Neutralizing antibody-dependent and -independent immune responses against SARS-CoV-2 in cynomolgus macaques
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Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infectious disease (COVID-19) has been threatening the world because of severe symptoms and relatively high mortality. To develop vaccines and antiviral drugs for COVID-19, an animal model of SARS-CoV-2 infection is required to evaluate the efficacy of prophylactics and therapeutics in vivo . Therefore, we examined the pathogenicity of SARS-CoV-2 in cynomolgus macaques until 28 days after virus inoculation in the present study. Cynomolgus macaques showed body temperature rises after infection and X-ray radiographic viral pneumonia was observed in one of three macaques. However, none of the macaques showed life-threatening clinical signs of disease corresponding that approximately 80% of human patients did not show a critical disease in COVID-19. A neutralizing antibody against SARS-CoV-2 and T-lymphocytes that produced interferon (IFN)-γ and interleukin (IL)-2 specifically for SARS-CoV-2 N protein were detected on day 14 in the macaque that showed viral pneumonia. On the other hand, in the other macaques, in which a neutralizing antibody was not detected, T-lymphocytes that produced IFN-γ specifically for SARS-CoV-2 N protein increased on day 7 to day 14 prior to an increase in the number of T-lymphocytes that produced IL-2. These results suggest that not only a neutralizing antibody but also cellular immunity augmented by IFN-γ has a role in the elimination of SARS-CoV-2. Thus, because of the mild clinical signs of disease and low/no antibody responses against SARS-CoV-2 in two thirds of the macaques, cynomolgus macaques are appropriate to extrapolate human responses in vaccine and drug development.
Author Summary
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infectious disease (COVID-19) has been threatening the world. To develop vaccines and antiviral drugs for COVID-19, an animal model of SARS-CoV-2 infection is required to evaluate their efficacy in vivo . Therefore, we examined the pathogenicity of SARS-CoV-2 in a non-human primate model until 28 days after virus inoculation. Cynomolgus macaques showed a fever after infection and X-ray radiographic viral pneumonia was observed in one of three macaques. However, none of the macaques showed life-threatening symptoms. A neutralizing antibody against SARS-CoV-2 and T-lymphocytes that produced interferon (IFN)-γ and interleukin (IL)-2 specifically for SARS-CoV-2 protein were detected on day 14 in the macaque that showed viral pneumonia. In the other macaques, in which a neutralizing antibody was not detected, T-lymphocytes that produced IFN-γ specifically for SARS-CoV-2 N protein increased on day 7 to day 14. These results suggest that not only a neutralizing antibody but also cellular immunity augmented by IFN-γ has a role in the elimination of SARS-CoV-2. Thus, because of the mild symptoms and low/no antibody responses against SARS-CoV-2 in two thirds of the macaques, cynomolgus macaques are appropriate to extrapolate human responses in vaccine and drug development.
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SciScore for 10.1101/2020.08.18.256446: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: The protocols were approved by the Shiga University of Medical Science Animal Experiment Committee (permit number: 2020-4-2). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Animals: Fifteen-year-old and ten-year-old female cynomolgus macaques (CE0324F and CE1242F) and a fifteen-year-old male cynomolgus macaque (CE0202M) born at Shiga University of Medical Science were used. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Thawed cells (5 × 105/well) were cultured with a peptide pool of SARS-CoV-2 N protein (0.6 nmol/mL) (PepTivator, Miltenyi Biotech, Bergisch Gladbach, … SciScore for 10.1101/2020.08.18.256446: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: The protocols were approved by the Shiga University of Medical Science Animal Experiment Committee (permit number: 2020-4-2). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Animals: Fifteen-year-old and ten-year-old female cynomolgus macaques (CE0324F and CE1242F) and a fifteen-year-old male cynomolgus macaque (CE0202M) born at Shiga University of Medical Science were used. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Thawed cells (5 × 105/well) were cultured with a peptide pool of SARS-CoV-2 N protein (0.6 nmol/mL) (PepTivator, Miltenyi Biotech, Bergisch Gladbach, Germany) in the presence of anti-CD28 antibody (0.1 µg/mL) overnight in ELISPOT plates coated with anti-IFN-γ and IL-2 antibodies (Cellular Technology Limited, Shaker Heights, OH). anti-CD28suggested: Noneanti-IFN-γsuggested: NoneIL-2suggested: NoneExperimental Models: Cell Lines Sentences Resources VeroE6 cells were grown in MEM supplemented with 10% inactivated fetal bovine serum (Capricorn Scientific GmbH), VeroE6suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)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: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.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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