The utilization of advance telemetry to investigate critical physiological parameters including electroencephalography in cynomolgus macaques following aerosol challenge with eastern equine encephalitis virus

Figures

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  Fig 1. Experimental design of the NHP study. All parameters were continuously and simultaneously measured throughout the study. Baseline was established for each parameter for an individual NHP by measuring 5 daytime (6 am-6 pm) and nighttime (6 pm– 6 am). All comparisons of post-challenge data was time-matched to pre-challenge baseline.

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  Fig 2. Administered aerosol EEEV dose (A) and survival of the NHPs (B).

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  Fig 3. Quantitation of infectious EEEV in NHP tissues collected at the time of euthanasia. The time of tissue collection for each NHP is provided in the x-axis. Limit of detection of plaque assay is 1.0 log10 PFU/mL or 1.0 log10 PFU/g and are indicated by a dashed line.

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  Fig 4. Body temperature of NHPs pre- and post-EEEV challenge. Data analysis is shown in 0.5- (A) and 12-hr (B) daytime/nighttime intervals. All NHPs were continuously monitored pre- and post-challenge. Pre-challenge baseline temperature was measured for five day/night cycles and a 0.5-hr interval baseline average was calculated by averaging raw data of five time-matched day or night time intervals. Forty-eight 0.5-hr interval averages were used to construct a baseline temperature for a day/night cycle and is shown as a grey line (A). Post-challenge values within ≤3 standard deviations (SD) are indicated with (), >3 SD above baseline are indicated with (), and >3 SD below baseline are indicated with (). Hyperpyrexia is indicated by () (A). Temperature change in a 12-hr interval is shown in grey (daytime) and in black (nighttime) (B). All raw data from either 12-hr daytime or nighttime intervals were averaged and normalized. The last data point for NHP #3 and #4 is an average of 8 hrs (B).

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  Fig 5. Respiration rate of NHPs pre- and post-EEEV challenge. Data analysis is shown in 0.5- (A) and 12-hr (B) daytime/nighttime intervals. All NHPs were continuously monitored pre- and post-challenge. Pre-challenge baseline respiration rate was measured for five day/night cycles and a 0.5-hr interval baseline average was calculated by averaging raw data of five time-matched day or night time intervals. Forty-eight 0.5-hr interval averages were used to construct a baseline respiration rate for a day/night cycle and is shown as a grey line (A).Post-challenge values within ≤3 standard deviations (SD) are indicated with (), >3 SD above baseline are indicated with (), and >3 SD below baseline are indicated with (). Percent change from baseline in 12-hr interval is shown in grey (daytime) and in black (nighttime) (B). All raw data from either 12-hr daytime or nighttime intervals were averaged and normalized. The last data point for NHP #3 and 4 is an average of 8 hrs (B). p-values ≤0.03 are indicated with *. bpm = breaths per minute.

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  Fig 6. Measurement of activity in NHPs pre- and post-EEEV challenge. Data analysis is shown in 6- (A) and 12-hr (B) daytime/nighttime intervals. All NHPs were continuously monitored pre- and post-challenge. Pre-challenge baseline activity was measured for five day/night cycles and a 6-hr interval baseline average was calculated by averaging raw data of five time-matched day or night time intervals. Four 6-hr interval averages were used to construct a baseline activity for a day/night cycle and is shown as a grey line (A). Post-challenge values within ≤3 standard deviations (SD) are indicated with (), >3 SD above baseline are indicated with (), and >3 SD below baseline are indicated with (). Percent change from baseline in 12-hr interval is shown in grey (daytime) and in black (nighttime) (B). All raw data from either 12-hr daytime or nighttime intervals were averaged and normalized. The last data point for NHP #3 and 4 is an average of 8 hrs (B). p-values ≤0.041 are indicated with *.

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  Fig 7. Heart rate of NHPs pre- and post-EEEV challenge. Data analysis is shown in 0.5- (A) and 12-hr (B) daytime/nighttime intervals. All NHPs were continuously monitored pre- and post-challenge. Pre-challenge baseline heart rate was measured for five day/night cycles and a 0.5-hr interval baseline average was calculated by averaging raw data of five time-matched day or night time intervals. Forty-eight 0.5-hr interval averages were used to construct a baseline heart rate for a day/night cycle and is shown as a grey line (A).Post-challenge values within ≤3 standard deviations (SD) are indicated with (), >3 SD above baseline are indicated with (), and >3 SD below baseline are indicated with (). Percent change from baseline in 12-hr interval is shown in grey (daytime) and in black (nighttime) (B). All raw data from either 12-hr daytime or nighttime intervals were averaged and normalized. The last data point for NHP #3 and 4 is an average of 8 hrs (B). p-values ≤0.035 are indicated with *. bpm = beats per minute.

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  Fig 8. Systolic blood pressure of NHPs pre- and post-EEEV challenge. Data analysis is shown in 0.5- (A) and 12-hr (B) daytime/nighttime intervals. All NHPs were continuously monitored pre- and post-challenge. Pre-challenge baseline systolic blood pressure was measured for five day/night cycles and a 0.5-hr interval baseline average was calculated by averaging raw data of five time-matched day or night time intervals. Forty-eight 0.5-hr interval averages were used to construct a baseline systolic blood pressure for a day/night cycle and is shown as a grey line (A). Post-challenge values within ≤3 standard deviations (SD) are indicated with (), >3 SD above baseline are indicated with (), and >3 SD below baseline are indicated with (). Percent change from baseline in 12-hr interval is shown in grey (daytime) and in black (nighttime) (B). All raw data from either 12-hr daytime or nighttime intervals were averaged and normalized. The last data point for NHP #3 and 4 is an average of 8 hrs (B). p-values ≤0.028 are indicated with *.

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  Fig 9. Diastolic blood pressure of NHPs pre- and post-EEEV challenge. Data analysis is shown in 0.5- (A) and 12-hr (B) daytime/nighttime intervals. All NHPs were continuously monitored pre- and post-challenge. Pre-challenge baseline diastolic blood pressure was measured for five day/night cycles and a 0.5-hr interval baseline average was calculated by averaging raw data of five time-matched day or night time intervals. Forty-eight 0.5-hr interval averages were used to construct a baseline diastolic blood pressure for a day/night cycle and is shown as a grey line (A). Post-challenge values within ≤3 standard deviations (SD) are indicated with (), >3 SD above baseline are indicated with (), and >3 SD below baseline are indicated with (). Percent change from baseline in 12-hr interval is shown in grey (daytime) and in black (nighttime) (B). All raw data from either 12-hr daytime or nighttime intervals were averaged and normalized. The last data point for NHP #3 and 4 is an average of 8 hrs (B). p-values ≤0.023 are indicated with *.

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  Fig 10. ECG abnormalities in EEEV infected NHPs 24 hrs prior to euthanasia. Representative ~5 second intervals of pre- and post-EEEV challenge are shown.

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  Fig 11. Pre- and post-EEEV challenge quantitative electroencephalography (qEEG) heat maps of NHPs #1 and #2. The top and bottom x-axes display brain waves [(delta (δ), theta (θ), alpha (α), sigma (σ), and gamma (γ)] and frequency in hertz (Hz), respectively. The left and right y-axes display time (hr) and 12-hr day/night time intervals, respectively. 12-hr nighttime is indicated by (❚) and daytime by a gap. Pre-EEEV challenge baseline (left) and post-EEEV challenge (right) heat maps are shown.

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  Fig 12. Pre- and post-EEEV challenge quantitative electroencephalography (qEEG) heat maps of NHPs #3 and #4. The top and bottom x-axes display brain waves [(delta (δ), theta (θ), alpha (α), sigma (σ), and gamma (γ)] and frequency in hertz (Hz), respectively. The left and right y-axes display time (hr) and 12-hr day/night time intervals, respectively. 12-hr nighttime is indicated by (❚) and daytime by a gap. Pre-EEEV challenge baseline (left) and post-EEEV challenge (right) heat maps are shown.