Detection of CD16^low neutrophil subpopulations

Abstract

Background The success of complex molecular cytogenetic studies depends on having properly spread chromosomes. However, inconsistency of optimum chromosome spreading remains a major problem in cytogenetic studies. Methods The metaphase spreading process was carefully timed to identify the most critical phase of chromosome spreading. The effects of dropping height of cell suspension, slide condition, drying time, fixative ratio, and relative humidity on the quality of metaphase spreads were studied by quantitative examination of metaphase chromosome spreads. Normal and immortalized human epithelial ovarian cells, neuroblastoma cells, and normal lymphocytes were tested. Results Humidity over the slide was the most important variable affecting the quality of chromosome spreads. Consistent improvement in chromosome spreading (larger metaphase area, less chromosome overlaps, or lower frequencies of broken metaphases) was obtained for all cell types if dynamic cell rehydration, occurring as fixative absorbs moisture from air, was made to coincide with the prompt fixation of spread chromosomes to the slide. This was achieved by dropping cells on dry glass slides placed in a shallow metal tray and then quickly lowering the tray into a covered 50°C water bath for slide drying. Conclusions A new and simple method for improving metaphase chromosome spreading was developed based on our study on the characteristics of chromosome spreading. Cytometry Part A 51A:46–51, 2003. © 2002 Wiley‐Liss, Inc.

Prevention of Graft-versus-Host Disease by Adoptive T Regulatory Therapy Is Associated with Active Repression of Peripheral Blood Toll-Like Receptor 5 mRNA Expression

“…Indeed, samples from conventionally treated patients contain more of these CD16 low cells (Supplementary Figure 3). The CD33 + CD16 low cells detected in our samples seem to be identical with the recently described CD16 low granulocytic subset (37-39). Indeed, it is known for some years that granulocytes reduce their CD16 surface expression during activation (40, 41).…”

Anti‐polymorphonuclear neutrophil antibodies in patients with leukopenia or neutropenia

“…CD16 b low was evaluated by direct staining with the following monoclonal antibodies CD49d‐FITC (44H6, Immunotech, Beckman Coulter Company), CD16‐PE (G38; Caltag‐Invitrogen, California, CA, USA), CD45‐TC (J33; Cytognos, Salamanca, Spain) or using the combination of CD16‐FITC (3G8; Caltag‐Invitrogen), CD56‐PE (B159; BD) and CD3‐PE‐Cy5 (UCHT1; e‐Bioscience, San Diego, CA, USA) and isotype‐matched antibodies. Samples were acquired and analyzed using R3 strategy of region intersections (Riera et al. , 2003).…”

In-depth immune cellular profiling reveals sex-specific associations with frailty

“…CD16 is usually expressed on the surface of neutrophils [17] and is seen as a neutrophil maturation marker [18]. Lower expression of CD16 by neutrophils was seen in several diseases and in states of neutropenia [19]. Monocytes were gated as SSC DIM CD45 DIM and, to ensure that B cells or T cells did not contaminate the gate, as CD3 -CD19 -.…”