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Assessing the Functional Heterogeneity of Monocytes in Human Septic Shock: a Proof-of-Concept Microfluidic Assay of TNFα Secretion
- Source :
- Frontiers in Immunology, Frontiers in Immunology, Vol 12 (2021)
- Publication Year :
- 2021
- Publisher :
- Frontiers Media SA, 2021.
-
Abstract
- ObjectiveThe development of advanced single-cell technologies to decipher inter-cellular heterogeneity has enabled the dynamic assessment of individual cells behavior over time, overcoming the limitation of traditional assays. Here, we evaluated the feasibility of an advanced microfluidic assay combined to fluorescence microscopy to address the behavior of circulating monocytes from septic shock patients.MethodsSeven septic shock patients and ten healthy volunteers were enrolled in the study. Using the proposed microfluidic assay we investigated the production over time of LPS-elicited TNFα by single monocytes encapsulated within droplets. Cellular endocytic activity was assessed by internalization of magnetic nanoparticles. Besides, we assessed HLA-DR membrane expression and LPS-induced TNFα production in monocytes through classical flow cytometry assays.ResultsConsistent with the flow cytometry results, the total number of TNFα molecules secreted by encapsulated single monocytes was significantly decreased in septic shock patients compared to healthy donors. TNFα production was dampened as soon as 30 and 60 minutes after LPS stimulation in monocytes from septic patients. Furthermore, the microfluidic assay revealed heterogeneous individual behavior of monocytes from septic shock patients. Of note, monocytes from both healthy donors and patients exhibited similar phagocytic activities over time.ConclusionThe microfluidic assay highlights the functional heterogeneity of monocytes, and provides in-depth resolution in assessing the hallmark monocyte deactivation encountered in post-septic immunosuppression.
- Subjects :
- Adult
Lipopolysaccharides
Male
0301 basic medicine
medicine.medical_treatment
media_common.quotation_subject
Microfluidics
Immunology
microfluidic
Proof of Concept Study
Monocytes
Flow cytometry
03 medical and health sciences
0302 clinical medicine
medicine
Fluorescence microscope
Humans
Immunology and Allergy
Secretion
Internalization
Original Research
Aged
media_common
Aged, 80 and over
Immunosuppression Therapy
tolerance
medicine.diagnostic_test
Tumor Necrosis Factor-alpha
Septic shock
Chemistry
Monocyte
Immunosuppression
HLA-DR Antigens
RC581-607
Middle Aged
Flow Cytometry
medicine.disease
Shock, Septic
030104 developmental biology
medicine.anatomical_structure
monocyte
septic shock
Female
Tumor necrosis factor alpha
immune suppression
Immunologic diseases. Allergy
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 16643224
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- Frontiers in Immunology
- Accession number :
- edsair.doi.dedup.....775ae8fb888e0da1520b6adbc0659040