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Vaccine Comparison

Dobrava-Belgrade virus S nucleocapsid protein vaccine Recombinant DOBV nucleocapsid protein (rDOBV N) Seoul virus vaccine rCAV-2-Gc Sin Nombre virus DNA vaccine encoding G1 Sin Nombre virus DNA vaccine encoding SNVsSgp1 (NP) Truncated Recombinant DoBV Nucleocapsid Proteins (Dob-N rNp)
Vaccine Information Vaccine Information Vaccine Information Vaccine Information Vaccine Information Vaccine Information
  • Vaccine Ontology ID: VO_0011512
  • Type: Subunit vaccine
  • Status: Research
  • Antigen: Dobrava-Belgrade virus S nucleocapsid protein
  • S nucleocapsid protein from Dobrava-Belgrade virus gene engineering:
    • 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.
  • Adjuvant: Freunds emulsified oil adjuvant
  • Immunization Route: Subcutaneous injection
  • Vaccine Ontology ID: VO_0004102
  • Type: Subunit vaccine
  • S nucleocapsid protein from Dobrava-Belgrade virus gene engineering:
    • Type: Recombinant protein preparation
    • Description: Dobrava hantavirus nucleocapsid protein was used for the vaccine development (Klingstrom et al., 2004).
    • Detailed Gene Information: Click Here.
  • Adjuvant: Freunds emulsified oil adjuvant
    • VO ID: VO_0000133
    • Description: Dobrava hantavirus nucleocapsid protein in Freund's adjuvant (Klingstrom et al., 2004).
  • Preparation: Recombinant DOBV nucleocapsid protein (rDOBV N) given with Alum or Freund's as adjuvant (Klingstrom et al., 2004).
  • Virulence: Not virulent
  • 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).
  • Vaccine Ontology ID: VO_0011402
  • Type: Recombinant vector vaccine
  • Status: Research
  • Antigen: Seoul virus Gc glycoprotein
  • Gc gene engineering:
    • 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.
  • Vector: replication-competent recombinant canine adenovirus type 2
  • Immunization Route: Intramuscular injection (i.m.)
  • Vaccine Ontology ID: VO_0011401
  • Type: DNA vaccine
  • Status: Research
  • Antigen: Sin Nombre virus envelope glycoprotein G1
  • G1 gene engineering:
    • 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.
  • Vector: CMV expression vector pCMVi (-H3) UBs
  • Vaccine Ontology ID: VO_0011543
  • Type: DNA vaccine
  • Status: Research
  • Antigen: Sin nombre virus nucleocapsid protein (SNVsSgp1)
  • SNVsSgp1 gene engineering:
    • 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.
  • Vector: CMV expression vector pCMVi (-H3) UBs
  • Vaccine Ontology ID: VO_0004103
  • Type: Subunit vaccine
  • Adjuvant: Alhydrogel
    • 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).
  • 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).
  • Virulence: Not virulent
Host Response Host Response Host Response Host Response Host Response Host Response

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 Il2 response
    • Description: 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 Il4 (interleukin 4) response
    • Description: 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.

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).

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).

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).

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).

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).
References References References References References References
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].
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].
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].
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].
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].
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].