DOI: 10.5061/dryad.vq83bk43q
title: Neonatal infection with Bordetella pertussis promotes autism-like phenotypes in mice

description: Autism spectrum disorder (ASD) has been linked with infections early in
 life. Here we demonstrate that infection of neonatal mice with the
 respiratory pathogen Bordetella pertussis leads to neuroinflammation,
 neurodevelopmental defects and ASD-like behaviours. Following respiratory
 challenge of neonatal mice with B. pertussis, multiple atypical CNS
 findings were observed, including blood-brain barrier disruption,
 dissemination of live B. pertussis bacteria to the brain with concomitant
 infiltration of inflammatory monocytes, neutrophils and activated IL-17A-
 and IFN-γ-producing CD4 T cells. Microglia from infected mice were
 activated, with impaired phagocytic function, resulting in defective
 synaptic pruning and disrupted neuronal circuit formation. Impaired
 neurodevelopment in B. pertussis-infected post-natal mice was associated
 with ASD-like behavioural abnormalities in young adulthood. Our data
 indicate that infection with virulent B. pertussis during infancy
 increases the risk of autism-like behaviour in young adult mice. Study
 into the potential role of B. pertussis in human ASD is warranted.
description: # Neonatal infection with Bordetella pertussis promotes autism-like
 phenotypes in mice
 [https://doi.org/10.5061/dryad.vq83bk43q](https://doi.org/10.5061/dryad.vq83bk43q) ## Description of the data and file structure The data are structured with treatment groups (Control vs. B. pertussis) vertically in column 1 and the headings for each metric analyzed are structured horizontally in row 1. For clarity, there is a separate excel sheet for raw data within each Figure. Empty cells are not applicable and are replaced with ''n/a'' PTx: Pertussis toxin concentration in Lung or Brain (ng/ml). MHCII LY6C Monocytes: Number of activated monocytes in Lung or Brain (# cells) LY6G Neutrophils: Number of neutrophils in Lung or Brain (# cells) IFN-G or IL-17A T-Cells: Number of IFN-G or IL-17A producing T-cells in Lung or Brain (# cells) MPO: Myeloperoxidase expression in brain tissue (integrated density) CXCL1: The chemokine (C-X-C motif) ligand 1 (CXCL1) CD44 T cell: type I transmembrane protein and member of the cartilage link protein family, marker for effector memory T cells Fibrinogen: Expression levels of Fibrinogen in brain tissue (% Area) IL17A: Interleukin-17A concentration in brain tissue analyzed by ELISA  IL-17A+: Number of Interleukin-17A-producing T cells IFN-γ+: Number of Interferon-gamma-producing T cells Microglia/Brain: Number of microglial cells in brain tissue (%) Branches/Microglia: Number of branches per microglia cell Junctions/Microglia: Number of junctions per microglia cell HPC CD68 Volume: Volume of CD68 protein expression inside microglia in the hippocampus CTX CD68 Volume: Volume of CD68 protein expression inside microglia in the cortex HPC Phagocytic index: Volume of CD68 in hippocampus divided by Iba1 x 100 (%) CTX Phagocytic index: Volume of CD68 in cortex divided by Iba1 x 100 (%) HPC VGLUT2/SMI32: Colocalization of VGLUT2 with SMI32+ neurons in hippocampus CTX VGLUT2/SMI32: Colocalization of VGLUT2 with SMI32+ neurons in cortex HPC VGLUT2 ENGULFMENT: Volume of internalized VGLUT2 by microglia in hippocampus  (μm^3) CTX VGLUT2 ENGULFMENT: Volume of internalized VGLUT2 by microglia in cortex  (μm^3) HPC IBA1 MFI: Mean fluorescence intensity of IBA1 in hippocampus (arbitrary units) CTX IBA1 MFI: Mean fluorescence intensity of IBA1 in cortex (arbitrary units) HPC MAP2: Volume of internalized MAP2 protein by microglia in hippocampus (μm^3) CTX MAP2: Volume of internalized MAP2 protein by microglia in cortex (μm^3) HPC %AREA: Area of Golgi+ neurons in hippocampus (%) HPC INTDEN: Area of Golgi+ neurons in hippocampus (integrated density) CTX %AREA: Area of Golgi+ neurons in cortex (%) CTX INTDEN: Area of Golgi+ neurons in cortex (integrated density) Marbles buried: Number of marbles buried over 30min trial  Grooming: Time spent grooming (minutes) Rears: Number of rears performed over a 5 minute trial NOR: Novel object recognition task exploration time (%) Latency to dig: Duration of time before digging behaviour commences (minutes)  Social interaction: Time spent interacting with another mouse (minutes) LUNG: Colony forming units (CFU) per lung homogenate Nasal wash: Colony forming units (CFU) per nasal wash DAY 15 NEUTROPHILS NOSE: Number of Neutrophils in nasal tissue at day 15 post-infection assessed by flow cytometry DAY 6 NEUTROPHILS NOSE: Number of Neutrophils in nasal tissue at day 6 post-infection assessed by flow cytometry DAY 15 MHCII MONOCYTES: Number of Monocytes in nasal tissue at day 15 post-infection assessed by flow cytometry DAY 6 MHCII MONOCYTES: Number of Monocytes in nasal tissue at day 6 post-infection assessed by flow cytometry IFN T-cells: Number of interferon-gamma producing T-cells in lung at day 15 post-infection assessed by flow cytometry IL17A T-cells: Number of interleukin-17 producing T-cells in lung at day 15 post-infection assessed by flow cytometry HPC LCN2: Relative mRNA expression of Lipocalin-2 in hippocampus CTX LCN2: Relative mRNA expression of Lipocalin-2 in cortex Squares crossed: Number of squares crossed in the open field over a 5 minute time period ### Files and variables #### File: Figure\_1\_raw\_data.xlsx #### File: Figure\_2\_raw\_data.xlsx #### File: Figure\_3\_raw\_data.xlsx #### File: Figure\_4\_raw\_data.xlsx #### File: Figure\_5\_raw\_data.xlsx #### File: Figure\_6\_raw\_data.xlsx #### File: Figure\_7\_raw\_data.xlsx #### File: Supplemental\_information.xlsx **Description:** We have submitted our raw data for our flow cytometry analyses, confocal immunofluorescence data and Behavioural data. Each excel file contains one sheet corresponding to Figures 1-7 in our paper. We have also supplied our raw data in supplemental information for supplementary figures S1, S2, S3, S4, S5, S7 & S9 with a separate excel sheet for the raw data contained in each figure. ##### Variables * The main variable is neonatal infection with B. pertussis. All data is analyzed via Students unpaired 2-tailed t-tests. ## Code/software GraphPad Prism version 9 & Microsoft Excel ## Access information Other publicly accessible locations of the data: n/a Data was derived from the following sources: n/a