VIOLIN: Vaccine Investigation and Online Information Network

Hantavirus

Table of Contents

General Information
Vaccine Related Pathogen Genes
1. Hanta_G2
2. ANDVsSgp1 (Protective antigen)
3. G1 (Protective antigen)
4. Gc (Protective antigen)
5. GP (Protective antigen)
6. HTNVsSgp1 (Protective antigen)
7. M Segment (Protective antigen)
8. PUUVsSgp1 (Protective antigen)
9. S nucleocapsid protein from Dobrava-Belgrade virus (Protective antigen)
10. S segment (Protective antigen)
11. SNVsSgp1 (Protective antigen)
Vaccine Related Host Genes
Vaccine Information
References

I. General Information

1. NCBI Taxonomy ID:

11598

2. Disease:

Hantavirus Pulmonary Syndrome

3. Introduction

Hantaviruses are rodent-borne agents that cause hemorrhagic fever with renal syndrome or hantavirus pulmonary syndrome (HPS) in humans (de et al., 2002). Annually, around 150,000 cases of HFRS are reported worldwide, caused by Hantaan (HTNV) and Seoul (SEOV) hantaviruses in Asia and by Puumala (PUUV), Dobrava (DOBV), and Saaremaa (SAAV) hantaviruses in Europe (Klingstrom et al., 2004). Hantavirus was first identified in the United States in 1993. HPS is a deadly disease from rodents. Although rare, Although rare, the consequences of getting it are serious. Early symptoms include fatigue, fever and muscle aches. There may also be headaches, dizziness, chills, and abdominal problems, such as nausea, vomiting, diarrhea, and abdominal pain. The major HPS symptoms may not appear until the illness becomes life-threatening. Rodent control in and around the home remains the primary strategy for preventing hantavirus infection (CDC Hantaviruses). Hantaviruses can also cause human hemorrhagic fever with renal syndrome (HFRS) (Lednicky, 2003).

Increased numbers of rodents in the household is the strongest risk factor for infection. Among documented U.S. cases of HPS, patients with potential occupational exposures have included grain farmers, an extension livestock specialist, field biologists, and agricultural, mill, construction, utility and feedlot workers (CDC Hantaviruses).

Hantaviruses contain a three-segmented ssRNA-genome of negative polarity encoding four proteins: the nucleocapsid protein (N), the envelope proteins (G1 and G2), and the RNA dependent RNA polymerase (Klingstrom et al., 2004).

4. Microbial Pathogenesis

Rodents (primarily deer mice) carry hantaviruses that cause hantavirus pulmonary syndrome in human. Rodents shed the virus in their urine, droppings, and saliva. The virus is mainly transmitted to people when they breathe in air contaminated with the virus. Functional impairment of vascular endothelium is central to the pathogenesis of HPS, and a myocardial depressant may contribute significantly to the mortality of this disease. Hantaviruses can infect endothelial cells, macrophages and dendritic cells. The mechanism of inflammatory cell recruitment in the lungs of HPS patients may result from specific attraction and adherence of a selective population of inflammatory cells to an activated pulmonary microvascular endothelium (CDC Hantaviruses).

5. Host Ranges and Animal Models

n the US, primarily deer mice, along with cotton rats and rice rats in the southeastern states and the white-footed mouse in the Northeast, carry hantaviruses that cause hantavirus pulmonary syndrome (CDC Hantaviruses).

6. Host Protective Immunity

Klingström et al. reported that the strong Th2-type of immune response induced by Alum against rDOBV N did not induce protection in mice (Klingstrom et al., 2004).

II. Vaccine Related Pathogen Genes

1. ANDVsSgp1

Gene Name : ANDVsSgp1
Sequence Strain (Species/Organism) : Andes orthohantavirus
VO ID : VO_0011092
NCBI Gene ID : 991232
NCBI Protein GI : 19744950
Locus Tag : ANDVsSgp1
Genbank Accession : AF004660
Protein Accession : NP_604471
Taxonomy ID : 1980456
Segment No : Segment S
Gene Starting Position : 42
Gene Ending Position : 1328
Gene Strand (Orientation) : +
Protein Name : nucleocapsid protein
Protein pI : 7.18
Protein Weight : 45461.89
Protein Length : 428

DNA Sequence : Show Sequence

>NC_003466.1:42-1328 Andes virus segment S, complete sequence
AATGAGCACCCTCCAAGAATTGCAGGAAAACATCACAGCACACGAACAACAGCTCGTGACTGCTCGGCAA
AAGCTTAAGGATGCCGAGAAGGCAGTGGAGGTGGACCCGGATGACGTTAACAAGAGCACACTACAAAGTA
GACGGGCAGCTGTGTCTACATTGGAGACCAAACTCGGAGAACTTAAGAGGCAACTTGCAGATTTGGTGGC
AGCTCAAAAATTGGCTACAAAACCAGTTGATCCAACAGGGCTTGAGCCTGATGATCATCTAAAGGAAAAA
TCATCTCTGAGATATGGGAATGTCCTGGATGTTAATTCAATTGATTTGGAAGAACCGAGTGGACAGACTG
CTGATTGGAAGGCTATAGGAGCATACATCTTAGGGTTTGCAATTCCGATCATCCTAAAGGCCTTATACAT
GCTGTCAACCCGTGGGAGACAAACTGTGAAAGACAACAAAGGGACCAGGATAAGGTTTAAGGATGATTCT
TCCTTTGAAGAAGTCAATGGGATACGTAAACCAAAACACCTTTACGTCTCAATGCCAACTGCACAGTCCA
CTATGAAGGCTGAAGAAATCACGCCAGGACGATTTAGGACAATTGCTTGTGGCCTTTTTCCAGCACAGGT
CAAAGCCCGAAATATAATAAGTCCTGTAATGGGAGTAATTGGATTTGGCTTCTTTGTAAAGGATTGGATG
GATCGGATAGAAGAGTTTCTGGCTGCAGAGTGTCCATTCTTACCTAAGCCAAAGGTCGCCTCAGAAGCCT
TCATGTCTACCAATAAGATGTATTTTCTGAACAGACAGAGACAAGTCAATGAATCTAAGGTTCAAGATAT
TATCGATTTGATAGACCATGCTGAGACCGAGTCTGCTACCTTGTTTACAGAGATTGCAACACCCCATTCA
GTCTGGGTGTTTGCATGTGCACCTGACCGGTGCCCTCCAACTGCATTGTATGTTGCAGGGGTACCGGAAC
TTGGTGCATTTTTTTCTATCCTTCAGGACATGCGTAATACCATCATGGCATCTAAATCTGTAGGGACTGC
AGAAGAGAAGCTAAAGAAAAAATCTGCCTTCTACCAATCATACCTAAGAAGGACACAATCTATGGGAATC
CAACTGGACCAGAAGATCATAATCCTTTACATGCTATCATGGGGTAAAGAAGCTGTGAATCACTTCCATC
TTGGTGATGATATGGACCCTGAACTCAGGCAGCTAGCACAATCTCTGATCGATACTAAGGTGAAGGAGAT
CTCCAACCAAGAGCCACTTAAGTTGTA

Protein Sequence : Show Sequence

>NP_604471.1 nucleocapsid protein [Andes orthohantavirus]
MSTLQELQENITAHEQQLVTARQKLKDAEKAVEVDPDDVNKSTLQSRRAAVSTLETKLGELKRQLADLVA
AQKLATKPVDPTGLEPDDHLKEKSSLRYGNVLDVNSIDLEEPSGQTADWKAIGAYILGFAIPIILKALYM
LSTRGRQTVKDNKGTRIRFKDDSSFEEVNGIRKPKHLYVSMPTAQSTMKAEEITPGRFRTIACGLFPAQV
KARNIISPVMGVIGFGFFVKDWMDRIEEFLAAECPFLPKPKVASEAFMSTNKMYFLNRQRQVNESKVQDI
IDLIDHAETESATLFTEIATPHSVWVFACAPDRCPPTALYVAGVPELGAFFSILQDMRNTIMASKSVGTA
EEKLKKKSAFYQSYLRRTQSMGIQLDQKIIILYMLSWGKEAVNHFHLGDDMDPELRQLAQSLIDTKVKEI
SNQEPLKL

Molecule Role : Protective antigen
Molecule Role Annotation : Nonreplicating adenovirus (Ad) vectors that express Andes hantavirus (ANDV) nucleocapsid protein (AdN, ANDVsSgp1) or glycoproteins (AdG(N) and AdG(C)) were constructed . When administered once, all three Ad vectors, individually or in combination, elicited a robust immune response that protected Syrian hamsters from a lethal ANDV infection that mimics the pulmonary disease seen in humans. No vaccinated animal died, and there were no obvious clinical signs of disease (Safronetz et al., 2009).
Related Vaccine(s): Andes virus vaccine using adenovirus viral vector expressing ANDV NP , rVSVΔG-ANDV-GPC

2. G1

Gene Name : G1
Sequence Strain (Species/Organism) : Sin Nombre virus
VO ID : VO_0011086
NCBI Protein GI : 1113859
3D structure: PDB ID : 2K9H
Other Database IDs : CDD:279853
Taxonomy ID : 1980491
Gene Strand (Orientation) : ?
Protein Name : envelope glycoprotein G1
Protein pI : 5.59
Protein Weight : 9420.59
Protein Length : 169
Protein Note : SNV

Protein Sequence : Show Sequence

>AAC54579.1 envelope glycoprotein G1, partial [Sin Nombre orthohantavirus]
FLVVLTTATAGLTRNLYELKIECPHTVGLGQGYVTGSVETTPVLFSQVADLKIESSCNFDLHVPATTTQK
YNQVDWTKKSSTTENTNAGAS

Molecule Role : Protective antigen
Molecule Role Annotation : Study used a deer mouse infection model to test the protective efficacy of genetic vaccine candidates for Sin Nombre (SN) virus that were known to provoke immunological responses in BALB/c mice. Protective epitopes were localized in each of four overlapping cDNA fragments that encoded portions of the SN virus G1 glycoprotein antigen; the nucleocapsid gene also was protective (Bharadwaj et al., 2002).
Related Vaccine(s): Sin Nombre virus DNA vaccine encoding G1

3. Gc

Gene Name : Gc
Sequence Strain (Species/Organism) : Seoul virus
VO ID : VO_0011085
NCBI Protein GI : 284157806
Other Database IDs : CDD:144961
Taxonomy ID : 1980490
Gene Strand (Orientation) : ?
Protein Name : Gc glycoprotein
Protein pI : 6.5
Protein Weight : 11999.31
Protein Length : 170
Protein Note : Hantavirus glycoprotein G2; pfam01561

Protein Sequence : Show Sequence

>ADB80030.1 Gc glycoprotein, partial [Seoul orthohantavirus]
VPLWTDNAHGVGSVPMHTDLELDFSLPSSSKYTYKRHLTNPVNDQQSVSLHIEIESQGIGADVHHLGHWY
DARLNLKTSFHCYGACTKYQYPWHTAKCHFEKDYEY

Molecule Role : Protective antigen
Molecule Role Annotation : Seoul virus (SEOV) is one of the four known hantaviruses causing hemorrhagic fever with renal syndrome (HFRS). A replication-competent recombinant canine adenovirus type 2 expressing the Gc protein of SEOV (rCAV-2-Gc) in BALB/c mice induced complete protection against a intensive infectious challenge with ~1,000 50% infective doses (ID50) for SEOV strain CC-2 (Yuan et al., 2010).
Related Vaccine(s): Seoul virus vaccine rCAV-2-Gc

4. GP

Gene Name : GP
Sequence Strain (Species/Organism) : Hantaan virus 76-118
NCBI Protein GI : 138339
Other Database IDs : CDD:279853
CDD:119058
CDD:144961
Taxonomy ID : 11602
Gene Strand (Orientation) : ?
Protein Name : Envelopment polyprotein
Protein pI : 7.1
Protein Weight : 122560.28
Protein Length : 1460
Protein Note : Glycoprotein precursor; M polyprotein

Protein Sequence : Show Sequence

>sp|P08668.1|GP_HANTV RecName: Full=Envelopment polyprotein; AltName: Full=Glycoprotein precursor; AltName: Full=M polyprotein; Contains: RecName: Full=Glycoprotein N; Short=Gn; AltName: Full=Glycoprotein G2; Contains: RecName: Full=Glycoprotein C; Short=Gc; AltName: Full=Glycoprotein G1; Flags: Precursor
MGIWKWLVMASLVWPVLTLRNVYDMKIECPHTVSFGENSVIGYVELPPVPLADTAQMVPESSCNMDNHQS
LNTITKYTQVSWRGKADQSQSSQNSFETVSTEVDLKGTCVLKHKMVEESYRSRKSVTCYDLSCNSTYCKP
TLYMIVPIHACNMMKSCLIALGPYRVQVVYERSYCMTGVLIEGKCFVPDQSVVSIIKHGIFDIASVHIVC
FFVAVKGNTYKIFEQVKKSFESTCNDTENKVQGYYICIVGGNSAPIYVPTLDDFRSMEAFTGIFRSPHGE
DHDLAGEEIASYSIVGPANAKVPHSASSDTLSLIAYSGIPSYSSLSILTSSTEAKHVFSPGLFPKLNHTN
CDKSAIPLIWTGMIDLPGYYEAVHPCTVFCVLSGPGASCEAFSEGGIFNITSPMCLVSKQNRFRLTEQQV
NFVCQRVDMDIVVYCNGQRKVILTKTLVIGQCIYTITSLFSLLPGVAHSIAVELCVPGFHGWATAALLVT
FCFGWVLIPAITFIILTVLKFIANIFHTSNQENRLKSVLRKIKEEFEKTKGSMVCDVCKYECETYKELKA
HGVSCPQSQCPYCFTHCEPTEAAFQAHYKVCQVTHRFRDDLKKTVTPQNFTPGCYRTLNLFRYKSRCYIF
TMWIFLLVLESILWAASASETPLTPVWNDNAHGVGSVPMHTDLELDFSLTSSSKYTYRRKLTNPLEEAQS
IDLHIEIEEQTIGVDVHALGHWFDGRLNLKTSFHCYGACTKYEYPWHTAKCHYERDYQYETSWGCNPSDC
PGVGTGCTACGLYLDQLKPVGSAYKIITIRYSRRVCVQFGEENLCKIIDMNDCFVSRHVKVCIIGTVSKF
SQGDTLLFFGPLEGGGLIFKHWCTSTCQFGDPGDIMSPRDKGFLCPEFPGSFRKKCNFATTPICEYDGNM
VSGYKKVMATIDSFQSFNTSTMHFTDERIEWKDPDGMLRDHINILVTKDIDFDNLGENPCKIGLQTSSIE
GAWGSGVGFTLTCLVSLTECPTFLTSIKACDKAICYGAESVTLTRGQNTVKVSGKGGHSGSTFRCCHGED
CSQIGLHAAAPHLDKVNGISEIENSKVYDDGAPQCGIKCWFVKSGEWISGIFSGNWIVLIVLCVFLLFSL
VLLSILCPVRKHKKS

Molecule Role : Protective antigen
Related Vaccine(s): Hantavirus DNA vaccine pWRG/HTN-M

5. Hanta_G2

Gene Name : Hanta_G2
Sequence Strain (Species/Organism) : Andes virus Case T isolate
NCBI Nucleotide GI : 2801763
NCBI Protein GI : 2801763
Genbank Accession : AAB97456
Other Database IDs : Accession:AAB97456; GI:2801763
Gene Strand (Orientation) : ?
Protein Name : Andes Hantavirus M segment polyprotein

DNA Sequence : Show Sequence

>gi|2801762|gb|AF042137.1|AF042137 Andes virus Case T M segment polyprotein gene, partial cds
AAGGAAATACCATTTCTGGATATAAAAGAATGATGGCAACAAAAGATTCATTCCAATCGTTTAACTTAAC
AGAACCTCACATCACAGCAAATAAGCTTGAATGGATTGACCCAGATGGAAATACAAGAGACCATGTAAAT
CTTGTCTTAAATAGAGATGTTTCATTT

Protein Sequence : Show Sequence

>gi|2801763|gb|AAB97456.1| M segment polyprotein [Andes virus]
GNTISGYKRMMATKDSFQSFNLTEPHITANKLEWIDPDGNTRDHVNLVLNRDVSF

Related Vaccine(s): DNA vaccine encoding ANDV glycoproteins

6. HTNVsSgp1

Gene Name : HTNVsSgp1
Sequence Strain (Species/Organism) : Hantaan orthohantavirus
VO ID : VO_0011091
NCBI Gene ID : 2943078
NCBI Protein GI : 38371711
Locus Tag : HTNVsSgp1
Genbank Accession : M14626
Protein Accession : NP_941977
Taxonomy ID : 1980471
Segment No : segment S
Gene Starting Position : 36
Gene Ending Position : 1325
Gene Strand (Orientation) : +
Protein Name : hypothetical protein
Protein pI : 7.11
Protein Weight : 45859.21
Protein Length : 429

DNA Sequence : Show Sequence

>NC_005218.1:36-1325 Hantaan virus, complete genome
GATGGCAACTATGGAGGAATTACAGAGGGAAATCAATGCCCATGAGGGTCAATTAGTGATAGCCAGGCAG
AAGGTGAGGGATGCAGAAAAACAGTATGAAAAGGATCCAGATGAGTTGAACAAGAGAACATTAACTGACC
GAGAGGGCGTTGCAGTATCTATCCAGGCAAAAATTGATGAGTTAAAAAGGCAACTGGCAGATAGGATTGC
AACTGGGAAAAACCTTGGGAAGGAACAAGATCCAACAGGGGTGGAGCCTGGAGACCATCTGAAAGAGAGG
TCAATGCTCAGTTATGGTAATGTGCTGGATTTAAACCATTTGGATATTGATGAACCTACAGGACAGACAG
CAGACTGGCTGAGCATCATCGTCTATCTTACATCCTTTGTCGTCCCGATACTTCTGAAAGCTCTGTATAT
GTTGACAACAAGGGGGAGGCAAACTACCAAGGATAATAAAGGGACCCGGATTCGATTTAAGGATGATAGC
TCGTTCGAGGATGTTAACGGTATCCGGAAACCAAAACATCTTTACGTGTCCTTGCCAAATGCACAGTCAA
GCATGAAGGCAGAAGAGATTACACCTGGTAGATATAGAACAGCAGTCTGTGGGCTCTACCCTGCACAGAT
TAAGGCACGGCAGATGATCAGTCCAGTTATGAGTGTAATTGGTTTTCTAGCATTAGCAAAGGACTGGAGT
GATCGTATCGAACAATGGTTAATTGAACCTTGCAAGCTTCTTCCAGATACAGCAGCAGTTAGCCTCCTTG
GTGGTCCTGCAACAAACAGGGACTACTTACGGCAGCGGCAAGTGGCATTAGGCAATATGGAGACAAAGGA
GTCAAAGGCTATACGCCAGCATGCAGAAGCAGCTGGCTGTAGCATGATTGAAGATATTGAGTCACCATCA
TCAATATGGGTTTTTGCTGGAGCACCAGACCGTTGTCCACCAACATGTTTGTTTATAGCAGGTATTGCTG
AGCTTGGGGCATTTTTTTCCATCCTGCAGGACATGCGAAATACAATCATGGCATCTAAGACAGTTGGAAC
ATCTGAGGAGAAGCTACGGAAGAAATCATCATTTTATCAGTCCTACCTCAGAAGGACACAATCAATGGGG
ATACAACTAGGCCAGAGAATTATTGTGCTCTTCATGGTTGCCTGGGGAAAGGAGGCTGTGGACAACTTCC
ACTTAGGGGATGATATGGATCCTGAGCTAAGGACACTGGCACAGAGCTTGATTGATGTCAAAGTGAAGGA
AATCTCCAACCAAGAGCCTTTGAAACTCTA

Protein Sequence : Show Sequence

>NP_941977.1 hypothetical protein [Hantaan orthohantavirus]
MATMEELQREINAHEGQLVIARQKVRDAEKQYEKDPDELNKRTLTDREGVAVSIQAKIDELKRQLADRIA
TGKNLGKEQDPTGVEPGDHLKERSMLSYGNVLDLNHLDIDEPTGQTADWLSIIVYLTSFVVPILLKALYM
LTTRGRQTTKDNKGTRIRFKDDSSFEDVNGIRKPKHLYVSLPNAQSSMKAEEITPGRYRTAVCGLYPAQI
KARQMISPVMSVIGFLALAKDWSDRIEQWLIEPCKLLPDTAAVSLLGGPATNRDYLRQRQVALGNMETKE
SKAIRQHAEAAGCSMIEDIESPSSIWVFAGAPDRCPPTCLFIAGIAELGAFFSILQDMRNTIMASKTVGT
SEEKLRKKSSFYQSYLRRTQSMGIQLGQRIIVLFMVAWGKEAVDNFHLGDDMDPELRTLAQSLIDVKVKE
ISNQEPLKL

Molecule Role : Protective antigen
Molecule Role Annotation : Passive transfer of spleen cells from mice immunized with rNP (HTNVsSgp1) conferred partial protection or prolongation of time to death from fatal Hantaan virus infection in suckling mice which were challenged with Hantaan virus at 40 LD50 (survival rate: 43%) or 4 LD50 (survival rate: 43%) (Yoshimatsu et al., 1993).

7. M Segment

Gene Name : M Segment
Sequence Strain (Species/Organism) : Hantaan virus
VO ID : VO_0011090
NCBI Protein GI : 1389703
Other Database IDs : CDD:144961
Taxonomy ID : 1980471
Gene Strand (Orientation) : ?
Protein Name : M polyprotein
Protein pI : 6.21
Protein Weight : 12313.73
Protein Length : 172
Protein Note : Hantavirus glycoprotein G2; pfam01561

Protein Sequence : Show Sequence

>AAB02907.1 M polyprotein, partial [Hantaan orthohantavirus]
TPLTPVWNDNAHGVGSVPMHTDLELDFSLTSSSKYTYRRKLTNPLEEAQSVDLHIEIEEQTIGVDVHALG
HWFDGRLNLKTSFHCYGACTKYDYPWHTAKCHYERDYQ

Molecule Role : Protective antigen
Molecule Role Annotation : Researchers vaccinated hamsters with a vaccinia virus-vectored vaccine expressing the M and the S segments of Hantaan (HTN) virus and challenged them with HTN, Seoul (SEO), or Puumala (PUU) virus. Study found that vaccinated hamsters, challenged with HTN or SEO virus, were neither viremic nor had evidence of virus in their lungs or kidneys (Chu et al., 1995).

8. PUUVsSgp1

Gene Name : PUUVsSgp1
Sequence Strain (Species/Organism) : Puumala orthohantavirus
VO ID : VO_0011088
NCBI Gene ID : 2943083
NCBI Protein GI : 38371721
Locus Tag : PUUVsSgp1
Genbank Accession : HE801633
Protein Accession : NP_941984
Taxonomy ID : 1980486
Gene Starting Position : 42
Gene Ending Position : 1343
Gene Strand (Orientation) : +
Protein Name : nucleocapsid protein
Protein pI : 6.05
Protein Weight : 46970.43
Protein Length : 433

DNA Sequence : Show Sequence

>NC_005224.1:42-1343 Puumala virus segment S, complete sequence
AATGAGTGACTTGACAGATATCCAAGAGGATATAACCCGCCATGAACAGCAACTTATTGTTGCCAGACAA
AAACTTAAGGATGCAGAGAGAGCAGTGGAAGTGGACCCAGATGACGTTAATAAAAACACACTGCAAGCCA
GGCAACAAACAGTGTCAGCACTGGAGGACAAACTCGCAGACTACAAGAGAAGGATGGCAGATGCTGTGTC
CAGGAAAAAAATGGATACTAAACCTACTGACCCGACTGGGATTGAACCTGATGACCACCTCAAGGAGAGA
TCAAGCCTTAGGTATGGAAATGTCCTTGATGTAAATGCAATTGACATTGAAGAACCAAGTGGCCAAACAG
CAGATTGGTATACAATTGGAGTGTATGTGATAGGGTTCACACTTCCTATCATCCTTAAAGCTTTATACAT
GCTCTCAACGCGTGGGAGACAGACTGTAAAGGAAAATAAGGGGACACGTATAAGGTTTAAGGATGATACA
TCATTTGAAGACATCAATGGCATAAGGAGACCAAAGCATTTATATGTTTCTATGCCTACTGCCCAGTCAA
CTATGAAAGCAGAAGAACTCACACCTGGCAGATTTCGCACAATAGTATGTGGTCTTTTTCCCACTCAGAT
CCAGGTTCGTAACATCATGAGTCCAGTTATGGGGGTCATTGGTTTTTCATTCTTTGTGAAGGATTGGTCT
GAGAGAATCAGAGAGTTCATGGAAAAAGAGTGCCCATTCATAAAGCCTGAAGTAAAACCAGGCACACCAG
CACAGGAGATTGAGATGTTAAAAAGAAATAAGATCTACTTTATGCAGCGCCAGGATGTGCTTGACAAAAA
TCATGTGGCAGACATTGACAAGTTAATTGACTATGCAGCCTCTGGAGACCCTACATCACCTGACAACATT
GATTCGCCTAATGCACCATGGGTCTTTGCATGTGCACCAGACCGATGCCCACCAACATGTATCTATGTTG
CAGGGATGGCAGAGCTTGGGGCCTTTTTTTCAATATTGCAGGACATGAGAAACACAATAATGGCATCTAA
GACTGTTGGCACAGCAGAAGAGAAATTAAAGAAAAAGTCTTCCTTTTACCAATCTTACCTGCGTCGAACT
CAATCAATGGGAATTCAGCTTGATCAAAGAATTATCCTCTTGTTTATGTTGGAATGGGGCAAAGAGATGG
TAGATCACTTCCATCTTGGTGATGATATGGATCCAGAGCTCAGAGGTCTTGCACAGGCACTGATAGATCA
AAAGGTCAAGGAAATATCGAACCAGGAACCACTAAAGATATA

Protein Sequence : Show Sequence

>NP_941984.1 nucleocapsid protein [Puumala orthohantavirus]
MSDLTDIQEDITRHEQQLIVARQKLKDAERAVEVDPDDVNKNTLQARQQTVSALEDKLADYKRRMADAVS
RKKMDTKPTDPTGIEPDDHLKERSSLRYGNVLDVNAIDIEEPSGQTADWYTIGVYVIGFTLPIILKALYM
LSTRGRQTVKENKGTRIRFKDDTSFEDINGIRRPKHLYVSMPTAQSTMKAEELTPGRFRTIVCGLFPTQI
QVRNIMSPVMGVIGFSFFVKDWSERIREFMEKECPFIKPEVKPGTPAQEIEMLKRNKIYFMQRQDVLDKN
HVADIDKLIDYAASGDPTSPDNIDSPNAPWVFACAPDRCPPTCIYVAGMAELGAFFSILQDMRNTIMASK
TVGTAEEKLKKKSSFYQSYLRRTQSMGIQLDQRIILLFMLEWGKEMVDHFHLGDDMDPELRGLAQALIDQ
KVKEISNQEPLKI

Molecule Role : Protective antigen
Molecule Role Annotation : To investigate the ability of recombinant N (rN, nucleocapsid proteins) from different hantaviruses to elicit cross-protection, we immunized bank voles with rN from Puumala (PUUV), Topografov (TOPV), Andes (ANDV), and Dobrava (DOBV) viruses and subsequently challenged them with PUUV. All animals immunized with PUUV (PUUVsSgp1) and TOPV rN were completely protected (de et al., 2002).
Related Vaccine(s): Puumala virus PUUVsSgp1 (NP) protein vaccine

9. S nucleocapsid protein from Dobrava-Belgrade virus

Gene Name : S nucleocapsid protein from Dobrava-Belgrade virus
Sequence Strain (Species/Organism) : Dobrava-Belgrade orthohantavirus
VO ID : VO_0010960
NCBI Gene ID : 2656266
NCBI Protein GI : 38505524
Locus Tag : DOBVsSp1
Genbank Accession : AJ410615
Protein Accession : NP_942553
Other Database IDs : GI:63030019; Accession:AY961618.1
Taxonomy ID : 1980467
Gene Starting Position : 35
Gene Ending Position : 1324
Gene Strand (Orientation) : +
Protein Name : nucleocapsid protein
Protein pI : 7.53
Protein Weight : 46162.49
Protein Length : 429

DNA Sequence : Show Sequence

>NC_005233.1:35-1324 Dobrava virus complete S segment gene for nucleocapsid protein, strain DOBV/Ano-Poroia/Afl9/1999
GATGGCAACATTAGAGGAACTCCAAAAAGAAATCAACAACCACGAGGGCCAATTGGTGATAGCCAGGCAG
AAGGTGAAAGATGCAGAAAAGCAGTATGAAAAGGATCCTGACGACCTGAATAAAAGGGCATTGAGTGACC
GGGAAAGCATTGCACAATCAATTCAGGGAAAAATAGATGAATTAAGGAGACAGCTGGCTGATCGTGTGGC
AGCAGGGAAAAACATTGGCAAAGAGAGGGACCCAACTGGACTAGACCCTGGAGATCACCTCAAAGAGAAG
TCAATGCTCAGTTATGGAAATGTCATTGACCTCAACCATCTTGACATTGATGAACCTACAGGGCAAACCG
CAGACTGGCTGAGCATTGTGGTCTACCTGACATCATTCGTGGTTCCAATATTGTTGAAGGCTCTTTACAT
GCTTACCACCAGAGGGAGACAAACTACTAAAGACAATAAGGGAATGAGAATTCGATTTAAGGATGACAGC
TCTTTTGAAGATGTGAATGGGATTAGAAAGCCAAAGCACCTGTTCTTGTCAATGCCCAATGCACAATCTA
GCATGAAGGCAGATGAGATTACACCAGGTCGGTTCAGGACTGCAATTTGTGGACTATACCCAGCCCAGGT
GAAGGCAAGGAACTTAATCAGTCCTGTGATGAGTGTGATTGGGTTTTTAGCCCTTGCAAAGAACTGGACA
GAGCGGGTTGAGGAATGGCTTGACCTCCCGTGCAAGCTACTATCTGAGCCATCTCCAACGTCTTTGACCA
AAGGCCCATCCACCAATCGTGACTACTTGAATCAAAGACAAGGAGCGCTTGCAAAAATGGAAACAAAAGA
AGCTCAGGCTGTGAGGAAACATGCCATAGATGCTGGCTGCAACCTTATCGACCATATAGACTCACCATCA
TCAATCTGGGTCTTTGCAGGAGCACCCGACAGATGCCCTCCTACCTGCCTGTTCATTGCAGGCATGGCAG
AGCTAGGTGCATTCTTTGCTGTTCTCCAGGATATGAGGAACACCATCATGGCGTCAAAAACTATCGGAAC
ATCTGAGGAGAAGCTAAAGAAGAAATCATCTTTTTACCAATCTTACCTACGGAGGACACAATCTATGGGG
ATACAACTGGACCAGCGCATCATCGTGCTCTTCATGGTTGACTGGGGAAAAGAGGCAGTTGATAGTTTTC
ATCTTGGTGATGATATGGATCCTGAGCTCCGGGGCCTGGCACAGGCATTAATTGATCAAAAAGTGAAGGA
AATATCTAATCAGGAACCGCTTAAGCTTTA

Protein Sequence : Show Sequence

>NP_942553.1 nucleocapsid protein [Dobrava-Belgrade orthohantavirus]
MATLEELQKEINNHEGQLVIARQKVKDAEKQYEKDPDDLNKRALSDRESIAQSIQGKIDELRRQLADRVA
AGKNIGKERDPTGLDPGDHLKEKSMLSYGNVIDLNHLDIDEPTGQTADWLSIVVYLTSFVVPILLKALYM
LTTRGRQTTKDNKGMRIRFKDDSSFEDVNGIRKPKHLFLSMPNAQSSMKADEITPGRFRTAICGLYPAQV
KARNLISPVMSVIGFLALAKNWTERVEEWLDLPCKLLSEPSPTSLTKGPSTNRDYLNQRQGALAKMETKE
AQAVRKHAIDAGCNLIDHIDSPSSIWVFAGAPDRCPPTCLFIAGMAELGAFFAVLQDMRNTIMASKTIGT
SEEKLKKKSSFYQSYLRRTQSMGIQLDQRIIVLFMVDWGKEAVDSFHLGDDMDPELRGLAQALIDQKVKE
ISNQEPLKL

Molecule Role : Protective antigen
Molecule Role Annotation : Study compared the immunogenicity and protective efficacy of recombinant DOBV nucleocapsid protein (rDOBV N, S) given with Alum or Freund's as adjuvant, or PBS, in C57/BL6 mice. Mice receiving rDOBV N with Freund's adjuvant were protected from challenge (75% protected) (Klingstrom et al., 2004).
Related Vaccine(s): Dobrava-Belgrade virus S nucleocapsid protein vaccine , Recombinant DOBV nucleocapsid protein (rDOBV N)

10. S segment

Gene Name : S segment
Sequence Strain (Species/Organism) : Topografov virus
VO ID : VO_0011089
NCBI Protein GI : 4757106
Other Database IDs : CDD:279218
GOA:Q9WMG3
InterPro: IPR002214
UniProtKB/TrEMBL: Q9WMG3
Taxonomy ID : 83192
Gene Strand (Orientation) : ?
Protein Name : N protein
Protein pI : 5.8
Protein Weight : 47447.65
Protein Length : 487
Protein Note : Hantavirus nucleocapsid protein; pfam00846

Protein Sequence : Show Sequence

>CAB42097.1 N protein [Topografov hantavirus]
MSNLKDIQDEITRYEQQLIVARQKLRDAEKTVEVDPDDVNKNTLQARRQTVSALEDKLADFKRQLADHVS
RQKMDEKPSDPTGIEPDDHLKERSSLRYGNVLDVNAIDIEEPSGQTADWFTIGVYIVSFTLPIILKALYM
LSTRGRQTVKENKGTRIRFKDDSSFEDVNGIRRPKHLYVSMPTAQSTMKAEELTPGRFRTIVCGLFPAQI
QARNIMSPVMGVIGFSFFVKDWPERIKNFLESKCPFIKPEVKPGAPAGEADFLSRNQIYFMRRQEVLEDN
HIPDIDKLLEYASSGDPTAPDSIESPYAPWVFACAPDRCPPTCIYIAGMAELGAFFSILQDMRNTIMASK
TVGTAEEKLKKKSSFYQSYLRRTQSMGIQLDQRIILLYMVEWGKEMVDHFHLGDDMDPDLRNLAQSLIDQ
KVKEISNQEPLKI

Molecule Role : Protective antigen
Molecule Role Annotation : To investigate the ability of recombinant N (rN, nucleocapsid proteins) from different hantaviruses to elicit cross-protection, we immunized bank voles with rN from Puumala (PUUV), Topografov (TOPV), Andes (ANDV), and Dobrava (DOBV) viruses and subsequently challenged them with PUUV. All animals immunized with PUUV and TOPV rN were completely protected (de Carvalho Nicacio et al., 2002).
Related Vaccine(s): Topografov virus N protein vaccine

11. SNVsSgp1

Gene Name : SNVsSgp1
Sequence Strain (Species/Organism) : Sin Nombre orthohantavirus
VO ID : VO_0011087
NCBI Gene ID : 2943142
NCBI Protein GI : 38371726
Locus Tag : SNVsSgp1
Genbank Accession : JQ690276
Protein Accession : NP_941975
Taxonomy ID : 1980491
Segment No : segment S
Gene Starting Position : 42
Gene Ending Position : 1328
Gene Strand (Orientation) : +
Protein Name : nucleocapsid protein
Protein pI : 6.64
Protein Weight : 45894.2
Protein Length : 428
Protein Note : putative

DNA Sequence : Show Sequence

>NC_005216.1:42-1328 Sin Nombre virus segment S, complete sequence
AATGAGCACCCTCAAAGAAGTGCAAGACAACATCACTCTCCACGAACAACAACTCGTGACTGCCAGGCAG
AAGCTCAAAGATGCAGAAAGAGCGGTGGAATTGGACCCCGATGATGTTAACAAAAGCACATTACAGAGCA
GACGGGCAGCTGTGTCTGCATTGGAGACCAAACTCGGAGAACTTAAGCGGGAACTGGCTGATCTTATTGC
AGCTCAGAAATTGGCTTCAAAACCTGTTGATCCAACAGGGATTGAACCTGATGACCATCTAAAGGAAAAG
TCATCATTGAGATATGGAAATGTCCTTGATGTAAATTCCATTGACTTAGAAGAGCCAAGTGGGCAAACAG
CTGATTGGAAATCCATCGGACTCTACATTCTAAGTTTCGCATTACCGATTATTCTTAAAGCCTTGTACAT
GTTATCTACTAGGGGCCGTCAAACAATCAAAGAAAACAAGGGAACAAGAATTCGATTCAAGGATGATTCA
TCTTATGAAGAAGTCAATGGGATACGTAAGCCAAGACATCTGTATGTTTCTATGCCAACTGCCCAGTCTA
CAATGAAAGCAGATGAGATTACTCCCGGGAGGTTCCGTACAATTGCTTGTGGATTATTCCCAGCCCAAGT
CAAAGCAAGGAATATTATCAGTCCTGTCATGGGTGTGATTGGCTTTAGTTTTTTTGTGAAAGATTGGATG
GAAAGGATTGATGACTTCCTGGCTGCACGTTGCCCATTTCTGCCTGAGCAGAAAGACCCTAGAGATGCTG
CATTGGCAACTAATAGAGCCTATTTTATAACACGTCAATTACAGGTTGATGAGTCAAAGGTTAGTGATAT
TGAGGACCTGATTGCTGATGCAAGGGCTGAGTCTGCCACTATATTCGCAGATATTGCTACTCCTCATTCA
GTTTGGGTCTTTGCATGTGCTCCAGATCGTTGTCCACCTACAGCATTATATGTGGCCGGGATGCCGGAAC
TGGGTGCATTTTTTGCTATTCTCCAGGATATGAGGAACACCATAATGGCATCCAAATCTGTGGGGACATC
TGAAGAGAAATTGAAGAAGAAATCAGCATTCTACCAGTCATACTTAAGACGTACTCAGTCAATGGGAATT
CAACTGGACCAGAAGATAATCATCTTATACATGAGCCATTGGGGAAGAGAGGCCGTGAATCACTTCCATC
TTGGAGATGATATGGATCCTGAGCTTAGGGAACTTGCCCAGACCCTTGTAGATATCAAGGTCAGGGAAAT
CTCTAACCAAGAACCACTTAAACTTTA

Protein Sequence : Show Sequence

>NP_941975.1 nucleocapsid protein [Sin Nombre orthohantavirus]
MSTLKEVQDNITLHEQQLVTARQKLKDAERAVELDPDDVNKSTLQSRRAAVSALETKLGELKRELADLIA
AQKLASKPVDPTGIEPDDHLKEKSSLRYGNVLDVNSIDLEEPSGQTADWKSIGLYILSFALPIILKALYM
LSTRGRQTIKENKGTRIRFKDDSSYEEVNGIRKPRHLYVSMPTAQSTMKADEITPGRFRTIACGLFPAQV
KARNIISPVMGVIGFSFFVKDWMERIDDFLAARCPFLPEQKDPRDAALATNRAYFITRQLQVDESKVSDI
EDLIADARAESATIFADIATPHSVWVFACAPDRCPPTALYVAGMPELGAFFAILQDMRNTIMASKSVGTS
EEKLKKKSAFYQSYLRRTQSMGIQLDQKIIILYMSHWGREAVNHFHLGDDMDPELRELAQTLVDIKVREI
SNQEPLKL

Molecule Role : Protective antigen
Molecule Role Annotation : Study used a deer mouse infection model to test the protective efficacy of genetic vaccine candidates for Sin Nombre (SN) virus that were known to provoke immunological responses in BALB/c mice. Protective epitopes were localized in each of four overlapping cDNA fragments that encoded portions of the SN virus G1 glycoprotein antigen; the nucleocapsid gene (SNVsSgp1) also was protective (Bharadwaj et al., 2002).
Related Vaccine(s): Sin Nombre virus DNA vaccine encoding SNVsSgp1 (NP)

III. Vaccine Related Host Genes

1. Ighg1

Gene Name : Ighg1
Sequence Strain (Species/Organism) : Mus musculus
NCBI Gene ID : 16017
Genbank Accession : AC160982
Molecule Role : Vaximmutor
Related Vaccine(s): Dobrava-Belgrade virus S nucleocapsid protein vaccine

2. Il2

Gene Name : Il2
Sequence Strain (Species/Organism) : Mus musculus
NCBI Gene ID : 16183
NCBI Protein GI : 7110653
Locus Tag : DN-144H19.3
Genbank Accession : AF195954
Protein Accession : NP_032392
Taxonomy ID : 10090
Chromosome No : 3
Gene Starting Position : 37120712
Gene Ending Position : 37125953
Gene Strand (Orientation) : -
Protein Name : interleukin 2
Protein pI : 4.63
Protein Weight : 18506.61
Protein Length : 169
Protein Note : Also known as Il-2

DNA Sequence : Show Sequence

>gi|372099107:37120712-37125953 Mus musculus strain C57BL/6J chromosome 3, GRCm38 C57BL/6J
ATTTTTTTTTAGAGGAGAGCTTTATTTCTTGAAAACACTGATTAACATAGAGTTCACAGGAATAACTGAG
AAGTTATTTTAGCGCTTACTTTGTGCTGTCCTAAAAATGACAGACATCTGAGCTTATTTATATTTGAATC
ATCTAAATACTTTTATTAATGAGTCTACCTACACATGATATTTAACAATTCAATATAATAAATAATTTCA
GATAAATAGTTTAAAACATTTTTGAGCCCTTGGGGCTTACAAAAAGAATCTTTAAAGATCCATATTTATC
ATCTGAAGACTAGTAGTTACAAAAGATAGTAAACAATACATCCAAAAATAAATTAAAGTTAAATATTTAA
ATAAATAGAGAGCCTTATGTGTTGTAAGCAGGAGGTACATAGTTATTGAGGGCTTGTTGAGATGATGCTT
TGACAGAAGGCTATCCATCTCCTCAGAAAGTCCACCACAGTTGCTGACTCATCATCGAATTGGCACTCAA
ATGTGTTGTCAGAGCCCTAAAAAAGAATATAAAATTGTAGGCTAAGGTAGCTTACTTTGCATATAATTAT
TCTTCACGGAAGCTTTTAGGCATTTATTTTAACACTGGTTAAATATATTGAACATTCACATTTCTGAAAT
GATCACAATTATTGGGTTATGTTTTATTATGATATCAAATGGTAAAGCATAAAAAATAAAGCTCTCCAGA
GTTGATTACCAACGAGCATTTGATGGGAACACTGAACACTTCTGTAGAATTCAGTTGCTTGACCACTTTT
TCTCTGGTAAGTGTACACAGCTATGCATCAATTTAGGCTTCAGGAATGGAGAAATGATACTTGTTATATG
TTCTTTGCTAAATCTTAAGAGTCTATTATTCAATTCTTTCCCCCCCAAGTCAGAATGGGAACAACCCAGA
GTAGGTTAGGACAAGCATCTATTGGGGGAAATTTGGAGAATTTTGTTATAGATATACCTACCCTGATACC
AGTGTAGATGAATCACAAAGTTTGTACATGTATAAATATTATAAATTTGTTCTGTGGATTAGCTTTTTGA
AAGCTAACCCTCTTTGCTAAAGGAGATATTTTAATTCTGTTTTTAGGTGACAAGCTGAGAAGGAAATATT
TTGACAAATTTATGGGATGGCTTTTCCAAATCTGACTAATTCCTTGTAAGTCAAATGAGGCAGATTATTC
CTTTTAAATAAATACTCCTTTTCAAATAAGCTGCTGTGGTCTTATATACCCTCTTTACATAGCTACTTCA
ATATTCTAATGTCTTGAATTTTTCATTTTTAAAAATATGTGGCCCTTGAAACATTTTTCTAGTTCTCATG
TAGCTCTAAAGTTTGTACCATTCATTAGGGGTGAACTTTCTGATGTTGGCCATAGATTTTAGAGCATCCA
TCTTTGCATACGTAAGTACTCGATTTTAGAGCATCCATCTTAGCATACGTAAGTACTCACACCTTTCAAC
GTCTAGATGATGTGTATTTCAGCCTGCTTTAAAAACCTCAAATGGAAGCTGGGCAGTGGTGGTGCAGGCC
TTTAATCCCAGCACTTGGGAGGCAGAGGCAGGCGGATTTCTGAGTTCGAGGCCAGCCTGGTCTACAGAGT
GAGTTCCAGGACAGCCAGGGCTACACAGAGAAATCCTGTCTCAAAAAAACAAACAAACAAACAAACAAGA
AACAAACAAACAAAAAAAACAAAAACAAACAAAACAAAACCTCAAATGGAAACAGTGCACTGAAATGTGA
CTTTGGTTTGGAAAGGACTAGCCCACACCCTCTTGGAGGCTTGCTGCTACTGTCTCACCAGAGTCCATGA
TTCTTGTGCATTTAAAAACAAAGCTCTGCAGTGGGCTCTCCCCGTCCTGCTCTGTCCATTTTAATGGCTG
CCATTTTTGGAGAGAAATGTCTGTTTTTCTACCAATACCAGCACAACTTCTCTGGAAAAACTTTTCAGAT
AATTTTTTCTGATCTGATGAATGTAACACCAGCAAGAGTTGCTTGTTTCTTGTGGAATTCTACTCCGTGC
TTTCTCTCACATCCAGTTCTATGCTGGTGTGGAGGGAGCAGAGTGTTCATGTTCCCAGTTTCCTTGCAGG
TGATGGTAGGTGGAAATTCTAGCATCATCCTACAGTGGAAGGATTCACTTGCACAGTGACTTTAAACTTT
GGCTGACTAAATGCACAGAACCCATCAAAGACCAGAAATGGCAAGCCACTTAAAAATGCATTTCTTCTCT
ATTTTATTTCCAGATTAGCAAATAAAGCAACACCTTACCTTTAGTTTTACAACAGTTACTCTGATATTGC
TGATGAAATTCTCAGCATCTTCCAATTGAAAGCTTTTGCTTTGAGTCAAATCCAGAACATGCCGCAGAGG
TCCAAGTTCATCTTCTAGGCACTGAAGATCTTTCAATTCTGTGGCCTAGAGGAGTAATAAGCTTAACCAT
CAGCTCAGCTCACCACATACTGAAGAGCCAGAAAGTTAGTCTGCAGTCTCTCTAATCAAGAGAAGGCAGC
ACATAGCTTTTACCACTCCCTGTCCTCTGTAAAGGGGGACTGAGGTAATCGATGCCATAAATCTAGAAGA
GCAAGAGCTCACAACCATAAGGGTCACCTTGACTGTTAGGCCACTCTAGTGAGCTCTTCTGGCTTCATTA
GACTTTGTAAAAAGTCTGTGTTTCTCTACCAATGCATAGCACAAGTTCAGACTATTGTTCCAATCTACAA
AGAAATCTGCAAGGTTCACATTCTAATATCTAATCGCAGAGTTGAGAATCACAGAAGGGTAAGGGACAGG
AATCCTTGGATGCCAACTGTCATAGGCCTAAATCTTACAGATTAGGTTATCATGGCCCTGACTGAGGGGT
GTCAAGATAGCCAGGAAGACACATGTAATATTTTTTAAATTTATACTTCTCTGCAATTACTAAAGATGAG
TTTTTTAAAAACCCATATGTCAAGCTAGGAAGATGGCTTTTAAGGTAAAGGTACCTGCCACCAAACTTGA
TGATTTGAGTTCAATTGCAGTGACCCACCCACATTGTGGAAAGAATGGCCCAACTTTCATATTTCGTCTT
CTGACCTCCACAGTTACACTGTGATACACATGCACCTCCACACATACATACATACATACATACATACATA
CATACATACATACATACATACATACATAAAATGTAAAATCTATTTGTCCCAATGAAAAAAGTCATAACAT
TTTGGCAGAACACTTGGGAGAATTTTTCTGATCCATGTATGAAATAATGTTTAAATACATCATGCATACT
TACATACATATACAGGGAAAAAGGATATTGTCTAGCTCTATAGTACACTATGATTTCCATAAAGTCAGCT
GCTTTAAGTCTACAGTGAACTGTGTTATCTCCATTTTACCAAGTCTCACAATCAGTGTCTTTCCCACAAC
CACATGAACGGGGAGCAAGGGTGGCCCGGTCGGTCGGTCGCTCCTTTCATTAACCACAGCCTTTGAATTA
CATGCTTCTATTGTTTTTAAAATCTTACATACCGAATGCAGGTTAAATGAAAACATCAGCAGAATTTACC
ACATTACTCTTGTTTTCAATAATTATTTTAAAACTTAGATTTCTAGGAATTTCACCACTCTCTAAATTAA
CATGATCTCTGTTGTTTCTCTAACAATCCTTAGAGGAAGAGGTTTGACAAGATAACACGTAGTTCCTGAT
TGGTCTGCTATTGACCTTCCAACTTGTTATGGTGTGTCTTGGGTATTCAGTGTGGAGCTCCTTCCCTCAC
AGAACTCTCCAGTTTGTTACTGTGGGAGCTGAACATTATGTGACTGTAATTAAGCTGGAAGAGCAACTGG
GGCCTTTCTTTTTCATCTCCTGTAGAACAGCTAGAGGCAAAGTTCCCCTGACTCAATAGGAATGGGTCAA
CAGCTGCCTCTTTGTCCCCTTAGATCCCATTTTGCTCTAGGCTTGGATTTACTTGGGACAAGCTCTTTCT
AGGACCAAAGTTATCCATTCTCTTTTGACTTTCGGGACTAGAGCAAATTTCTACCCTTGCTGGAGTTCTA
AGAGCTTGTGTAAGCTGTTTTGTATCACACAACATTCTGCCTCCAAAGGAGACCAAAAAACCTTTCAATA
TTGAGAGATAGTATCATGTAGTAATATTACAGCTCTCTAGTTCCACTAGTTAAAAGAGACTGTATAAATC
CAAAATTGTTTTATACATCTCGGACTTCTTCTCTAGCACTAATAGTGTGTTTATGTGCTGTTGGCCAGAC
ACTGTTCTGAAACCTGACAAGTGATAAGCATAGCAATGATCATTTATGTAAGTGGGTGAATGAGCATAGA
CAACTTCCTGAGGCCATGAGAGCTAGGCCCTAGCAGGGCTGGACTCCACTGCAGATCATTGGCCCTAGAA
TCTTAACATTGGTTATGCTGCTCCTCAAAAATGCAGTCATAGGAATCATTCTTAAAACAGTAAAGTGTGT
GTAAAATGAGACCCTCTCAAGTCAATTTTTACATTTAAGAAGCTGAAATAATGCACCTCTCTCAAATATC
TGTAGGACAAGCATAAAACAGAAATCAACAGTCTTTAAATTATCCCAGCATCAAAATGCAATCATCTTGC
ATTTTCTTCTCATTCCCTTTCCCAACACATAAATACACAGGAAAAATGTAAAGAAAACCATTTAAAAGAA
GTACCATTGTCAAGATTTTTTTACAGAGAGATATCAGAGAAACTTATTTTCTTCAGATGAGGTACTATTC
ATCTGGTCACATGTTATTTACACGTTATACACATGTATGTTACTTTGAATTCATTAGAGCACCAGTTAAA
CACAGAAACTCACTCACCTGCTTGGGCAAGTAAAATTTGAAGGTGAGCATCCTGGGGAGTTTCAGGTTCC
TGTAATTCTGAGAAAGCGTAACACATTATTATTAAAGGTTATTAAACACAGCCTTTGGCAAGAAAGCTAA
AGGTATTGCCTATAGATGGGATGGCTGTGCACTTACCTCCATCCTGCTCAGGAGCTCCTGTAGGTCCATC
AACAGCTGCTCCAGGTGCTGCTGCTGCTGCTGCTGCTGCTGCTGCTGCTGCTGTGCTTCCGCTGTAGAGC
TTGAAGTGGAGCTTGAAGTGGGTGCGCTGTTGACAAGGAGCACAAGTGTCAATGTGACACAGGATGCGAG
CTGCATGCTGTACATGCCTGCAGGACTTGAGGTCACTGTGAGGAGTGATTAGCAAGGGTGAT

Protein Sequence : Show Sequence

>gi|7110653|ref|NP_032392.1| interleukin-2 precursor [Mus musculus]
MYSMQLASCVTLTLVLLVNSAPTSSSTSSSTAEAQQQQQQQQQQQQHLEQLLMDLQELLSRMENYRNLKL
PRMLTFKFYLPKQATELKDLQCLEDELGPLRHVLDLTQSKSFQLEDAENFISNIRVTVVKLKGSDNTFEC
QFDDESATVVDFLRRWIAFCQSIISTSPQ

Molecule Role : Vaximmutor
Related Vaccine(s): Dobrava-Belgrade virus S nucleocapsid protein vaccine

3. Il4 (interleukin 4)

Gene Name : Il4 (interleukin 4)
Sequence Strain (Species/Organism) : Mus musculus
NCBI Gene ID : 16189
NCBI Protein GI : 10946584
Locus Tag : RP23-188H3.4
Genbank Accession : NM_021283.1
Protein Accession : NM_021283.1
Taxonomy ID : 10090
Gene Strand (Orientation) : ?
Protein Name : IL-4, Interleukin 4

DNA Sequence : Show Sequence

>gi|10946583|ref|NM_021283.1| Mus musculus interleukin 4 (Il4), mRNA
GGATCCCCGGGCAGAGCTGGGGGGGGATTTGTTAGCATCTCTTGATAAACTTAATTGTCTCTCGTCACTG
ACGGCACAGAGCTATTGATGGGTCTCAACCCCCAGCTAGTTGTCATCCTGCTCTTCTTTCTCGAATGTAC
CAGGAGCCATATCCACGGATGCGACAAAAATCACTTGAGAGAGATCATCGGCATTTTGAACGAGGTCACA
GGAGAAGGGACGCCATGCACGGAGATGGATGTGCCAAACGTCCTCACAGCAACGAAGAACACCACAGAGA
GTGAGCTCGTCTGTAGGGCTTCCAAGGTGCTTCGCATATTTTATTTAAAACATGGGAAAACTCCATGCTT
GAAGAAGAACTCTAGTGTTCTCATGGAGCTGCAGAGACTCTTTCGGGCTTTTCGATGCCTGGATTCATCG
ATAAGCTGCACCATGAATGAGTCCAAGTCCACATCACTGAAAGACTTCCTGGAAAGCCTAAAGAGCATCA
TGCAAATGGATTACTCGTAGTACTGAGCCACCATGCTTTAACTTATGAATTTTTAATGGTTTTATTTTAA
TATTTATATATTTATAATTCATAAAATAAAATATTTGTATAATGT

Protein Sequence : Show Sequence

>gi|10946584|ref|NP_067258.1| interleukin 4 [Mus musculus]
MGLNPQLVVILLFFLECTRSHIHGCDKNHLREIIGILNEVTGEGTPCTEMDVPNVLTATKNTTESELVCR
ASKVLRIFYLKHGKTPCLKKNSSVLMELQRLFRAFRCLDSSISCTMNESKSTSLKDFLESLKSIMQMDYS

Molecule Role : Vaximmutor
Molecule Role Annotation : IL-4 plays an important role in Th2 immune response.
Related Vaccine(s): Dobrava-Belgrade virus S nucleocapsid protein vaccine

IV. Vaccine Information

1. Andes virus vaccine using adenovirus viral vector expressing ANDV NP

a. Vaccine Ontology ID:

VO_0011561

b. Type:

Recombinant vector vaccine

c. Status:

Research

d. Antigen

Andes virus nucleocapsid protein (ANDVsSgp1)

e. Gene Engineering of ANDVsSgp1

Type: Recombinant vector construction
Description: Nonreplicating (E1− E3−) Ad vectors expressing ANDV proteins were constructed using the AdMax HI-IQ system (Microbix, Toronto, Canada). The ANDV GN, GC, GPC, and N open reading frames were PCR amplified from plasmids containing the appropriate cDNAs derived from a Chilean ANDV isolate, strain 9717869. AdEmpty was constructed using plasmid pDC316(io) containing no ANDV sequences. These pDC316(io)-based ANDV plasmids were cotransfected with plasmid pBHGloxΔE1,3Cre, containing the remainder of the Ad5 genomic plasmid, into 293 IQ cells. Supernatants were collected after 6 to 8 days, and the presence of Ad vectors expressing the ANDV proteins (designated AdN, AdGN, AdGC, and AdGPC) was confirmed by immunoprecipitation. Ad vectors were plaque purified, propagated in large-scale infections in 293 IQ cells, and purified using standard CsCl gradient methods (Safronetz et al., 2009).
Detailed Gene Information: Click here.

f. Vector:

Nonreplicating adenovirus (Ad) vectors

g. Immunization Route

Intraperitoneal injection (i.p.)

h. Hamster Response

Host Strain: Syrian gold
Vaccination Protocol: Syrian golden hamsters (Mesocricetus auratus) (4- to 6-week-old males; Charles River, Pointe Claire, Canada) were group housed in microisolator units situated in the biosafety level 4 area of the National Microbiology Laboratory, Public Health Agency of Canada. Prior to vaccine experiments, the 50% lethal dose (LD50) for i.p. injections of ANDV in hamsters was established by inoculating anesthetized animals with 0.8 to 80,000 FFU (using 10-fold dilutions) of ANDV. Hamsters were monitored twice daily for clinical signs of illness according to an approved scoring sheet (ruffled fur, lethargy, inappetence, and labored breathing). For vaccination, hamsters were anesthetized with isoflurane, a preimmunization blood sample was collected, and the animals were immunized with the Ad vectors using 108 (293 cell) PFU of each vector diluted in 100 μl phosphate-buffered saline at two sites in the hind-leg musculature (Safronetz et al., 2009).
Challenge Protocol: After 28 days, a second blood sample was collected and hamsters were challenged with ANDV by i.p. injection of 100 LD50s (equivalent to 154 FFU). Hamsters were examined twice daily for signs of illness. Survivors were monitored for 40 to 45 days and then anesthetized and exsanguinated via cardiac puncture (Safronetz et al., 2009).
Efficacy: Nonreplicating adenovirus (Ad) vectors that express Andes hantavirus (ANDV) nucleocapsid protein (AdN, ANDVsSgp1) or glycoproteins (AdG(N) and AdG(C)) were constructed . When administered once, all three Ad vectors, individually or in combination, elicited a robust immune response that protected Syrian hamsters from a lethal ANDV infection that mimics the pulmonary disease seen in humans. No vaccinated animal died, and there were no obvious clinical signs of disease (Safronetz et al., 2009).

2. Chimaeric HBV core particles carrying Puumala virus nucleocapsid protein fragments

a. Vaccine Ontology ID:

VO_0004120

b. Type:

Recombinant vector vaccine

c. Antigen

The Puumala virus nucleocapsid protein (Ulrich et al., 1998).

d. Preparation

Chimaeric hepatitis B virus (HBV) core particles were constructed to carry defined fragments of the Puumala virus nucleocapsid protein. Puumala is a European hantavirus (Ulrich et al., 1998).

3. DNA vaccine encoding ANDV glycoproteins

a. Vaccine Ontology ID:

VO_0004119

b. Type:

DNA vaccine

c. Antigen

Both the G1 and G2 glycoproteins are expressed (Custer et al., 2003).

d. Gene Engineering of Hanta_G2

Type: Recombinant protein preparation
Description: The Hantavirus G2 glycoprotein was expressed in the DNA vaccine (Custer et al., 2003).
Detailed Gene Information: Click here.

e. Vector:

pWRG/HTN-M(x), a customized plasmid built from pWRG7077 (Custer et al., 2003)

f. Preparation

An expression plasmid, pWRG/AND-M, was constructed that contains the full-length M genome segment of Andes virus (ANDV), a South American hantavirus (Custer et al., 2003).

g. Macaque Response

Vaccine Immune Response Type: VO_0000286
Efficacy: Rhesus macaques vaccinated by gene gun with pWRG/AND-M developed remarkably high levels of neutralizing antibodies that not only neutralized ANDV (a South American hantavirus) but also cross-neutralized other HPS-associated hantaviruses, including Sin Nombre virus (Custer et al., 2003).

4. Dobrava-Belgrade virus S nucleocapsid protein vaccine

a. Vaccine Ontology ID:

VO_0011512

b. Type:

Subunit vaccine

c. Status:

Research

d. Antigen

Dobrava-Belgrade virus S nucleocapsid protein

e. Gene Engineering of S nucleocapsid protein from Dobrava-Belgrade virus

Type: Recombinant protein preparation
Description: His-tagged rDOBV N and recombinant mouse dihydrofolate reductase (rDHFR) was prepared. Freund’s complete adjuvant (FCA) or incomplete (FIA) adjuvant (Sigma, St. Louis, MO) or PBS. A total of 50 μg of recombinant protein was mixed with Alum and FCA/FIA, according to the manufacturers’ descriptions, or PBS, in a total volume of 200 μl per dosage (Klingstrom et al., 2004).
Detailed Gene Information: Click here.

f. Adjuvant:

VO ID: VO_0000133

g. Immunization Route

Subcutaneous injection

h. Mouse Response

Host Strain: C57/BL6
Vaccination Protocol: A total of 50 μg of recombinant protein was mixed with Alum and FCA/FIA, according to the manufacturers’ descriptions, or PBS, in a total volume of 200 μl per dosage. All immunizations and boosters were administered subcutaneously. rDOBV N or rDHFR in Alum, FCA or PBS were administered at day 0. At days 21 and 92, mice were boosted with rDOBV N or rDHFR in Alum, FIA or PBS (Klingstrom et al., 2004).
Challenge Protocol: At day 118, all mice were challenged with 10 mouse ID50 of DOBV and 21 days later, all mice were sacrificed. Serum, plasma, and EDTA-blood were drawn at the time points indicated below (Klingstrom et al., 2004).
Efficacy: Study compared the immunogenicity and protective efficacy of recombinant DOBV nucleocapsid protein (rDOBV N, S) given with Alum or Freund's as adjuvant, or PBS, in C57/BL6 mice. Mice receiving rDOBV N with Freund's adjuvant were protected from challenge (75% protected) (Klingstrom et al., 2004).
Host Gene Response of Il2
- Gene Response: Significantly higher levels of IL-2 producing cells were found in the group given rDOBV N with Alum as compared to the rDHFR vaccinated groups and PBS vaccinated groups. These results were found in mice peripheral blood mononuclear cells (PBMCs) 113 days after vaccination (Klingstrom et al., 2004).
- Detailed Gene Information: Click here.
Host Gene Response of Il4 (interleukin 4)
- Gene Response: Significantly higher numbers of IL-4 producing cells were found in the groups given rDOBV N with Freund’s adjuvant as compared to rDOBV N with PBS. These reactions were found in peripheral blood mononuclear cells (PBMCs) 113 days after vaccination (Klingstrom et al., 2004).
- Detailed Gene Information: Click here.

5. Hantavirus DNA vaccine pWRG/HTN-M

a. Vaccine Ontology ID:

VO_0004576

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Hamsters

e. Gene Engineering of GP

Type: DNA vaccine construction
Description:
Detailed Gene Information: Click here.

f. Vector:

pWRG7077 (Hooper et al., 2001)

g. Immunization Route

Gene gun

h. Hamster Response

Vaccine Immune Response Type: VO_0003057
Efficacy: All of the hamsters that were vaccinated with pWRG/HTN-M were protected against infection as defined by an absence of a postchallenge N-specific antibody response. In addition, the pre- and postchallenge PRNT titers differed by ≤4-fold. In contrast, all of the negative control hamsters, whether they were vaccinated with pWRG7077 or remained unvaccinated, were infected, as evidenced by the development of N-specific antibodies and neutralizing antibodies postchallenge (Hooper et al., 2001).

6. Puumala virus PUUVsSgp1 (NP) protein vaccine

a. Vaccine Ontology ID:

VO_0011400

b. Type:

Subunit vaccine

c. Status:

Research

d. Antigen

Puumala virus nucleocapsid protein (PUUVsSgp1)

e. Gene Engineering of PUUVsSgp1

Type: Recombinant protein preparation
Description: N proteins from PUUV, TOPV, ANDV, and DOBV, carrying a polyhistidine tag to facilitate protein purification, were produced in Escherichia coli cells. The N ORF of PUUV, TOPV, and DOBV were cloned into the pQE-32 vector (Qiagen, Hilden, Germany), while the N ORF of ANDV was cloned into pRSET (R&D Systems Europe, Oxford, United Kingdom). An irrelevant protein, mouse dihydrofolate reductase (DHFR) (Qiagen), provided by the manufacturer and expressed in the same system, was used as a negative control protein (de et al., 2002).
Detailed Gene Information: Click here.

f. Adjuvant:

VO ID: VO_0000139

g. Adjuvant:

VO ID: VO_0000142

h. Bank vole Response

Vaccination Protocol: To asses the immunogenicity and protective capacity of the expressed rN proteins, 4- to 10-week-old bank voles, derived from a PUUV-free colony established with animals captured in Sweden, were immunized with purified PUUV, TOPV, ANDV, or DOBV rN or DHFR control protein. Bank voles were immunized three times with 50 μg of protein at intervals of 3 weeks. The animals were injected with protein emulsified in Freund's complete adjuvant, incomplete Freund's adjuvant, and phosphate-buffered saline (PBS), respectively (de et al., 2002).
Challenge Protocol: Bank voles were challenged subcutaneously 2 weeks after the last immunization with approximately 20 50% infective doses of wild-type PUUV (strain Kazan) (de et al., 2002).
Efficacy: To investigate the ability of recombinant N (rN, nucleocapsid proteins) from different hantaviruses to elicit cross-protection, we immunized bank voles with rN from Puumala (PUUV), Topografov (TOPV), Andes (ANDV), and Dobrava (DOBV) viruses and subsequently challenged them with PUUV. All animals immunized with PUUV (PUUVsSgp1) and TOPV rN were completely protected (de et al., 2002).

7. Recombinant DOBV nucleocapsid protein (rDOBV N)

a. Vaccine Ontology ID:

VO_0004102

b. Type:

Subunit vaccine

c. Gene Engineering of S nucleocapsid protein from Dobrava-Belgrade virus

Type: Recombinant protein preparation
Description: Dobrava hantavirus nucleocapsid protein was used for the vaccine development (Klingstrom et al., 2004).
Detailed Gene Information: Click here.

d. Adjuvant:

VO ID: VO_0000133
Description: Dobrava hantavirus nucleocapsid protein in Freund's adjuvant (Klingstrom et al., 2004).

e. Preparation

Recombinant DOBV nucleocapsid protein (rDOBV N) given with Alum or Freund's as adjuvant (Klingstrom et al., 2004).

f. Virulence

Not virulent

g. Description

Dobrava hantavirus (DOBV) causes a severe form of hemorrhagic fever with renal syndrome (HFRS). Currently there is no therapy or vaccine available for HFRS (Klingstrom et al., 2004).

h. Mouse Response

Host Strain: C57/BL6 mice
Vaccination Protocol: His-tagged rDOBV N and recombinant mouse dihydrofolate reductase (rDHFR) was emulsified with Imject® Alum (Pierce, Rockford, IL), Freund’s complete adjuvant (FCA) or incomplete (FIA) adjuvant (Sigma, St. Louis, MO) or PBS. A total of 50 μg of recombinant protein was mixed with Alum and FCA/FIA, or PBS, in a total volume of 200 μl per dosage. All immunizations and boosters were administered subcutaneously. rDOBV N or rDHFR in Alum, FCA or PBS were administered at day 0. At days 21 and 92, mice were boosted with rDOBV N or rDHFR in Alum, FIA or PBS. At day 118, all mice were challenged with 10 mouse ID50 of DOBV and 21 days later, all mice were sacrificed. Serum, plasma, and EDTA-blood were drawn at the time points indicated below (Klingstrom et al., 2004).
Side Effects: No side effects.
Challenge Protocol: All vaccinated mice were subcutaneously challenged with 10 mouse ID50 DOBV at day 118. The challenge did not kill mice. Three weeks after challenge, the mice were sacrificed and serum tested for the presence of neutralizing antibodies (Klingstrom et al., 2004).
Efficacy: The immunogenicity and protective efficacy of recombinant DOBV nucleocapsid protein (rDOBV N) given with Alum or Freund’s as adjuvant, or PBS, in C57/BL6 mice, were compared. All mice given Alum or Freund’s seroconverted as did 6/8 mice given rDOBV N with PBS. Reciprocal geometric mean total IgG-titers were 5380, 18,100, and 800, respectively, while the mean IgG1/IgG2a ratios were 17.5, 9.25, and 12, respectively. Furthermore, ELIspot assays showed higher levels of IL-4 producing peripheral blood mononuclear cells (PBMCs) in the group given Alum as compared to the other groups. Interestingly, only mice receiving rDOBV N with Freund’s adjuvant were protected from challenge (75% protected), indicating that the strong Th2-type of immune response induced by Alum against rDOBV N did not induce protection in mice (Klingstrom et al., 2004).

8. rVSVΔG-ANDV-GPC

a. Vaccine Ontology ID:

VO_0004662

b. Type:

Recombinant vector vaccine

c. Status:

Research

d. Host Species for Licensed Use:

Baboon

e. Immunization Route

Intramuscular injection (i.m.)

f. Hamster Response

Vaccination Protocol: Syrian hamsters were immunized with a single injection of VSVΔG/ANDVGPC (Brown et al., 2011).
Vaccine Immune Response Type: VO_0000287
Challenge Protocol: After immunization, the hamsters were challenged at 28, 14, 7, or 3 days postimmunization with a lethal dose of ANDV (Brown et al., 2011).
Efficacy: The hamsters were fully protected against the disease; however, the mechanism of protection seems to differ depending on when the immunization occurs. Administration of the vaccine at 7 or 3 days before challenge also resulted in full protection but with no specific neutralizing humoral immune response, suggesting a possible role of innate responses in protection against challenge virus replication. Administration of the vaccine 24 h postchallenge was successful in protecting 90% of hamsters and again suggested the induction of a potent antiviral state by the recombinant vector as a potential mechanism (Brown et al., 2011).

9. Seoul virus vaccine rCAV-2-Gc

a. Vaccine Ontology ID:

VO_0011402

b. Type:

Recombinant vector vaccine

c. Status:

Research

d. Antigen

Seoul virus Gc glycoprotein

e. Gene Engineering of Gc

Type: Recombinant protein preparation
Description: The Gc protein expressed by rCAV-2-Gc in MDCK cells was evaluated by a SEOV-specific indirect IFA. MDCK cells grown on 15 mm glass coverslips in 12-well culture plates were infected with rCAV-2-Gc or CAV-2 at an m.o.i. of 20. After 48 h infection, the coverslips were rinsed once with PBS (pH 7.4), fixed with acetone for 10 min at room temperature and then reacted with rabbit anti-SEOV polyclonal antiserum and washed three times with PBS. The fixed monolayers were incubated at 37 °C for 30 min in a moist chamber with donkey anti-rabbit IgG labelled with fluorescence isothiocyanate (Amersham). The coverslips were rinsed three times with PBS. Cell monolayers that bound the antibody were covered with glycerine and examined for specific fluorescence under a Zeiss Axioplan fluorescence microscope. Expression of SEOV Gc in MDCK cells infected with rCAV-2-Gc was identified by Western blotting. rCAV-2-Gc-infected cell lysates were separated by 12 % SDS-PAGE, and the proteins transferred to nitrocellulose membrane (Pall Corporation) and probed with positive serum against SEOV and HRP-labelled goat anti-mouse IgG antibody (Sigma) (Yuan et al., 2010).
Detailed Gene Information: Click here.

f. Vector:

replication-competent recombinant canine adenovirus type 2

g. Immunization Route

Intramuscular injection (i.m.)

h. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Mice were randomly assigned to four experimental groups (20 mice per group). Group I was intramuscularly inoculated once with 0.1 ml rCAV-2-Gc (108.0 p.f.u. ml–1); group II received 0.1 ml CAV-2 (108.2 p.f.u. ml–1) intramuscularly as a negative control; group III were inoculated intramuscularly with one dose of HFRS bivalent purified vaccine Youerjian (0.5 ml per dose; GuangDong HongMing Biological Science and Technology Co.) as a positive-control; and group IV were injected with 0.1 ml PBS as a negative control (Yuan et al., 2010).
Challenge Protocol: At the end of the trial, all mice were injected intramuscularly with SEOV strain CC-2 diluted in 0.2 ml PBS. The challenge dose for each virus was 2000 p.f.u. This dose is 1000 50 % infective doses for SEOV. At 14 days after challenge, the mice were sacrificed by CO2 asphyxiation, as approved by the China Small Animal Protection Association. Pre- and post-challenge sera were evaluated for the presence of N-specific antibodies by ELISA and for the presence of neutralizing antibodies by FRNT. Detecting post-challenge N-specific antibody indicated that the mice were infected with the challenge virus (Yuan et al., 2010).
Efficacy: eplication-competent recombinant canine adenovirus type 2 expressing the Gc protein of SEOV (rCAV-2-Gc) in BALB/c mice induced complete protection against a intensive infectious challenge with ~1,000 50% infective doses (ID50) for SEOV strain CC-2 (Yuan et al., 2010).

10. Sin Nombre virus DNA vaccine encoding G1

a. Vaccine Ontology ID:

VO_0011401

b. Type:

DNA vaccine

c. Status:

Research

d. Antigen

Sin Nombre virus envelope glycoprotein G1

e. Gene Engineering of G1

Type: DNA vaccine construction
Description: Cloned the G1 and G2 glycoprotein genes of SN virus strain CC107 into the CMV expression vector pCMVi (-H3) UBs (Bharadwaj et al., 1999 ). The M segment fragments 3' of the first fragment were prepared in a similar manner, such that each expression construct shared 100 nt of sequence at the 5' end with the 3' end of the fragment that preceded it. The coordinates of each of the ten glycoprotein fragments, designated M-CMV-A thorough -I. We also cloned the entire SN virus N gene in a single fragment in a separate expression construct. The same viral cDNA fragments were cloned into bacterial expression vectors to allow bacterial synthesis of the cognate antigens as fusion proteins, as described (Bharadwaj et al., 1997 ; Yamada et al., 1995 ) (Bharadwaj et al., 2002).
Detailed Gene Information: Click here.

f. Vector:

CMV expression vector pCMVi (-H3) UBs

g. Mouse Response

Vaccination Protocol: We purified plasmid DNA with an endotoxin-free kit (EndoFree, Qiagen), and dissolved DNA to a concentration of 1 mg/ml in 0·9% NaCl. Five to twelve mice were immunized with each construct three times at 4 week intervals, using 50 µg of plasmid into each set of quadriceps muscles for a total of 100 µg. No adjuvants were used (Bharadwaj et al., 2002).
Challenge Protocol: Challenged the mice in the challenge replicate with 5 ID50 of SN77734 by the i.m. route 2 weeks after the third vaccination, a dose that corresponds roughly to 50–200 focus-forming units (Bharadwaj et al., 2002).
Efficacy: Study used a deer mouse infection model to test the protective efficacy of genetic vaccine candidates for Sin Nombre (SN) virus that were known to provoke immunological responses in BALB/c mice. Protective epitopes were localized in each of four overlapping cDNA fragments that encoded portions of the SN virus G1 glycoprotein antigen; the nucleocapsid gene also was protective (Bharadwaj et al., 2002).

11. Sin Nombre virus DNA vaccine encoding SNVsSgp1 (NP)

a. Vaccine Ontology ID:

VO_0011543

b. Type:

DNA vaccine

c. Status:

Research

d. Antigen

Sin nombre virus nucleocapsid protein (SNVsSgp1)

e. Gene Engineering of SNVsSgp1

Type: DNA vaccine construction
Description: Cloned the G1 and G2 glycoprotein genes of SN virus strain CC107 into the CMV expression vector pCMVi (-H3) UBs (Bharadwaj et al., 1999 ). The M segment fragments 3' of the first fragment were prepared in a similar manner, such that each expression construct shared 100 nt of sequence at the 5' end with the 3' end of the fragment that preceded it. The coordinates of each of the ten glycoprotein fragments, designated M-CMV-A thorough -I. We also cloned the entire SN virus N gene in a single fragment in a separate expression construct. The same viral cDNA fragments were cloned into bacterial expression vectors to allow bacterial synthesis of the cognate antigens as fusion proteins (Bharadwaj et al., 2002).
Detailed Gene Information: Click here.

f. Vector:

CMV expression vector pCMVi (-H3) UBs

g. Mouse Response

Vaccination Protocol: Five to twelve mice were immunized with each plasmid DNA construct three times at 4 week intervals, using 50 µg of plasmid into each set of quadriceps muscles for a total of 100 µg. No adjuvants were used (Bharadwaj et al., 2002).
Challenge Protocol: Challenged the mice in the challenge replicate with 5 ID50 of SN77734 by the i.m. route 2 weeks after the third vaccination, a dose that corresponds roughly to 50–200 focus-forming units (Bharadwaj et al., 2002).
Efficacy: Study used a deer mouse infection model to test the protective efficacy of genetic vaccine candidates for Sin Nombre (SN) virus that were known to provoke immunological responses in BALB/c mice. Protective epitopes were localized in each of four overlapping cDNA fragments that encoded portions of the SN virus G1 glycoprotein antigen; the nucleocapsid gene (SNVsSgp1) also was protective (Bharadwaj et al., 2002).

12. Topografov virus N protein vaccine

a. Vaccine Ontology ID:

VO_0011403

b. Type:

Subunit vaccine

c. Status:

Research

d. Antigen

Topografov virus Nucleocapsid protein

e. Gene Engineering of S segment

Type: Recombinant protein preparation
Description:
Detailed Gene Information: Click here.

f. Adjuvant:

VO ID: VO_0000139

g. Adjuvant:

VO ID: VO_0000142

h. Bank vole Response

Vaccination Protocol: To asses the immunogenicity and protective capacity of the expressed rN proteins, 4- to 10-week-old bank voles, derived from a PUUV-free colony established with animals captured in Sweden, were immunized with purified PUUV, TOPV, ANDV, or DOBV rN or DHFR control protein. Bank voles were immunized three times with 50 μg of protein at intervals of 3 weeks. The animals were injected with protein emulsified in Freund's complete adjuvant, incomplete Freund's adjuvant, and phosphate-buffered saline (PBS), respectively (de et al., 2002).
Challenge Protocol: Bank voles were challenged subcutaneously 2 weeks after the last immunization with approximately 20 50% infective doses of wild-type PUUV (strain Kazan) (de et al., 2002).
Efficacy: To investigate the ability of recombinant N (rN, nucleocapsid proteins) from different hantaviruses to elicit cross-protection, we immunized bank voles with rN from Puumala (PUUV), Topografov (TOPV), Andes (ANDV), and Dobrava (DOBV) viruses and subsequently challenged them with PUUV. All animals immunized with PUUV and TOPV rN were completely protected (de et al., 2002).

13. Truncated Recombinant DoBV Nucleocapsid Proteins (Dob-N rNp)

a. Vaccine Ontology ID:

VO_0004103

b. Type:

Subunit vaccine

c. Adjuvant:

VO ID: VO_0001241
Description: Dob-N rNp was emulsified in 2% Alhydrogel (Accurate Chemical & Scientific Corp., Westbury, N.Y., USA). The rNp P40-Dob-N, P40-Dob118 and P40p-Dob118 were administered in sterile PBS (Maes et al., 2006).

d. Preparation

DOBV RNA (strain DOB-90/5) was extracted from infected Vero E6 cells (CRL 1586; ATCC, USA). The genomic RNA of DOBV was reverse-transcribed and PCR amplified in order to generate the entire S-segment, using following oligonucleotide primers: 5'-GCGAATTCGCAACACTAGAGGAACTCCAAAAGG-3' and 5'-CGAAGCTTAGTGGTGGTGGTGGTGGTGAAGTTTGAGCGGCTCC-3'. The amino-terminal part encoding the first 118 amino acids was generated using 5'-CTGGCGCCTAACCGACGTGGTGGTGGTGGTGGTGATTCGAAGC-3' as reverse primer. In all constructs, a histidine (His) tag was introduced at the C-terminal end. PCR fragments were cloned in plasmids pTEX(rP40) or pTEXmp18 respectively with or without the inclusion of the P40 sequence in the construct. Following transformation of the E. coli ICONE 200 strain, the recombinant proteins were produced as intracellular inclusion bodies, recovered, and renatured. The recombinant proteins were purifi ed by metal chelate affinity chromatography using a HisTrap kit (Pharmacia, Puurs, Belgium). Using this protocol, four DOBV rNp constructs were expressed and purified. The complete nucleocapsid protein of DOBV was expressed with or without the addition of the rP40 protein (constructs P40-Dob-N and Dob-N). The amino-terminal part of the DOBV nucleocapsid protein was expressed with the addition of the rP40 protein (construct P40-Dob118) or with the addition of only the periplasmic part of the rP40 protein (construct P40p-Dob118) (Maes et al., 2006).

e. Virulence

Not virulent

f. Mouse Response

Host Strain: NMRI mice (Elevage Janvier, Le Genest Saint Isle, France)
Vaccination Protocol: Groups of ten 6-week-old NMRI mice (Elevage Janvier, Le Genest Saint Isle, France) were immunized three times subcutaneously with three different concentrations (0.2, 2 and 10 ug) of rNp with intervals of 2 weeks. The animals were injected with Dob-N rNp emulsifi ed in 2% Alhydrogel (Accurate Chemical & Scientific Corp., Westbury, N.Y., USA). The rNp P40-Dob-N, P40-Dob118 and P40p-Dob118 were administered in sterile PBS. Blood was drawn 14 days after each immunization (Maes et al., 2006).
Persistence: N/A
Side Effects: None.
Challenge Protocol: Groups of 10 NMRI mice were immunized three times subcutaneously with 10 ug of the different constructs with intervals of 2 weeks, and were challenged intraperitoneal with DOBV 2 weeks after the last immunization. Three weeks later, all mice were sacrificed and serum samples were collected. Mice immunized three times subcutaneously with 10 ug of rP40 were used as a control group (Maes et al., 2006).
Efficacy: All recombinant proteins were found to be highly immunogenic after three immunizations of rNp. The immunizations resulted in the induction of a strong Np-specific IgG response with a predominance of IgG1 over IgG2b and IgG2a, suggesting a mixed Th1/Th2 cell involvement. A specific IgG3 response could not be detected. Mice immunized with recombinant DOBV rNp without rP40 showed lower nucleocapsid-specific antibody responses in comparison with the rP40-conjugated constructs, but all mice were found to be protected against DOBV challenge. The results indicate that the rNp constructs coupled to rP40, represent promising vaccine candidates (Maes et al., 2006).

V. References

1. Bharadwaj et al., 2002: Bharadwaj M, Mirowsky K, Ye C, Botten J, Masten B, Yee J, Lyons CR, Hjelle B. Genetic vaccines protect against Sin Nombre hantavirus challenge in the deer mouse (Peromyscus maniculatus). The Journal of general virology. 2002; 83(Pt 7); 1745-1751. [PubMed: 12075094].

2. Brown et al., 2011: Brown KS, Safronetz D, Marzi A, Ebihara H, Feldmann H. Vesicular stomatitis virus-based vaccine protects hamsters against lethal challenge with Andes virus. Journal of virology. 2011; 85(23); 12781-12791. [PubMed: 21917979].

3. CDC Hantaviruses: CDC Hantaviruses [http://0-www.cdc.gov.mill1.sjlibrary.org/ncidod/diseases/hanta/hps/]

4. Chu et al., 1995: Chu YK, Jennings GB, Schmaljohn CS. A vaccinia virus-vectored Hantaan virus vaccine protects hamsters from challenge with Hantaan and Seoul viruses but not Puumala virus. Journal of virology. 1995; 69(10); 6417-6423. [PubMed: 7666542].

5. Custer et al., 2003: Custer DM, Thompson E, Schmaljohn CS, Ksiazek TG, Hooper JW. Active and passive vaccination against hantavirus pulmonary syndrome with Andes virus M genome segment-based DNA vaccine. Journal of virology. 2003 Sep; 77(18); 9894-905. [PubMed: 12941899 ].

6. de Carvalho Nicacio et al., 2002: de Carvalho Nicacio C, Gonzalez Della Valle M, Padula P, Bjorling E, Plyusnin A, Lundkvist A. Cross-protection against challenge with Puumala virus after immunization with nucleocapsid proteins from different hantaviruses. Journal of virology. 2002 Jul; 76(13); 6669-77. [PubMed: 12050380].

7. Hooper et al., 2001: Hooper JW, Custer DM, Thompson E, Schmaljohn CS. DNA vaccination with the Hantaan virus M gene protects Hamsters against three of four HFRS hantaviruses and elicits a high-titer neutralizing antibody response in Rhesus monkeys. Journal of virology. 2001; 75(18); 8469-8477. [PubMed: 11507192].

8. Klingstrom et al., 2004: Klingstrom J, Maljkovic I, Zuber B, Rollman E, Kjerrstrom A, Lundkvist A. Vaccination of C57/BL6 mice with Dobrava hantavirus nucleocapsid protein in Freund's adjuvant induced partial protection against challenge. Vaccine. 2004 Sep 28; 22(29-30); 4029-34. [PubMed: 15364453].

9. Kruger et al., 2001: Kruger DH, Ulrich R, Lundkvist A A. Hantavirus infections and their prevention. Microbes and infection / Institut Pasteur. 2001 Nov; 3(13); 1129-44. [PubMed: 11709294 ].

10. Lednicky, 2003: Lednicky JA. Hantaviruses. a short review. Archives of pathology & laboratory medicine. 2003 Jan; 127(1); 30-5. [PubMed: 12521363].

11. Maes et al., 2006: Maes P, Keyaerts E, Bonnet V, Clement J, Avsic-Zupanc T, Robert A, Van Ranst M. Truncated recombinant Dobrava hantavirus nucleocapsid proteins induce strong, long-lasting immune responses in mice. Intervirology. 2006; 49(5); 253-60. [PubMed: 16714853].

12. Safronetz et al., 2009: Safronetz D, Hegde NR, Ebihara H, Denton M, Kobinger GP, St Jeor S, Feldmann H, Johnson DC. Adenovirus vectors expressing hantavirus proteins protect hamsters against lethal challenge with andes virus. Journal of virology. 2009; 83(14); 7285-7295. [PubMed: 19403663].

13. Ulrich et al., 1998: Ulrich R, Lundkvist A, Meisel H, Koletzki D, Sjolander KB, Gelderblom HR, Borisova G, Schnitzler P, Darai G, Kruger DH. Chimaeric HBV core particles carrying a defined segment of Puumala hantavirus nucleocapsid protein evoke protective immunity in an animal model. Vaccine. 1998 Jan-Feb; 16(2-3); 272-80. [PubMed: 9607042 ].

14. Yoshimatsu et al., 1993: Yoshimatsu K, Yoo YC, Yoshida R, Ishihara C, Azuma I, Arikawa J. Protective immunity of Hantaan virus nucleocapsid and envelope protein studied using baculovirus-expressed proteins. Archives of virology. 1993; 130(3-4); 365-376. [PubMed: 8517793].

15. Yuan et al., 2010: Yuan ZG, Luo SJ, Xu HJ, Wang XH, Li J, Yuan LG, He LT, Zhang XX. Generation of E3-deleted canine adenovirus type 2 expressing the Gc glycoprotein of Seoul virus by gene insertion of deletion of related terminal region sequences. The Journal of general virology. 2010; ; . [PubMed: 20181748].