In vivo quantification of rolling and adhered leukocytes in human sepsis

Abstract

Background: The use of in vivo videomicroscopy at the bedside has demonstrated microcirculatory flow disturbances in sepsis. The ability of in vivo videomicroscopy to detect changes in the prevalence of rolling and adhered leukocytes that occur in sepsis is not well-described in humans. We sought to (1) develop methodology for accessing and quantifying sublingual leukocyte rolling and adherence with sidestream dark field (SDF) imaging; (2) compare the number of rolling and adhered leukocytes between patients with septic shock and non-infected controls; and (3) compare the number of rolling and adhered leukocytes between survivors and non-survivors of septic shock.

Methods: We included adult (age > 18 years) patients in the emergency department presenting with septic shock prospectively enrolled in the ProCESS trial. We recruited comparison non-infected patients as emergency department controls. Using a SDF videomicroscope, we obtained image sequences from the sublingual mucosa, quantifying rolling and adhered leukocytes per 1 mm × 1 mm visual field in a standardized 3-s clip. We report data as median and interquartile range and depicted as box plots. We compared groups using the Mann-Whitney U test, considering a p value < 0.05 significant.

Results: We included a total of 64 patients with septic shock and 32 non-infected controls. The median number of adhered leukocytes per field in the sepsis group was 1.0 (IQR 0-3.5) compared to 0 (0-0) in the non-infected group (p < 0.001). The median number of rolling leukocytes was 26 (10.3-42) in the sepsis group and 9.8 (4.8-17.3) in the non-infected group (p < 0.001) per field. Among the patients with sepsis (n = 64), there was an increased number of adhered leukocytes in non-survivors compared to survivors (3.0 (1-5.5) vs. 1.0 (0-3.0)) (p < 0.05); however, there was no difference in rolling leukocytes (35 (20-48) vs. 26 (10-41)) (p = 0.31).

Conclusions: Our results demonstrated a higher number of rolling and adhered leukocytes in patients with septic shock when compared to non-infected controls, and an increased number of adhered leukocytes in non-survivors.

Trial registration: ClinicalTrials.gov , NCT00793442 ; Registered on 19 November 2008 PG0GM076659 (US NIH Grant/Contract). First submitted 18 July 2007. First posted 2 August 2007.

Keywords: Leukocyte; Microcirculation; SDF; Sepsis; Septic shock; Sidestream dark field.

Conflict of interest statement

Ethics approval and consent to participate

The Beth Israel Deaconess Medical Center Committee for Clinical Investigations and each site’s review board approved the design. Each subject or legal representative gave written informed consent.

Consent for publication

Not applicable.

Competing interests

NIS received funding from the National Institutes of Health; research funding from Cheetah Medical, Thermo-Fisher, Siemens, and Rapid Pathogen Screening. He was on an advisory board/consultant for Baxter and Cyon. The MicroScan equipment was provided by MicroVision Medical BV.

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Figures

Fig. 1

The figure shows the same vessel visualized by sidestream dark field imaging at different time points. The black arrows show rolling leukocytes with a shift towards the right throughout the time course. The red arrow shows an adhered leukocyte

Fig. 2

Comparison of adhered leukocytes in patients with sepsis and in controls. The boxes delineate the interquartile range, the median is shown as a line in the middle of the box, and tails show the 95% range; ***p < 0.001

Fig. 3

Comparison of rolling leukocytes in patients with sepsis and in controls. The boxes delineate the interquartile range, the median is shown as a line in the middle of the box, and tails show the 95% range; ***p < 0.001

Fig. 4

Comparison of adhered leukocytes in survivors and non-survivors in the sepsis group. The boxes delineate the interquartile range, the median is shown as a line in the middle of the box, and tails show the 95% range; *p < 0.05

Fig. 5

Comparison of rolling leukocytes in survivors and non-survivors in the sepsis group. The boxes delineate the interquartile range, the median is shown as a line in the middle of the box, and tails show the 95% range. The groups were not significantly different, p = 0.31