Protocol for large scale whole blood immune monitoring by mass cytometry and Cyto Quality Pipeline

Rybakowska, Paulina; Gassen, Sofie Van; Martorell-Marugán, Jordi; Quintelier, Katrien; Saeys, Yvan; Alarcón‐Riquelme, Marta E.; Marañón, Concepción

doi:10.1016/j.xpro.2022.101697

Cited by 7 publications

(8 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cells were acquired on a mass cytometer (HELIOS, Standard BioTools) at an event rate of 400 cells/s together with EQ Four Element Calibration Beads (Standard BioTools). Raw data were normalized using CytoQP function bead_normalize ( 26 ). FlowJo software v10.0 was used to analyse normalized data by manual gating.…”

Section: Methodsmentioning

confidence: 99%

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Van Breedam,

Buyle-Huybrecht,

Govaerts

et al. 2023

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

With Varicella-Zoster Virus (VZV) being an exclusive human pathogen, human induced pluripotent stem cell (hiPSC)-derived neural cell culture models are an emerging tool to investigate VZV neuro-immune interactions. Using a compartmentalized hiPSC-derived neuronal model allowing axonal VZV infection, we previously demonstrated that paracrine interferon (IFN)-α2 signalling is required to activate a broad spectrum of interferon-stimulated genes able to counteract a productive VZV infection in hiPSC-neurons. In this new study, we now investigated whether innate immune signalling by VZV-challenged macrophages was able to orchestrate an antiviral immune response in VZV-infected hiPSC-neurons. In order to establish an isogenic hiPSC-neuron/hiPSC-macrophage co-culture model, hiPSC-macrophages were generated and characterised for phenotype, gene expression, cytokine production and phagocytic capacity. Even though immunological competence of hiPSC-macrophages was shown following stimulation with the poly(dA:dT) or treatment with IFN-α2, hiPSC-macrophages in co-culture with VZV-infected hiPSC-neurons were unable to mount an antiviral immune response capable of suppressing a productive neuronal VZV infection. Subsequently, a comprehensive RNA-Seq analysis confirmed the lack of strong immune responsiveness by hiPSC-neurons and hiPSC-macrophages upon, respectively, VZV infection or challenge. This may suggest the need of other cell types, like T-cells or other innate immune cells, to (co-)orchestrate an efficient antiviral immune response against VZV-infected neurons.

show abstract

Section: Methodsmentioning

confidence: 99%

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Van Breedam,

Buyle-Huybrecht,

Govaerts

et al. 2023

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Blood samples were thawed, lysed and stained for surface markers as described 21 . Briefly, 1.5*10 6 cells/sample were used for the barcoding with Cell-ID 20-Plex Pd Barcoding Kit (Standard BioTools).…”

Section: Sample Staining and Acquisition On Cytof/heliosmentioning

confidence: 99%

Systemic autoimmune disease patients’ blood immunome reveals specificities and commonalities among different diagnostic entities

Rybakowska,

Van Gassen,

Barturen

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

1AbstractBackgroundSystemic autoimmune diseases (SADs) are characterized by internal heterogeneity, overlapping clinical symptoms, and shared molecular pathways. Therefore, they are difficult to diagnose and new tools allowing precise diagnosis are needed. Molecular-based reclassification studies enable to find patterns in a diagnosis-independent way.ObjectiveTo evaluate the possibility of using high-content immunophenotyping for detecting patient subgroups in the context of precise treatment.MethodsWhole blood high-content immunophenotyping of 101 patients with 7 systemic autoimmune diseases and 22 controls was performed using 36-plex mass cytometry panel. Patients were compared across diagnostic entities and re-classified using Monte Carlo reference-based consensus clustering. Levels of 45-plex multiplexed cytokine were measured and used for cluster characterization.ResultsDifferential analysis by diagnosis did not reveal any disease-specific pattern in the cellular compositions and phenotypes but rather their relative similarities. Accordingly, patients were classified into phenotypically distinct groups composed of different diagnostic entities sharing common immunophenotypes and cytokine signatures. These features were mainly based on granulocyte activation and CD38 expression in discrete lymphocyte populations and were related to Th17 or IFN-dependent cytokines.ConclusionsOur data indicate that specific individuals could potentially benefit from the same line of treatment independently of their diagnosis and emphasize the possibility of using immunophenotyping as a stratification tool in precision rheumatology.2Graphical abstractKey messagesWhole blood immmunophenotyping could be used to stratify systemic autoimmune patients, thus it is a useful tool in precision medicine.Patients’ groups could benefit from the same line of treatment.

show abstract

“…Epigenetic modification by environmental signals is implicated in the pathogenesis of SSc in genetically susceptible individuals [ 144 ]. Advances in technologies, such as cytometry by time of flight (CyTOF), are promising with the possibility to now look at cellular markers in whole blood, including platelet and red blood cells [ 145 , 146 , 147 ]. Furthermore, the Extended Polydimensional Immunome Characterization (EPIC), a web-based tool, could be used for the analysis of high-dimensional biomarkers in SSc patients compared with datasets of healthy controls [ 148 ].…”

Section: Unmet Needs and Future Studiesmentioning

confidence: 99%

The Pathogenesis of Systemic Sclerosis: The Origin of Fibrosis and Interlink with Vasculopathy and Autoimmunity

Ko,

Noviani,

Chellamuthu

et al. 2023

IJMS

View full text Add to dashboard Cite

Systemic sclerosis (SSc) is an autoimmune disease associated with increased mortality and poor morbidity, impairing the quality of life in patients. Whilst we know that SSc affects multiple organs via vasculopathy, inflammation, and fibrosis, its exact pathophysiology remains elusive. Microvascular injury and vasculopathy are the initial pathological features of the disease. Clinically, the vasculopathy in SSc is manifested as Raynaud’s phenomenon (reversible vasospasm in reaction to the cold or emotional stress) and digital ulcers due to ischemic injury. There are several reports that medications for vasculopathy, such as bosentan and soluble guanylate cyclase (sGC) modulators, improve not only vasculopathy but also dermal fibrosis, suggesting that vasculopathy is important in SSc. Although vasculopathy is an important initial step of the pathogenesis for SSc, it is still unclear how vasculopathy is related to inflammation and fibrosis. In this review, we focused on the clinical evidence for vasculopathy, the major cellular players for the pathogenesis, including pericytes, adipocytes, endothelial cells (ECs), and myofibroblasts, and their signaling pathway to elucidate the relationship among vasculopathy, inflammation, and fibrosis in SSc.

show abstract

Protocol for large scale whole blood immune monitoring by mass cytometry and Cyto Quality Pipeline

Cited by 7 publications

References 26 publications

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Systemic autoimmune disease patients’ blood immunome reveals specificities and commonalities among different diagnostic entities

The Pathogenesis of Systemic Sclerosis: The Origin of Fibrosis and Interlink with Vasculopathy and Autoimmunity

Contact Info

Product

Resources

About