Automated bedside flow cytometer for mHLA-DR expression measurement: a comparison study with reference protocol

Zouiouich, Mehdi; Gossez, Morgane; Venet, Fabienne; Rimmelé, Thomas; Monneret, Guillaume

doi:10.1186/s40635-017-0156-z

Cited by 20 publications

(20 citation statements)

References 12 publications

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“…To circumvent this drawback, a fully automated tabletop cytometer with simple operating procedures (injection of patient's sample into a cartridge) was developed for use at the bedside or in emergency labs. Beta site evaluation for mHLA‐DR evaluation showed convincing preliminary results .…”

Section: Landmarks Of Flow Cytometry In Sepsismentioning

confidence: 86%

“…As an example, FOXP3‐lyophilized tubes are currently used for regulatory T cells determination in an RCT evaluating low‐dose IL‐2 treatment for type 1 diabetes (DIABIL‐2 study, NCT02411253). Furthermore, new technical developments (low cost compact portable flow cytometer, bedside flow cytometry or chip‐based flow cytometry) are now being proposed that will facilitate the use of Flow at the bedside in ICU (see above in mHLA‐DR paragraph ). For now, such techniques remain devoted to very simple applications but we may expect major developments in the forthcoming years, including multicolor flow cytometry.…”

Section: Current Multicenter Studies and Clinical Trialsmentioning

confidence: 99%

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How Clinical Flow Cytometry Rebooted Sepsis Immunology

et al. 2019

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On May 2017, the World Health Organization (WHO) recognized sepsis as a global health priority by adopting a resolution to improve the prevention, diagnosis, and management of this deadly disease. While it has long been known that sepsis deeply perturbs immune homeostasis by inducing a tremendous systemic inflammatory response, pivotal observations based on clinical flow cytometry indicate that sepsis indeed initiates a more complex immune response that varies over time, with the concomitant occurrence of both pro-and anti-inflammatory mechanisms. As a resultant, some septic patients enter a stage of protracted immunosuppression. This paved the way for immunostimulation approaches in sepsis. Clinical flow cytometry permitted this evolution by drawing a new picture of pathophysiology and reshaping immune trajectories in patients. Additional information from cytometry by time of flight mass cytometry and other highdimensional flow cytometry platform should rapidly enrich our understanding of this complex disease. This review reports on landmarks of clinical flow cytometry in sepsis and how this single-cell analysis technique permitted to breach the wall of decades of unfruitful anti-inflammatory-based clinical trials in sepsis.On May 2017, the World Health Organization (WHO) recognized sepsis as a global health priority by adopting a resolution to improve the prevention, diagnosis, and management of this deadly disease (1). Sepsis is defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection (2). Septic shock is the most severe form of sepsis in which hypotension persists despite adequate volume resuscitation thus requiring the use of vasopressors. According to recent definitions, in response to an infectious trigger, the unbalanced host (immune) response is at the center of sepsis pathophysiology (2).Sepsis represents a major healthcare problem worldwide. As an example, in the US, sepsis is more frequent than myocardial infarction, breast, or colon cancers (3-6). Recent modeling of available data determined the number of cases of sepsis in the entire world to over 20-25 million corresponding to >6 million deaths per annum (7,8). Over the years, 28-day mortality has remained high, ranging from 20% for sepsis to over 40% for septic shock despite improvement in patients' care (fluid resuscitation, source control, antimicrobial therapy). Whereas few therapeutic options evaluated in phase II clinical trials have demonstrated the potential efficacy of immunostimulation in sepsis, to date, no therapeutic intervention targeting host response has specifically been approved and sepsis remains the leading cause of mortality in intensive care units (ICU)(9,10). In addition, sepsis incidence has dramatically increased over the past decade and is expected to further augment. This rising

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Section: Landmarks Of Flow Cytometry In Sepsismentioning

confidence: 86%

Section: Current Multicenter Studies and Clinical Trialsmentioning

confidence: 99%

How Clinical Flow Cytometry Rebooted Sepsis Immunology

et al. 2019

Self Cite

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“…In the phase of severe immunosuppression, viral reactivations or opportunistic infections occur frequently and account for the majority of all sepsis-related deaths, which can also occur years after hospitalization [2,9]. As a reliable biomarker for immune function and immunosuppression serves the human leucocyte antigen-D related on monocytes (mHLA-DR), as it correlates with immune competence, occurrence of nosocomial infections and mortality [2,6,9,10].…”

Section: Introductionmentioning

confidence: 99%

Circadian Rhythms in Septic Shock Patients

Lachmann

Ananthasubramaniam

Wünsch

et al. 2020

Preprint

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Background: Although intensive efforts to improve diagnosis and therapy of sepsis over the last decade, the mortality of septic shock remains high and causes substantial socioeconomical burden of disease. The function of immune cells is time-of-day-dependent and is regulated by several circadian clock genes. This study aims to investigate whether the rhythmicity of clock gene expression is altered in patients with septic shock.Methods: This prospective pilot study was performed at the university hospital Charité – Universitätsmedizin Berlin, Department of Anesthesiology and Operative Intensive Care Medicine (CCM, CVK). We included 20 patients with septic shock between May 2014 and January 2018, from whom blood was drawn every 4 hours over a 24-hour period to isolate CD14-positive monocytes and to measure expression of 17 clock and clock-associated genes. Of these patients, 3 whose samples expressed fewer than 8 clock genes were excluded from the final analysis. Expression data were compared to data of a healthy study population and a rhythmicity score SP was calculated, which comprises values between -1 (arrhythmic) and 1 (rhythmic). Results: Overall, the rhythmicity scores for septic shock patients were significantly (p < 0.0001) lower (0.23 ± 0.26) compared to the control group (12 healthy young men, 0.70 ± 0.18). 77% of the measured clock genes were classified as having inconclusive rhythms, i.e. neither rhythmic nor arrhythmic. The clock genes NR1D1, NR1D2 and CRY2 were the most rhythmic, while CLOCK and ARNTL were the least rhythmic. In addition, the expression of clock genes CRY1, NR1D1, NR1D2, DBP, and PER2 was suppressed in septic shock patients and CRY2 was significantly upregulated compared to controls.Conclusion: Compared to young healthy men, molecular rhythms in immune cells of septic shock patients were substantially decreased. The decrease in rhythmicity was clock gene-dependent. The loss of rhythmicity and downregulation of clock gene expression might be caused by sepsis and might further deteriorate immune responses and organ injury, but further studies are necessary to understand underlying pathophysiological mechanisms.Clinical trial registered with www.ClinicalTrials.gov (NCT02044575) on 24 January 2014.

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“…Firstly, the choice of HLA-DR as the sole, surrogate, marker of antigen presentation capacity. Although the use of functional assays to assess APC-effector T lymphocyte interactions as well as detecting the antigen density of co-stimulatory molecules (CD80, CD86, and CD40) on the monocyte cell surface would be ideal, in clinical practice mHLA-DR analysis is well established and a standardised [ 25 ] bedside assay [ 48 ] that has been previously utilised in clinical trials of sepsis immuno-modulation [ 26 ]. It is also known to correlate with antigen presentation capacity as well as pro-inflammatory cytokine release [ 49 ].…”

Section: Discussionmentioning

confidence: 99%

Post-operative immune suppression is mediated via reversible, Interleukin-10 dependent pathways in circulating monocytes following major abdominal surgery

et al. 2018

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IntroductionPost-operative infections occur frequently following major surgery. The magnitude of the post-operative immune response is associated with an increased risk of post-operative infections, although the mechanisms driving post-operative immune-dysfunction and the potential reversibility of this response with immune stimulants are not well understood. This study aims to describe the immediate immune response to major surgery and establish links to both post-operative infection and functional aspects of immune dysregulation. We also investigate the potential of clinically available immune stimulants to reverse features of post-operative immune-dysfunction.MethodsPatients over 45 years old undergoing elective gastro-intestinal surgery with planned post-operative surgical ICU admission were recruited. The expression of selected genes was determined pre-operatively and at 2, 24 and 48 hours post-operatively using qRT-PCR. Circulating levels of Interleukin-10 protein were determined by ELISA. Peri-operative cell surface monocyte HLA-DR (mHLA-DR) expression was determined using flow cytometry. Gene expression and mHLA-DR levels were determined in healthy monocytes cultured in peri-operative serum with and without neutralising antibodies and immune stimulants.Results119 patients were recruited; 44 developed a post-operative infection. Interleukin-10 mRNA and protein increased 4-fold post-operatively (P<0.0001), peaking within 2 hours of the procedure. Higher post-operative Interleukin-10 mRNA (P = 0.007) and protein (P = 0.001) levels were associated with an increased risk of infection. Cell surface mHLA-DR expression fell post-operatively (P<0.0001). Reduced production, rather than intracellular sequestration, accounted for the post-operative decline in cell surface mHLA-DR expression. Interleukin-10 antibody prevented the decrease in mHLA-DR expression observed when post-operative serum was added to healthy monocytes. GM-CSF and IFN-γ prevented the decline in mHLA-DR production through distinct pathways.ConclusionsMonocyte dysfunction and features of immune suppression occur frequently after major surgery. Greater post-operative Interleukin-10 production is associated with later infection. Interleukin-10 is an important mediator of post-operative reductions in mHLA-DR expression, while clinically available immune stimulants can restore mHLA-DR levels.

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Automated bedside flow cytometer for mHLA-DR expression measurement: a comparison study with reference protocol

Cited by 20 publications

References 12 publications

How Clinical Flow Cytometry Rebooted Sepsis Immunology

How Clinical Flow Cytometry Rebooted Sepsis Immunology

Circadian Rhythms in Septic Shock Patients

Post-operative immune suppression is mediated via reversible, Interleukin-10 dependent pathways in circulating monocytes following major abdominal surgery

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