Marburg was identified in 1967 in Central Africa. It has been known to infect bats, humans, and non-human primates. The virus has caused severe outbreaks of, usually, fatal hemorrhagic fever. In primates, the symtoms are known to be fever, diarrhea, vomiting, coagulation deficits, and severe liver damage. The virus causes a profuse release of inflammatory cytokines, which affects vascular permeability. The excessive bleeding involved sometimes leads to the activation of tissue factor in macrophages and monocytes, as well as a drop in platlet numbers (Mohamadzadeh et al., 2007).
4. Microbial Pathogenesis
The initial targets of infection are generally macrophages, monocytes, and DCs. Filoviruses generally bind to the TREM or C-type lectin receptors on myloid cells. However, since some cells do not express C-lectin or TREM receptors, viral entry may be due to molecules such as heparin-sulphate proteoglycan and folate receptor-α. Data also indicate that the expression of TLR1 (whose signal converges with that of TLR2) was increased on MARV-activated neutrophils (Mohamadzadeh et al., 2007).
5. Host Ranges and Animal Models
The natural host for MARV is known to be bats of Central African origin (Mohamadzadeh et al., 2007).
6. Host Protective Immunity
Filoviruses have been found to disable some of the host IFN pathway, such as the VP35, which prevents the production of type I IFNs, and VP24, which interferes with the abilities of certain molecules to induce antiviral states. IFN is crucial to immune response, as indicated by elevated levels of IFN in the blood even during acute infection. It had also been seen that there is a deficit (virus-induced) in the co-stimulatory properties of infected Dcs, identified by a supressed capacity of the DCs to stimulate allogenic T cells. By affecting either viral exit or entry, cellular proteases could influence viral cellular tropisms and interactions between antibodies and glycoproteins. All host protective immunity observations are also tied with the prevalence of an abundance of cathepsins (Mohamadzadeh et al., 2007).
Molecule Role Annotation :
Study developed replication-competent vaccines against EBOV and MARV based on attenuated recombinant vesicular stomatitis virus vectors expressing either the EBOV glycoprotein or MARV glycoprotein from Lake Victoria marburgvirus - Musoke. A single intramuscular injection of the EBOV or MARV vaccine elicited completely protective immune responses in nonhuman primates against lethal EBOV or MARV challenges (Jones et al., 2005).
Protein Note :
small non-structural secreted glycoprotein; forms dimers linked by disulfide bonds (parallel orientation); processed by furin to yield sGP and delta peptide; sGP
Molecule Role Annotation :
Nonhuman primates (cynomolgus macaques) were inoculated with VEE replicons expressing MBGV GP and/or NP and challenged with MGBV. MBGV NP afforded incomplete (partial) protection, sufficient to prevent death but not disease in two of three macaques (Hevey et al., 1998).
Molecule Role Annotation :
Guinea pigs were vaccinated with recombinant VEE replicons (packaged into VEE-like particles), inoculated with MBGV, and evaluated for viremia and survival after challenge with typical MGBV disease. Results indicated that VP35 afforded incomplete protection (Hevey et al., 1998).
Glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
d. Gene Engineering of
GP from Marburg virus Ci67
Type:
Recombinant protein preparation
Description:
This Ci67 GP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Ci67strain of MARV (Wang et al., 2006).
Description:
This Ravn GP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Ci67strain of MARV (Wang et al., 2006).
f. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant protein preparation
Description:
This Musoke GP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Musoke strain of MARV (Wang et al., 2006).
The Ci67 GP (Genbank accession number AF005735; Protein ID AAC40460.1; a.a. 1–681), Ravn GP (Genbank accession number AF005734; Protein ID AAC40459.1; a.a. 1–681), and Musoke GP (Genbank accession number DQ217792; Protein ID ABA87127; a.a. 1–681) genes were amplified by PCR. Each MARV antigen was subcloned into the pLAd and pRAd shuttle vectors to create a series of MARV adenovirus vaccines targeted against the Ci67, Ravn, and Musoke strains of MARV. All cAdVax vector genomes were based on a modified Ad5sub360 vector backbone, which contains deletions in E1, E3 and almost all E4 ORFs with the exception of ORF6 (Wang et al., 2006).
h. Virulence
Although Ad vectors have a high affinity for the liver and may potentially cause inflammation in the liver, there is not pathology indicative of inflammation or cytotoxicity as a result of vaccination in mice (Wang et al., 2006).
i.
Mouse Response
Host Strain:
C57BL/6 mice
Vaccination Protocol:
Four groups of 28 C57BL/6 mice were immunized intra-peritoneally (i.p.) at weeks 0, 4 and 8 with 1 × 10^8 pfu of either cAdVaxM(ci), cAdVaxM(ra), cAdVaxM(mu), or HC4 control vector prepared in 100 μl PBS/10% glycerol. HC4, an unrelated adenovirus-based Hepatitis C vaccine, served as a negative control vaccine. Four mice per group were sacrificed at weeks 2, 4, 6, 8, 10, 12, and 24 (Wang et al., 2006).
Persistence:
The persistence of these three vaccine candidates in mice was not reported.
Immune Response:
Immunization of mice with complex adenovirus (Ad)-based vaccine candidates (cAdVax vaccines) induced efficient production of both antibodies and cytotoxic T lymphocytes (CTL) specific to Musoke strain GP and Ci67 strain GP, respectively. Antibody responses were also cross-reactive across the MARV strains, but not cross-reactive to Ebola virus, a related filovirus (Wang et al., 2006).
Side Effects:
Three 1 × 10^8 pfu doses of vaccine vector did not lead to any detectable toxicity in liver or spleen, so it appears to be safe (Wang et al., 2006).
Challenge Protocol:
No challenge experiment was conducted.
e. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant vector construction
Description:
Novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
Description:
Novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
Description:
Novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
Three novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
i. Immunization Route
Intramuscular injection (i.m.)
j.
Mouse Response
Vaccination Protocol:
Immunization of mice with complex adenovirus (Ad)-based vaccine candidates (cAdVax vaccines) (Wang et al., 2006).
Vaccine Immune Response Type:
VO_0003057
Efficacy:
Vaccination led to efficient production of both antibodies and cytotoxic T lymphocytes (CTL) specific to Musoke strain GP and Ci67 strain GP, respectively. Antibody responses were also shown to be cross-reactive across the MARV strains, but not cross-reactive to Ebola virus, a related filovirus. Additionally, three 1 x 10(8)pfu doses of vaccine vector were demonstrated to be safe in mice, as this did not lead to any detectable toxicity in liver or spleen (Wang et al., 2006).
e. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant vector construction
Description:
Novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
Description:
Novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
Description:
Novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
Three novel gene-based vaccine candidates which express the viral glycoprotein (GP) from either the Ci67, Ravn or Musoke strain of MARV (Wang et al., 2006).
i. Immunization Route
Intramuscular injection (i.m.)
j.
Mouse Response
Vaccination Protocol:
Immunization of mice with complex adenovirus (Ad)-based vaccine candidates (cAdVax vaccines) (Wang et al., 2006).
Vaccine Immune Response Type:
VO_0003057
Description:
Vaccination led to efficient production of both antibodies and cytotoxic T lymphocytes (CTL) specific to Musoke strain GP and Ci67 strain GP, respectively. Antibody responses were also shown to be cross-reactive across the MARV strains, but not cross-reactive to Ebola virus, a related filovirus. Additionally, three 1 x 10(8)pfu doses of vaccine vector were demonstrated to be safe in mice, as this did not lead to any detectable toxicity in liver or spleen (Wang et al., 2006).
d. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant protein preparation
Description:
MBGV gene clone pGem-GP was provided by Heinz Feldmann and Anthony Sanchez (Centers for Disease Control and Prevention, Atlanta, GA). The MBGV GP gene from pGem-GP was excised with SalI and subcloned into the SalI site of a shuttle vector. A clone with the MBGV GP gene in the correct orientation was excised with ApaI and NotI, and this fragment was cloned into the ApaI and NotI sites of a VEE replicon plasmid (Hevey et al., 1998).
Description:
An RNA replicon based on VEEV was used as the vector, with the VEE structural genes replaced by VP40. VP40 seems to serve as a matrix protein, affecting interactions between the nucleoprotein complex and lipid membrane. It is also the most abundant part of the virion (Hevey et al., 1998).
Expression plasmids were made through the use of MARV-Musoke strain. To produce the actual viral preparations, MARV-infected cell supernatents were clarified at 1500×g and then pelleted at 9500×g for 4 h in a Sorvall GSA rotor (Swenson et al., 2005).
h. Virulence
GP vaccination formed sufficient protection against homologous filovirus challenge, yet heterologous wild-type VLPs without GP failed to protect. Our data indicate that vaccination with GP was required and sufficient to form an immune response as heterologous wild-type VLPs or hybrid VLPs that did not contain the homologous GP failed. Vaccination with a mixture of EBOV and MARV VLPs was successful in forming an immune response (Swenson et al., 2005).
i.
Guinea pig Response
Host Strain:
Strain 13
Vaccination Protocol:
Strain 13 guinea pigs were vaccinated once with mVLPs, eVLPs, or an equal mixture of eVLPs and mVLPs in RIBI adjuvant, and the serum antibody levels against MARV and EBOV were measured (via ELISA) prior to challenge (Swenson et al., 2005).
Control guinea pigs were vaccinated with RIBI adjuvant alone. Serum samples from the guinea pigs were
obtained immediately before (PRE) or 28 days post-challenge (POST).Guinea pigs were vaccinated
Persistence:
Not noted.
Immune Response:
Animals vaccinated with the wild-type eVLP or e/m-VLPs did not have high serum antibody titers against MARV, but did have them against EBOV. Injection with mVLP and m/e-VLP vaccination resulted in high titers against MARV but not against EBOV. The vaccination containing EBOV GPin the form of eVLPor e/m-VLP resulted in antibodies against EBOV but not MARV. Finally, animals vaccinated with mVLP orm/e-VLP did not develop significant antibody titers against EBOV or MARV (Swenson et al., 2005).
Side Effects:
None noted.
Challenge Protocol:
Guinea pigs were challenged 28 days after a single vaccination with 1000 pfu of guinea pig-adapted MARV or EBOV (Swenson et al., 2005).
Efficacy:
Guinea pigs challenged wuth VLPs containing homologous GP were protected from a lethal filovirus, and a eVLP or e/m-VLP vaccination yielded protection against EBOV infection. Vaccines containing heterologous proteins or homologous VP40 did not protect against lethal challenge (Swenson et al., 2005).
Vaccination Protocol:
Animals were immunized with irradiated MBGV (strains Musoke and Ravn)(Hevey et al., 1997).
Persistence:
None noted.
Immune Response:
Not noted for irradiated MBGV specifically.
Side Effects:
None noted.
Challenge Protocol:
All animals challenged with strains Musoke and Ravn survived without regard to challenge virus (Hevey et al., 1997).
Efficacy:
Gradient-purified, irradiated virus was able to completely protect strain 13 guinea pigs from challenge with either MBGV (strain Musoke) or
indicated that the product immunoprecipitated from GP- MBGV (strain Ravn)(Hevey et al., 1997).
6. Marburg virus DNA prime/boost vaccine DNA/rAd5-GP encoding GP from strain Angola
Immune Response:
The vaccine induced humoral responses , as well as CD4(+) and CD8(+) cellular immune responses, with skewing toward CD4(+) T-cell activity against MARV GP. The highest antibody titers were achieved with a heterologous prime-boost vaccine. rAd5-GP boosted titers in DNA-primed animals more than 2 orders of magnitude to a final prechallenge GP ELISA IgG titer of 1:237,167 (Geisbert et al., 2010).
Efficacy:
Heterologous prime-boost with DNA/rAd vectors generated protective immunity in all subjects after challenge with a lethal dose of MARV Angola (Geisbert et al., 2010).
Immune Response:
The DNA/DNA vaccine induced humoral responses comparable to those induced by a single inoculation with rAd5-GP, as well as CD4(+) and CD8(+) cellular immune responses, with skewing toward CD4(+) T-cell activity against MARV GP (Geisbert et al., 2010).
Efficacy:
The DNA-GP-only vaccine prevented death in all vaccinated subjects after challenge with a lethal dose of MARV Angola (Geisbert et al., 2010).
Immune Response:
All of the MARV GP DNA-vaccinated guinea pigs developed antibodies to MARV (Riemenschneider et al., 2003).
Efficacy:
In both studies (two different strains - Musoke and Ravn), two of three GP DNA-vaccinated monkeys were aviremic on the days assayed, and survived challenge, while one monkey in each study developed viremia levels similar to those of control monkeys and died. These results indicate that DNA vaccination alone is able to offer immunity to nonhuman primates, but suggest that the protective effect is near the threshold of vaccine efficacy (Riemenschneider et al., 2003).
9. Marburg virus glycoprotein expressed by baculovirus recombinants
The full-length and truncated GP were expressed by baculovirus recombinants (Hevey et al., 1997). Both antigens were abundantly glycosylated with both N- and O-linked glycans.
d. Gene Engineering of
NP from Marburg virus Musoke
Type:
Recombinant protein preparation
Description:
This Musoke NP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Musoke strain of MARV (Wang et al., 2006).
e. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant protein preparation
Description:
This Musoke GP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Musoke strain of MARV (Wang et al., 2006).
MBGV glycoprotein (GP) was expressed in Baculovirus recombinants either as a slightly truncated product secreted into medium or a complete, cell-associated molecule(Hevey et al., 1997).
g. Virulence
Irradiated MBGV antigen was protective against two MBGV strains (Musoke and Ravn). The recombinant truncated glycoprotein did elicit protection against challenge with the MBGV isolate from which it was taken(Hevey et al., 1997).
h.
Guinea pig Response
Host Strain:
Strain 13 and Hartley
Vaccination Protocol:
Groups of animals were challenged with Ravn and Musoke strains, and ELISA titers were use to measure response 28 days after challenge (Hevey et al., 1997).
Immune Response:
Irradiated, gradient-purified virus completely protected Strain 13 from both Ravn and Musoke MBGV strains (Hevey et al., 1997).
Challenge Protocol:
Guinea pigs (Hartley and Strain 13) were divided into groups and injected with irradiated GP or recombinant GP. ELISA titers measureing response were taken 2 days before challenge and 28 days after for comparison (Hevey et al., 1997).
Description:
Animals that recieved the MBGV antigen (strainRavn) had a lower survival rate than those challenged with the Musoke strain(Hevey et al., 1997).
An RNA replicon, based upon Venezuelan equine encephalitis (VEE) virus, was used as a vaccine vector (Hevey et al., 1998).
f. Immunization Route
Subcutaneous injection
g.
Macaque Response
Host Strain:
Macaca fascicularis
Vaccination Protocol:
Twelve cynomolgus macaques (Macaca fascicularis), 11 females and 1 male, ranging from 2.8 to 4.5 kg, were inoculated subcutaneously with 10^7 FFU of VRP in a total volume of 0.5 ml at one site. Monkeys were anesthetized with ketamine, bled, and inoculated (as described for the first vaccine dose) 28 days after the primary injection, and again 28 days after the second (Hevey et al., 1998).
Challenge Protocol:
Animals were challenged 14 days after 3rd vaccine dose with 10^3.9 PFU MBGV subcutaneously (Hevey et al., 1998).
Efficacy:
MBGV NP afforded incomplete (partial) protection, sufficient to prevent death but not disease in two of three macaques (Hevey et al., 1998).
Immune Response:
All vaccinated animals from groups 1(challenged with 1,000 PFU MARV Musoke) and 2 (challenged with the same dose of ZEBOV) mounted strong antibody titers against all five filoviruses with similar kinetics (Swenson et al., 2005).
Efficacy:
Vaccination of NHP with CAdVax-Panfilo was 100% protective against challenge with multiple filovirus species, including ZEBOV, SEBOV, MARV Musoke, and MARV Ci67 (Swenson et al., 2005).
12. Marburg virus recombinant vector vaccine MBGV GP
Efficacy:
Three monkeys vaccinated with replicons which expressed MBGV GP, and three others vaccinated with both replicons that expressed GP or NP, remained aviremic and were completely protected from disease (Hevey et al., 1998).
Efficacy:
Three monkeys vaccinated with replicons which expressed MBGV GP, and three others vaccinated with both replicons that expressed GP or NP, remained aviremic and were completely protected from disease (Hevey et al., 1998).
This vaccine includes two doses with two different vaccine formulae - Ad26.Filo and MVA-BN-Filo. For Ad26.Filo , ..... Replication-incompetent, E1/E3-deleted recombinant adenoviral vectors based on Ad26 were engineered using the AdVac® system with the humanized GP DNA sequences for EBOV Mayinga (Ad26.ZEBOV), SUDV Gulu (Ad26.SUDV), and MARV Angola (Ad26.MARVA). The combination of these three Ad26 vectors in a 1:1:1 ratio will be further referred to as Ad26.Filo; MVA-BN-Filo is a recombinant, modified vaccinia Ankara-vectored vaccine, non-replicating in human cells, encoding the EBOV Mayinga, SUDV Gulu, and MARV Musoke GPs as well as the nucleoprotein of the Tai Forest virus. Adenovirus vaccines were given at 1.2 × 10^11 viral particles (vp; 4 × 10^10 vp/vector) and the MVA-BN-Filo vector at a dose of 5 × 10*8 infectious units (infU). MVA-BN-Filo is composed of the Tai Forest Virus. (Tiemessen et al., 2022).
k. Immunization Route
Intramuscular injection (i.m.)
l. Description
Two-dose regimen, consisting of Ad26.Filo followed by MVA-BN-Filo with an 8-week interval (Tiemessen et al., 2022)
m.
Macaque Response
Vaccination Protocol:
For that purpose, 16 cynomolgus macaques were assigned to four groups: one control group (n = 2) and three groups each receiving a different vaccine regimen (Table 2). The negative control group received an Ad26 empty vector followed by Tris buffer as dose 2. The three groups immunized with vaccine regimens received either Ad26.Filo, Ad35.Filo (n = 4); Ad26.Filo, MVA-BN-Filo (n = 5); or MVA-BN-Filo, Ad26.Filo regimen (n = 5). Adenovirus vaccines were given at 1.2 × 10^11 vp (4 × 10^10 vp/vector) and the MVA-BN-Filo vector at a dose of 5 × 108 infU. (Tiemessen et al., 2022)
Immune Response:
The MARV GP–specific antibody levels induced by the three different heterologous vaccine regimens were very similar 3 weeks post-dose 2 (study week 11). However, the MARV GP–specific antibody levels induced post-dose 1, at study weeks 4 and 8, were lower post-MVA compared to post-Ad26. Additionally, MARV-specific neutralizing antibodies could be detected in all immunized animals except for two animals in the MVA-BN-Filo, Ad26.Filo group.
Side Effects:
ne animal of the Ad26.Filo, MVA-BN-Filo regimen group (NHP 33841), that experienced transient severe clinical symptoms from day 8 until day 12 after infection, showed the lowest MARV GP–specific antibody concentration in serum at weeks 10 and 11 from the 5 animals in that group (2.45 and 2.26 EU/mL [log10]). MARV GP–specific binding antibody concentrations are good predictors for survival outcome after challenge, even when analyzed across various vaccination regimens. (Tiemessen et al., 2022)
Challenge Protocol:
Four weeks after dose 2, the animals were challenged i.m. with 1000 pfu MARV Angola. (Tiemessen et al., 2022)
Efficacy:
21 days post-dose 2, 100% of participants on the active regimen responded to vaccination and exhibited binding antibodies against EBOV, SUDV, and MARV GPs. (Tiemessen et al., 2022)
15. Marburg Virus Vaccine mVLP Poly I:C Adjuvant
a. Type:
Recombinant vector vaccine
b. Status:
Research
c. Host Species for Licensed Use:
None
d. Antigen
Marburg Virus (MARV) glycoprotein, MARV nucleoprotein, MARV Matrix Protein VP40 (Dye et al., 2016)
For a generation of the MARV VLPs, the MARV GP, NP and VP40 genes were inserted into a single baculovirus vector system for expression in insect cells Sf9 insect cells were infected with the single recombinant baculovirus, and the VLPs were recovered from the culture supernatants by high-speed centrifugation, purified on sucrose gradients, and resuspended in phosphate buffered saline (PBS), as previously described. Total proteins in the VLP preparations were determined and the VLPs were analyzed by SDS-PAGE/Western blotting and ELISA for filovirus protein content and identity, immunogenicity in mice and endotoxin levels. (Dye et al., 2016)
i. Immunization Route
subcutaneous injection
j.
Macaque Response
Vaccination Protocol:
Thirteen macaques received intramuscular injections in the caudal thigh muscle containing 3 mg (total protein) of MARV VLPs and 0.5 mg/kg of polyI:C adjuvant. The three control animals received injections of one of the adjuvants only. Immunizations were performed on study days 0, 42, and 84. (Dye et al., 2016)
Immune Response:
Cynomolgus macaques were vaccinated with MARV VLPs on study days 0, 42, and 84 and serum antibody titers against purified MARV GP and VP40 were determined for each animal every two to three weeks (study days 0, 14, 28, 42, 56, 70, 84 and 105). Control animals, which were vaccinated with polyI:C adjuvant alone, did not generate any antibody responses to either MARV GPdTM or VP40 (below the limit of detection at a 1:100 dilution of serum). The vaccinated animals exhibited similar kinetics of antibody responses to both antigens, with detectable antibody titers at day 14, which waned slightly at day 42 and increased again after the second and third vaccinations were administered on study days 42 and 84. Animals vaccinated with MARV VLPs with polyI:C exhibited similar responses to the protective GP antigens (p = 0.6006). Animals vaccinated with MARV VLPs and polyI:C had higher responses to the VP40 antigen than those vaccinated with MARV VLP and QS-21, specifically at the later time points of days 70, 84 and 105 post vaccination (p = 0.0057) (Dye et al., 2016)
Side Effects:
In Groups 1, 2 and 4, the macaques vaccinated with MARV VLPs and challenged via either aerosol or SQ route, there were no animals with visible clinical signs of filovirus infection. The adjuvant only control macaques presented with typical clinical signs of filovirus infection in macaques. Animal 16-P-S, which was vaccinated with polyI:C alone and challenged by the SQ route exhibited severe depression, moderate rash, and no food intake at day 10 post challenge. (Dye et al., 2016)
Challenge Protocol:
MARV was grown on Vero cells (6 total passages) and enumerated using standard plaque assay. Administration of the virus challenge was performed on study day 112 with the challenge material administered in the subcutaneous (SQ) tissues of the left thigh of each animal or via the aerosol route, as previously described. The target challenge dose for both SQ and aerosol exposure was 1000 plaque-forming units (pfu)/mL. For the SQ challenge, each macaque received 0.5 mL of challenge stock MARV-Musoke, which upon back-titration revealed an actual challenge dose of 315 pfu/macaque. For aerosol challenge, each macaque was exposed to 10 mL of challenge stock MARV-Musoke using a previously described methodology, and the air in the aerosolization chamber was sampled during each exposure to calculate the actual inhaled dose of virus that each animal received. This revealed that the inhaled dose ranged between 40–135 pfu/macaque. (Dye et al., 2016)
Efficacy:
Vaccination of cynomolgus macaques with MARV VLP with polyI:C adjuvant provided complete protection against challenge with aerosolized MARV-Musoke. (Dye et al., 2016)
16. Marburg Virus Vaccine mVLP QS-21 Adjuvant
a. Type:
Virus Like Particle
b. Status:
Research
c. Host Species for Licensed Use:
None
d. Antigen
Marburg Virus (MARV) glycoprotein, MARV nucleoprotein, MARV Matrix Protein VP40 (Dye et al., 2016)
For a generation of the MARV VLPs, the MARV GP, NP and VP40 genes were inserted into a single baculovirus vector system for expression in insect cells Sf9 insect cells were infected with the single recombinant baculovirus, and the VLPs were recovered from the culture supernatants by high-speed centrifugation, purified on sucrose gradients, and resuspended in phosphate buffered saline (PBS), as previously described. Total proteins in the VLP preparations were determined and the VLPs were analyzed by SDS-PAGE/Western blotting and ELISA for filovirus protein content and identity, immunogenicity in mice and endotoxin levels. (Dye et al., 2016)
i. Immunization Route
subcutaneous injection
j.
Macaque Response
Vaccination Protocol:
Thirteen macaques received intramuscular injections in the caudal thigh muscle containing 3 mg (total protein) of MARV VLPs and 0.1 mg of QS-21 adjuvant. The three control animals received injections of one of the adjuvants only. Immunizations were performed on study days 0, 42, and 84.
Immune Response:
Cynomolgus macaques were vaccinated with MARV VLPs on study days 0, 42, and 84 and serum antibody titers against purified MARV GP and VP40 were determined for each animal every two to three weeks (study days 0, 14, 28, 42, 56, 70, 84 and 105). Control animals, which were vaccinated with QS-2 alone, did not generate any antibody responses to either MARV GPdTM or VP40 (below the limit of detection at a 1:100 dilution of serum). The vaccinated animals exhibited similar kinetics of antibody responses to both antigens, with detectable antibody titers at day 14, which waned slightly at day 42 and increased again after the second and third vaccinations were administered on study days 42 and 84. Animals vaccinated with MARV VLPs with QS-21 exhibited similar responses to the protective GP antigens (p = 0.6006). Animals vaccinated with MARV VLPs and polyI:C had higher responses to the VP40 antigen than those vaccinated with MARV VLP and QS-21, specifically at the later time points of days 70, 84 and 105 post vaccination (p = 0.0057) (Dye et al., 2016)
Side Effects:
In Groups 1, 2 and 4, the macaques vaccinated with MARV VLPs and challenged via either aerosol or SQ route, there were no animals with visible clinical signs of filovirus infection. The adjuvant only control macaques presented with typical clinical signs of filovirus infection in macaques. Both of the macaques that received QS-21 only (animals 11-Q-A and 15-Q-S) exhibited severe depression, widespread rash, and no food intake at day 10 post challenge. (
Challenge Protocol:
MARV was grown on Vero cells (6 total passages) and enumerated using standard plaque assay. Administration of the virus challenge was performed on study day 112 with the challenge material administered in the subcutaneous (SQ) tissues of the left thigh of each animal or via the aerosol route, as previously described. The target challenge dose for both SQ and aerosol exposure was 1000 plaque-forming units (pfu)/mL. For the SQ challenge, each macaque received 0.5 mL of challenge stock MARV-Musoke, which upon back-titration revealed an actual challenge dose of 315 pfu/macaque. For aerosol challenge, each macaque was exposed to 10 mL of challenge stock MARV-Musoke using a previously described methodology, and the air in the aerosolization chamber was sampled during each exposure to calculate the actual inhaled dose of virus that each animal received. This revealed that the inhaled dose ranged between 40–135 pfu/macaque. (Dye et al., 2016)
Efficacy:
Vaccination of cynomolgus macaques with MARV VLP with either QS-21 provided complete protection against challenge with aerosolized MARV-Musoke.(Dye et al., 2016)
The virus was generated via reverse genetics rescue essentially as described previously for a similar construct expressing the MARV variant Musoke GP [12]. A passage level 2 (P2) stock of virus was derived from the rescued virus (considered P0) via amplification on Vero E6 cells at the Public Health Agency of Canada. The P2 stock virus was subsequently amplified and plaque purified in qualified CGMP Vero cells in proprietary serum-free tissue culture media to generate premaster viral seeds at P8. A single P8 clone (Clone 5) was chosen based on plaque morphology, growth kinetics and productivity in Vero cells, and MARV GP transgene sequence fidelity. This clone was amplified to P9 under CGMP conditions to generate the PHV01 MVS. PHV01 MVS release criteria included the well-defined requirements for GMP viral seed manufacturing for purity (including adventitious agents), potency/strength, identity, and sterility. PHV01 MVS was further amplified to P10 under CGMP conditions to generate the PHV01 Working Viral Seed (WVS). The PHV01 Formulated Drug Substance (FDS) at P11 was produced from the PHV01 WVS in a disposable bioreactor using the same CGMP Vero cells and proprietary media described above. PHV01 FDS was harvested from cell culture medium containing virus, clarified, and further purified by nuclease digestion, depth filtration, and tangential flow ultrafiltration/diafiltration in a recombinant human albumin and Tris buffer formulation. Separately, an rVSVΔG-MARV P3 virus was generated from the initial P2 stock virus for research purposes, and PHV01 MVS was passaged three additional times (PHV01 MVS+3) to P12. (Zhu et al., 2022)
g. Immunization Route
Intramuscular injection (i.m.)
h.
Guinea pig Response
Vaccination Protocol:
This study was composed of two experiments, termed Experiment #1 and Experiment #2. Both were designed to test the ability of PHV01 to protect female Hartley guinea pigs (Charles River Laboratories) from morbidity and mortality resulting from inoculation with a lethal dose of GPA-MARV/Ang. In Experiment #1, rVSVΔG-MARV research virus P3 and PHV01 MVS were evaluated at three different dose levels: 2 x 10^6 plaque-forming units (PFU; high), 2 × 10^4 PFU (medium), and 2 × 102 PFU (low). Groups of 6 guinea pigs were immunized with one of the two vaccine preparations at one of the three specified doses (prepared with sterile, nontoxic, nonpyrogenic 0.9% saline as diluent), while a control group of 6 animals received an equivalent volume of 0.9% saline. In Experiment #2, the PHV01 FDS was evaluated at two different doses, high and medium (as described above), and PHV01 MVS was evaluated again at the high dose for comparative purposes. Groups of 6 guinea pigs were immunized with one of the two vaccine preparations at the specified doses (as described above), and a control group of 6 animals receiving 0.9% saline was also included. All vaccines were administered intramuscularly (IM) in a total volume of 300 µL, with 150 µL delivered to each of the rear quadriceps muscles. Twenty-eight (28) days postvaccination (DPV), animals were inoculated with 1000 times the median lethal dose (LD50) of GPA-MARV/Ang via intraperitoneal (IP) injection. Animals were monitored for disease and survival up to 29 days postinfection (DPI), equivalent to 57 DPV in Experiment #1 and up to 28 DPI (56 DPV) in Experiment #2. EDTA blood and plasma (Experiment #1) or serum (Experiment #2) samples were obtained at 0 DPV (prior to vaccination), 2 DPV, 27 DPV, 5 DPI (33 DPV), and 29 or 28 DPI (57 or 56 DPV). (Zhu et al., 2022)
Immune Response:
Vaccinemia: At 2 days postvaccination (DPV), the majority of vaccinated animals showed robust levels of RNA that loosely correlated with the dose level of vaccine administered. As expected, unvaccinated control animals exhibited no detectable levels of MARV GP RNA during the vaccination phase. With the exception of two guinea pigs, the three remaining vaccinated, non-surviving guinea pigs showed no detectable levels of MARV GP RNA postvaccination. By 27 DPV, MARV GP RNA was no longer detectable in any animal.
Viremia: MARV GP-specific RNA in the blood of animals was quantified via RT-qPCR. At 5 DPI, all control animals exhibited very high levels of MARV RNA in the blood, with an average of approximately 9–10 Log10 GEQ/mL, indicating abundant MARV replication concomitant with severe infection. Conversely, the majority of vaccinated animals exhibited no viral RNA in the blood at 5 DPI, suggesting that vaccine-elicited immunity prevented MARV replication. Indeed, all vaccinated groups of animals showed significantly lower mean MARV RNA levels compared with the control animals.
Humoral Response: The geometric mean endpoint titers for all vaccinated groups were significantly higher than that of the unvaccinated animals. Of the vaccinated, non-surviving guinea pigs, most had low or undetectable IgG endpoint titers, which exhibited moderate IgG levels. Pooling the data from the PHV01 MVS and FDS groups showed no significant difference in mean IgG titers between the medium and high dose levels, suggesting that both doses of vaccine elicited similarly robust immune responses. Moreover, logistic regression analysis comparing IgG endpoint titers and survival for all animals immunized with PHV01 MVS or FDS revealed a 90% probability of surviving infection with an IgG endpoint titer of 1600 (~Log10 3.2), which was achieved in almost all vaccinated animals from the medium- and high-dose groups. At 28 or 29 DPI (56 or 57 DPV), IgG endpoint titers increased to similar levels for all surviving animals, signifying an enhancement in the immune response to MARV following infection.
Neutralizing Antibody Response: Pooling the data from the clonal PHV01-vaccinated animals revealed that the high-dose groups had statistically significant higher geometric mean PRNT50 endpoint titers compared to both the control animals and the animals vaccinated with the medium dose. Together, these data suggest that immunization with PHV01 elicits a potent and dose-dependent neutralizing antibody response. (Zhu et al., 2022)
Side Effects:
All vaccinated, surviving animals gained weight following GPA-MARV/Ang infection, and the vast majority exhibited no clinical signs of disease. Indeed, only 3 of the 32 vaccinated, surviving animals showed any clinical signs, which were all extremely mild (e.g., minor weight loss, ruffling of fur, and/or moderately reduced activity) and resolved completely within a few days and before 14 DPI. (Zhu et al., 2022)
Challenge Protocol:
1000 LD50 dose of GPA-MARV/Ang (Zhu et al., 2022)
Efficacy:
In general, vaccination with rVSVΔG-MARV P3 and PHV01 prevented severe disease and death due to MARV infection in nearly all animals. Animals vaccinated with the high dose of rVSVΔG-MARV P3, PHV01 MVS, or PHV01 FDS showed 100% protection from MARV, as did animals vaccinated with the medium dose of PHV01 MVS. The medium and low dose of rVSV-MARV P3, as well as the medium dose of PHV01 FDS, resulted in 83% survival, with one of six guinea pigs in each group (animals 9, 16, and 60) succumbing to MARV infection. The low dose of PHV01 MVS resulted in 67% survival, with two of six guinea pigs (animals 31 and 32) succumbing to infection. In total, of the 54 animals that were vaccinated with any vaccine, 49 survived GPA-MARV/Ang challenge, giving an overall vaccine efficacy of ~91%. To specifically assess the level of protection conferred by the clonal PHV01, we pooled the data from the PHV01 MVS and FDS medium- and high-dose groups. All animals (n = 18) that received the high dose of these vaccines survived, while only 1 of 12 animals that received the medium dose succumbed. Of the 30 animals vaccinated with either the high or medium dose of PHV01 MVS or FDS, 29 survived, giving an overall PHV01 vaccine efficacy of 97%. (Zhu et al., 2022)
Description:
PHV01 provided high survival rates against homologous MARV/Ang challenge, protecting 100% of the animals at the highest dose level tested and offering significant protection even at very low dose levels. (Zhu et al., 2022)
18. Marburg Virus Vaccine pVAKS-GPVM
a. Type:
DNA vaccine
b. Status:
Research
c. Host Species for Licensed Use:
None
d. Antigen
Marburgvirus Glycoprotein
e. Gene Engineering of
GP protein [Marburg marburgvirus]
Type:
DNA vaccine construction
Description:
pVAKS-GPVM DNA vaccine contains a gene encoding Marburgvirus glycoprotein.
To construct DNA immunogen, a nucleotide sequence encoding MARV glycoprotein (GP) (GenBank CAA82539.1) was used. Surface GP of the virus con- sisting of GP1 and GP2 subunits (170 and 46 kDa, respectively) linked by disulfide bonds was chosen as the immunogen for immunity development. This GP plays the key role in the tropism of the virus to the target cells. GP1 contains a receptor-binding domain, and numerous sugar residues; glycosylation sites are concentrated in the mucin-like domain (MLD) [8]. Anti-MLD antibodies do not play a role in the protective immunity against MARV [5]. The nucleotide fragment corresponding to the MLD (from 290Leu to 422Asn) was deleted from the MARV GP sequence. The gene in the plasmid pGH and primers for PCR were synthesized by the DNA Synthesis Company. The amplified PCR product corresponding to MARV GP without a MLD was cleaved with restriction enzymes AsuNHI and BseX3I (SibEnzyme) and the vector pVAKS was cleaved with restriction enzymes AsuNHI and PspOMI (SibEnzyme). Then, the restriction products were mixed and ligated using bacteriophage T4 DNA ligase (SibEnzyme) for 30 min at room temperature. Transformation of competent cells of E. coli strain NEB Stable (New England Biolabs) with ligation products was performed using the heat- shock technique. The presence of the insert was confirmed by restriction analysis and Sanger sequencing. DNA for immunization was prepared in 2 liters ofLB nutrient medium (lysogeny broth) supplemented with ampicillin sodium salt (Sintez) at a working con- centration of 25 μg/ml. Plasmid DNA pVAKS-GPVM for immunization of guinea pigs was isolated using an EndoFree Plasmid GigaKit kit (Qiagen) according to manufacturer’s recommendations. (Volkova et al., 2021)
h. Immunization Route
Intramuscular injection (i.m.)
i.
Guinea pig Response
Vaccination Protocol:
Intramuscular immunization of guinea pigs weighing 180-200 g was carried out 3 times with an interval of 28 days. The dose of the immunogen for both pVAKS-GPVM and negative control pVAKS was 600 μg per animal. 3 antigens were used: recombinant MARV GP, virus-like particles based on the recombinant vesicular stomatitis virus, and inactivated MARV [3](Volkova et al., 2021)
Immune Response:
When determining the neutralization titer of serum from guinea pigs against MARV, the sera neutralized the virus in titers from 1:20 to more than 1:40, exhibiting potent neutralizing activity (Volkova et al., 2021)
Side Effects:
The animals have no rash, but develop coagulation defects, including a decrease in platelet count and an increase in coagulation time. (Volkova et al., 2021)
An expression cassette encoding the full-length MARV-Angola GP was cloned into a plasmid containing the full-length VSV genome. This plasmid encodes for a VSV N1 to N4 gene translocation and VSV G CT1 truncation; the MARV-Angola GP or HIV gag gene is expressed from the first genomic position from the single 3’-proximal promoter site to maximize GP antigen expression. Vectors were then recovered from Vero cells following electroporation with the resulting plasmids along with VSV helper plasmids. The rescued virus was plaque purified and amplified to produce virus seed stocks.(Woolsey et al., 2022)
g. Immunization Route
Intramuscular injection (i.m.)
h.
Macaque Response
Vaccination Protocol:
Eighteen adult (9 females and 9 males) cynomolgus macaques (Macaca fascicularis) of Chinese origin (PreLabs, Worldwide Primates) ranging in age from 3 to 8 years and weighing 2.86 to 7.60 kg were used for three separate studies at the GNL. Macaques were immunized with a single 10 million PFU intramuscular (i.m.) injection of rVSV-N4CT1-MARV-GP at 7 (N = 5), 5 (N = 5), or 3 (N = 5) days prior to MARV exposure. Three animals were immunized with an identical dose of rVSVN4CT1-HIVgag at each respective time point to serve as non-specific controls. The inoculation was equally distributed between the left and right quadriceps. (Woolsey et al., 2022)
Immune Response:
Vaccinated survivors expressed greater development of MARV GP-specific antibodies and early expression of predicted NK cell-, B-cell-, and cytotoxic T-cell-type quantities. (Woolsey et al., 2022)
Side Effects:
Regardless of the rVSV vaccine vector administered, all fatal cases presented with typical MVD clinical signs such as fever, anorexia, dyspnea, macular rash, and/or depression. Specifically vaccinated survivors remained healthy and did not display clinical signs of disease other than anorexia at 5 DPI in one subject in the -7 group and transient anorexia and a mild petechial rash in the sole survivor in the -3 group. However, all survivors exhibited various hematological changes over the course of the study. Postmortem gross examination of fatal cases in both specifically and non-specifically vaccinated macaques revealed lesions consistent with MVD including subcutaneous hemorrhage; necrotizing hepatitis; splenomegaly; lymphadenitis; and hemorrhagic interstitial pneumonia (characterized as failure to completely collapse and multifocal reddening of the lungs). No significant lesions were detected in examined tissues of vaccinated survivors at the study endpoint. (Woolsey et al., 2022)
Challenge Protocol:
All macaques were challenged i.m. in the left quadriceps with a uniformly lethal 1000 PFU target dose of MARV-Angola (actual doses were 1475, 1475, and 1300 PFU, respectively). An internal scoring protocol was implemented to track disease progression in challenged animals. (Woolsey et al., 2022)
Efficacy:
Survival rates of groups immunized with rVSVN4CT1-MARV-GP were significantly different than the vector control group with 100% (log-rank test, p = 0.0046) and 80% (p = 0.0153) efficacy for -7 DPI and -5 DPI groups, respectively. No statistical difference (p = 0.5110) was noted for the -3 DPI vaccination group, although a sole subject (20%) survived. (Woolsey et al., 2022)
Description:
rVSV-N4CT1-MARV-GP-mediated protection appears to be at least partially attributed to tight control of virus replication and rapid stimulation of innate immunity. Resolution of the innate immune response coincided with development of adaptive immunity including the generation of MARV GP-specific immunoglobulins, and transcriptional evidence of recruitment of cytotoxic and effector cells. In contrast, non-specific vaccination led to the development of MVD with characteristic uncontrolled virus replication and transcriptional evidence of sustained innate immunity, complement dysregulation, and immune checkpoint expression. (Woolsey et al., 2022)
d. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant protein preparation
Description:
This Musoke GP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Musoke strain of MARV (Wang et al., 2006).
Description:
An RNA replicon based on VEEV was used as the vector, with the VEE structural genes replaced by VP40. VP40 seems to serve as a matrix protein, affecting interactions between the nucleoprotein complex and lipid membrane. It is also the most abundant part of the virion (Hevey et al., 1998).
To generate mVLPs, 293T cells were co-transfected with pWRG 7077 vectors encoding for MARV VP40 and GP using Lipofectamine 2000 (Invitrogen, Carlsbad, CA). To purify the VLPs, the cell supernatants were cleared from cellular debris and subsequently pelleted at 9500×g for 4 h. The crude VLP preparations were then separated on a 20â��60% continuous sucrose gradient centrifuged. The VLPs were concentrated by a second centrifugation and resuspended in endotoxin-free phosphate-buffered saline (PBS) (Warfield et al., 2004).
h. Virulence
Guinea pigs that were vaccinated with inactivated MARV or mVLPs developed MARV-specific antibody titers(Warfield et al., 2004).
i.
Guinea pig Response
Host Strain:
Strain 13
Vaccination Protocol:
Guinea pigs were vaccinated intramuscularly with 50 μg of mVLPs, eVLPs, or iMARV with 200 μl of RIBI monophosphoryl lipid+synthetic trehalose dicorynomycolate+cell wall skeleton emulsion diluted in endotoxin-free PBS (Warfield et al., 2004).
Persistence:
None noted.
Immune Response:
Both inactivated MARV and mVLP induced maximal humoral responses to MARV after only two vaccinations (Warfield et al., 2004) .
Side Effects:
Not noted.
Challenge Protocol:
Thirty days after the third vaccination, the guinea pigs were challenged subcutaneously with 1000 plaque-forming units (pfu) or 2000 LD50 of guinea pig-adapted MARV diluted in PBS [18](Warfield et al., 2004).
Efficacy:
Strong filovirus-specific antibody responses correlate with vaccine protective efficacy in guinea pigs(Warfield et al., 2004).
21. Multivalent DNA vaccine for B. anthracis, Ebola virus, Marburg virus, and VEE virus
Multiple antigens from B. anthracis, Ebola virus, Marburg virus, and VEE virus were used. Specifically, this DNA vaccine includes Protective Antigen (PA) from B. anthracis, Glycoprotein (GP) and Nucleoprotein (NP) from Ebloa virus, Glycoprotein (GP) from Marburg virus strain Ravn, and 26S from VEE virus (Riemenschneider et al., 2003).
d. Gene Engineering of
PagA from Bacillus anthracis
Type:
Recombinant protein preparation
Description:
A DNA vaccine for the anthrax was made by PCR-amplifying the PA gene (Riemenschneider et al., 2003).
Description:
Ebola NP genes were cloned and the vaccine was produced without additional signal sequence with the use of plasmid pWRG7077 (Riemenschneider et al., 2003).
Description:
Ebola NP genes were cloned and the vaccine was produced without additional signal sequence with the use of plasmid pWRG7077 (Riemenschneider et al., 2003).
Description:
This Ravn GP gene was amplified by PCR, and subcloned to create MARV adenovirus vaccine targeted against the Ci67strain of MARV (Wang et al., 2006).
The necessary genes were inserted into expression plasmids following a cytomegalovirus immediate early promotor. MARV was procured through experiementally infected monkeys, then passed three times (Ravn) or one time(Musoke) in Vero cells. Inbred Strain 13 and outbred Hartley guinea pigs were injected subcutaneously with the vaccine. Responses were measured by IgG antibody ELISA with the use of cobalt-irradiated purified MARV in both strains. A study also included non-human primates, which underwent serum tests for viremia determination and blood chemistry(Riemenschneider et al., 2003).
k.
Guinea pig Response
Host Strain:
Strain 13 and Hartley
Vaccination Protocol:
Gun-vaccinated guinea pigs were gene gun-vaccinated three (Musoke) or four (Ravn) times at 4-week intervals with approximately 2.5 μg of the MARV GP DNA (Riemenschneider et al., 2003).
Persistence:
Not noted.
Immune Response:
All of the MARV GP DNA-vaccinated guinea pigs developed antibodies to MARV(Riemenschneider et al., 2003).
Side Effects:
Not noted.
Challenge Protocol:
The challenge was a subcutaneous injection of 1000 plaque forming units (pfu) of homologous virus 4 weeks after the final vaccination for each guinea pig (Riemenschneider et al., 2003).
Efficacy:
Guinea pigs vaccinated with control DNA were viremic at day 7 post-challenge, as measured by plaque assay, and were infected by day 9. All guinea pigs vaccinated with the GP DNA vaccines were aviremic at day 7 and appreared healthy throughout the observation period(Riemenschneider et al., 2003).
Marburg virus (MBGV) GP (Hevey et al., 1998). An RNA replicon from Venezuelan equine encephalitis (VEE) virus was used as a vaccine vector. The VEE structural genes were replaced by genes for MBGV GP, nucleoprotein (NP), VP30, VP35, VP40, or VP24 (Hevey et al., 1998).
d. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant vector construction
Description:
MBGV gene clone pGem-GP was provided by Heinz Feldmann and Anthony Sanchez (Centers for Disease Control and Prevention, Atlanta, GA). The MBGV GP gene from pGem-GP was excised with SalI and subcloned into the SalI site of a shuttle vector. A clone with the MBGV GP gene in the correct orientation was excised with ApaI and NotI, and this fragment was cloned into the ApaI and NotI sites of a VEE replicon plasmid (Hevey et al., 1998).
Description:
An RNA replicon based on VEEV was used as the vector, with the VEE structural genes replaced by VP30. Function has yet to be determined; VP30 is found on the VP40 virion (Hevey et al., 1998).
Description:
An RNA replicon based on VEEV was used as the vector, with the VEE structural genes replaced by VP35. Function has yet to be determined; VP35 is found on the VP40 virion (Hevey et al., 1998).
Description:
An RNA replicon based on VEEV was used as the vector, with the VEE structural genes replaced by VP40. VP40 seems to serve as a matrix protein, affecting interactions between the nucleoprotein complex and lipid membrane. It is also the most abundant part of the virion (Hevey et al., 1998).
Description:
An RNA replicon based on VEEV was used as the vector, with the VEE structural genes replaced by VP24. Function has yet to be determined; VP24 is found on the VP40 virion (Hevey et al., 1998).
Guinea pigs were inoculated with packaged recombinant VEE replicons expressing individual MBGV proteins and later injected with 10^3.3 LD50 guinea pig-adapted MBGV subcutaneously(Hevey et al., 1998).
k. Virulence
MBGV NP protected all vaccinated guinea pigs from both death and viremia, while MBGV VP35 vaccination resulted in a majority of the animals surviving. Four of the five survivors were viremic 7 days after infection(Hevey et al., 1998).
l. Description
Results indicated that VP35 afforded incomplete protection while either GP or NP were protective antigens(Hevey et al., 1998).
m.
Monkey Response
Host Strain:
cynomolgus
Vaccination Protocol:
Several groups of macaques were tested: a group received VRPs that expressed MBGV NP; a group received VRPs that expressed MBGV GP; a group received a mixture of MBGV GP and MBGV NP VRPs; and a group received VRPs that expressed a control antigen (influenza HA)Anti-MBGV ELISA antibody titers were monitored throughout the experiment(Hevey et al., 1998).
Persistence:
Not noted.
Immune Response:
Animals inoculated with replicons that expressed MBGV proteins demonstrated prechallenge ELISA titers to purified MBGV antigen. Of the GP-vaccinated animals that survived challenge, a few demonstrated a modest boost in ELISA antibody titer (10- to 30-fold) when pre and postchallenge samples were compared. The surviving NP-inoculated macaques had larger boosts in ELISA antibody titers when pre- and
postchallenge samples were compared. Some animals vaccinated with both GP and NP also demonstrated 100- to 300-fold rise in ELISA titers(Hevey et al., 1998).
Side Effects:
Not noted.
Challenge Protocol:
Monkeys recieved injections that expressed VEE replicons with either MBGV GP or MBGV NPor both(Hevey et al., 1998).
Efficacy:
All animals that received VEE replicons expressing MBGV GP, either alone or in combination with MBGV NP, survived challenge with 8000 PFU MBGV without any observed signs of illness(Hevey et al., 1998).
A gene coding for a protein of interest is cloned in place of the VEE virus structural genes (Hevey et al., 1998).
f. Virulence
Without any observed signs of illness, all animals that received VEE replicons expressing MBGV GP, either alone or in combination with MBGV NP, survived challenge with 8000 PFU MBGV(Hevey et al., 1998).
g.
Monkey Response
Host Strain:
cynomolgus
Vaccination Protocol:
Several groups of macaques were tested: a group received VRPs that expressed MBGV NP; a group received VRPs that expressed MBGV GP; a group received a mixture of MBGV GP and MBGV NP VRPs; and a group received VRPs that expressed a control antigen (influenza HA)Anti-MBGV ELISA antibody titers were monitored throughout the experiment
Persistence:
Not noted.
Immune Response:
Animals inoculated with replicons that expressed MBGV proteins demonstrated prechallenge ELISA titers to purified MBGV antigen. Of the GP-vaccinated animals that survived challenge, a few demonstrated a modest boost in ELISA antibody titer (10- to 30-fold) when pre and postchallenge samples were compared. The surviving NP-inoculated macaques had larger boosts in ELISA antibody titers when pre- and
postchallenge samples were compared. Some animals vaccinated with both GP and NP also demonstrated 100- to 300-fold rise in ELISA titers(Hevey et al., 1998).
Side Effects:
Not noted.
Challenge Protocol:
Monkeys recieved injections that expressed VEE replicons with either MBGV GP or MBGV NPor both(Hevey et al., 1998).
Efficacy:
All animals that received VEE replicons expressing MBGV GP, either alone or in combination with MBGV NP, survived challenge with 8000 PFU MBGV without any observed signs of illness(Hevey et al., 1998).
Marburg virus (MBGV) glycoprotein (GP) was used. A recombinant vesicular stomatitis virus (VSV) was used as a vector. The vaccine utilizes the VSVΔG/MARVGP-Musoke strain (Daddario-DiCaprio et al., 2006).
d. Gene Engineering of
GP from Musoke Marburgvirus
Type:
Recombinant protein preparation
Description:
The ORF for the glycoproteins were generated by PCR and cloned into GP-lacking VSV vectors (Daddario-DiCaprio et al., 2006).
The ORF for the glycoproteins for the MARV-Musoke and the ZEBOV were generated by PCR and cloned into GP-lacking VSV vectors. Infectious clones for the VSV Indiana serotype were used (Daddario-DiCaprio et al., 2006).
f. Virulence
All animals developed high anti-MARV IgG antibody levels by the challenge time, while low levels of anti-MARV neutralizing antibodies were observed for a large percentage of animals vaccinated with VSVΔG/MARVGP-Musoke at the challenge day. Protection of the host subjects injected with the VSVΔG/MARVGP-Musoke vaccine appears to be associated with humoral response, as opposed to cellular immune response (Daddario-DiCaprio et al., 2006).
g.
Monkey Response
Host Strain:
cynomolgus macaques
Vaccination Protocol:
Nine adult macaques were used. Seven were injected intramuscularly with the VSVΔG/MARVGP-Musoke vaccine, and two recieved VSVΔG/ZEBOVGP as experimental controls (Daddario-DiCaprio et al., 2006).
Persistence:
Not noted.
Immune Response:
With the use of purified virus particles for an antigen source, immunoglobulin G (IgG) antibodies against MARV were detected through an enzyme-linked immunosorbent assay (ELISA). A transient and low-level recombinant VSV viremia was detected through virus isolation on the third day after vaccination in plasma from four of the VSVΔG/MARVGP-Musoke vaccinated animals. Both the MARV-Angola-challenged control animal and the MARV-Ravn-challenged control animal developed high titers in the blood, detected by plaque assay (Daddario-DiCaprio et al., 2006).
Side Effects:
After either vaccination with VSVΔG/MARVGP-Musoke or after the MARV challenge, none of the animals showed any evidence of clinical illness (Daddario-DiCaprio et al., 2006).
Challenge Protocol:
All animals were challenged with either MARV-Angola, MARV-Musoke, or MARV-Ravn 28 days after immunization (Daddario-DiCaprio et al., 2006).
Efficacy:
VSVΔG/MARVGP-Musoke vector does protect nonhuman primates against a lethal challenge with both Ravn and Angola strains of MBGV. This approach seems almost as successful as the use of VEEV MARV GP and/or VEEV MARV NP, which protected NHP against a lethal homologous challenge, but did not protect against a lethal heterologous Ravn challenge (Daddario-DiCaprio et al., 2006).
IV. References
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15. Warfield et al., 2004: Warfield KL, Swenson DL, Negley DL, Schmaljohn AL, Aman MJ, Bavari S. Marburg virus-like particles protect guinea pigs from lethal Marburg virus infection. Vaccine. 2004 Sep 3; 22(25-26); 3495-502. [PubMed: 15308377 ].
