An mRNA Vaccine Encoding Rabies Virus Glycoprotein Induces Protection against Lethal Infection in Mice and Correlates of Protection in Adult and Newborn Pigs

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Death commonly follows within an average survival time between 6 and 11 days after first symptomatic onset for furious or paralytic forms, leaving little time and extremely limited therapeutic options.

“While incubation phases vary, death commonly follows within an average survival time between 6 and 11 days after first symptomatic onset for furious or paralytic forms, respectively, thus leaving little time and extremely limited therapeutic options [1,2].”

• cites [1] T Hemachudha, 2013, Human rabies: neuropathogenesis, diagnosis, and management, Lancet Neurology sourceNot checked — no full text of the source
• cites [2] S Pathak, 2014, Diagnosis, management and post-mortem findings of a human case of rabies imported into the United Kingdom from India: a case report, Virology Journal sourcePartly supported

  The Background says clinical rabies is invariably fatal, and the Conclusion describes limited evidence for the Milwaukee Protocol’s efficacy. The Case presentation records one patient’s deterioration through day 10, but the paper does not give average survival times of 6 and 11 days after symptom onset for furious and paralytic rabies, respectively. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Novel vaccine formats such as subunit, DNA or viral vector vaccines have been assessed successfully for protective capacity against rabies infection in preclinical settings, but none resulted in a licensed product for human use yet.

“Novel vaccine formats such as subunit, DNA or viral vector vaccines have been assessed successfully for their protective capacity against rabies infection in preclinical settings [3,13] but none of these resulted in a licensed product for human use yet.”

• cites [3] HCJ Ertl, 2009, Novel vaccines to human rabies, PLoS neglected tropical diseases sourceSupported
• cites [13] R Amann, 2013, A new rabies vaccine based on a recombinant ORF virus (parapoxvirus) expressing the rabies virus glycoprotein, Journal of Virology sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Benefits of mRNA vaccines are the induction of balanced and enduring immunity as demonstrated for anti-tumor and prophylactic vaccination, simple supply, and storage at elevated temperature.

“Benefits are the induction of balanced and enduring immunity as demonstrated for anti-tumor and prophylactic vaccination [14–16], simple supply, and storage at elevated temperature, as reviewed elsewhere [17,18].”

• cites [14] I Hoerr, 2000, In vivo application of RNA leads to induction of specific cytotoxic T lymphocytes and antibodies, Eur J Immunol sourceNot checked — no full text of the source
• cites [15] M Fotin-Mleczek, 2011, Messenger RNA-based vaccines with dual activity induce balanced TLR-7 dependent adaptive immune responses and provide antitumor activity, J Immunother sourceNot checked — no full text of the source
• cites [16] EJ Solari, 2012, Differentiating Literacy Growth of ELL Students With LD From Other High-Risk Subgroups and General Education Peers: Evidence From Grades 3–10, J Learn Disabil sourceNot checked — no full text of the source
• cites [17] K-J Kallen, 2013, A novel, disruptive vaccination technology: self-adjuvanted RNActive vaccines, Human Vaccines & Immunotherapeutics sourceSupported
• cites [18] T Kramps, 2013, Messenger RNA-based vaccines: progress, challenges, applications, Wiley interdisciplinary reviews RNA sourceNot checked — no full text of the source

May combine two claims.

After vaccination with 20 μg and 80 μg RABV-G mRNA or with Rabipur, antibody titers remained stable throughout the observation period with mean titers between 17 and 65 IU/ml.

“We found that after vaccination with 20 μg and 80 μg RABV-G mRNA or with Rabipur antibody titers remained stable throughout the observation period with mean titers between 17 and 65 IU/ml (Fig 1E).”

May combine two claims.

In C57BL/6 mice, immunization with 10 μg RABV-G mRNA vaccine revealed comparable immunogenicity to HDC, and significantly higher titers were achieved at mRNA doses of ≥20 μg.

“In C57BL/6 mice immunization with a dose of 10 μg RABV-G mRNA vaccine (range: 0.5 to 40.6 IU/ml) revealed comparable immunogenicity to HDC (range: 1.5 to 13.5 IU/ml) and significantly higher titers were achieved at mRNA doses of ≥20 μg (range: 17.8 to 92.4 IU/ml).”

May combine two claims.

In both mouse strains, a dose-response correlation was seen and high neutralizing antibody titers were induced at doses ≥10 μg mRNA.

“Thus, in both mouse strains, a dose-response correlation was seen and high neutralizing antibody titers were induced at doses ≥10 μg mRNA.”

May combine two claims.

mRNA vaccination led to early, efficient viral clearance and brain homeostasis upon intracerebral infection.

“Moreover, we demonstrate the early, efficient viral clearance and brain homeostasis by mRNA vaccination upon intracerebral infection.”

May combine two claims.

In splenocytes of RABV-G mRNA vaccinated mice, IFN-γ secreting cells were detected upon Rabipur stimulation, but not after stimulation with BSA or in splenocytes of buffer treated control mice.

“This analysis revealed that in splenocytes of RABV-G mRNA vaccinated mice, IFN-γ secreting cells were detected upon Rabipur stimulation, but not after stimulation with BSA (negative control) or in splenocytes of buffer treated control mice (Fig 1B).”

May combine two claims.

For the 1.25 μg and 5 μg mRNA vaccine doses, response onset was delayed, but caught up by week 12 and titers remained at a high level of ≥10 IU/ml.

“For the 1.25 μg and 5 μg mRNA vaccine doses, response onset was delayed, but caught up by week 12 of the experiment and titers remained at a high level of ≥ 10 IU/ml.”

May combine two claims.

In the CD4-depleted mRNA-vaccinated group, all mice had to be sacrificed within 11 days after infection with similar kinetics as the negative control group.

“In the CD4-depleted, mRNA vaccinated group, all mice had to be sacrificed within 11 days after infection with similar kinetics as in the negative control group”

May combine two claims.

All Rabipur-vaccinated undepleted positive control mice survived lethal i.c. CVS-11 challenge, while buffer-injected negative control mice had to be sacrificed by day nine post challenge due to dramatic weight loss.

“All mice vaccinated with Rabipur (undepleted positive control) survived the challenge infection while buffer injected mice (negative controls) had to be sacrificed by day nine post challenge due to dramatic weight loss”

May combine two claims.

In buffer-treated BALB/c mice at day 3 after i.c. CVS-11 infection, low N-protein mRNA was detectable in the telencephalon (mean ct 31.4) but not the cerebellum; by day 6 high viral loads were detected in both telencephalon (mean ct 23.1) and cerebellum (mean ct 24.7).

“At day 3 after infection a low quantity of N-protein mRNA was detectable in the telencephalon of animals treated with buffer (mean: 31.4 cycle threshold (ct)-value) whereas the cerebellum was negative for viral N-protein mRNA (mean above detection limit). Ct-values, however, decreased over the period of infection as at day 6 high viral loads were detected in both analyzed parts of the brains (mean”

May combine two claims.

In buffer-treated mice, viral replication was found in the telencephalon and spread to the cerebellum, and both regions showed increased TNFα expression.

“In buffer-treated mice we found viral replication in the telencephalon (the site of injection) and spreading to the cerebellum. In addition, both telencephalon and cerebellum of these animals showed increased TNFα expression”

Probably not a substantial claim: may not pass S3 (load-bearing).

Inter-individual variability was low in all test groups.

“Inter-individual variability was low in all test groups.”

May combine two claims.

At day 9 after infection, first buffer treated mice had to be sacrificed due to pronounced weight loss (25% of initial body weight) and none in this group survived the infection.

“In contrast, at day 9 after infection, first buffer treated mice had to be sacrificed due to pronounced weight loss (25% of initial body weight) and none in this group survived the infection.”

May combine two claims.

In CD4-depleted mRNA-immunized mice, VN titers were strongly reduced: 3/8 had titers below 0.5 IU/ml and the mean titer was about 3 IU/ml, more than 15-fold lower than the CD4-intact mRNA-vaccinated group.

“In the CD4-depleted, mRNA immunized group VN titers were strongly reduced: 3 out of 8 mice showed VN titers below 0.5 IU/ml and the mean titer of the groups was about 3 IU/ml, i.e. more than 15-fold reduced compared to the CD4-intact, mRNA-vaccinated group.”

May combine two claims. The quote only partly states this claim.

In RABV-G mRNA and Rabipur vaccinated mice, TNFα mRNA transcript levels were reduced compared to mock-vaccinated controls at both analysis time points and brain sections, though not statistically significant.

“In mRNA and Rabipur vaccinated mice TNFα mRNA transcript level was reduced compared to mock vaccinated control group at both analysis time points and different brain sections”

• Also stated in: § Discussion · hedged

  “Though not statistically significant, the data indicate a reduced innate inflammatory reaction”

May combine two claims.

In groups with an intact CD4+ T cell compartment during prophylactic immunization, more than 87% of mRNA-vaccinated mice survived infection; one of eight mRNA-immunized mice without detectable VN antibodies after immunization was sacrificed on day ten due to pronounced weight loss.

“In groups with an intact CD4+ T cell compartment during prophylactic immunization, more than 87% of mRNA-vaccinated mice survived infection (one out of eight mRNA-immunized mice without detectable virus neutralizing antibodies after immunization had to be sacrificed on day ten due to pronounced weight loss)”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Two mice treated with HDC showed a transient weight loss (10-20% of initial body weight), but survived the experiment.

“Two mice treated with HDC showed a transient weight loss (10–20% of initial body weight), but survived the experiment (Fig 3B).”

May combine two claims.

Eight weeks post second boost, RABV-G specific neutralizing antibodies in adult pigs had a mean titer of 2.9 IU/ml and were still above 0.5 IU/ml.

“Eight weeks post second boost RABV-G specific neutralizing antibodies had a mean titer of 2.9 IU/ml and were still above the 0.5 IU/ml limit.”

May combine two claims.

In newborn piglets, neutralizing antibody titers after first vaccination were in the protective range and further increased after booster vaccination.

“Furthermore, immunogenicity of the RABV-G mRNA was also investigated in newborn piglets, again showing neutralizing antibody titers after the first vaccination in the protective range that were further increased after the booster vaccination.”

May combine two claims.

In adult pigs, titers reached the protective limit of ≥0.5 IU/ml after the first immunization and could be further increased by booster vaccination, then remaining stable throughout the experiment.

“In adult pigs, titers reached the protective limit of ≥0.5 IU/ml after the first immunization and could be further increased by booster vaccination, then remaining stable throughout the experiment.”

May combine two claims.

The RABV-G mRNA vaccine was immunogenic upon two injections at microgram doses and induced virus neutralizing antibodies, antigen-specific CD4+ T cells and CD8+ T cells.

“This mRNA vaccine encoding the Rabies virus glycoprotein was immunogenic upon two injections at microgram doses and resulted in the induction of virus neutralizing antibodies, antigen-specific CD4+ T cells and CD8+ T cells (Fig 1).”

May combine two claims.

Piglets receiving 80 μg RABV-G mRNA developed mean protective titers of 11.0 IU/ml at day 28 and 2.5 IU/ml at day 70, while piglets immunized with 240 μg had higher mean antibody titers of 56.8 IU/ml at day 28 and 8.3 IU/ml at day 70, comparable to full doses of licensed human vaccine.

“Piglets receiving 80 μg RABV-G mRNA developed mean protective titers of 11.0 IU/ml at day 28 and 2.5 IU/ml at day 70, while piglets immunized with the 240 μg dose revealed higher mean antibody titers of 56.8 IU/ml at day 28 and 8.3 IU/ml at day 70 that were comparable to those induced by full doses of the licensed human vaccine”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

In India, more human fatalities are caused by rabies than in any other country, with an estimated 16,000 deaths in 2010.

“In India, more human fatalities are caused by rabies than in any other country of the world with an estimated 16,000 deaths in the year 2010 alone [10].”

• cites [10] World Health O (2013) WHO Expert Consultation on Rabies. Second report. World Health Organization Technical Report Series: 1–139, back cover.Not checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Despite the availability of effective vaccines, rabies infection continues to claim at least 55,000 human lives per year.

“Despite the availability of effective vaccines, rabies infection continues to claim at least 55,000 human lives per year (WHO Weekly epidemiological record, No. 49/50, 2007), not counting the great excess of fatal infections in animals.”

• cites WHO Weekly epidemiological record, No. 49/50, 2007Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S3 (load-bearing).

All animal experiments were conducted according to German and Hungarian laws and guidelines for animal protection.

“All animal experiments were conducted according to German and Hungarian laws and guidelines for animal protection.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Experiments in Germany were approved by the regional council Tübingen (reference numbers FLI238/08, FLI242/09, FLI246/09, CUR 4-13).

“Experiments in Germany were approved by the regional council Tübingen (reference numbers FLI238/08, FLI242/09, FLI246/09, CUR 4–13).”

Classified as probably not a substantial claim. May combine two claims.

In case of initiation of vaccination after exposure, protection has first to be built up, which requires time, as done by PEP vaccination.

“In contrast thereto, protection has first to be built up in case of initiation of vaccination after exposure to the rabies virus, which requires time, as it is done by PEP vaccination.”

Classified as probably not a substantial claim.

Anti-rabies serum antibodies were analyzed by FAVN test according to WHO protocol.

“Anti-rabies serum antibodies were analyzed by FAVN test by Eurovir Hygiene-Institut, Luckenwalde, Germany according to WHO protocol [30].”

• cites [30] F Cliquet, 1998, Development of a fluorescent antibody virus neutralisation test (FAVN test) for the quantitation of rabies-neutralising antibody, Journal of Immunological Methods sourceNot checked — no full text of the source

Classified as probably not a substantial claim.

Effective immunological defense against rabies must intercept virus before productive neuronal infection.

“As a consequence, effective immunological defense against rabies must intercept virus before productive neuronal infection.”

May combine two claims.

These experiments indicate comparable immunogenicity of the mRNA vaccine with licensed rabies vaccine and the general induction of specific immune response in mice.

“These experiments, therefore, indicate comparable immunogenicity of the mRNA vaccine with licensed rabies vaccine and the general induction of specific immune response in mice.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Rabies immunoglobulin (RIG) is administered together with active PEP vaccination in suspected rabies exposure in rabies naïve individuals because protection takes time to build up.

“That is the reason for the administration of rabies immunoglobulin (RIG) together with active PEP vaccination in the case of suspected rabies exposure in rabies naïve individuals.”

May combine two claims.

Protection is most effective when neutralizing antibodies and clearing T cells are already present in situ at the time of infection, which is the case after prophylactic vaccination.

“This is most effective when the neutralizing antibodies and clearing T cells are already present in situ at the time of infection, which is the case after prophylactic vaccination.”

May combine two claims.

Depletion of CD4+ T cells during immunization results in loss of protection against lethal challenge, indicating a need for T cell help in mounting a protective neutralizing antibody response.

“Our finding that depletion of CD4+ T cells during the immunization phase results in a loss of protection against lethal challenge infection further indicates the need for T cell help in mounting a protective neutralizing antibody response.”

May combine two claims.

The reduced TNFα mRNA expression in vaccinated mice indicates a reduced innate inflammatory reaction which might result in maintenance of brain homeostasis upon vaccination with RABV-G mRNA or licensed rabies vaccine.

“Though not statistically significant, the data indicate a reduced innate inflammatory reaction which might result in the maintenance of brain homeostasis upon vaccination with RABV-G mRNA or licensed rabies vaccine.”

May combine two claims.

These results indicate efficient RABV-G-specific T cell induction by RABV-G mRNA and better induction of CD4+ T cells by the mRNA vaccine compared to Rabipur.

“These results indicate efficient RABV-G-specific T cell induction by RABV-G mRNA and better induction of CD4+ T cells by the mRNA vaccine compared to Rabipur.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Intracerebral infection forms the basis of the widely used NIH potency test for rabies vaccines.

“Importantly, it also forms the basis of the widely used National Institutes of Health (NIH) potency test for rabies vaccines [40].”

• cites [40] F.-X. KMM Meslin, 1996Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

T cells do not prevent initial viral infection of host cells, but provide important impact on rabies virus clearance in mice.

“Although T cells do not prevent initial viral infection of host cells, they provide important impact on rabies virus clearance in mice [38].”

• cites [38] TJ Wiktor, 1978, Cell-mediated immunity and postexposure protection from rabies by inactivated vaccines of tissue culture origin, Developments in Biological StandardizationNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Rabies-specific T cell clones have been isolated from human vaccinees.

“Rabies-specific T cell clones have also been isolated from human vaccinees [39].”

• cites [39] E Celis, 1988, Genetic restriction and fine specificity of human T cell clones reactive with rabies virus, Journal of Immunology (Baltimore, Md: 1950) sourceNot checked — no full text of the source

May combine two claims.

The study further substantiated and extended previously published work on non-replicating influenza-specific mRNA vaccines, proving protective efficacy of mRNA vaccination in a second, harsh viral challenge model.

“In summary, we further substantiated and extended previously published work on non-replicating influenza-specific mRNA vaccines [16], proving the protective efficacy of mRNA vaccination in a second, harsh viral challenge model.”

• cites [16] EJ Solari, 2012, Differentiating Literacy Growth of ELL Students With LD From Other High-Risk Subgroups and General Education Peers: Evidence From Grades 3–10, J Learn Disabil sourceNot checked — no full text of the source

May combine two claims.

The mRNA-based rabies vaccine is immunogenic and protective in animals.

“In the present study, we demonstrated the immunogenicity and protective efficacy of an experimental mRNA-based rabies vaccine in animals.”

May combine two claims.

A third vaccination did not result in a further strong titer increase, suggesting a maximal humoral immune response by the prime-boost strategy in pigs.

“Interestingly, a third vaccination did not result in a further strong titer increase, suggesting a maximal humoral immune response by the prime-boost strategy in pigs.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The data support and extend previous findings for mRNA vaccines against influenza in adult and newborn mice.

“These data support and extend previous findings for mRNA vaccines against influenza in adult and newborn mice [16].”

• cites [16] EJ Solari, 2012, Differentiating Literacy Growth of ELL Students With LD From Other High-Risk Subgroups and General Education Peers: Evidence From Grades 3–10, J Learn Disabil sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Rabies virus-induced encephalitis results in elevation of pro-inflammatory cytokines, and increased TNFα and IFN-γ have been causally linked to increased BBB permeability allowing strong influx of immune factors.

“It has been reported that rabies virus-induced encephalitis results in an elevation of different pro-inflammatory cytokines. Increased levels of TNFα and IFN-γ have been causally linked to increased BBB permeability which also allows the strong influx of immune factors [42,43].”

• cites [42] TW Phares, 2006, Regional differences in blood-brain barrier permeability changes and inflammation in the apathogenic clearance of virus from the central nervous system, Journal of Immunology (Baltimore, Md: 1950) sourceNot checked — no full text of the source
• cites [43] A Solanki, 2009, Correlation of cytokine expression with rabies virus distribution in rabies encephalitis, Journal of Neuroimmunology sourceNot checked — no full text of the source

May combine two claims.

RABV-G vaccine is immunogenic in newborn piglets, leading to rapid seroconversion and indicating comparable VN titer kinetics for mRNA vaccine compared to licensed Rabipur vaccine.

“These data demonstrate that RABV-G vaccine is immunogenic in newborn piglets, leading to rapid seroconversion and indicating comparable VN titer kinetics for mRNA vaccine compared to the licensed Rabipur vaccine.”