New flow cytometric method for surface phenotyping basophils from peripheral blood

Takahashi, Kiyoshi; Takata, Minoru; Suwaki, Toshimitsu; Kawata, Noriko; Tanimoto, Yasushi; Soda, Ryo; Kimura, Ikuo

doi:10.1016/0022-1759(93)90402-s

Cited by 22 publications

(16 citation statements)

References 15 publications

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“…First, human basophils are considered as CD16-negative cells (13)(14)(15). We show here that blood basophils detectably expressed Fc␥RIII in all donors, whether normal individuals or patients.…”

Section: Discussionmentioning

confidence: 78%

“…Basophil enrichment techniques use magnetic beads and a mixture of mAbs against membrane molecules that are not expressed by basophils. As basophils were described to express CD13, CD22, CD32, CD33, CD123 and CD203c, but not CD2, CD3, CD14, CD15, CD16, CD19, CD21, CD64, and HLA-DR (13-15), negative selection procedures used to enrich basophils from white blood cells include anti-CD16, i.e., anti-Fc␥RIII (13,39), among other mAbs. Although Fc␥RIII expression is low on basophils, one cannot exclude that enrichment procedures in which CD16 ϩ cells are discarded may remove a significant proportion of basophils.…”

Section: Discussionmentioning

confidence: 99%

“…Using this nomenclature, basophils were described to express CD32, but not CD16 (13)(14)(15). CD32 refers to Fc␥RII whereas CD16 refers to Fc␥RIII.…”

mentioning

confidence: 99%

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Human Basophils Express the Glycosylphosphatidylinositol-Anchored Low-Affinity IgG Receptor FcγRIIIB (CD16B)

Meknache

Jönsson

Laurent

et al. 2009

The Journal of Immunology

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Basophils express not only high-affinity IgE receptors, but also low-affinity IgG receptors. Which, among these receptors, are expressed by human basophils is poorly known. Low-affinity IgG receptors comprise CD32 (FcγRIIA, FcγRIIB, and FcγRIIC) and CD16 (FcγRIIIA and FcγRIIIB). FcγRIIA, FcγRIIC, and FcγRIIIA are activating receptors, FcγRIIB are inhibitory receptors, FcγRIIIB are GPI-anchored receptors whose function is poorly understood. Basophils were reported to express FcγRII, but not FcγRIII. We aimed at further identifying basophil IgG receptors. Basophils from normal donors and from patients suffering from an allergic skin disease (atopic dermatitis), allergic respiratory diseases (allergic rhinitis and asthma), or a nonallergic skin disease (chronic urticaria) were examined. We found that normal basophils contain FcγRIII transcripts and express FcγRIIIB, but not FcγRIIIA, which were detected on 24–81% basophils from normal donors and on 12–100% basophils from patients. Noticeably, the proportion of FcγRIIIB+ basophils was significantly lower in atopic dermatitis patients than in other subjects. This decreased FcγRIII expression was not correlated with an activated phenotype of basophils in atopic dermatitis patients, although FcγRIIIB expression was down-regulated upon basophil activation by anti-IgE. Our results challenge the two dogmas 1) that basophils do not express FcγRIII and 2) that FcγRIIIB is exclusively expressed by neutrophils. They suggest that a proportion of basophils may be lost during enrichment procedures in which FcγRIII+ cells are discarded by negative sorting using anti-CD16 Abs. They unravel an unexpected complexity of IgG receptors susceptible to modulate basophil activation. They identify a novel systemic alteration in atopic dermatitis.

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Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 99%

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Human Basophils Express the Glycosylphosphatidylinositol-Anchored Low-Affinity IgG Receptor FcγRIIIB (CD16B)

Meknache

Jönsson

Laurent

et al. 2009

The Journal of Immunology

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“…It has already been shown that human basophils selec tively express FcyRll among IgG receptors [10][11][12][13] al though which isoform of FcyRll is expressed remains to be clarified. In other cell systems, FcyRIIA is known to induce various biological reactions such as phagocytosis or release ofproinflammatory mediators [6,14,15].…”

Section: Introductionmentioning

confidence: 99%

“…On contact with allergen, recruited baso phils are activated through a cross-linking of surface bound IgE molecules and high-affinity IgE receptors, FceRI, lead ing to biochemical signaling events and chemical mediator release [6][7][8][9], Besides FceRI, human basophils also express a number of surface molecules and receptors including Fc receptors K Ä R G E R ' 1996 S. Karger AG. Basel l \ W I \ V J C I \ 1018 2438/96/1091 -0027 $8.00/0 H-Mail karger«/ kargcr.ch Fax +41 61 306 12 34 forlgG [10][11][12][13], Based on the reactivity o f monoclonal anti bodies (rnAbs) and molecular cloning, three different IgG receptors, namely FcyRI (CD64), FcyRll (CDw32) and FcyRIll (CD16) have been identified to date [6.14.15]. Each of them has distinct isoforms that show cell-type-specific expression.…”

Section: Introductionmentioning

confidence: 99%

Signal Transduction by IgG Receptors Induces Calcium Mobilization, but not Histamine Release, in the Human Basophilic Cell Line KU812F

Kawata

Takata²,

Suwaki³

et al. 1996

Int Arch Allergy Immunol

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Human blood basophils selectively express FcγRII (CDw32) among IgG receptor subtypes, but its functional role in allergic reactions remains unknown. Using the human basophilic leukemia cell line KU812F as a model system, we investigated cellular signaling events mediated through IgG receptor stimulation. KU812F cells express FcγRII on their surface. mRNAs for both FcγRIIA and IIB subtypes were detected by reverse transcriptase-PCR analysis. In this cell line, FcγRII stimulation induced mobilization of free intracellular calcium and actin polymerization. Yet, no significant histamine release was observed, nor did blood basophils stimulated by anti-FcγRII monoclonal antibody IV.3 and by a secondary antibody release histamine. These data indicate that FcγRII stimulation induces cellular signaling events such as calcium mobilization in human basophils. However, these events do not lead to histamine release.

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Flow cytometric monitoring of allergen induced basophil activation

et al. 1995

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Human basophils express many clustered differentiation antigens (CD), including CD45; however, none are speciac for these cells. In a previous study, we described a two-color immunofluorescence procedure, employing antibodies to CD45 and I@ for the cytometric evaluation of basophils. In Human basophils are quantitatively the least important population of circulating leukocytes. Nevertheless, they are specialized effector cells of the immune system and play an important role in host defense mechanisms and allergic reactions. The binding of allergens to specific IgE fixed on the basophil membrane via the FceRI receptor triggers cellular activation, degranulation, and the release of potent mediators, including histamine (4,6,7). Basophil activation is usually quantified by the measurement of secreted histamine (4,16) and by direct microscopic examination of the percentage of degranulated cells (6). This latter method is very simple but, due to the low frequency of circulating basophils, it is not of current use as it is poorly reliable.In a previous study (5) we showed that the CD45dimIgEb'igh' phenotype could be used for the flow cytometric evaluation of human basophils. However, when these cells were incubated in the presence of calcium and magnesium, three anti-IgE concentration-dependent modifications were observed: (i) a decrease in the number of detectable basophils, (ii) a decrease in IgE mean fluorescence intensity (MFI), and (iii) an increase in CD45 MFI. In the present work, the modulation of CD45 and IgE antigen expression induced on basophils by allergen and calcium ionophore is studied and a flow cytometric approach is described that can be used for the quantification of basophil activation. MATERIAL AND METHODSCell Preparation Peripheral blood from 10 nonatopic and 25 atopic adult volunteers was collected on EDTA. Atopic individuals were selected based on clinical allergic symptoms, positivity to at least one common aeroallergen (skin test), and the presence of specific IgE to this allergen. Seven patients were allergic to Dermutopbagoides pteronyssintcs (DP), 1 1 were allergic to Ductylis glomautu (DG), 4 were allergic to cat dander, and 3 were allergic to birch pollen. The blood, diluted 1/2 in phosphate buffer saline (PBS pH 7.2, BioMerieux, MarcyI'Etoile, France), was layered over a cushion of Percoll (Pharmacia, Uppsala, Sweden) adjusted to a specific gravity of 1.080 g/ml and centrifuged at 300 X g for 20 min at room temperature. The leukocyte-rich layer was recovered and washed in PBS, and the cells were finally resuspended in PBS supplemented with 0.5% bovine serum albumin

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New flow cytometric method for surface phenotyping basophils from peripheral blood

Cited by 22 publications

References 15 publications

Human Basophils Express the Glycosylphosphatidylinositol-Anchored Low-Affinity IgG Receptor FcγRIIIB (CD16B)

Human Basophils Express the Glycosylphosphatidylinositol-Anchored Low-Affinity IgG Receptor FcγRIIIB (CD16B)

Signal Transduction by IgG Receptors Induces Calcium Mobilization, but not Histamine Release, in the Human Basophilic Cell Line KU812F

Flow cytometric monitoring of allergen induced basophil activation

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