Evidence for anti-viral effects of complete Freund’s adjuvant in the mouse model of enterovirus infection
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Abstract
Group B Coxsackieviruses belonging to the genus, Enterovirus, contain six serotypes that induce various diseases, whose occurrence may involve the mediation of more than one serotype. We recently identified immunogenic epitopes within CVB3 viral protein 1 that induce anti-viral T cell responses in mouse models of CVB infections. In our investigations to determine the protective responses of the viral epitopes, we unexpectedly noted that animals immunized with complete Freund’s adjuvant (CFA) alone and later challenged with CVB3 were completely protected against myocarditis. Similarly, the pancreatitis-inducing ability of CVB3 was remarkably reduced to only 10% in the CFA group as opposed to 73.3% in the control group that received no CFA. Additionally, no mortalities were noted in the CFA group, whereas 40% of control animals died during the course of 21 days post-infection with CVB3. Taken together, our data suggest that the adjuvant effects of CFA may be sufficient for protection against CVB infections. These observations may provide new insights into our understanding of the occurrence of viral infections. One example is Coronavirus disease-19 (COVID-19) as individuals suffering from COVID-19 who have been vaccinated with Bacillus Calmette–Guérin appear to have fewer morbidities and mortalities than unvaccinated individuals.
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SciScore for 10.1101/2020.05.27.120121: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Animals were maintained according to the institutional guidelines of the University of Nebraska-Lincoln (UNL), Lincoln, NE, and approval for animal studies was granted by the Institutional Animal Care and Use Committee, UNL (protocol #1904, approved January 2, Randomization not detected. Blinding The analysis was performed by a board-certified pathologist blinded to treatment, and total number of inflammatory foci were obtained as reported previously [10, 16]. Power Analysis not detected. Sex as a biological variable Mice: Six-to-eight-week old, female A/J mice (H-2a) were procured from the Jackson Laboratory (Bar Harbor, ME, USA). Cell Line Authentication not … SciScore for 10.1101/2020.05.27.120121: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Animals were maintained according to the institutional guidelines of the University of Nebraska-Lincoln (UNL), Lincoln, NE, and approval for animal studies was granted by the Institutional Animal Care and Use Committee, UNL (protocol #1904, approved January 2, Randomization not detected. Blinding The analysis was performed by a board-certified pathologist blinded to treatment, and total number of inflammatory foci were obtained as reported previously [10, 16]. Power Analysis not detected. Sex as a biological variable Mice: Six-to-eight-week old, female A/J mice (H-2a) were procured from the Jackson Laboratory (Bar Harbor, ME, USA). Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources The adherent Vero cells were grown to 80 to 90% confluence in 75cm2 flasks in EMEM/10% fetal bovine serum (FBS) and were later infected with CVB3 with multiplicity of infection 1 in EMEM containing no FBS. Verosuggested: NoneExperimental Models: Organisms/Strains Sentences Resources Mice: Six-to-eight-week old, female A/J mice (H-2a) were procured from the Jackson Laboratory (Bar Harbor, ME, USA). A/Jsuggested: RRID:IMSR_JAX:000646)Software and Algorithms Sentences Resources Statistics: Generalized linear mixed models were used to analyze the data pertaining to body weights and survival curves using Proc Glimmix in SAS (Version 9.3, SAS Institute Inc. SAS Institutesuggested: (Statistical Analysis System, RRID:SCR_008567)Graphs were prepared by GraphPad Prism software Version 8.0 GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)(GraphPad Software, Inc. La Jolla, CA, USA). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:One limitation of our study is that we did not investigate the presence of virus in tissues of CVB3-infected mice. Similarly, it is unknown whether CFA administration can potentiate the production of protective neutralizing antibodies to CVB3, and if so, how long such an effect would last against different doses of virus. Likewise, whether administration of CFA in the face of CVB3 infection can mitigate the disease process is also unknown. At the time of this writing, we could not execute these experiments since our institutional guidelines do not allow any new animal experiments because of the ongoing threat of COVID-19 to the public. Nonetheless, our data may provide insights into our understanding of the occurrence of viral infections in the face of pre-existing, non-antigen-specific immune responses generated in response to a potentially wide range of environmental pathogens/microbes or gut microbiota over a period of time, which also may include formation of virtual memory cells [57, 58].
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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