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Vaccine Comparison

Lassa fever virus recombinant vector vaccine YFV17D/LASV-GPC encoding the glycoprotein precursor Lassa Virus Nucleoprotein Subunit Vaccine ML29 V-LSGPC YFV17D/LAS-GPC
Vaccine Information Vaccine Information Vaccine Information Vaccine Information Vaccine Information
  • Vaccine Ontology ID: VO_0004380
  • Type: Recombinant vector vaccine
  • Status: Research
  • Host Species as Laboratory Animal Model: Guinea pig
  • GPC gene engineering:
    • Type: Recombinant vector construction
    • Description: Vector Yellow Fever Vaccine 17D (YFV17D) expressed the Lassa virus glycoprotein precursor (LASV-GPC) (Bredenbeek et al., 2006).
    • Detailed Gene Information: Click Here.
  • Vector: Yellow Fever Vaccine 17D (YFV17D) (Bredenbeek et al., 2006)
  • Immunization Route: Intramuscular injection (i.m.)
  • Vaccine Ontology ID: VO_0011441
  • Type: Recombinant vector vaccine
  • Status: Research
  • LASVsSgp1 nucleoprotein gene engineering:
    • Type: Recombinant vector construction
    • Detailed Gene Information: Click Here.
  • Vector: Vaccinia virus (Morrison et al., 1989).
  • Immunization Route: Intradermal injection (i.d.)
  • Vaccine Ontology ID: VO_0004085
  • Type: Attenuated
  • Preparation: Clone ML29 is selected and triple plaque purified. The viruses are grown on Vero E6 cells cultured in Dulbecco's modified minimum Eagle's medium with 2% fetal calf serum, 1% penicillin-streptomycin, and l-glutamine (2 mM) at 37°C in 5% CO2. Cells and virus stocks were free of mycoplasma contamination (Lukashevich et al., 2005).
  • Description: ML29 is a clone that has been isolated from a Mopeia virus (MOPV) and Lassa virus (LASV) reassortant. It contains the L RNA from MOPV and the S RNA segment from LASV (Lukashevich et al., 2005).
  • Vaccine Ontology ID: VO_0004086
  • Type: Vaccina virus
  • LASVsSgp2 glycoprotein gene engineering:
    • Type: Protein
    • Detailed Gene Information: Click Here.
  • Preparation: The Lassa virus GPC gene was assembled and ligated into the unique SmaI site of the vaccinia virus expression vector pSCl1. The products of this reaction were transfected into competent Escherichia coli MC1061 cells, and a transformant containing the proper orientation of the Lassa GPC gene was identified by restriction enzyme digestion of plasmid DNA and confirmed by nucleotide sequence analysis (Auperin et al., 1988).
  • Description: V-LSGPC is a cloned cDNA containing the complete glycoprotein gene of the Josiah strain of Lassa virus was inserted into the thymidine kinase (TK) gene of the New York Board of Health (WYETH) strain of vaccinia virus (Auperin et al., 1988).
  • Vaccine Ontology ID: VO_0004088
  • Type: Live Attenuated
  • Preparation: The YFV17D/LASV-GPC plasmid is constructed in the background of the full-length YFV17D cDNA clone by fusion PCR mutagenesis. The LASV-GPC gene of the AV strain is amplified by RT/PCR and cloned into pcDNA. The nucleotide sequences of PCR-derived DNA fragments and gene fusions are confirmed by sequencing. The recombinant YFV17D/LASV-GPC plasmid is linearized by XhoI and used for in vitro RNA transcription (Bredenbeek et al., 2006).
  • Virulence:
  • Description: YFV17D/LAS-GPC is a Yellow Fever Vaccine 17D (YFV17D) that has been used as a vector for the Lassa virus glycoprotein precursor (LASV-GPC) resulting in construction of YFV17D/LASV-GPC recombinant virus. The virus is replication-competent and processes the LASV-GPC in cell cultures (Bredenbeek et al., 2006).
Host Response Host Response Host Response Host Response Host Response

Monkey Response

  • Host Strain: Rhesus macaques
  • Vaccination Protocol: Two adult rhesus macaques were s.c. injected with 103 PFU of ML29. Blood samples were taken weekly and submitted to the clinical laboratory for complete blood counts and standard 20-assay chemistry panels. At days 14 and 28, the monkeys were euthanized and total blood and tissues were collected. A portion of each tissue was submerged in MEM with 10% FCS (for plaque titration) and in RNAlater (for RNA isolation). The remaining tissue portions were fixed in 10% neutral formalin for the preparation of standard histological sections and stained with hematoxylin-eosin (Lukashevich et al., 2005).
  • Efficacy: This data indicates that ML29 vaccination of rhesus macaques results in a short, inapparent, self-limited infection (Lukashevich et al., 2005).
  • Description: The ML29-vaccinated animals were afebrile throughout the experiment and had no clinical manifestations. Hematological and chemical parameters were in the normal ranges, as was gross appearance at necropsy. Detailed histological examination of rhesus macaques infected with the ML29 reassortant revealed no tissue lesions. The ML29 virus replicated poorly in monkeys and was not detectable in the plasma and tissues by conventional infectious plaque assay. The only organ from which the virus was recovered was the spleen. RT/PCR with LASV GPC-derived primers was transiently positive with RNA plasma and tissue samples (Lukashevich et al., 2005).

Guinea pig Response

  • Vaccine Immune Response Type: VO_0000286
  • Immune Response: The YFV17D/LAS-GPC vaccination elicited humoral immune responses against YFV17D and LASV antigens. Neutralizing antibodies play the major role in protection against yellow fever (Bredenbeek et al., 2006).
  • Efficacy: A single subcutaneous injection of the recombinant vaccine protected 13 guinea pigs against fatal Lassa Fever (Bredenbeek et al., 2006).

Guinea pig Response

  • Vaccination Protocol: Control animals received either no vaccine or 108 PFU of NYBH vaccinia; vaccinated animals received108 PFU of V-LSGPC or V-LSN, or a simultaneous injection with both recombinant vaccinia viruses at separate sites, by intradermal injection of 0.1 ml on the back (Morrison et al., 1989).
  • Challenge Protocol: Animals were challenged with 104 PFU of guinea-pig-passaged Lassa virus by intraperitoneal inoculation 3 to 4 weeks postvaccination (Morrison et al., 1989).
  • Efficacy: Ninety-four percent of the animals vaccinated with V-LSN survived a Lassa virus challenge in which only 14% of unvaccinated animals and 39% of animals vaccinated with the New York Board of Health (NYBH) strain of vaccinia virus survived (Morrison et al., 1989).

Guinea pig Response

  • Host Strain: 13
  • Vaccination Protocol: Ten animals were inoculated subcutaneously (s.c.) with the ML29 clone, and 10 guinea pigs received the same dose of MOPV. Eight animals were used as negative controls. At day 30 after vaccination, the animals were s.c. challenged with 103 PFU of LASV (Josiah) and followed for 70 days. Liver enzymes were measured in plasma. Vaccinated animals were euthanized on day 70 after challenge, and tissues were removed (Lukashevich et al., 2005).
  • Persistence: None noted
  • Side Effects: In vaccinated animals, LASV infection did not induce alterations in target tissues. The lungs and livers of vaccinated animals looked essentially like normal tissues. There were also no lesions in other major organs (Lukashevich et al., 2005).
  • Efficacy: All strain 13 guinea pigs vaccinated with clone ML29 survived at least 70 days after LASV challenge without either disease signs or histological lesions (Lukashevich et al., 2005).
  • Description: Infection of strain 13 guinea pigs with MOPV or with the ML29 reassortant was not lethal for the animals and did not induce clinical or biochemical signs of the disease. All animals survived after challenge and had no clinical manifestations. All measured parameters were in normal ranges in ML29-vaccinated guinea pigs. In MOPV-vaccinated animals, a transient elevation of AST and AlkPh in plasma was observed at week 3 after challenge (Lukashevich et al., 2005).

Guinea pig Response

  • Host Strain: Hartley
  • Vaccination Protocol: Animals were vaccinated with l0^8 plaque-forming units (PFUs) of V-LSGPC recombinant virus using a single intradermal injection of 0.1 ml on the back. l04 PFUs of guinea of pig-cultured Lassa virus were given by intraperitoneal innoculation 21 days post-vaccination.
  • Persistence: All surviving animals were completely free of virus in their blood and tissues by 61 days postchallenge, when the experiment was terminated (Auperin et al., 1988).
  • Side Effects: Two to three days after vaccination, vesicles developed at the site of inoculation on each animal. The vesicles arising from V-LSGPC were approximately 2-3 mm. All vesicles scabbed and completely healed by 19 days postvaccination. vaccinated animals developed mild symptoms of Lassa fever following challenge. The animals vaccinated with V-LSGPC virus developed low-grade fevers, which began about 8 days post-challenge and lasted approximately 4 days. The unprotected animals, however, developed significantly higher fevers, which persisted until
    they died (Auperin et al., 1988).
  • Efficacy: Vaccination with V-LSGPC effectively limited the replication of Lassa virus in comparison to the unvaccinated animals. The ability to construct recombinant vaccinia viruses that express heterologous genes offers great potential for vaccine development (Auperin et al., 1988).
  • Description: 21 days post vaccination the animals were challenged with Lassa virus. All animals that received V-LSGPC recombinant virus survived the lethal Lassa virus challenge (Auperin et al., 1988).

Guinea pig Response

  • Host Strain: 13
  • Vaccination Protocol: Sixteen outbred guinea pigs were subcutaneously inoculated with 1 × 105 PFU/0.5 ml of the recombinant YFV17D/LASV-GPC virus and two animals were sacrificed at days 0, 4, 7, 10 and 14 to track the virus distribution in blood and tissues. At day 14, six animals were boosted with the same dose of the recombinant virus and plasma samples were collected on days 8, 15 and 24 after to measure antibody responses against YFV17D and LASV-GPC in IgG ELISA. Antigens were prepared from serum-free virus stocks of YFV17D and MOP/LAS (Lukashevich et al., 2005) by ultracentrifugation on sucrose cushion. Concentrated viruses were suspended in carbonate–bicarbonate buffer, briefly sonicated and used to cover wells of microtitration plates overnight at 4 °C. After blocking, 1:100 dilutions of guinea pig sera were added and incubated for 2 h at room temperature. Challenge experiments were preformed (Bredenbeek et al., 2006).
  • Persistence: None noted
  • Side Effects: None noted
  • Efficacy: 80% of animals were protected against the fatal challenge. This study demonstrates the potential to develop an YFV17D-based bivalent vaccine against Lassa Virus (Bredenbeek et al., 2006).
  • Description: 80% of animals were protected against the fatal challenge. Incomplete protection could be explained by differences in vaccine formulation (GPC + NP vs. GPC) and by GPC sequence differences between AV and Josiah strains of LASV. Blood and tissue samples were collected from vaccinated animals at different time points for hematology, blood chemistry, RNA extraction, plaque assay, virus isolation and ELISA. As expected, the inoculated animals had no clinical manifestations and all standard measurable blood and chemistry parameters were in normal ranges. In plasma, the recombinant virus was not detectable by plaque assay or by biological amplification. Recombinant viral RNA was not detectable by RT/PCR in 140 μl of plasma extracted on days 4, 10, and 21. However, when RNA samples were prepared from 0.5 ml of total blood, an RT/PCR assay gave a strong positive signal on day 4 after YFV17D/ LAS-GPC inoculation. Still, blood samples collected on day 10 and 21 were PCR negative. Viral RNA sequences were only transiently detectable on days 7–14 in spleen and liver. Nucleotide sequence analysis of PCR products confirmed their derivation from YFV17D/LAS-GPC. Taken together, these data confirm that recombinant YF17D/LAS-GPC replicated poorly in tissues of vaccinated guinea pigs. Interestingly, in clinical trials in individuals vaccinated with chimeric YFV17D-based vaccines, viremia levels were even lower than the levels of YFV17D determined to be safe. This suggests that insertion of a foreign gene can affect in vivo viral replication and make recombinant YFV17D-based vaccines even safer than the parental vaccine, YFV17D (Bredenbeek et al., 2006).
References References References References References
Bredenbeek et al., 2006: Bredenbeek PJ, Molenkamp R, Spaan WJ, Deubel V, Marianneau P, Salvato MS, Moshkoff D, Zapata J, Tikhonov I, Patterson J, Carrion R, Ticer A, Brasky K, Lukashevich IS. A recombinant Yellow Fever 17D vaccine expressing Lassa virus glycoproteins. Virology. 2006 Feb 20; 345(2); 299-304. [PubMed: 16412488].
Morrison et al., 1989: Morrison HG, Bauer SP, Lange JV, Esposito JJ, McCormick JB, Auperin DD. Protection of guinea pigs from Lassa fever by vaccinia virus recombinants expressing the nucleoprotein or the envelope glycoproteins of Lassa virus. Virology. 1989; 171(1); 179-188. [PubMed: 2741340].
Lukashevich et al., 2005: Lukashevich IS, Patterson J, Carrion R, Moshkoff D, Ticer A, Zapata J, Brasky K, Geiger R, Hubbard GB, Bryant J, Salvato MS. A live attenuated vaccine for Lassa fever made by reassortment of Lassa and Mopeia viruses. Journal of virology. 2005 Nov; 79(22); 13934-42. [PubMed: 16254329].
Auperin et al., 1988: Auperin DD, Esposito JJ, Lange JV, Bauer SP, Knight J, Sasso DR, McCormick JB. Construction of a recombinant vaccinia virus expressing the Lassa virus glycoprotein gene and protection of guinea pigs from a lethal Lassa virus infection. Virus research. 1988 Feb; 9(2-3); 233-48. [PubMed: 3354260].
Bredenbeek et al., 2006: Bredenbeek PJ, Molenkamp R, Spaan WJ, Deubel V, Marianneau P, Salvato MS, Moshkoff D, Zapata J, Tikhonov I, Patterson J, Carrion R, Ticer A, Brasky K, Lukashevich IS. A recombinant Yellow Fever 17D vaccine expressing Lassa virus glycoproteins. Virology. 2006 Feb 20; 345(2); 299-304. [PubMed: 16412488].