willey&SonsUS (1993)), literature (Sambrook, J).&Russel, d. Molecular Cloning, a LABORATORY MANUAL, Cold Spring Harbor LABORATORY Press, first and second volumes released on 15/01/2001), and the like.

Effects of the invention

As described above, the present invention can provide a vaccine composition against SFTS virus comprising, as an active ingredient, the extracellular domain of Gc protein, the extracellular domain of Gn protein, the full-length NP protein, or a mixture thereof.

Drawings

Fig. 1 is a schematic diagram illustrating the structure and molecular weight of Gc protein, Gn protein, or NP protein, which is an active ingredient of the vaccine composition of the present invention.

FIG. 2 shows the results of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the active ingredients of the vaccine composition of the present invention, i.e., Gc protein, Gn protein or NP protein.

FIG. 3 shows the results of specific antibody induction based on Gn and Gc antigen immunization and virus neutralization test using immune serum (a: antibody titer to Gn and Gn proteins, b and c: neutralization assay test to SFTS virus, FRNT: lesion reduction neutralization titer).

FIG. 4 shows the results of measuring the survival rate and the change in body weight of a first type interferon receptor KO mouse immunized with Gc protein, Gn protein or NP protein.

FIG. 5 shows the results of immunizing a first type interferon receptor KO mouse with a mixture of Gc protein, Gn protein and/or NP protein and measuring the survival rate and the change in body weight.

Detailed Description

Next, the present invention will be described in detail with reference to examples. However, the scope of the present invention is not limited by the examples.

< example 1> production of viral protein for use as vaccine antigen

< examples 1-1> amino acid sequences of viral proteins used as vaccine antigens

Using the amino acid information on GenBank, Gn protein, Gc protein and NP protein used as vaccine antigens were produced. Amino acid information on GenBank is GenBank ID: ATW63054.1(Gn protein), GenBank ID: AOO85591.1(Gc protein) and GenBank ID: AJO16084.1(NP protein).

The Gn and Gc proteins use only the extracellular domain (extracellular domain) except for the signal peptide (signal peptide), transmembrane domain (transmembrane domain) and cytoplasmic domain (cytoplasmic domain) (seq id nos. 1 and 2, respectively), while the NP protein uses the full-length sequence (seq id No. 3).

< example 1-2> cloning and expression of viral protein Gene

After synthesizing genes encoding Gn and Gc proteins (extracellular domains) (seq id nos. 4 and 5, respectively), a signal peptide (signal peptide) gene (seq id No. 6) of Ig kappa light chain (light chain) was fused to the N-terminus (N-terminal) and a recombinant gene in which a human (human) IgG1 CH2-CH3 site Fc domain (domain) gene (seq id No. 7) was fused to the C-terminus (C-terminal) via a hinge sequence (sequence No. 8) was cloned into pCEP4 mammalian expression vector (mammalin expression vector) in the manner shown in fig. 1. The gene (SEQ ID NO: 9) encoding the NP protein with 6His (used for purification purposes) ligated to each of the N-and C-termini was cloned into pET28a vector (vector). Protein expression was induced by introducing the cloned genes into HEK93T cells (Gn, Gc) and e.coli BL21(NP), respectively, and purified using Protein a/G columns (Protein a/G columns) and nickel-nitrilotriacetic acid columns (Ni-NTA columns), respectively. The size and purity of the purified protein was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (FIG. 2).

< example 2> immunization of SFTS viral protein and antibody analysis

Two weeks after the third immunization, blood was collected and the antibody titers of Gn, Gc and NP were measured by an enzyme-linked immunosorbent assay (ELISA) method using Gn-Fc (seq id No. 10, the amino acid sequence of DVAQAA at the N-terminus in seq id No. 10 is a sequence inserted for cloning), Gc-Fc (seq id No. 11, the amino acid sequence of DVAQAA at the N-terminus in seq id No. 10 is a sequence inserted for cloning), NP (seq id No. 12) and human Fc in first type interferon receptor KO mouse (N ═ 4) three times at two week intervals (20ug/mouse + Alum, Gn-Fc, Gc-Fc, Fc; 10ug/mouse + Alum, NP), respectively (a of fig. 3). It was confirmed that a higher level of antibody could be produced for each antigen (anti-Gn or anti-Gc IgG mean titer 1:4,096, anti-NP IgG mean titer 1:24,576; n: 4/group), and in particular, a higher antibody titer induced for NP antigen was observed. To confirm the virus neutralizing ability of the antibodies generated against the viral glycoproteins (Gn and Gc), focus reduction analysis (focus reduction assay) was performed after reacting STFS virus with immune serum and infecting it into Vero cells (b and c of fig. 3). It was confirmed that, although almost similar levels of antibodies were produced, the sera obtained by immunization with Gn-Fc formed relatively high neutralizing antibody titers compared to the sera obtained by immunization with Gc-Fc, and had significantly higher levels of virus neutralizing ability compared to the sera obtained by immunization with only Fc in both of the immune sera (neutralizing antibody titers: Gn-Fc: 929. + -. 1134, Gc-Fc: 209. + -. 140, Fc: 42. + -. 29; n ═ 4/group, mean titer. + -. S.D.).

< example 3> protective immune Effect against SFTS viral protein immunization

Three immunizations (20ug/mouse + Alum, Gn-Fc, Gc-Fc, Fc; 10ug/mouse + Alum, NP) were performed at two week intervals in first type interferon receptor KO mice (n ═ 4-6) using Gn-Fc, Gc-Fc, NP, and Fc, respectively, and 10ug/mouse + Alum, NP, two weeks after the third immunization using 10³SFTS virus of focus-forming units (FFU) was infected subcutaneously. In addition, daily weight change and mortality were observed (fig. 4).

Mice immunized with phosphate Buffer (BPS) had gradually lost body weight and all died by day six of infection (control). Mice immunized with Fc (control group) lost weight gradually starting from the day of infection and all died until the fifth day of infection. Mice immunized with Gc-Fc lost weight gradually from the fourth day of infection and all died until the eighth day of infection. Mice immunized with Gn-Fc gradually lost weight from the third day of infection and died half by the eighth day of infection, but the remaining mice gradually recovered in weight and survived thereafter. Mice immunized with NP lost weight gradually starting on day three of infection and died 40% by day eight, but the remaining 60% of mice had weight gradually recovered and survived thereafter. It was confirmed by statistical analysis that (Log-rank (Mantel-Cox) Test), the survival prolongation effect (Gc-Fc group: p ═ 0.0046) or the mortality reduction effect (Gn-Fc group: p ═ 0.0054, NP group: p ═ 0.0023) based on the vaccine exhibited a statistically significant improvement effect compared to the control group.

< example 4> protective immune Effect against SFTS Virus protein Mixed immunization

Three immunizations (20 or 30ug/mouse + Alum) were performed in two weeks at intervals in first type interferon receptor KO mice (n ═ 7) using a mixture of Gn-Fc and NP (Gn + NP, mixing ratio: 10ug +10ug), a mixture of Gc-Fc and NP (Gc + NP, mixing ratio: 10ug +10ug), a mixture of Gn-Fc and Gc-Fc and NP (Gn + Gc + NP), and phosphate Buffer (BPS) (control), respectively, and two weeks after the third immunization, using 10ug/mouse + Alum³SFTS virus of focus-forming units (FFU) was infected subcutaneously. In addition, daily weight change and mortality were observed (fig. 5).

Mice immunized with phosphate Buffer (BPS) were gradually lost in body weight and all died until the sixth day of infection (control group). Mice immunized with Gn + NP lost weight gradually starting on day four of infection, died 14.3% on day five and 28.6% until day six of infection, but the remaining mice recovered weight gradually and survived thereafter. Mice immunized with Gc + NP gradually lost weight starting on the fifth day of infection and died 14.3% on the sixth day of infection, but the remaining mice gradually recovered in weight and survived thereafter. Mice immunized with Gn + Gc + NP gradually lost weight starting on day four of infection and died 42.9% on day seven of infection, but the remaining mice gradually recovered in weight and survived thereafter.

It was confirmed by statistical analysis that (Log-rank (Mantel-Cox) Test), the survival prolongation effect (Gc-Fc group: p ═ 0.0046) or the mortality reduction effect (Gn-Fc group: p ═ 0.0054, NP group: p ═ 0.0023) based on the vaccine exhibited a statistically significant improvement effect compared to the control group.

SEQUENCE LISTING

<110> Seoul university industry cooperative group

<120> vaccine composition for severe fever with thrombocytopenia syndrome virus

<130> P22111831WP

<160> 12

<170> PatentIn version 3.5

<210> 1

<211> 438

<212> PRT

<213> Unknown (Unknown)

<220>

<223> SFTS Virus Gn protein extracellular domain

<400> 1

Asp Ser Gly Pro Ile Ile Cys Ala Gly Pro Ile His Ser Asn Lys Ser

1 5 10 15

Ala Asp Ile Pro His Leu Leu Gly Tyr Ser Glu Lys Ile Cys Gln Ile

20 25 30

Asp Arg Leu Ile His Val Ser Ser Trp Leu Arg Asn His Ser Gln Phe

35 40 45

Gln Gly Tyr Val Gly Gln Arg Gly Gly Arg Ser Gln Val Ser Tyr Tyr

50 55 60

Pro Ala Glu Asn Ser Tyr Ser Arg Trp Ser Gly Leu Leu Ser Pro Cys

65 70 75 80

Asp Ala Asp Trp Leu Gly Met Leu Val Val Lys Lys Ala Lys Gly Ser

85 90 95

Asp Met Ile Val Pro Gly Pro Ser Tyr Lys Gly Lys Val Phe Phe Glu

100 105 110

Arg Pro Thr Phe Asp Gly Tyr Val Gly Trp Gly Cys Gly Ser Gly Lys

115 120 125

Ser Arg Thr Glu Ser Gly Glu Leu Cys Ser Ser Asp Ser Gly Thr Ser

130 135 140

Ser Gly Leu Leu Pro Ser Asp Arg Val Leu Trp Ile Gly Asp Val Ala

145 150 155 160

Cys Gln Pro Met Thr Pro Ile Pro Glu Glu Thr Phe Leu Glu Leu Lys

165 170 175

Ser Phe Ser Gln Ser Glu Phe Pro Asp Ile Cys Lys Ile Asp Gly Ile

180 185 190

Val Phe Asn Gln Cys Glu Gly Glu Ser Leu Pro Gln Pro Phe Asp Val

195 200 205

Ala Trp Met Asp Val Gly His Ser His Lys Ile Ile Met Arg Glu His

210 215 220

Lys Thr Lys Trp Val Gln Glu Ser Ser Ser Lys Asp Phe Val Cys Tyr

225 230 235 240

Lys Glu Gly Thr Gly Pro Cys Ser Glu Ser Glu Glu Lys Thr Cys Lys

245 250 255

Thr Ser Gly Ser Cys Arg Gly Asp Met Gln Phe Cys Lys Val Ala Gly

260 265 270

Cys Glu His Gly Glu Glu Ala Ser Glu Ala Lys Cys Arg Cys Ser Leu

275 280 285

Val His Lys Pro Gly Glu Val Val Val Ser Tyr Gly Gly Met Arg Val

290 295 300

Arg Pro Lys Cys Tyr Gly Phe Ser Arg Met Met Ala Thr Leu Glu Val

305 310 315 320

Asn Gln Pro Glu Gln Arg Ile Gly Gln Cys Thr Gly Cys His Leu Glu

325 330 335

Cys Ile Asn Gly Gly Val Arg Leu Ile Thr Leu Thr Ser Glu Leu Lys

340 345 350

Ser Ala Thr Val Cys Ala Ser His Phe Cys Ser Ser Ala Thr Ser Gly

355 360 365

Lys Lys Ser Thr Glu Ile Gln Phe His Ser Gly Ser Leu Val Gly Lys

370 375 380

Thr Ala Ile His Val Lys Gly Ala Leu Val Asp Gly Thr Glu Phe Thr

385 390 395 400

Phe Glu Gly Ser Cys Met Phe Pro Asp Gly Cys Asp Ala Val Asp Cys

405 410 415

Thr Phe Cys Arg Glu Phe Leu Lys Asn Pro Gln Cys Tyr Pro Ala Lys

420 425 430

Lys Gly Gln Ala Gly Gln

435

<210> 2

<211> 473

<212> PRT

<213> Unknown (Unknown)

<220>

<223> SFTS virus Gc protein extracellular domain

<400> 2

Cys Asp Glu Met Val His Ala Asp Ser Lys Leu Val Ser Cys Arg Gln

1 5 10 15

Gly Ser Gly Asn Met Lys Glu Cys Val Thr Thr Gly Arg Ala Leu Leu

20 25 30

Pro Ala Val Asn Pro Gly Gln Glu Ala Cys Leu His Phe Thr Ala Pro

35 40 45

Gly Ser Pro Asp Ser Lys Cys Leu Lys Ile Lys Val Lys Arg Ile Asn

50 55 60

Leu Lys Cys Lys Lys Ser Ser Ser Tyr Phe Val Pro Asp Ala Arg Ser

65 70 75 80

Arg Cys Thr Ser Val Arg Arg Cys Arg Trp Ala Gly Asp Cys Gln Ser

85 90 95

Gly Cys Pro Pro His Phe Thr Ser Asn Ser Phe Ser Asp Asp Trp Ala

100 105 110

Gly Lys Met Asp Arg Ala Gly Leu Gly Phe Ser Gly Cys Ser Asp Gly

115 120 125

Cys Gly Gly Ala Ala Cys Gly Cys Phe Asn Ala Ala Pro Ser Cys Ile

130 135 140

Phe Trp Arg Lys Trp Val Glu Asn Pro His Gly Ile Ile Trp Lys Val

145 150 155 160

Ser Pro Cys Ala Ala Trp Val Pro Ser Ala Val Ile Glu Leu Thr Met

165 170 175

Pro Ser Gly Glu Val Arg Thr Phe His Pro Met Ser Gly Ile Pro Thr

180 185 190

Gln Val Phe Lys Gly Val Ser Val Thr Tyr Leu Gly Ser Asp Met Glu

195 200 205

Val Ser Gly Leu Thr Asp Leu Cys Glu Ile Glu Glu Leu Lys Ser Lys

210 215 220

Lys Leu Ala Leu Ala Pro Cys Asn Gln Ala Gly Met Gly Val Val Gly

225 230 235 240

Lys Val Gly Glu Ile Gln Cys Ser Ser Glu Glu Ser Ala Arg Thr Ile

245 250 255

Lys Lys Asp Gly Cys Ile Trp Asn Ala Asp Leu Val Gly Ile Glu Leu

260 265 270

Arg Val Asp Asp Ala Val Cys Tyr Ser Lys Ile Thr Ser Val Glu Ala

275 280 285

Val Ala Asn Tyr Ser Ala Ile Pro Thr Thr Ile Gly Gly Leu Arg Phe

290 295 300

Glu Arg Ser His Asp Ser Gln Gly Lys Ile Ser Gly Ser Pro Leu Asp

305 310 315 320

Ile Thr Ala Ile Arg Gly Ser Phe Ser Val Asn Tyr Arg Gly Leu Arg

325 330 335

Leu Ser Leu Ser Glu Ile Thr Ala Thr Cys Thr Gly Glu Val Thr Asn

340 345 350

Val Ser Gly Cys Tyr Ser Cys Met Thr Gly Ala Lys Val Ser Ile Lys

355 360 365

Leu His Ser Ser Lys Asn Ser Thr Ala His Val Arg Cys Lys Gly Asp

370 375 380

Glu Thr Ala Phe Ser Val Leu Glu Gly Val His Ser Tyr Thr Val Ser

385 390 395 400

Leu Ser Phe Asp His Ala Val Val Asp Glu Gln Cys Gln Leu Asn Cys

405 410 415

Gly Gly His Glu Ser Gln Val Thr Leu Lys Gly Asn Leu Ile Phe Leu

420 425 430

Asp Val Pro Lys Phe Val Asp Gly Ser Tyr Met Gln Thr Tyr His Ser

435 440 445

Thr Val Pro Thr Gly Ala Asn Ile Pro Ser Pro Thr Asp Trp Leu Asn

450 455 460

Ala Leu Phe Gly Asn Gly Leu Ser Arg

465 470

<210> 3

<211> 245

<212> PRT

<213> Unknown (Unknown)

<220>

<223> NP protein of SFTS virus

<400> 3

Met Ser Glu Trp Ser Arg Ile Ala Val Glu Phe Gly Glu Gln Gln Leu

1 5 10 15

Asn Leu Ser Glu Leu Glu Asp Phe Ala Arg Glu Leu Ala Tyr Glu Gly

20 25 30

Leu Asp Pro Ala Leu Ile Ile Lys Lys Leu Lys Glu Thr Gly Gly Asp

35 40 45

Asp Trp Val Lys Asp Thr Lys Phe Ile Ile Val Phe Ala Leu Thr Arg

50 55 60

Gly Asn Lys Ile Val Lys Ala Ser Gly Lys Met Ser Asn Ser Gly Ser

65 70 75 80

Lys Arg Leu Met Ala Leu Gln Glu Lys Tyr Gly Leu Val Glu Arg Ala

85 90 95

Glu Thr Arg Leu Ser Ile Thr Pro Val Arg Val Ala Gln Ser Leu Pro

100 105 110

Thr Trp Thr Cys Ala Ala Ala Ala Ala Leu Lys Glu Tyr Leu Pro Val

115 120 125

Gly Pro Ala Val Met Asn Leu Lys Val Glu Asn Tyr Pro Pro Glu Met

130 135 140

Met Cys Met Ala Phe Gly Ser Leu Ile Pro Thr Ala Gly Val Ser Glu

145 150 155 160

Ala Thr Thr Lys Thr Leu Met Glu Ala Tyr Ser Leu Trp Gln Asp Ala

165 170 175

Phe Thr Lys Thr Ile Asn Val Lys Met Arg Gly Ala Ser Lys Thr Glu

180 185 190

Val Tyr Asn Ser Phe Arg Asp Pro Leu His Ala Ala Val Asn Ser Val

195 200 205

Phe Phe Pro Asn Asp Val Arg Val Lys Trp Leu Lys Ala Lys Gly Ile

210 215 220

Leu Gly Pro Asp Gly Val Pro Ser Arg Ala Ala Glu Val Ala Ala Ala

225 230 235 240

Ala Tyr Arg Asn Leu

245

<210> 4

<211> 1299

<212> DNA

<213> Unknown (Unknown)

<220>

<223> Gene encoding extracellular domain of SFTS Virus Gn protein

<400> 4

gactctggac ctattatctg tgctggacct attcattcaa acaaatctgc tgacatccct 60

catctgctgg gctactccga aaaaatctgc cagattgatc gactgatcca cgtgagctcc 120

tggctgcgga accatagcca gttccaggga tacgtcggac agaggggcgg gcgctctcag 180

gtgagttact atccagccga gaacagctac agccggtggt ccggactgct gtctccatgt 240

gacgcagatt ggctgggaat gctggtggtc aagaaagcta agggctcaga tatgattgtc 300

cccggcccta gctacaaggg gaaagtgttc tttgaacgcc ccaccttcga cggctatgtg 360

ggctgggggt gcggatcagg caaaagccga acagagtccg gagaactgtg cagcagcgac 420

tctggaactt caagcggcct gctgcctagc gatagggtcc tgtggattgg cgacgtggcc 480

tgccagccaa tgaccccaat ccccgaggaa acatttctgg agctgaagag tttctcacag 540

agcgaatttc cagacatctg caaaattgac ggcatcgtgt tcaaccagtg tgagggggaa 600

agcctgcctc agccatttga tgtcgcttgg atggacgtgg gccactccca taagatcatt 660

atgcgcgagc acaagactaa atgggtccag gaatcctcta gtaaggattt cgtgtgctac 720

aaagagggga ccggaccctg ttccgaatct gaggaaaaga cttgcaaaac cagtgggtca 780

tgtcggggag acatgcagtt ttgcaaggtg gctggctgtg agcacgggga ggaagcctct 840

gaagctaaat gcagatgtag tctggtccat aaacccggcg aggtggtcgt gagttacgga 900

ggcatgcggg tgagacctaa atgctatggg ttcagcagaa tgatggcaac cctggaggtg 960

aaccagcctg aacagaggat cggacagtgc acaggctgtc acctggagtg tattaatggg 1020

ggagtccgac tgatcaccct gacaagtgaa ctgaagtcag caacagtgtg cgcctctcat 1080

ttctgttcaa gcgccactag cggcaagaaa tccaccgaga ttcagtttca cagcgggtcc 1140

ctggtgggaa agactgctat ccatgtcaaa ggagcactgg tggatggcac agagttcact 1200

tttgaagggt cctgcatgtt tccagacgga tgtgatgcag tggactgcac attctgtcgc 1260

gagtttctga agaatcccca gtgctatcct gccaagaaa 1299

<210> 5

<211> 1419

<212> DNA

<213> Unknown (Unknown)

<220>

<223> Gene encoding extracellular domain of Gc protein of SFTS virus

<400> 5

tgcgatgaaa tggtccacgc cgatagcaaa ctggtcagct gtaggcaggg aagcgggaac 60

atgaaggaat gcgtcacaac cggaagagct ctgctgccag cagtgaaccc tggacaggag 120

gcctgcctgc acttcacagc tcctggaagt ccagactcaa aatgcctgaa gatcaaagtg 180

aagcggatta atctgaagtg taagaaaagc tcctcttact ttgtgcccga cgcaaggtca 240

cgctgcacta gcgtccggag atgtagatgg gcaggcgatt gccagagcgg atgtccacct 300

catttcacct ctaacagttt ttcagacgat tgggcaggca agatggacag ggcaggactg 360

gggttcagcg gatgctccga tggatgtgga ggagcagctt gcggatgttt caacgcagcc 420

ccctcctgca tcttttggcg caaatgggtg gagaatcctc acggcatcat ttggaaggtc 480

tccccttgtg ctgcatgggt gccatctgcc gtcatcgagc tgacaatgcc cagcggcgaa 540

gtgagaactt tccatccaat gtccgggatt cccacccagg tctttaaggg agtgtctgtc 600

acatatctgg gcagcgacat ggaggtgtcc gggctgactg atctgtgcga aatcgaggaa 660

ctgaaatcca agaaactggc cctggctcct tgtaatcagg caggaatggg agtggtcggc 720

aaggtcgggg agattcagtg cagttcagag gaaagcgcac gcaccatcaa gaaagacggg 780

tgtatttgga acgccgatct ggtgggaatc gagctgcgag tggacgatgc tgtctgctac 840

tccaaaatta catctgtgga agcagtcgcc aattattccg ctatccctac cacaattggc 900

ggcctgcgat tcgaacggtc tcacgacagt caggggaaga tcagcggcag ccccctggac 960

atcaccgcca ttcgaggatc ttttagtgtg aactacagag gcctgaggct gtcactgagc 1020

gagatcactg ctacctgcac aggagaagtg actaatgtca gtggctgcta ttcatgtatg 1080

accggggcaa aagtgagtat taagctgcac agctccaaaa actcaacagc tcatgtgcgc 1140

tgtaagggag acgagactgc attctctgtg ctggaaggcg tccacagtta cacagtgtcc 1200

ctgtcttttg accatgccgt ggtcgatgag cagtgccagc tgaactgtgg gggacacgaa 1260

agccaagtga ctctgaaagg caatctgatc ttcctggacg tgccaaagtt tgtcgatggg 1320

tcatacatgc agacctatca tagcactgtg cccaccggcg ccaatattcc atctcccaca 1380

gattggctga acgctctgtt tgggaatgga ctgagtcgg 1419

<210> 6

<211> 60

<212> DNA

<213> Unknown (Unknown)

<220>

<223> Gene encoding Signal peptide

<400> 6

atggagacag acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt 60

<210> 7

<211> 654

<212> DNA

<213> Unknown (Unknown)

<220>

<223> Gene encoding human Fc

<400> 7

gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc 60

ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac 120

cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 180

ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac 240

caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc 300

cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc 360

ctgcccccat cccgggatga gctgaccaag aaccaggtca gcctgacctg cctggtcaaa 420

ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac 480

tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaagctc 540

accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 600

gctctgcaca accactacac gcagaagagc ctctccctgt ccccgggtaa atga 654

<210> 8

<211> 60

<212> DNA

<213> Unknown (Unknown)

<220>

<223> Gene encoding hinge region

<400> 8

ggccaggccg gccaggagcc caaatctagc gacaaaactc acacaagccc accgagccca 60

<210> 9

<211> 738

<212> DNA

<213> Unknown (Unknown)

<220>

<223> Gene encoding SFTS Virus NP protein

<400> 9

atgtcggagt ggtccaggat tgcagtggag tttggtgagc agcagctcaa tttgtctgag 60

cttgaggatt tcgcgagaga actggcctat gaaggccttg accctgcttt gatcatcaag 120

aagctgaagg agacaggtgg agatgattgg gtgaaggata caaagttcat cattgtcttt 180

gccctgactc gaggcaacaa gatcgtcaag gcatcaggga aaatgtcaaa ctcagggtcc 240

aagaggttga tggcactcca agagaaatat ggactggttg agagggcaga gaccaggctc 300

tcaatcactc ctgtgagggt tgcgcagagc cttcccactt ggacatgtgc tgcagcagca 360

gccttgaagg agtatctccc tgtggggcca gccgtcatga acctgaaggt cgagaattat 420

ccccctgaga tgatgtgcat ggcctttgga tccctgattc caactgcggg ggtatctgaa 480

gccacaacca agaccctgat ggaggcctac tctctgtggc aagatgcctt caccaagact 540

atcaatgtga agatgcgcgg agccagcaag acagaagttt acaactcctt cagggaccct 600

cttcatgctg ctgtgaactc tgtcttcttt cccaatgatg ttcgggtaaa gtggctgaag 660

gccaagggga tccttggccc agatggggtc cccagcagag ctgctgaggt tgctgctgct 720

gcttacagaa acctgtaa 738

<210> 10

<211> 676

<212> PRT

<213> Unknown (Unknown)

<220>

<223> Gn-Fc

<400> 10

Asp Val Ala Gln Ala Ala Asp Ser Gly Pro Ile Ile Cys Ala Gly Pro

1 5 10 15

Ile His Ser Asn Lys Ser Ala Asp Ile Pro His Leu Leu Gly Tyr Ser

20 25 30

Glu Lys Ile Cys Gln Ile Asp Arg Leu Ile His Val Ser Ser Trp Leu

35 40 45

Arg Asn His Ser Gln Phe Gln Gly Tyr Val Gly Gln Arg Gly Gly Arg

50 55 60

Ser Gln Val Ser Tyr Tyr Pro Ala Glu Asn Ser Tyr Ser Arg Trp Ser

65 70 75 80

Gly Leu Leu Ser Pro Cys Asp Ala Asp Trp Leu Gly Met Leu Val Val

85 90 95

Lys Lys Ala Lys Gly Ser Asp Met Ile Val Pro Gly Pro Ser Tyr Lys

100 105 110

Gly Lys Val Phe Phe Glu Arg Pro Thr Phe Asp Gly Tyr Val Gly Trp

115 120 125

Gly Cys Gly Ser Gly Lys Ser Arg Thr Glu Ser Gly Glu Leu Cys Ser

130 135 140

Ser Asp Ser Gly Thr Ser Ser Gly Leu Leu Pro Ser Asp Arg Val Leu

145 150 155 160

Trp Ile Gly Asp Val Ala Cys Gln Pro Met Thr Pro Ile Pro Glu Glu

165 170 175

Thr Phe Leu Glu Leu Lys Ser Phe Ser Gln Ser Glu Phe Pro Asp Ile

180 185 190

Cys Lys Ile Asp Gly Ile Val Phe Asn Gln Cys Glu Gly Glu Ser Leu

195 200 205

Pro Gln Pro Phe Asp Val Ala Trp Met Asp Val Gly His Ser His Lys

210 215 220

Ile Ile Met Arg Glu His Lys Thr Lys Trp Val Gln Glu Ser Ser Ser

225 230 235 240

Lys Asp Phe Val Cys Tyr Lys Glu Gly Thr Gly Pro Cys Ser Glu Ser

245 250 255

Glu Glu Lys Thr Cys Lys Thr Ser Gly Ser Cys Arg Gly Asp Met Gln

260 265 270

Phe Cys Lys Val Ala Gly Cys Glu His Gly Glu Glu Ala Ser Glu Ala

275 280 285

Lys Cys Arg Cys Ser Leu Val His Lys Pro Gly Glu Val Val Val Ser

290 295 300

Tyr Gly Gly Met Arg Val Arg Pro Lys Cys Tyr Gly Phe Ser Arg Met

305 310 315 320

Met Ala Thr Leu Glu Val Asn Gln Pro Glu Gln Arg Ile Gly Gln Cys

325 330 335

Thr Gly Cys His Leu Glu Cys Ile Asn Gly Gly Val Arg Leu Ile Thr

340 345 350

Leu Thr Ser Glu Leu Lys Ser Ala Thr Val Cys Ala Ser His Phe Cys

355 360 365

Ser Ser Ala Thr Ser Gly Lys Lys Ser Thr Glu Ile Gln Phe His Ser

370 375 380

Gly Ser Leu Val Gly Lys Thr Ala Ile His Val Lys Gly Ala Leu Val

385 390 395 400

Asp Gly Thr Glu Phe Thr Phe Glu Gly Ser Cys Met Phe Pro Asp Gly

405 410 415

Cys Asp Ala Val Asp Cys Thr Phe Cys Arg Glu Phe Leu Lys Asn Pro

420 425 430

Gln Cys Tyr Pro Ala Lys Lys Gly Gln Ala Gly Gln Glu Pro Lys Ser

435 440 445

Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu

450 455 460

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

465 470 475 480

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

485 490 495

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

500 505 510

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

515 520 525

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

530 535 540

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

545 550 555 560

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

565 570 575

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

580 585 590

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

595 600 605

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

610 615 620

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

625 630 635 640

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

645 650 655

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

660 665 670

Ser Pro Gly Lys

675

<210> 11

<211> 711

<212> PRT

<213> Unknown (Unknown)

<220>

<223> Gc-Fc

<400> 11

Asp Val Ala Gln Ala Ala Cys Asp Glu Met Val His Ala Asp Ser Lys

1 5 10 15

Leu Val Ser Cys Arg Gln Gly Ser Gly Asn Met Lys Glu Cys Val Thr

20 25 30

Thr Gly Arg Ala Leu Leu Pro Ala Val Asn Pro Gly Gln Glu Ala Cys

35 40 45

Leu His Phe Thr Ala Pro Gly Ser Pro Asp Ser Lys Cys Leu Lys Ile

50 55 60

Lys Val Lys Arg Ile Asn Leu Lys Cys Lys Lys Ser Ser Ser Tyr Phe

65 70 75 80

Val Pro Asp Ala Arg Ser Arg Cys Thr Ser Val Arg Arg Cys Arg Trp

85 90 95

Ala Gly Asp Cys Gln Ser Gly Cys Pro Pro His Phe Thr Ser Asn Ser

100 105 110

Phe Ser Asp Asp Trp Ala Gly Lys Met Asp Arg Ala Gly Leu Gly Phe

115 120 125

Ser Gly Cys Ser Asp Gly Cys Gly Gly Ala Ala Cys Gly Cys Phe Asn

130 135 140

Ala Ala Pro Ser Cys Ile Phe Trp Arg Lys Trp Val Glu Asn Pro His

145 150 155 160

Gly Ile Ile Trp Lys Val Ser Pro Cys Ala Ala Trp Val Pro Ser Ala

165 170 175

Val Ile Glu Leu Thr Met Pro Ser Gly Glu Val Arg Thr Phe His Pro

180 185 190

Met Ser Gly Ile Pro Thr Gln Val Phe Lys Gly Val Ser Val Thr Tyr

195 200 205

Leu Gly Ser Asp Met Glu Val Ser Gly Leu Thr Asp Leu Cys Glu Ile

210 215 220

Glu Glu Leu Lys Ser Lys Lys Leu Ala Leu Ala Pro Cys Asn Gln Ala

225 230 235 240

Gly Met Gly Val Val Gly Lys Val Gly Glu Ile Gln Cys Ser Ser Glu

245 250 255

Glu Ser Ala Arg Thr Ile Lys Lys Asp Gly Cys Ile Trp Asn Ala Asp

260 265 270

Leu Val Gly Ile Glu Leu Arg Val Asp Asp Ala Val Cys Tyr Ser Lys

275 280 285

Ile Thr Ser Val Glu Ala Val Ala Asn Tyr Ser Ala Ile Pro Thr Thr

290 295 300

Ile Gly Gly Leu Arg Phe Glu Arg Ser His Asp Ser Gln Gly Lys Ile

305 310 315 320

Ser Gly Ser Pro Leu Asp Ile Thr Ala Ile Arg Gly Ser Phe Ser Val

325 330 335

Asn Tyr Arg Gly Leu Arg Leu Ser Leu Ser Glu Ile Thr Ala Thr Cys

340 345 350

Thr Gly Glu Val Thr Asn Val Ser Gly Cys Tyr Ser Cys Met Thr Gly

355 360 365

Ala Lys Val Ser Ile Lys Leu His Ser Ser Lys Asn Ser Thr Ala His

370 375 380

Val Arg Cys Lys Gly Asp Glu Thr Ala Phe Ser Val Leu Glu Gly Val

385 390 395 400

His Ser Tyr Thr Val Ser Leu Ser Phe Asp His Ala Val Val Asp Glu

405 410 415

Gln Cys Gln Leu Asn Cys Gly Gly His Glu Ser Gln Val Thr Leu Lys

420 425 430

Gly Asn Leu Ile Phe Leu Asp Val Pro Lys Phe Val Asp Gly Ser Tyr

435 440 445

Met Gln Thr Tyr His Ser Thr Val Pro Thr Gly Ala Asn Ile Pro Ser

450 455 460

Pro Thr Asp Trp Leu Asn Ala Leu Phe Gly Asn Gly Leu Ser Arg Glu

465 470 475 480

Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro

485 490 495

Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

500 505 510

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

515 520 525

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

530 535 540

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

545 550 555 560

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

565 570 575

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

580 585 590

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

595 600 605

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys

610 615 620

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

625 630 635 640

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

645 650 655

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

660 665 670

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

675 680 685

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

690 695 700

Leu Ser Leu Ser Pro Gly Lys

705 710

<210> 12

<211> 245

<212> PRT

<213> Unknown (Unknown)

<220>

<223> NP

<400> 12

Met Ser Glu Trp Ser Arg Ile Ala Val Glu Phe Gly Glu Gln Gln Leu

1 5 10 15

Asn Leu Ser Glu Leu Glu Asp Phe Ala Arg Glu Leu Ala Tyr Glu Gly

20 25 30

Leu Asp Pro Ala Leu Ile Ile Lys Lys Leu Lys Glu Thr Gly Gly Asp

35 40 45

Asp Trp Val Lys Asp Thr Lys Phe Ile Ile Val Phe Ala Leu Thr Arg

50 55 60

Gly Asn Lys Ile Val Lys Ala Ser Gly Lys Met Ser Asn Ser Gly Ser

65 70 75 80

Lys Arg Leu Met Ala Leu Gln Glu Lys Tyr Gly Leu Val Glu Arg Ala

85 90 95

Glu Thr Arg Leu Ser Ile Thr Pro Val Arg Val Ala Gln Ser Leu Pro

100 105 110

Thr Trp Thr Cys Ala Ala Ala Ala Ala Leu Lys Glu Tyr Leu Pro Val

115 120 125

Gly Pro Ala Val Met Asn Leu Lys Val Glu Asn Tyr Pro Pro Glu Met

130 135 140

Met Cys Met Ala Phe Gly Ser Leu Ile Pro Thr Ala Gly Val Ser Glu

145 150 155 160

Ala Thr Thr Lys Thr Leu Met Glu Ala Tyr Ser Leu Trp Gln Asp Ala

165 170 175

Phe Thr Lys Thr Ile Asn Val Lys Met Arg Gly Ala Ser Lys Thr Glu

180 185 190

Val Tyr Asn Ser Phe Arg Asp Pro Leu His Ala Ala Val Asn Ser Val

195 200 205

Phe Phe Pro Asn Asp Val Arg Val Lys Trp Leu Lys Ala Lys Gly Ile

210 215 220

Leu Gly Pro Asp Gly Val Pro Ser Arg Ala Ala Glu Val Ala Ala Ala

225 230 235 240

Ala Tyr Arg Asn Leu

245

Claims (8)

1. A vaccine composition against fever with thrombocytopenia syndrome (STFS) virus,

the active ingredients include the extracellular domain of Gc protein (SEQ ID NO: 1), the extracellular domain of Gn protein (SEQ ID NO: 2), the full-length NP protein (SEQ ID NO: 3), or a mixture thereof.

2. The vaccine composition of claim 1, characterized in that:

the extracellular domain of Gc protein and the extracellular domain of Gn protein are fused to the Fc portion of human antibodies, respectively.

3. The vaccine composition of claim 1, wherein:

the mixture is (i) a mixture of the extracellular domain of the Gc protein and the full-length NP protein, (ii) a mixture of the extracellular domain of the Gn protein and the full-length NP protein, or (iii) a mixture of the extracellular domain of the Gc protein, the extracellular domain of the Gn protein, and the full-length NP protein.

4. The vaccine composition of claim 3, wherein:

the extracellular domain of the Gc protein and the extracellular domain of the Gn protein are fused to the Fc region of the human antibody, respectively.

5. The vaccine composition of claim 1, wherein:

the composition comprises a pharmaceutically acceptable carrier.

6. The vaccine composition of claim 5, wherein:

the pharmaceutically acceptable carrier includes one or more selected from the group consisting of a diluent, an excipient, a stabilizer, and a preservative.

7. The vaccine composition of claim 1, wherein:

the composition further comprises an active ingredient adjuvant.

8. The vaccine composition of claim 7, wherein:

the adjuvant is aluminum salt with gel property.

Images