Monoclonal antibody (MAB) BH2C6 recognizes a plasma membrane antigen, the BH2-Ag, specifically expressed by human neutrophils. While studies with peripheral blood and bone marrow from healthy adults clearly demonstrate the absence of BH2-Ag from other cellular components except neutrophils, they also indicate that the BH2-Ag is expressed more strongly by mature than immature neutrophils. The purpose of this study was to determine the expression of the BH2-Ag by peripheral blood neutrophils from premature newborns to adults. Seventy-two donors were studied in six age groups: newborns <36 weeks of gestational age; newborns >36 weeks of gestational age; 0.5-2 years; 4-8 years; 12-17 years; >30 years. Expression of the BH2-Ag by peripheral blood neutrophils was examined by cytofluorography using MAB BH2-C6 directly labeled with fluorescein isothiocyanate (FITC). Neutrophils were reacted in parallel with FITC-MAB directed against CD11b, the alpha-chain of the CD11b/CD18 antigen (CR3). BH2-Ag is expressed by 98.3-99.6% of the neutrophils in all groups, and is absent on other blood cells, including those of very premature newborns. Statistical comparisons with respect to the mean fluorescence intensity of the FITC-MAB BH2C6 bound did not support a significant difference in the expression of BH2-Ag in any age group. CD11b expression was also detected in every individual studied and its mean fluorescence intensity correlated significantly with that of BH2Ag (p <0.001). The uniform presence of BH2Ag in every individual including a very premature infant suggests that BH2-Ag is likely to be an essential component of neutrophil development in humans. A highly significant correlation between the mean fluorescence intensity obtained with MAB BH2C6 and MAB CD11b suggests a possible interactive role of the two antigens in neutrophil development and/or function.
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Cytofluorometric analysis of the neutrophil-specific BH2-Ag on cells from neonates to adults.
J Investig Allergol Clin Immunol
March 2001
Department of Pediatrics, State University of New York, Health Science Center at Brooklyn, New York, USA.
Monoclonal antibody (MAB) BH2C6 recognizes a plasma membrane antigen, the BH2-Ag, specifically expressed by human neutrophils. While studies with peripheral blood and bone marrow from healthy adults clearly demonstrate the absence of BH2-Ag from other cellular components except neutrophils, they also indicate that the BH2-Ag is expressed more strongly by mature than immature neutrophils. The purpose of this study was to determine the expression of the BH2-Ag by peripheral blood neutrophils from premature newborns to adults.
View Article and Find Full Text PDFExpression of Fc(gamma)r1 (CD64) on polymorphonuclear leucocytes during progression to acquired immunodeficiency syndrome in perinatally human immunodeficiency virus-infected children.
Scand J Immunol
August 2000
Department of Pediatrics, State University of New York, Health Science Center at Brooklyn, NY 11203, USA.
CD64, the high-affinity receptor in the family of FCgamma receptors, is not expressed constitutively in polymorphonuclear leucocytes (PMN). CD64 is expressed by PMNs in the late stages of human immunodeficiency virus (HIV) infection in adults. We followed the expression of CD64 on PMNs in perinatally HIV-infected children during disease progression.
View Article and Find Full Text PDFSelective binding of a monoclonal antibody to Aspergillus niger glucose oxidase by formaldehyde fixed human polymorphonuclear leukocytes.
Cytometry
April 2000
Department of Pediatrics, State University of New York, Health Science Center at Brooklyn, Brooklyn, New York, USA.
Background: Many of the procedures used in handling neutrophils may affect the expression of surface antigens, and hence their quantitation by flow cytometry.
Methods: Because the enzyme glucose oxidase of Aspergillus niger is absent in human tissues, an IgM against it (mAb GO) was used as negative control in a study involving the normal expression of neutrophil specific BH2-Ag in different age groups.
Results: When peripheral blood leukocytes (PBL) were freshly prepared, processed and stained with FITC-mAb GO without fixation or when the cells were stained with FITC-mAb GO prior to fixation with 2% formaldehyde, both median fluorescent intensity (MFI) and per cent of positively stained polymorphonuclear leukocytes (PMN) were similar to that obtained with a background sample without any antibody.
A monoclonal antibody to a human neutrophil-specific plasma membrane antigen. Effect of the antibody on the C3bi-mediated adherence by neutrophils and expression of the antigen during myelopoiesis.
J Exp Med
February 1988
Department of Pathology, SUNY-Health Sciences Center, Brooklyn 11203.
We have used mice selectively tolerized to antigens of human lymphocytes by treatment with cyclophosphamide to raise an mAb, BH2-C6, that reacts with a plasma membrane antigen specific for human neutrophils. This specificity is demonstrated by indirect immunofluorescence microscopy, cytochemical analysis of fluorescence-positive and -negative cell populations separated by flow cytometry, and by the selective, complement-mediated killing of mAb BH2-C6-treated neutrophils. Additional evidence for the neutrophil specificity of mAb BH2-C6 is shown by immunoelectron microscopy, which demonstrates a lack of reactivity with human eosinophils.
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