1. Immunology and Inflammation
  2. Microbiology and Infectious Disease

Vaccination induces rapid protection against bacterial pneumonia via training alveolar macrophage in mice

  1. Hao Gu
  2. Xi Zeng
  3. Liusheng Peng
  4. Chuanying Xiang
  5. Yangyang Zhou
  6. Xiaomin Zhang
  7. Jixin Zhang
  8. Ning Wang
  9. Gang Guo
  10. Yan Li
  11. Kaiyun Liu
  12. Jiang Gu
  13. Hao Zeng
  14. Yuan Zhuang
  15. Haibo Li
  16. Jinyong Zhang
  17. Weijun Zhang
  18. Quanming Zou  Is a corresponding author
  19. Yun Shi  Is a corresponding author
  1. West China Biopharmaceutical Research Institute,West China Hospital, Sichuan University, China
  2. National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University, China
  3. Department of Clinical Laboratory, 971st Hospital of People's Liberation Army, China
  4. Department of Phamacy, The 78 th Group Army Hospital of Chinese People's Liberation Army, China
https://doi.org/10.7554/eLife.69951
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  1. Hao Gu
  2. Xi Zeng
  3. Liusheng Peng
  4. Chuanying Xiang
  5. Yangyang Zhou
  6. Xiaomin Zhang
  7. Jixin Zhang
  8. Ning Wang
  9. Gang Guo
  10. Yan Li
  11. Kaiyun Liu
  12. Jiang Gu
  13. Hao Zeng
  14. Yuan Zhuang
  15. Haibo Li
  16. Jinyong Zhang
  17. Weijun Zhang
  18. Quanming Zou
  19. Yun Shi
(2021)
Vaccination induces rapid protection against bacterial pneumonia via training alveolar macrophage in mice
eLife 10:e69951.
https://doi.org/10.7554/eLife.69951
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8 figures, 1 table and 2 additional files

Figures

Figure 1 with 2 supplements
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Rapid protection against A. baumannii pneumonia by a single intranasal vaccination.

(A) Schematic diagram of the experimental procedure. C57BL/6 mice were immunized intranasally (i.n.) with inactivated whole cell (IWC) of A. baumannii strain LAC-4 and challenged intratracheally …

Figure 1—source data 1

Raw data for Figure 1.

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Figure 1—figure supplement 1
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The histopathology images of lungs for each mouse in Figure 1.
Figure 1—figure supplement 2
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Gating strategy for detecting neutrophil, monocyte, and alveolar macrophages in lungs.
Figure 2
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Broad protection against infection of clinical strains by intranasal immunization of inactivated whole cell (IWC) of A. baumannii.

(A) Mice were immunized intranasally (i.n.) with IWC of LAC-4 on day 0. On day 7, mice were infected intratracheally (i.t.) with lethal dose of clinical strains of A. baumannii SJZ24 or SJZ26 (n = …

Figure 2—source data 1

Raw data for the survival rate.

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Figure 3
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Rapid immune memory induced by a single intranasal vaccination.

(A) Dynamic responses of TNF-α and IL-6 in the lungs of A. baumannii inactivated whole cell (IWC) (LAC-4)-immunized mice (n = 3 per timepoint). (B–D) IWC (LAC-4)-immunized mice were challenged …

Figure 3—source data 1

Raw data for Figure 3.

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Figure 4 with 1 supplement
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Trained innate immunity mediates vaccination-induced rapid protection.

(A) Survival of WT and Rag1-/- mice immunized intranasally (i.n.) with inactivated whole cell (IWC) (LAC-4) or PBS, challenged intratracheally (i.t.) by lethal LAC-4 7 days later (n = 5 for WT mice, …

Figure 4—source data 1

Raw data for Figure 4A,E.

https://cdn.elifesciences.org/articles/69951/elife-69951-fig4-data1-v1.xlsx
Figure 4—source data 2

RNA-seq results of reads per kilobase per million reads (RPKM) for all samples and all genes.

https://cdn.elifesciences.org/articles/69951/elife-69951-fig4-data2-v1.xlsx
Figure 4—figure supplement 1
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Supplementary RNA-seq results.

(A–C) RNA-seq data related to Figure 4B. (A) The heatmap of 2401 differentially expressed genes (DEGs) in lungs. (B) Numbers of upregulated and downregulated DEGs. (C) Top 10 Gene Ontology (GO) …

Figure 5 with 1 supplement
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Trained immunity of alveolar macrophages (AMs) mediates rapid protection induced by vaccination.

(A) Heatmap of differentially expressed genes (DEGs) in lungs of inactivated whole cell (IWC) (LAC-4)-immunized and control Rag1-/- mice at day 7 after immunization. (B) Top 10 Gene Ontology (GO) …

Figure 5—source data 1

Raw data for Figure 5.

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Figure 5—figure supplement 1
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Transcriptional difference at day 7 after immunization of inactivated whole cell (IWC) (LAC-4) in Rag1-/- mice.

(A) Numbers of upregulated and downregulated differentially expressed genes (DEGs) of lungs in IWC-7d and control mice. (B) Top 10 mouse gene atlas terms of upregulated genes in lungs of IWC-7d vs. …

Figure 6 with 1 supplement
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Higher TLR4 expression on inactivated whole cell (IWC)-trained alveolar macrophages (AMs) mediates rapid protection.

(A) Heatmap of differentially expressed genes (DEGs) associated with myeloid leukocyte activation at day 7 after A. baumannii IWC (LAC-4) immunization in Rag1-/- mice. (B) Reads per kilobase per …

Figure 6—source data 1

Raw data for Figure 6.

https://cdn.elifesciences.org/articles/69951/elife-69951-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
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Upregulated differentially expressed genes (DEGs) at day 7 after inactivated whole cell (IWC) immunization in Rag1-/- mice.

(A) Top 20 KEGG terms of upregulated DEGs in IWC-7d vs. control mice. (B) Heatmap of DEGs related to defense response (GO: 0006952) and innate immune response (GO0045087).

Figure 7
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A rapid protection induced by intranasal vaccination against other bacteria.

C57BL/6 mice were immunized intranasally (i.n.) with inactivated whole cell (IWC) of (A) P. aeruginosa (IWC(P.a)), n = 5, (B) K. pneumoniae (IWC(K.p)), n = 5, (C) S. aureus (IWC(S.a)), n = 10, or (D)…

Figure 7—source data 1

Raw data for Figure 7.

https://cdn.elifesciences.org/articles/69951/elife-69951-fig7-data1-v1.xlsx
Figure 8
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Cross-protection induced by intranasal vaccination of inactivated whole cell (IWC) of A. baumannii.

C57BL/6 mice were immunized intranasally (i.n.) with IWC of A. baumannii LAC-4 (IWC(A.b)) and challenged intratracheally (i.t.) with (A) P. aeruginosa (1 × 107 CFU, n = 10), (B) K. pneumoniae (1 × …

Figure 8—source data 1

Raw data for Figure 8.

https://cdn.elifesciences.org/articles/69951/elife-69951-fig8-data1-v1.xlsx

Tables

Appendix 1—key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Genetic reagent (Mus musculus)C57BL/6JBeijing HFK Bioscience Limited CompanyN/A
Genetic reagent (Mus musculus)Rag1-/-: B6.129S7-Rag1tm1/NjuModel Animal Research Center of Nanjing UniversityStock no :N000088
Genetic reagent (Mus musculus)Tlr4-/- mice: C57BL/10ScNJNjuModel Animal Research Center of Nanjing UniversityStock no: N000192
Genetic reagent (Mus musculus)C57BL/10JNjuModel Animal Research Center of Nanjing UniversityStock no: N000023
Strain, strain background (Acinetobacter baumannii)LAC-4Gift from Professor Chen, Harris et al., 2013N/A
Strain, strain background (Acinetobacter baumannii)ATCC-17978ATCCATCC-17978
Strain, strain background (Acinetobacter baumannii)SJZ-09Clinical isolate, Zeng et al., 2019

Strain, strain background (Acinetobacter baumannii)SJZ-24Clinical isolate, Zeng et al., 2019

Strain, strain background (Acinetobacter baumannii)SJZ-26Clinical isolate, Zeng et al., 2019

Strain, strain background (Pseudomonas aeruginosa)XN-1Clinical isolate, Gao et al., 2020

Strain, strain background (Klebsiella pneumoniae)YBQClinical isolate, Liu et al., 2020

Strain, strain background (Staphylococcus aureus)KM-22Clinical isolate, Zeng et al., 2020

Strain, strain background (Streptococcus pneumoniae)S. pneumoniaeClinical isolate

AntibodyRat monoclonal anti-mouse CD45-PE/Cy7 (clone 30-F11)BioLegendCat# 103114 RRID:AB_312979(1:100)
AntibodyRat monoclonal anti-mouse/human CD11b-PerCP/Cy5.5 (clone M1/70)BioLegendCat# 101228 RRID:AB_893232(1:100)
AntibodyRat monoclonal anti-mouse F4/80-APC (clone BM8)BioLegendCat# 123116 RRID:AB_893481(1:100)
AntibodyRat monoclonal anti-mouse Ly-6C-PE (clone HK1.4)BioLegendCat# 128008 RRID:AB_1186132(1:100)
AntibodyRat monoclonal anti-mouse Ly-6G-FITC (clone 1A8)BioLegendCat# 127605 RRID:AB_1236488(1:100)
AntibodyHamster monoclonal anti-mouse CD11c-PE (clone N418)BioLegendCat# 117307 RRID:AB_313776(1:100)
AntibodyRat monoclonal anti-mouse CD11c-FITC (clone N418)BioLegendCat# 117305 RRID:AB_313774(1:100)
AntibodyMouse monoclonal anti-mouse CD284(TLR4)-PE (clone UT41)Thermo FisherCat# 12-9041-80 RRID:AB_466236(1:100)
AntibodyRat monoclonal rat IgG1 isotype control (clone HRPN)Bio X CellCat# BE0088, RRID:AB_1107775200 μg/per mouse
AntibodyRat monoclonal anti-mouse TNF-α (clone XT3.11)Bio X CellCat# BE0058, RRID:AB_1107764200 μg/per mouse
Sequence-based reagentPrimers for qPCRThis paperN/ASee Supplementary file 1
Commercial assay or kitRNAiso PlusTakaraCat# 9109
Commercial assay or kitPrimeScript RT reagent kitTakaraCat# RR037A
Commercial assay or kitTB Green Premix Ex Taq IITakaraCat# RR820A
Commercial assay or kitIL-6 ELISA kitBioLegendCat# 431304
Commercial assay or kitTNF-α ELISA kitBioLegendCat# 430904
Commercial assay or kitIL-1β ELISA kitBioLegendCat# 432604
Commercial assay or kitAnti-mouse CD11c Microbeads kitMiltenyi BiotecCat# 130-125-835
Software, algorithmGraphPad PrismGraphPad SoftwareRRID:SCR_002798
software, algorithmFlowJoFlowJoRRID:SCR_008520

Additional files

Transparent reporting form
https://cdn.elifesciences.org/articles/69951/elife-69951-transrepform1-v1.docx
Supplementary file 1

Primers used in real-time PCR.

https://cdn.elifesciences.org/articles/69951/elife-69951-supp1-v1.docx

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