Impaired immune response drives age-dependent severity of COVID-19
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Abstract
Severity of COVID-19 shows an extraordinary correlation with increasing age. We generated a mouse model for severe COVID-19 and show that the age-dependent disease severity is caused by the disruption of a timely and well-coordinated innate and adaptive immune response due to impaired interferon (IFN) immunity. Aggravated disease in aged mice was characterized by a diminished IFN-γ response and excessive virus replication. Accordingly, adult IFN-γ receptor-deficient mice phenocopied the age-related disease severity, and supplementation of IFN-γ reversed the increased disease susceptibility of aged mice. Further, we show that therapeutic treatment with IFN-λ in adults and a combinatorial treatment with IFN-γ and IFN-λ in aged Ifnar1−/− mice was highly efficient in protecting against severe disease. Our findings provide an explanation for the age-dependent disease severity and clarify the nonredundant antiviral functions of type I, II, and III IFNs during SARS-CoV-2 infection in an age-dependent manner. Our data suggest that highly vulnerable individuals could benefit from immunotherapy combining IFN-γ and IFN-λ.
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SciScore for 10.1101/2022.04.21.489072: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Euthanasia Agents: Experimental end points when mice were euthanized by cervical dislocation were defined as ≥25 % weight loss or ≥20 % weight loss for longer than two days.
IACUC: All animal work conducted at the University Medical Center Freiburg and the Francis Crick Institute followed the German animal protection law, or the Animals (Scientific Prpcedures) Act 1986, respectively, and was approved by the respective local animal welfare committee (Regierungspräsidium Freiburg #35-9185.81/G-20/91) or the UK Home Office London
Consent: Written informed consent was obtained from participants and the study was conducted according to federal guidelines and local ethics committee regulations …SciScore for 10.1101/2022.04.21.489072: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Euthanasia Agents: Experimental end points when mice were euthanized by cervical dislocation were defined as ≥25 % weight loss or ≥20 % weight loss for longer than two days.
IACUC: All animal work conducted at the University Medical Center Freiburg and the Francis Crick Institute followed the German animal protection law, or the Animals (Scientific Prpcedures) Act 1986, respectively, and was approved by the respective local animal welfare committee (Regierungspräsidium Freiburg #35-9185.81/G-20/91) or the UK Home Office London
Consent: Written informed consent was obtained from participants and the study was conducted according to federal guidelines and local ethics committee regulations (Albert-Ludwigs-Universität, Freiburg, Germany: No. F-2020-09-03-160428 and no. 322/20).
IRB: Written informed consent was obtained from participants and the study was conducted according to federal guidelines and local ethics committee regulations (Albert-Ludwigs-Universität, Freiburg, Germany: No. F-2020-09-03-160428 and no. 322/20).Sex as a biological variable Animals of both sexes were used. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The primary polyclonal serum was applied overnight at 4 °C (1:3000, diluted in TRIS buffer), the secondary biotinylated goat anti-rabbit antibody was applied for 30 minutes at room temperature (Vector Laboratories, Burlingame, CA, USA, 1:200). anti-rabbitsuggested: NoneExperimental Models: Cell Lines Sentences Resources For virus growth curves, Vero E6 or Calu-3 cells seeded in 24-well plates were washed with PBS and then infected with the respective virus strain by incubating the cells with virus containing Opti-MEM 0.3 % BSA for 2 h at 37 °C with 5 % CO2 using an MOI of 0.001. Calu-3suggested: NoneThe mixture was then added to Vero E6 cells and incubated for 1.5 h at 37 °C with 5 % CO2. Vero E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Mice: C57BL/6JRj, BALB/cJRj and 129S2/SvPasOrlRj mice were purchased from Janvier Labs. C57BL/6JRjsuggested: None129S2/SvPasOrlRjsuggested: NoneB6.A2G-Mx1, B6.A2G-Mx1-Ifnar1−/−, B6.A2G-Mx1-Ifnlr1−/−, B6.AG2-Mx1-Ifnar1−/−Ifnlr1−/−, B6.Ifngr1-/-, B6.Ifnar1-/-Ifngr1-/-, and B6.A2G-Mx1-Stat1-/- were bred and kept at the animal facilities of the University Medical Center Freiburg. B6.A2G-Mx1suggested: NoneB6.A2G-Mx1-Ifnar1−/−suggested: NoneB6.A2G-Mx1-Ifnlr1−/−suggested: NoneB6.AG2-Mx1-Ifnar1−/−Ifnlr1−/−suggested: NoneB6.Ifngr1-/-suggested: RRID:IMSR_JAX:007077)B6.Ifnar1-/-Ifngr1-/-suggested: NoneB6.A2G-Mx1-Stat1-/-suggested: NoneIn addition, to identify subtle inflammation in brain and heart samples from infected 10-week (n=10) and 40-week-old (n=6) C57BL/6, hearts were evaluated for the presence of CD3-positive T cell infiltrates and brains evaluated for the presence of CD3-positive T cells and Iba-1-positive microglial cells/macrophages as described in 80. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources De-multiplexed raw reads were subjected to a customized Galaxy pipeline based on bioinformatics pipelines on usegalaxy.eu68. Galaxysuggested: (Galaxy, RRID:SCR_006281)0.20.169 and mapped to the SARS-CoV-2 Wuhan-Hu-1 reference genome (Genbank: NC_045512) using BWA-MEM v. BWA-MEMsuggested: NoneVariants (SNPs and INDELs) were called using the ultrasensitive variant caller LoFreq v2.1.571, demanding a minimum base quality of 30 and a coverage ≥ 10-fold. LoFreqsuggested: (LoFreq, RRID:SCR_013054)Effects of mutations were automatically annotated in vcf files using SnpEff v.4.3.172. SnpEffsuggested: (SnpEff, RRID:SCR_005191)Read quality trimming and adaptor removal was carried out using Trimmomatic (version 0.36). Trimmomaticsuggested: NoneAll data was processed relative to the mouse GRCm38 genome downloaded from Ensembl. Ensemblsuggested: NoneGene counts per gene per sample were obtained using the RSEM-STAR (77, 78) option of the pipeline and they were imported on DESeq (v1.28.0, 79) within R environment v4.0.2 for differential expression analysis. DESeqsuggested: (DESeq, RRID:SCR_000154)Gene Ontology and Gene Set Enrichment analysis (GSEA) were carried out using R package Cluster Profiler (v3.16). Cluster Profilersuggested: (clusterProfiler, RRID:SCR_016884)Gene signatures were considered significant if FDR q-value ≤ 0.05. ggplot2, RColorBrewer, ComplexHeatmap were used for plotting purposes. ComplexHeatmapsuggested: (ComplexHeatmap, RRID:SCR_017270)Ingenuity Pathway Analysis was performed using differentially expressed genes (fold change ≥ 1.5, padj ≤ 0.05). Ingenuity Pathway Analysissuggested: NoneStatistical analyses: Data visualization and analyses were performed using GraphPad Prism 9.0 and R version 3.5.1. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: Thank you for sharing your code and data.
Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:Small animal models faithfully recapitulating characteristics of human disease are pivotal to overcome these limitations. In this study, we generated a highly pathogenic mouse-adapted SARS-CoV-2 variant that can be used to model mild, moderate or severe COVID-19 as well as the age-associated aggravation of disease in standard inbred mice. Using this small animal model, we found that the age-dependent increase in disease severity is driven by an impaired interferon response which causes a delayed, insufficient and dysregulated innate and adaptive immune response in the aged host. Transcriptome analyses of infected lungs from mature adult and middle-aged C57BL/6 mice revealed that adult mice initiated a rapid and well-coordinated innate and adaptive immune response, which was associated with high IFN-γ and low IL-10 expression levels. This effective and timely immune response in adults limited viral loads, mediated rapid viral clearance, and efficiently prevented the development of severe disease. In aged mice, by contrast, virus replication was markedly increased which correlated with the absence of effective antiviral immune responses. Instead, aged mice showed strong IL-6- and IL-1-mediated responses associated with low IFN-γ and high IL-10 expression levels. The markedly different IFN-γ to IL-10 ratio likely explains the effective immune response, including NK cell mediated immunity, efficient antigen presentation, lymphocyte activation, and immunoglobulin production in adu...
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.
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Results from scite Reference Check: We found no unreliable references.
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