In addition, our technique allows direct ex vivo visualisation without any need for further processing of the tissues, in contrast to immunohistochemistry

and MPO analysis. Histology is labour-intensive and tedious, while MPO assays can be problematic and do not distinguish between neutrophils and macrophages. In conclusion, this study presents a robust model to track neutrophil recruitment which can be used to complement other available BGJ398 methods traditionally used for tracking neutrophils. In addition to experimental models of IBD, this versatile technique will be useful for monitoring neutrophil trafficking during inflammatory responses in a range of disease settings and constitutes a novel approach for the assessment of potential therapeutics that aim to reduce neutrophil infiltration. Thus, it can be used as an informative and specific tool for both the pharmaceutical industry and the basic research community. We thank

Grainne Hurley for her excellent technical assistance. NVP-BEZ235 The authors are supported in part by Science Foundation Ireland and by a research grant from GlaxoSmithKline. None of the co-authors have any conflict of interest to declare in connection to the paper. The work described has not been published or submitted elsewhere. S.M. and G.M. are employees of GlaxoSmithKline. “
“B1 B cells represent a unique subset of B lymphocytes distinct from conventional B2 B cells, and are important in the production of natural antibodies. A potential human homologue of murine B1 cells was defined recently as a CD20+CD27+CD43+ cell. Common variable immunodeficiency (CVID) is a group of heterogeneous conditions linked by symptomatic primary antibody failure. In this preliminary report, we examined the potential clinical utility of introducing CD20+CD27+CD43+ B1 cell immunophenotyping as a routine assay in a diagnostic clinical laboratory. Using a whole blood assay, putative B1 B cells in healthy controls and in CVID patients were measured. Peripheral blood from 33 healthy donors and 16 CVID

patients were stained with relevant monoclonal antibodies and underwent flow cytometric evaluation. We established a rapid, pheromone whole blood flow cytometric assay to investigate putative human B1 B cells. Examination of CD20+CD27+CD43+ cells is complicated by CD3+CD27+CD43hi T cell contamination, even when using stringent CD20 gating. These can be excluded by gating on CD27+CD43lo–int B cells. Although proportions of CD20+CD27–CD43lo–int cells within B cells in CVID patients were decreased by 50% compared to controls (P < 0·01), this was not significant when measured as a percentage of all CD27+ B cells (P = 0·78). Immunophenotypic overlap of this subset with other innate-like B cells described recently in humans is limited. We have shown that putative B1 B cell immunophenotyping can be performed rapidly and reliably using whole blood. CD20+CD27+CD43lo–int cells may represent a distinct B1 cell subset within CD27+ B cells.

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