Web‐Based Analysis and Publication of Flow Cytometry Experiments

Kotecha, Nikesh; Krutzik, Peter O.; Irish, Jonathan M.

doi:10.1002/0471142956.cy1017s53

Cited by 468 publications

(390 citation statements)

References 9 publications

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“…For viSNE analysis, raw fcs files were uploaded to Cytobank. 17 Live events were gated based on exclusion of the DAPI dye, and viSNE analysis 16 was conducted clustering on the expression of ST2, TSLPR, and IL17BR. Analysis was repeated 3 times in order to confirm the reproducibility and stability of clustering.…”

Section: Flow Cytometrymentioning

confidence: 99%

IL-33, IL-25, and TSLP induce a distinct phenotypic and activation profile in human type 2 innate lymphoid cells

Camelo¹,

Rosignoli²,

Ohne³

et al. 2017

Blood Advances

146

138

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Section: Flow Cytometrymentioning

confidence: 99%

IL-33, IL-25, and TSLP induce a distinct phenotypic and activation profile in human type 2 innate lymphoid cells

Camelo¹,

Rosignoli²,

Ohne³

et al. 2017

Blood Advances

146

138

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“…Data were collected on the Accuri C6 flow cytometer or Influx cell sorter instruments (BD Biosciences) and analyzed using FlowJo software (Tree Star). Visualized t-distributed stochastic neighbor embedding (viSNE) (14) was performed on arcsinh-transformed fluorescent parameters using Cytobank (15).…”

Section: Flow Cytometrymentioning

confidence: 99%

Characterization of the Expression and Function of the C-Type Lectin Receptor CD302 in Mice and Humans Reveals a Role in Dendritic Cell Migration

Silveira

Fromm

et al. 2016

The Journal of Immunology

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C-type lectin receptors play important roles in immune cell interactions with the environment. We described CD302 as the simplest, single domain, type I C-type lectin receptor and showed it was expressed mainly on the myeloid phagocytes in human blood. CD302 colocalized with podosomes and lamellopodia structures, so we hypothesized that it played a role in cell adhesion or migration. In this study, we used mouse models to obtain further insights into CD302 expression and its potential immunological function. Mouse CD302 transcripts were, as in humans, highest in the liver, followed by lungs, lymph nodes (LN), spleen, and bone marrow. In liver, CD302 was expressed by hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells. A detailed analysis of CD302 transcription in mouse immune cells revealed highest expression by myeloid cells, particularly macrophages, granulocytes, and myeloid dendritic cells (mDC). Interestingly, 2.5-fold more CD302 was found in migratory compared with resident mDC populations and higher CD302 expression in mouse M1 versus M2 macrophages was also noteworthy. CD302 knockout (CD302KO) mice were generated. Studies on the relevant immune cell populations revealed a decrease in the frequency and numbers of migratory mDC within CD302KO LN compared with wild-type LN. In vitro studies showed CD302KO and wild-type DC had an equivalent capacity to undergo maturation, prime T cells, uptake Ags, and migrate toward the CCL19/CCL21 chemokines. Nevertheless, CD302KO migratory DC exhibited reduced in vivo migration into LN, confirming a functional role for CD302 in mDC migration.

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“…Fluorescence was measured on 10,000 cells/sample using an FACS (FACScantoII; Becton Dickinson, Mountain View, CA) and analyzed using FlowJo software (Tree Star, Inc., Ashland, OR) and Cytobank. 50 Data are depicted in histograms plotting median ou geomean fluorescence intensity (MFI) on a fourdecade logarithmic scale (x-axis) versus cell number (y-axis).…”

Section: Immunoblottingmentioning

confidence: 99%

MICA Variant Promotes Allosensitization after Kidney Transplantation

Tonnerre

Gérard

Chatelais

et al. 2013

Journal of the American Society of Nephrology

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MHC class I-related chain A (MICA) antigens are surface glycoproteins strongly implicated in innate immunity, and the MICA gene is highly polymorphic. Clinical observations suggest a role for donor MICA antigens expressed on transplant endothelial cells in the alloimmune response, but the effect of MICA genotype is not well understood. Here, we investigated the immunologic effect of the A5.1 mutation, related to the common MICA*008 allele. Compared with wild-type endothelial cells (ECs), homozygosity for MICA A5.1 associated with an endothelial phenotype characterized by 7-to 10-fold higher levels of MICA mRNA and MICA proteins at the cell surface, as well as exclusive release in exosomes instead of enzymatic cleavage. Mechanistically, we did not detect quantitative changes in regulatory microRNAs. Functionally, A5.1 ECs enhanced NKG2D interaction and natural killer cell activation, promoting NKG2D-dependent lysis of ECs. In kidney transplant recipients, polyreactive anti-MICA sera bound preferentially to ECs from MICA A5.1 donors, suggesting that MICA*008(A5.1) molecules are the preferential antigenic determinants on ECs of grafts. Furthermore, the incidence of MICA A5.1 mismatch revealed a statistically significant association between donor MICA A5.1 and both anti-MICA sensitization and increased proteinuria in kidney recipients. Taken together, these results identify the A5.1 mutation as an immunodominant factor and a potential risk factor for transplant survival.

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Web‐Based Analysis and Publication of Flow Cytometry Experiments

Cited by 468 publications

References 9 publications

IL-33, IL-25, and TSLP induce a distinct phenotypic and activation profile in human type 2 innate lymphoid cells

IL-33, IL-25, and TSLP induce a distinct phenotypic and activation profile in human type 2 innate lymphoid cells

Characterization of the Expression and Function of the C-Type Lectin Receptor CD302 in Mice and Humans Reveals a Role in Dendritic Cell Migration

MICA Variant Promotes Allosensitization after Kidney Transplantation

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