Dataset DOI: 10.5061/dryad.0p2ngf2c1

Description of the data and file structure

This dataset was collected to investigate the role of capsular polysaccharide (CPS) dynamic synthesis in the pathogenesis of Streptococcus suis serotype 2 (SS2) strain 05ZYH33, using a combination of in vitro assays and an intranasal mouse infection model. In vitro experiments included bacterial growth curve measurements, hydrophobicity (BATH) assays to quantify CPS thickness, adherence assays to epithelial and brain microvascular endothelial cells, and bactericidal assays in mouse blood. In vivo experiments involved intranasal inoculation of mice, followed by quantification of bacterial burden (CFU) in nasal-associated lymphoid tissue, cerebrospinal fluid, lung, blood, spleen, and heart at various time points. Re-isolated bacteria from different body compartments were further analyzed for CPS thickness by BATH assay and transmission electron microscopy. Passive immunization with anti-CPS and anti-V5 (non-CPS virulence factors) sera was performed to evaluate their protective effects at different infection stages. Histological and immunofluorescence analyses were conducted to trace the bacterial invasion route through the olfactory nerve pathway. All raw data supporting the findings are provided in the files listed below.

Files and variables

File: Figure_1A-source_data_1.xlsx

Description: Raw data for Figure 1A, showing the growth curve of S. suis 05ZYH33 (serotype 2) cultured in tryptic soy broth (TSB) containing 5% newborn bovine serum (FBS). The optical density at 600 nm (OD600) was measured at hourly intervals from 1 to 14 hours post-inoculation to monitor bacterial growth. These data were used to define the lag, logarithmic (log), and stationary growth phases for subsequent experiments. Five technical replicates (1#–5#) were performed.

Variables

• Hour – Time post-inoculation (hours).
• 1# – OD600 (optical density at 600 nm) of replicate 1.
• 2# – OD600 (optical density at 600 nm) of replicate 2.
• 3# – OD600 (optical density at 600 nm) of replicate 3.
• 4# – OD600 (optical density at 600 nm) of replicate 4.
• 5# – OD600 (optical density at 600 nm) of replicate 5.

File: Figure_1A-source_data_2.xlsx

Description: Raw data for Figure 1A, representing an independent repeat of the growth curve experiment for S. suis 05ZYH33 (serotype 2) cultured in TSB containing 5% FBS. OD600 measurements were taken at hourly intervals from 1 to 14 hours post-inoculation. This dataset, together with source_data_1, comprises the three independent experiments (n = 10 technical replicates total) shown in Figure 1A. Five technical replicates (1#–5#) were performed.

Variables

• Hour – Time post-inoculation (hours).
• 1# – OD600 (optical density at 600 nm) of replicate 1.
• 2# – OD600 (optical density at 600 nm) of replicate 2.
• 3# – OD600 (optical density at 600 nm) of replicate 3.
• 4# – OD600 (optical density at 600 nm) of replicate 4.
• 5# – OD600 (optical density at 600 nm) of replicate 5.

File: Figure_1B-source_data_2.xlsx

Description: Raw data for Figure 1B, representing an independent repeat of the BATH assay measuring hydrophobicity of S. suis 05ZYH33 in different growth phases (lag, log, and stationary phases) and the CPS-deficient strain Δ2BSS2 (CPS⁻). PBS and NC (nasal cavity, i.e., bacterial suspension without hydrocarbon) served as negative controls. PC refers to Δ2BSS2 (positive control for high hydrophobicity due to lack of CPS). Data represent four independent replicates.

Variables

• PBS – Negative control (phosphate-buffered saline without bacteria), BATH value (%).
• NC – Negative control (bacterial suspension without hydrocarbon addition), BATH value (%).
• Δ2BSS2 (CPS⁻) – CPS-deficient mutant strain Δ2BSS2 in log phase (positive control for high hydrophobicity), BATH value (%).
• Lag – 05ZYH33 in lag phase, BATH value (%).
• Log – 05ZYH33 in logarithmic (log) phase, BATH value (%).
• Sta – 05ZYH33 in stationary phase, BATH value (%).

Note: BATH value (%) is calculated as [(Ac - Ab) / Ac] × 100, where Ac is the OD600 of the bacterial control and Ab is the OD600 of the aqueous layer after hydrocarbon mixing.

File: Figure_1B-source_data_3.png

Description: Source image for Figure 1B, showing a representative photograph of the BATH assay tubes. The top layer (hydrocarbon phase) contains adherent hydrophobic bacteria, while the lower aqueous layer contains less hydrophobic bacteria. Lanes are labeled as PBS (negative control), NC (bacterial suspension without hydrocarbon, negative control), PC (Δ2BSS2, positive control), Lag (05ZYH33 lag phase), Log (05ZYH33 log phase), and Sta (05ZYH33 stationary phase).

File: Figure_1C-source_data_3.png

Description: Source image for Figure 1C, showing a representative photograph of the BATH assay tubes for different S. suis serotypes (SS2/05ZYH33, SS3, SS7, SS9, and SS1/2) in both log and stationary phases. PBS served as the negative control. The upper hydrocarbon layer contains adherent hydrophobic bacteria, while the lower aqueous layer contains less hydrophobic bacteria.

File: Figure_4A-source_data_3.png

Description: Source image for Figure 4A, showing a representative photograph of the BATH assay tubes for 05ZYH33 re-isolated from different body compartments (NALT, blood, and CSF) of infected mice. PBS served as the negative control, NC (bacterial suspension without hydrocarbon) served as the no-hydrocarbon control, and Δ2BSS2 (CPS-deficient mutant) served as the positive control. The upper hydrocarbon layer contains adherent hydrophobic bacteria, while the lower aqueous layer contains less hydrophobic bacteria.

File: Figure_4B_source_date_1.tif

Description: Source image for Figure 4B, showing transmission electron microscopy (TEM) images of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2. The image contains six panels (a–f) arranged as follows: Panel a: Δ2BSS2 (CPS-deficient mutant) cultured in vitro in stationary phase – shows no visible CPS layer on the bacterial surface (negative control).

File: Figure_4B_source_date_2.tif

Description: Source image for Figure 4B, showing transmission electron microscopy (TEM) images of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2. The image contains six panels (a–f) arranged as follows: Panel b: 05ZYH33 cultured in vitro in log phase – shows no CPS on the dividing cell (left, larger cell) and only a thin, partial CPS layer on the smaller dividing cell (right).

File: Figure_4B_source_date_3.tif

Description: Source image for Figure 4B, showing transmission electron microscopy (TEM) images of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2. The image contains six panels (a–f) arranged as follows: Panel c: 05ZYH33 cultured in vitro in stationary phase – shows a thicker and more uniform CPS layer on the bacterial surface.

File: Figure_4B_source_date_4.tif

Description: Panel d: 05ZYH33 re-isolated from NALT (nasal-associated lymphoid tissue) of infected mice – shows a thinner and sparser CPS layer on the bacterial surface.

File: Figure_4B_source_date_5.tif

Description: Panel e: 05ZYH33 re-isolated from CSF (cerebrospinal fluid) of infected mice – shows a highly dense outermost layer without an apparent CPS layer.

File: Figure_4B_source_date_6.tif

Description: Panel f: The image contains six panels (a–f) arranged as follows: 05ZYH33 re-isolated from blood of infected mice – shows a much thicker, denser, and uniformly distributed CPS layer compared to NALT and CSF isolates.

File: Figure_6.tif

Description: Source image for Figure 6, showing the detection of S. suis 05ZYH33 in the olfactory nerve pathway and histological analysis of brain inflammation. This file contains the composite figure with six panels (A–F) as described below. Scale bars and labels are as indicated in the figure.

File: Figure_6A_source_date_1.tif

Description: Source image for Figure 6A panel a, showing a sagittal section of the distal nasal cavity and olfactory epithelium from an uninfected (control) mouse. Immunofluorescence staining was performed with anti-S. suis antibody (red/orange), anti-β-tubulin III (green, neuronal marker), and DAPI (blue, DNA). No bacterial signal is detected in the control tissue. Scale bar = 100 μm.

File: Figure_6A_source_date_3.tif

Description: Source image for Figure 6A panel c, showing a higher magnification sagittal section of the olfactory epithelium region from an uninfected (control) mouse. Immunofluorescence staining was performed with anti-S. suis antibody (red/orange), anti-β-tubulin III (green, neuronal marker), and DAPI (blue, DNA). The tissue architecture remains intact with no bacterial signal. Scale bar = 100 μm. NC, nasal cavity; OE, olfactory epithelium; LP, lamina propria.

File: Figure_6A_source_date_2.tif

Description: Source image for Figure 6A panel b, showing a sagittal section of the distal nasal cavity and olfactory epithelium from a mouse infected with 05ZYH33 (1 hour post-inoculation). Immunofluorescence staining was performed with anti-S. suis antibody (red/orange), anti-β-tubulin III (green, neuronal marker), and DAPI (blue, DNA). Scattered and grouped bacteria (orange signal, arrowheads) are observed on the surface of and within the olfactory epithelium (OE), accompanied by tissue destruction. Scale bar = 100 μm. NC, nasal cavity; OE, olfactory epithelium.

File: Figure_6A_source_date_4.tif

Description: Source image for Figure 6A panel d, showing a higher magnification sagittal section of the olfactory epithelium region from a mouse infected with 05ZYH33 (1 hour post-inoculation). Immunofluorescence staining was performed with anti-S. suis antibody (red/orange), anti-β-tubulin III (green, neuronal marker), and DAPI (blue, DNA). Bacteria (orange signal, arrowheads) are observed invading the olfactory epithelium, accompanied by severe tissue destruction (indicated by loss of immunofluorescence signal). Scale bar = 100 μm. NC, nasal cavity; OE, olfactory epithelium; LP, lamina propria.

File: Figure_6A_source_date_5.tif

Description: Source image for Figure 6A panel e, showing a sagittal section of the olfactory bulb region from an uninfected (control) mouse. Immunofluorescence staining was performed with anti-S. suis antibody (red/orange), anti-β-tubulin III (green, neuronal marker), and DAPI (blue, DNA). No bacterial signal is detected in the olfactory bulb of the control tissue. Scale bar = 100 μm. OB, olfactory bulb; CP, cribriform plate.

File: Figure_6A_source_date_6.tif

Description: Source image for Figure 6A panel f, showing a sagittal section of the olfactory bulb region from a mouse infected with 05ZYH33 (1 hour post-inoculation). Immunofluorescence staining was performed with anti-S. suis antibody (red/orange), anti-β-tubulin III (green, neuronal marker), and DAPI (blue, DNA). Large groups of bacteria (orange signal) are observed within the olfactory bulb (OB), indicating nasal-CNS transmigration along the olfactory nerve pathway. Scale bar = 100 μm. OB, olfactory bulb; CP, cribriform plate.

File: Figure_6C_source_date_2.tif

Description: Source image for Figure 6C panel b, showing a coronal brain section from an uninfected (control) mouse stained with H&E (higher magnification view of panel a). The section displays the region of the ventral striatum and basal forebrain. No pathological changes are observed. Scale bar as indicated in the figure.

File: Figure_6C_source_date_1.tif

Description: Source image for Figure 6C panel a, showing a coronal brain section from an uninfected (control) mouse stained with hematoxylin and eosin (H&E). The section displays the region of the ventral striatum and basal forebrain located behind the anterior olfactory nucleus. No inflammatory cell infiltration is observed in the control tissue. Scale bar as indicated in the figure.

File: Figure_6C_source_date_3.tif

Description: Source image for Figure 6C panel c, showing a coronal brain section from a mouse infected with 05ZYH33 (1 hour post-inoculation) stained with H&E. The section displays the region of the ventral striatum and basal forebrain. Arrows indicate infiltrated inflammatory cells in the lower areas of the ventral striatum or basal forebrain, with surrounding edema. Scale bar as indicated in the figure.

File: Figure_6C_source_date_4.tif

Description: Source image for Figure 6C panel d, showing a coronal brain section from a mouse infected with 05ZYH33 (1 hour post-inoculation) stained with H&E (higher magnification view of panel c). Arrows indicate infiltrated inflammatory cells. Early brain inflammation is observed at this early time point post-infection. Scale bar as indicated in the figure.

File: Figure_6C_source_date_5.tif

Description: Source image for Figure 6C panel e, showing a coronal brain section from a mouse infected with 05ZYH33 (9 days post-inoculation) stained with H&E. The section displays the region of the ventral striatum and basal forebrain. More severe pathological changes, including extensive inflammatory cell infiltration and tissue damage, are observed compared to the 1-hour time point. Scale bar as indicated in the figure.

File: Figure_6C_source_date_6.tif

Description: Source image for Figure 6C panel f, showing a coronal brain section from a mouse infected with 05ZYH33 (9 days post-inoculation) stained with H&E (higher magnification view of panel e). Arrows indicate infiltrated inflammatory cells. Severe inflammation and tissue destruction are observed in the distal part of the brain tissue. Scale bar as indicated in the figure.

File: Figure_1C-source_data_1.xlsx

Description: Raw data for Figure 1C, showing the hydrophobicity of five different S. suis serotype strains (05ZYH33 serotype 2, SS3, SS7, SS9, and SS1/2) in both logarithmic (log) and stationary (Sta) growth phases, as measured by bacterial adhesion to hydrocarbon (BATH) assay. Higher BATH values indicate greater surface hydrophobicity, which reflects lower levels of CPS on the bacterial surface. PBS and NC served as negative controls. Data represent three independent replicates (1#–3#).

Variables

• Phase – Designation of bacterial growth phase and strain identity for each column.
• 05ZYH33-Log – 05ZYH33 (serotype 2) in log phase, BATH value (%).
• 05ZYH33-Sta – 05ZYH33 (serotype 2) in stationary phase, BATH value (%).
• SS3-Log – Serotype 3 strain in log phase, BATH value (%).
• SS3-Sta – Serotype 3 strain in stationary phase, BATH value (%).
• SS7-Log – Serotype 7 strain in log phase, BATH value (%).
• SS7-Sta – Serotype 7 strain in stationary phase, BATH value (%).
• SS9-Log – Serotype 9 strain in log phase, BATH value (%).
• SS9-Sta – Serotype 9 strain in stationary phase, BATH value (%).
• SS1/2-Log – Serotype 1/2 strain in log phase, BATH value (%).
• SS1/2-Sta – Serotype 1/2 strain in stationary phase, BATH value (%).
• PBS – Negative control (phosphate-buffered saline without bacteria), BATH value (%).
• NC – Negative control (bacterial suspension without hydrocarbon addition), BATH value (%).

Note: BATH value (%) is calculated as [(Ac - Ab) / Ac] × 100, where Ac is the OD600 of the bacterial control and Ab is the OD600 of the aqueous layer after hydrocarbon mixing.

File: Figure_1C-source_data_2.xlsx

Description: Raw data for Figure 1C, representing an independent repeat of the BATH assay measuring hydrophobicity of five different S. suis serotype strains (05ZYH33 serotype 2, SS3, SS7, SS9, and SS1/2) in both logarithmic (log) and stationary (Sta) growth phases. PBS and NC served as negative controls. Data represent three independent replicates (1#–3#).

Variables

• Phase – Designation of bacterial growth phase and strain identity for each column.
• 05ZYH33-Log – 05ZYH33 (serotype 2) in log phase, BATH value (%).
• 05ZYH33-Sta – 05ZYH33 (serotype 2) in stationary phase, BATH value (%).
• SS3-Log – Serotype 3 strain in log phase, BATH value (%).
• SS3-Sta – Serotype 3 strain in stationary phase, BATH value (%).
• SS7-Log – Serotype 7 strain in log phase, BATH value (%).
• SS7-Sta – Serotype 7 strain in stationary phase, BATH value (%).
• SS9-Log – Serotype 9 strain in log phase, BATH value (%).
• SS9-Sta – Serotype 9 strain in stationary phase, BATH value (%).
• SS1/2-Log – Serotype 1/2 strain in log phase, BATH value (%).
• SS1/2-Sta – Serotype 1/2 strain in stationary phase, BATH value (%).
• PBS-Log – Negative control (PBS without bacteria) processed with log phase samples, BATH value (%).
• NC-Log – Negative control (bacterial suspension without hydrocarbon addition), BATH value (%).

Note: BATH value (%) is calculated as [(Ac - Ab) / Ac] × 100, where Ac is the OD600 of the bacterial control and Ab is the OD600 of the aqueous layer after hydrocarbon mixing.

File: Figure_2A-source_data_1.xlsx

Description: Raw data for Figure 2A, showing the adherence rate of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2 to human laryngeal epithelial (HEp-2) cells. Bacteria were tested in both logarithmic (log) and stationary growth phases at an MOI of 10. Adherence rate is expressed as the percentage of bacteria associated with epithelial cells, calculated as (CFU in wells with cells / CFU in wells without cells) × 100. This dataset represents one independent experiment with multiple technical replicates per group.

Variables

• Strain-Log / Strain-Stationary – Row headers indicating the bacterial strain and growth phase (log phase on the left, stationary phase on the right) and the number of technical replicates (1#–11# for log phase; 1–13 for stationary phase).
• 05ZYH33-Log – 05ZYH33 (wild-type serotype 2) in log phase, adherence rate (%).
• Δ2BSS2-Log – CPS-deficient mutant Δ2BSS2 in log phase, adherence rate (%).
• 05ZYH33-Stationary – 05ZYH33 (wild-type serotype 2) in stationary phase, adherence rate (%).
• Δ2BSS2-Stationary – CPS-deficient mutant Δ2BSS2 in stationary phase, adherence rate (%).

File: Figure_2A-source_data_2.xlsx

Description: Raw data for Figure 2A, representing an independent repeat of the adherence assay measuring the association of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2 with HEp-2 cells. Bacteria were tested in both logarithmic (log) and stationary growth phases at an MOI of 10. Adherence rate is expressed as the percentage of bacteria associated with epithelial cells, calculated as (CFU in wells with cells / CFU in wells without cells) × 100.

Variables

• Strain-Log / Strain-Stationary – Row headers indicating the bacterial strain and growth phase (log phase on the left, stationary phase on the right) and the number of technical replicates (1#–5# for log phase; 1–3 for stationary phase).
• 05ZYH33-Log – 05ZYH33 (wild-type serotype 2) in log phase, adherence rate (%).
• Δ2BSS2-Log – CPS-deficient mutant Δ2BSS2 in log phase, adherence rate (%).
• 05ZYH33-Stationary – 05ZYH33 (wild-type serotype 2) in stationary phase, adherence rate (%).
• Δ2BSS2-Stationary – CPS-deficient mutant Δ2BSS2 in stationary phase, adherence rate (%).

File: Figure_2B-source_data_1.xlsx

Description: Raw data for Figure 2B, showing the adherence rate of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2 to human brain microvascular endothelial cells (hBMECs), which constitute the major component of the blood-brain barrier (BBB). Bacteria were tested in both logarithmic (log) and stationary growth phases at an MOI of 10. Adherence rate is expressed as the percentage of bacteria associated with hBMECs, calculated as (CFU in wells with cells / CFU in wells without cells) × 100. Data represent one independent experiment with multiple technical replicates per group.

Variables

• Strain-Log / Strain-Stationary – Row headers indicating the bacterial strain and growth phase (log phase on the left, stationary phase on the right) and the number of technical replicates (1#–4# for log phase; 1–3 for stationary phase).
• 05ZYH33-Log – 05ZYH33 (wild-type serotype 2) in log phase, adherence rate (%).
• Δ2BSS2-Log – CPS-deficient mutant Δ2BSS2 in log phase, adherence rate (%).
• 05ZYH33-Stationary – 05ZYH33 (wild-type serotype 2) in stationary phase, adherence rate (%).
• Δ2BSS2-Stationary – CPS-deficient mutant Δ2BSS2 in stationary phase, adherence rate (%).

Note: Adherence rate (%) = (CFU in wells with cells / CFU in wells without cells) × 100.

File: Figure_2B-source_data_2.xlsx

Description: Raw data for Figure 2B, representing an independent repeat of the adherence assay measuring the association of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2 with hBMECs. Bacteria were tested in both logarithmic (log) and stationary growth phases at an MOI of 10. Adherence rate is expressed as the percentage of bacteria associated with hBMECs, calculated as (CFU in wells with cells / CFU in wells without cells) × 100.

Variables：

• Strain-Log / Strain-Stationary – Row headers indicating the bacterial strain and growth phase (log phase on the left, stationary phase on the right) and the number of technical replicates (1#–12# for log phase; 1–11 for stationary phase).
• 05ZYH33-Log – 05ZYH33 (wild-type serotype 2) in log phase, adherence rate (%).
• Δ2BSS2-Log – CPS-deficient mutant Δ2BSS2 in log phase, adherence rate (%).
• 05ZYH33-Stationary – 05ZYH33 (wild-type serotype 2) in stationary phase, adherence rate (%).
• Δ2BSS2-Stationary – CPS-deficient mutant Δ2BSS2 in stationary phase, adherence rate (%).

File: Figure_2C-source_data_1.xlsx

Description: Raw data for Figure 2C, showing the survival rate of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2 after incubation with whole blood from naive mice for 3 hours. Bacteria were tested in both logarithmic (log) and stationary growth phases. The anti-killing capacity (survival rate) is calculated as (CFU in blood / CFU in serum) × 100%. Higher values indicate greater resistance to bactericidal activity in blood. Data represent one independent experiment with multiple technical replicates per group.

Variables

• Strain-Log / Strain-Stationary – Row headers indicating the bacterial strain and growth phase (log phase on the left, stationary phase on the right) and the number of technical replicates (1#–6# for log phase; 1–5 for stationary phase).
• 05ZYH33-Log – 05ZYH33 (wild-type serotype 2) in log phase, survival rate (%).
• Δ2BSS2-Log – CPS-deficient mutant Δ2BSS2 in log phase, survival rate (%).
• 05ZYH33-Stationary – 05ZYH33 (wild-type serotype 2) in stationary phase, survival rate (%).
• Δ2BSS2-Stationary – CPS-deficient mutant Δ2BSS2 in stationary phase, survival rate (%).

Note: Survival rate (%) = (CFU in blood after 3 h incubation / CFU in serum control) × 100%.

File: Figure_2C-source_data_2.xlsx

Description: Raw data for Figure 2C, representing an independent repeat of the bactericidal assay measuring the survival of S. suis 05ZYH33 and the CPS-deficient mutant Δ2BSS2 after incubation with whole blood from naive mice for 3 hours. Bacteria were tested in both logarithmic (log) and stationary growth phases. The anti-killing capacity (survival rate) is calculated as (CFU in blood / CFU in serum) × 100%.

Variables

• Strain-Log / Strain-Stationary – Row headers indicating the bacterial strain and growth phase (log phase on the left, stationary phase on the right) and the number of technical replicates (1#–3# for log phase; 1–4 for stationary phase).
• 05ZYH33-Log – 05ZYH33 (wild-type serotype 2) in log phase, survival rate (%).
• Δ2BSS2-Log – CPS-deficient mutant Δ2BSS2 in log phase, survival rate (%).
• 05ZYH33-Stationary – 05ZYH33 (wild-type serotype 2) in stationary phase, survival rate (%).
• Δ2BSS2-Stationary – CPS-deficient mutant Δ2BSS2 in stationary phase, survival rate (%).

Note: This file uses a compact matrix format where column "E" contains the stationary phase group labels, and columns "F–I" contain the corresponding data. Survival rate (%) = (CFU in blood after 3 h incubation / CFU in serum control) × 100%.

File: Figure_3A-source_data_1.xlsx

Description: Raw data for Figure 3A, showing the bacterial burden (total colony-forming units, CFU) of S. suis 05ZYH33 in nasal-associated lymphoid tissue (NALT) of mice at various time points following intranasal inoculation. Mice were inoculated intranasally with 05ZYH33 (stationary phase, 2 × 10⁸ CFU), and NALT was collected at 0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation. Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in NALT of mouse 1.
• 2# – Total CFU in NALT of mouse 2.
• 3# – Total CFU in NALT of mouse 3.

Note: CFU values are expressed as total CFU per NALT organ. Values of "0" indicate that no bacteria were detected (below the detection limit).

File: Figure_3A-source_data_2.xlsx

Description: Raw data for Figure 3A, representing an independent repeat of the time-course bacterial burden assay measuring total CFU of S. suis 05ZYH33 in NALT of mice at various time points following intranasal inoculation (0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation). Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in NALT of mouse 1.
• 2# – Total CFU in NALT of mouse 2.
• 3# – Total CFU in NALT of mouse 3.

Note: CFU values are expressed as total CFU per NALT organ. Values of "0" indicate that no bacteria were detected (below the detection limit).

File: Figure_3B-source_data_2.xlsx

Description: Raw data for Figure 3B, representing an independent repeat of the time-course bacterial burden assay measuring total CFU of S. suis 05ZYH33 in CSF of mice at various time points following intranasal inoculation (0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation). Total CFU in CSF was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in CSF of mouse 1.
• 2# – Total CFU in CSF of mouse 2.
• 3# – Total CFU in CSF of mouse 3.

File: Figure_3C-source_data_1.xlsx

Description: Raw data for Figure 3C, showing the bacterial burden (total colony-forming units, CFU) of S. suis 05ZYH33 in the lungs of mice at various time points following intranasal inoculation. Mice were inoculated intranasally with 05ZYH33 (stationary phase, 2 × 10⁸ CFU), and lung tissue was collected at 0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation. Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in lung of mouse 1.
• 2# – Total CFU in lung of mouse 2.
• 3# – Total CFU in lung of mouse 3.

File: Figure_3C-source_data_2.xlsx

Description: Raw data for Figure 3C, representing an independent repeat of the time-course bacterial burden assay measuring total CFU of S. suis 05ZYH33 in the lungs of mice at various time points following intranasal inoculation (0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation). Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in lung of mouse 1.
• 2# – Total CFU in lung of mouse 2.
• 3# – Total CFU in lung of mouse 3.

File: Figure_3D-source_data_1.xlsx

Description: Raw data for Figure 3D, showing the bacterial burden (total colony-forming units, CFU) of S. suis 05ZYH33 in the blood of mice at various time points following intranasal inoculation. Mice were inoculated intranasally with 05ZYH33 (stationary phase, 2 × 10⁸ CFU), and blood samples were collected at 0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation. Total CFU in blood was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent six biological replicates (1#–6#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in blood of mouse 1.
• 2# – Total CFU in blood of mouse 2.
• 3# – Total CFU in blood of mouse 3.
• 4# – Total CFU in blood of mouse 4.
• 5# – Total CFU in blood of mouse 5.
• 6# – Total CFU in blood of mouse 6 ("no" indicates no sample collected for this mouse at any time point).

File: Figure_3D-source_data_2.xlsx

Description: Raw data for Figure 3D, representing an independent repeat of the time-course bacterial burden assay measuring total CFU of S. suis 05ZYH33 in the blood of mice at various time points following intranasal inoculation (0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation). Total CFU in blood was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent six biological replicates (1#–6#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in blood of mouse 1.
• 2# – Total CFU in blood of mouse 2.
• 3# – Total CFU in blood of mouse 3.
• 4# – Total CFU in blood of mouse 4.
• 5# – Total CFU in blood of mouse 5.
• 6# – Total CFU in blood of mouse 6 ("no" indicates no sample collected for this mouse at any time point).

File: Figure_3E-source_data_1.xlsx

Description: Raw data for Figure 3E, showing the bacterial burden (total colony-forming units, CFU) of S. suis 05ZYH33 in the spleen of mice at various time points following intranasal inoculation. Mice were inoculated intranasally with 05ZYH33 (stationary phase, 2 × 10⁸ CFU), and spleen tissue was collected at 0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation. Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in spleen of mouse 1.
• 2# – Total CFU in spleen of mouse 2.
• 3# – Total CFU in spleen of mouse 3.

File: Figure_3E-source_data_2.xlsx

Description: Raw data for Figure 3E, representing an independent repeat of the time-course bacterial burden assay measuring total CFU of S. suis 05ZYH33 in the spleen of mice at various time points following intranasal inoculation (0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation). Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in spleen of mouse 1.
• 2# – Total CFU in spleen of mouse 2.
• 3# – Total CFU in spleen of mouse 3.

File: Figure_3F-source_data_1.xlsx

Description: Raw data for Figure 3F, showing the bacterial burden (total colony-forming units, CFU) of S. suis 05ZYH33 in the heart of mice at various time points following intranasal inoculation. Mice were inoculated intranasally with 05ZYH33 (stationary phase, 2 × 10⁸ CFU), and heart tissue was collected at 0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation. Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in heart of mouse 1.
• 2# – Total CFU in heart of mouse 2.
• 3# – Total CFU in heart of mouse 3.

File: Figure_3F-source_data_2.xlsx

Description: Raw data for Figure 3F, representing an independent repeat of the time-course bacterial burden assay measuring total CFU of S. suis 05ZYH33 in the heart of mice at various time points following intranasal inoculation (0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation). Total CFU per organ was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in heart of mouse 1.
• 2# – Total CFU in heart of mouse 2.
• 3# – Total CFU in heart of mouse 3.

File: Figure_4A-source_data_1.xlsx

Description: Raw data for Figure 4A, showing the hydrophobicity (BATH value) of S. suis 05ZYH33 re-isolated from different body compartments of infected mice 12 hours post-intranasal inoculation. Bacteria were re-isolated from NALT (nasal-associated lymphoid tissue), blood, and CSF (cerebrospinal fluid), then cultured to logarithmic phase for BATH assay. Higher BATH values indicate greater surface hydrophobicity, which reflects lower levels of CPS on the bacterial surface. The CPS-deficient mutant Δ2BSS2 served as a positive control for high hydrophobicity, while PBS and NC (bacterial suspension without hydrocarbon) served as negative controls. Data represent three independent replicates from one independent experiment.

Variables

• PBS-BATH(%) – Negative control (phosphate-buffered saline without bacteria), BATH value (%).
• NC-BATH(%) – Negative control (bacterial suspension without hydrocarbon addition), BATH value (%).
• Δ2BSS2-BATH(%) – CPS-deficient mutant Δ2BSS2 (positive control for high hydrophobicity), BATH value (%).
• NALT-BATH(%) – 05ZYH33 re-isolated from NALT, BATH value (%).
• Blood-BATH(%) – 05ZYH33 re-isolated from blood, BATH value (%).
• CSF-BATH(%) – 05ZYH33 re-isolated from CSF, BATH value (%).

Note: BATH value (%) is calculated as [(Ac - Ab) / Ac] × 100, where Ac is the OD600 of the bacterial control and Ab is the OD600 of the aqueous layer after hydrocarbon mixing.

File: Figure_4A-source_data_2.xlsx

Description: Raw data for Figure 4A, representing an independent repeat of the BATH assay measuring hydrophobicity of S. suis 05ZYH33 re-isolated from NALT, blood, and CSF of infected mice 12 hours post-intranasal inoculation. The CPS-deficient mutant Δ2BSS2 served as a positive control, while PBS and NC served as negative controls. Data represent four independent replicates (one replicate has missing values) from one independent experiment.

Variables

• PBS-BATH(%) – Negative control (PBS without bacteria), BATH value (%).
• NC-BATH(%) – Negative control (bacterial suspension without hydrocarbon addition), BATH value (%).
• Δ2BSS2-BATH(%) – CPS-deficient mutant Δ2BSS2 (positive control), BATH value (%).
• NALT-BATH(%) – 05ZYH33 re-isolated from NALT, BATH value (%).
• Blood-BATH(%) – 05ZYH33 re-isolated from blood, BATH value (%).
• CSF-BATH(%) – 05ZYH33 re-isolated from CSF, BATH value (%).

Note: BATH value (%) is calculated as [(Ac - Ab) / Ac] × 100. Empty cells indicate no data collected for that replicate.

File: Figure_5B-source_data_2.xlsx

Description: Raw data for Figure 5B, representing an independent repeat of the passive immunization experiment measuring bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments. Mice received anti-CPS or anti-V5 serum via the i.n. route, were challenged i.n. with 05ZYH33, and CFU were determined in NALT, lung, blood, and CSF 12 hours post-challenge. Negative serum served as the control. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, Lung, Blood, CSF.
• Negative serum1 to -5 – Total CFU in mice receiving naive (negative) serum (five individual mice).
• CPS Ab1 to -5 – Total CFU in mice receiving anti-CPS serum (five individual mice).
• V5 Ab1 to -5 – Total CFU in mice receiving anti-V5 serum (five individual mice).

Note: CFU values are expressed as total CFU per organ or per sample. Values of "0" indicate that no bacteria were detected (below the detection limit).

File: Figure_5C-source_data_1.xlsx

Description: Raw data for Figure 5C, showing the bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments of mice following passive immunization with anti-CPS or anti-V5 serum administered via the intravenous (i.v.) route. Mice received antisera i.v., were challenged i.n. with 05ZYH33 6 hours later, and were euthanized 12 hours post-challenge. CFU were determined in NALT (nasal-associated lymphoid tissue), lung, blood, and CSF (cerebrospinal fluid). Negative serum (naive serum) served as the control. Data represent four biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, Lung, Blood, CSF.
• Negative serum 1 to -4 – Total CFU in mice receiving naive (negative) serum (four individual mice).
• CPS Ab1 to -4 – Total CFU in mice receiving anti-CPS serum (four individual mice).
• V5 Ab1 to -4 – Total CFU in mice receiving anti-V5 serum (four individual mice).

Note: CFU values are expressed as total CFU per organ or per sample. Values of "0" indicate that no bacteria were detected (below the detection limit).

File: Figure_5C-source_data_2.xlsx

Description: Raw data for Figure 5C, representing an independent repeat of the passive immunization experiment measuring bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments following i.v. administration of anti-CPS or anti-V5 serum. Mice received antisera i.v., were challenged i.n. with 05ZYH33, and CFU were determined in NALT, lung, blood, and CSF 12 hours post-challenge. Negative serum served as the control. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, Lung, Blood, CSF.
• Negative serum1 to -5 – Total CFU in mice receiving naive (negative) serum (five individual mice).
• CPS Ab1 to -5 – Total CFU in mice receiving anti-CPS serum (five individual mice).
• V5 Ab1 to -5 – Total CFU in mice receiving anti-V5 serum (five individual mice).

File: Figure_5D-source_data_1.xlsx

Description: Raw data for Figure 5D, showing the bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments of mice 1 hour post-infection following passive immunization with anti-CPS or anti-V5 serum administered via the intranasal (i.n.) route. Mice received antisera i.n., were challenged i.n. with 05ZYH33 6 hours later, and were euthanized 1 hour post-challenge (before bacteria could enter the bloodstream). CFU were determined in NALT (nasal-associated lymphoid tissue), CSF (cerebrospinal fluid), and blood. Negative serum (naive serum) served as the control. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, CSF, Blood.
• Negative serum 1 to -5 – Total CFU in mice receiving naive (negative) serum (five individual mice).
• CPS Ab1 to -5 – Total CFU in mice receiving anti-CPS serum (five individual mice).
• V5 Ab1 to -5 – Total CFU in mice receiving anti-V5 serum (five individual mice).

File: Figure_5D-source_data_2.xlsx

Description: Raw data for Figure 5D, representing an independent repeat of the passive immunization experiment measuring bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments 1 hour post-infection. Mice received anti-CPS or anti-V5 serum via the i.n. route, were challenged i.n. with 05ZYH33, and CFU were determined in NALT, CSF, and blood 1 hour post-challenge. Negative serum served as the control. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, CSF, Blood.
• Negative serum1 to -5 – Total CFU in mice receiving naive (negative) serum (five individual mice).
• CPS Ab1 to -5 – Total CFU in mice receiving anti-CPS serum (five individual mice).
• V5 Ab1 to -5 – Total CFU in mice receiving anti-V5 serum (five individual mice).

File: Figure_5E-source_data_1.xlsx

Description: Raw data for Figure 5E, showing the bacterial burden (total CFU) of the CPS-deficient mutant Δ2BSS2 in various body compartments of mice 1 hour post-infection following passive immunization with anti-V5 serum administered via the intranasal (i.n.) route. Mice received naive serum (negative control) or anti-V5 serum i.n., were challenged i.n. with Δ2BSS2 6 hours later, and were euthanized 1 hour post-challenge. CFU were determined in NALT (nasal-associated lymphoid tissue), CSF (cerebrospinal fluid), and blood. This experiment confirms that nasal-CNS transmigration occurs independently of CPS. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, CSF, Blood.
• Negative serum1 to -5 – Total CFU in mice receiving naive (negative) serum (five individual mice).
• V5 Ab1 to -5 – Total CFU in mice receiving anti-V5 serum (five individual mice).

File: Figure_5E-source_data_2.xlsx

Description: Raw data for Figure 5E, representing an independent repeat of the passive immunization experiment measuring bacterial burden (total CFU) of the CPS-deficient mutant Δ2BSS2 in various body compartments 1 hour post-infection. Mice received naive serum or anti-V5 serum via the i.n. route, were challenged i.n. with Δ2BSS2, and CFU were determined in NALT, CSF, and blood 1 hour post-challenge. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

File: Figure_6D-source_data_2.xlsx

Description: Raw data for Figure 6D, representing an independent repeat of the experiment measuring bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments 1 hour post-infection, comparing acetic acid pretreatment versus PBS pretreatment. Mice were pretreated with 1% acetic acid or PBS i.n., challenged with 05ZYH33, and CFU were determined in NALT, CSF, and blood 1 hour post-challenge. Data represent three biological replicates for each treatment group from one independent experiment.

Variables

File: Figure_6D-source_data_1.xlsx

Description: Raw data for Figure 6D, showing the bacterial burden (total CFU) of S. suis 05ZYH33 in various body compartments of mice 1 hour post-intranasal infection, comparing the effects of acetic acid pretreatment versus PBS pretreatment. Mice were pretreated with either 1% acetic acid or PBS intranasally, then challenged i.n. with 05ZYH33 1 hour later. CFU were determined in NALT (nasal-associated lymphoid tissue), CSF (cerebrospinal fluid), and blood 1 hour post-challenge. This experiment demonstrates that nasal-CSF transmission of 05ZYH33 occurs without acetic acid pretreatment, though acetic acid facilitates nasal-CNS transmission. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, CSF, Blood.
• Acidic acid1 to -3 – Total CFU in mice pretreated with 1% acetic acid (three individual mice).
• PBS1 to -3 – Total CFU in mice pretreated with PBS (three individual mice).

File: Figure_6E-source_data_1.xlsx

Description: Raw data for Figure 6E, showing the bacterial burden (total CFU) of the CPS-deficient mutant Δ2BSS2 in various body compartments of mice 1 hour post-intranasal infection, comparing the effects of acetic acid pretreatment versus PBS pretreatment. Mice were pretreated with either 1% acetic acid or PBS (phosphate-buffered saline) intranasally, then challenged i.n. with Δ2BSS2 1 hour later. CFU were determined in NALT (nasal-associated lymphoid tissue), CSF (cerebrospinal fluid), and blood 1 hour post-challenge. This experiment demonstrates that nasal-CSF transmission occurs independently of CPS and without acetic acid pretreatment. Data represent five biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, CSF, Blood.
• Acidic acid1 to -5 – Total CFU in mice pretreated with 1% acetic acid (five individual mice).
• PBS1 to -5 – Total CFU in mice pretreated with PBS (five individual mice).

Note: CFU values are expressed as total CFU per organ or per sample. Values of "0" indicate that no bacteria were detected (below the detection limit). Blood samples showed no detectable bacteria in all treatment groups.

File: Figure_6E-source_data_2.xlsx

Description: Raw data for Figure 6E, representing an independent repeat of the experiment measuring bacterial burden (total CFU) of the CPS-deficient mutant Δ2BSS2 in various body compartments 1 hour post-infection, comparing acetic acid pretreatment versus PBS pretreatment. Mice were pretreated with 1% acetic acid or PBS i.n., challenged with Δ2BSS2, and CFU were determined in NALT, CSF, and blood 1 hour post-challenge. Data represent three biological replicates for each treatment group from one independent experiment.

Variables

• Organ – Body compartment/tissue examined: NALT, CSF, Blood.
• Acidic acid1 to -3 – Total CFU in mice pretreated with 1% acetic acid (three individual mice).
• PBS1 to -3 – Total CFU in mice pretreated with PBS (three individual mice).

Note: CFU values are expressed as total CFU per organ or per sample. Values of "0" indicate that no bacteria were detected (below the detection limit). Blood samples showed no detectable bacteria in all treatment groups.

File: Figure1B-source_date_1.xlsx

Description: Raw data for Figure 1B, showing the hydrophobicity of S. suis 05ZYH33 in different growth phases (lag, log, and stationary phases) and the CPS-deficient strain Δ2BSS2 (log phase), as measured by bacterial adhesion to hydrocarbon (BATH) assay. Higher BATH values indicate greater surface hydrophobicity, which reflects lower levels of CPS on the bacterial surface. PBS and NC (nasal cavity, i.e., bacterial suspension without hydrocarbon) served as negative controls. Data represent three independent replicates.

Variables

• PBS – Negative control (phosphate-buffered saline without bacteria), BATH value (%).
• NC – Negative control (bacterial suspension without hydrocarbon addition), BATH value (%).
• Lag – 05ZYH33 in lag phase, BATH value (%).
• Log – 05ZYH33 in logarithmic (log) phase, BATH value (%).
• Sta – 05ZYH33 in stationary phase, BATH value (%).

Note: BATH value (%) is calculated as [(Ac - Ab) / Ac] × 100, where Ac is the OD600 of the bacterial control and Ab is the OD600 of the aqueous layer after hydrocarbon mixing.

File: Figure_3B-source_data_1.xlsx

Description: Raw data for Figure 3B, showing the bacterial burden (total colony-forming units, CFU) of S. suis 05ZYH33 in the cerebrospinal fluid (CSF) of mice at various time points following intranasal inoculation. Mice were inoculated intranasally with 05ZYH33 (stationary phase, 2 × 10⁸ CFU), and CSF was collected at 0.5, 1, 3, 5, 24, 48, 72, 96, and 120 hours post-inoculation. Total CFU in CSF was calculated as (CFU on plate × dilution factor × total volume of sample). Data represent three biological replicates (1#–3#) from one independent experiment.

Variables

• Hour – Time post-inoculation (hours): 0.5h, 1h, 3h, 5h, 24h, 48h, 72h, 96h, 120h.
• 1# – Total CFU in CSF of mouse 1.
• 2# – Total CFU in CSF of mouse 2.
• 3# – Total CFU in CSF of mouse 3.

Code/software

All data files are provided in standard formats (.xlsx, .png, .tif) that can be opened with Microsoft Excel (2016 or later), LibreOffice Calc, or any common image viewer. No custom code or scripts were used for data analysis; all statistical analyses (Student's t-test, one-way ANOVA with Tukey's post hoc test, and Mann-Whitney U test) were performed using GraphPad Prism version 7.0. The Excel files contain raw data that can be directly imported into any statistical software (e.g., R, SPSS, or Python) for re-analysis.

Access information

The data presented here are original and have not been published elsewhere. No external data sources were used in this study. The related peer-reviewed article is available at: https://doi.org/10.7554/eLife.101760