Flow cytometric study of neutrophilic granulopoiesis in normal bone marrow using an expanded panel of antibodies: Correlation with morphologic assessments

Abstract

Flow cytometry studies of surface markers of neutrophils have been performed mostly on peripheral blood, and for a limited spectrum of diseases. Studying maturation defects on developing neutrophils in the bone marrow (BM) may be helpful in BM diseases, such as myelodysplastic syndromes and Shwachman-Diamond syndrome. We applied an expanded panel of antibodies to examine normal maturation patterns in 26 control samples of BM together with microscopic correlation. Promyelocytes correlated well with the CD24(-) and CD11b(-) populations, and metamyelocytes correlated well with the CD16(+) population (intermediate positivity). An excellent correlation was also identified between the sum of bands and segmented neutrophils and each of the following: CD16(++) (strong positivity), CD35(+), CD87(+), and CD64(-). Although visually identified segmented neutrophils paralleled CD10 positivity, there was an appreciable difference between both methods. We conclude that neutrophilic granulocyte maturation in the BM is accompanied by a change in surface antigens that reflects certain stages of development. A successful strategy for detecting maturation defects is to include several antibodies that are known to be expressed or absent at the same stage of maturation, such as CD16, CD35, CD64, and CD87.

Four‐ and five‐color flow cytometry analysis of leukocyte differentiation pathways in normal bone marrow: A reference document based on a systematic approach by the GTLLF and GEIL

“…Data reported in several other papers from Stelzer et al (19) or Borowitz et al (20) on CD45 expression and Fujimoto et al (21) for CD11b/CD16 expression in granulocytic maturation were also considered, as well as other more recent proposals (22)(23)(24). Data from these reports were confirmed and refined by this new FCM analysis procedure.…”

Quantitative analysis of the expression of glycosylphosphatidylinositol‐anchored proteins during the maturation of different hematopoietic cell compartments of normal bone marrow

“…Up till now, the reactivity for a high number of proteins has been studied along the maturation of different BM hematopoietic cell lineages (7,12,(17)(18)(19). Despite this, the information currently …”

“…at the myelocyte and band maturation stages, respectively (5,6); in turn CD66b and CD16 would be expressed at earlier stages of maturation, the highest expression of CD16 being observed on bands and mature neutrophils (7)(8)(9)(10)(11). CD14 expression would be restricted to the latest stages of the maturation of normal monocytic cells (12).…”

A Systemic Neutrophil Response Precedes Robust CD8+T-Cell Activation during Natural Respiratory Syncytial Virus Infection in Infants

“…The population of cells characterized by CD16 int was a heterogeneous population of neutrophil precursors consisting of myelocytes, metamyelocytes, and banded neutrophils, which are neutrophil precursors normally residing in the bone marrow ( Fig. 3C) (13,14). The same cell types were also observed in direct blood films from RSV patients (data not shown).…”