ArticleClinical and vaccine immunology : CVI2012
Impact of human granulocyte and monocyte isolation procedures on functional studies.
Article in Clinical and vaccine immunology : CVI, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers, 1 of them a synthesis that pooled it.
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Who cites it
57 citing papers in PubMed, 1 synthesis or guideline pooled it.
- The Potential Use of Peripheral Blood Mononuclear Cells as Biomarkers for Treatment Response and Outcome Prediction in Psychiatry: A Systematic Review.Molecular diagnosis & therapy · 2021Pooled it
- Increased Circulating Epithelial Tumor Cells (CETC/CTC) over the Course of Adjuvant Radiotherapy Is a Predictor of Less Favorable Outcome in Patients with Early-Stage Breast Cancer.Current oncology (Toronto, Ont.) · 2022Trial
- Tumor-associated neutrophils stimulate T cell responses in early-stage human lung cancer.The Journal of clinical investigation · 2014Trial
- Protein S Enhances the Phagocytosis of Phosphatidylserine-Exposing Erythrocytes: Implications in Sickle Cell Disease.American journal of hematology · 2026Article
- Dendritic Cell Therapy in Immuno-Oncology: A Potentially Key Component of Anti-Cancer Immunotherapies.Cancers · 2025Review
- Advances in machine learning-enhanced microfluidic cell sorting.Science advances · 2025Review
- "Monocytes in B-Cell Malignancies: Their Role in Disease Progression and Therapy Resistance".Current oncology reports · 2025Review
- Microgravity activates monocyte ERK1/2 signaling and modulates the response to lipopolysaccharide.Molecular medicine (Cambridge, Mass.) · 2025Article
- A critical guideline for controlling monocyte-derived macrophages phenotypes.Frontiers in immunology · 2025Review
- Methods integrating innate and adaptive immune responses in humanFrontiers in immunology · 2025Review
- A protocol to isolate and characterize pure monocytes and generate monocyte-derived dendritic cells through FBS-Coated flasks.Scientific reports · 2024Article
- UK5099 Inhibits the NLRP3 Inflammasome Independently of its Long-Established Target Mitochondrial Pyruvate Carrier.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Transcriptomics analysis highlights potential ways in human pathogenesis in Leishmania braziliensis infected with the viral endosymbiont LRV1.PLoS neglected tropical diseases · 2024Article
- Analysis of Neutrophil Responses to Biological Exposures.Current protocols · 2023Article
- Monocyte-neutrophil entanglement in glioblastoma.The Journal of clinical investigation · 2023Review
- Mechanism of action of IC 100, a humanized IgG4 monoclonal antibody targeting apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC).Translational research : the journal of laboratory and clinical medicine · 2023Article
- Neutrophil degranulation and severely impaired extracellular trap formation at the basis of susceptibility to infections of hemodialysis patients.BMC medicine · 2022Article
- Article
- Quiescence of Human Monocytes after Affinity Purification: A Novel Method Apt for Monocyte Stimulation Assays.Biomolecules · 2022Article
- MyD88-dependent signaling drives toll-like receptor-induced trained immunity in macrophages.Frontiers in immunology · 2022Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
One of the first lines of defense against infection is the activation of the innate immune system. It is becoming clear that autoimmune diseases, such as rheumatoid arthritis and Crohn's disease, may be caused by disturbed innate immunity, and relating granulocyte and monocyte functions to the patient genotype has become an important part of contemporary research. Although it is essential to move this field forward, a systematic study comparing the efficacy and suitability for functional studies of the various available protocols for the isolation of these immune cells has not been performed. Here, we compare human granulocyte functionality under three enrichment protocols: (i) Ficoll density gradient centrifugation, (ii) anti-CD15 antibody-conjugated microbeads (positive selection), and (iii) Polymorphoprep. Primary monocytes were isolated in parallel using (i) anti-CD14 magnetic microbeads, (ii) non-monocyte depletion by antibody-conjugated magnetic microbeads (negative selection), (iii) RosetteSep antibody cocktail, and (iv) the classical adherence protocol. The best results in terms of purity and cell functionality were obtained with positive selection by magnetic microbeads for both human granulocytes and monocytes. Whereas phagocytosis of Escherichia coli bacteria was identical in all isolation procedures tested, the granulocyte respiratory burst was higher in positively selected cells. In addition, different granulocyte enrichment procedures affect cell surface receptor expression to different extents. In toto, we propose that positive selection of granulocytes and monocytes be adopted as the procedure of choice for studies of human granulocyte and monocyte functions but caution investigators to be aware of possible alterations in cell phenotypes with different isolation procedures.
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