Assessment of alkaline phosphatase on the surface membrane of neutrophils by immunofluorescence.

Abstract:

:Expression of alkaline phosphatase (ALP) on the surface membrane of neutrophils (mNAP) was studied by immunofluorescence using an anti-ALP monoclonal antibody. Fluorescent intensity distribution of mNAP was analyzed using FACS (fluorescence-activated cell sorter). The mean fluorescent intensity (MFI) of the mNAP in this assay was well correlated with the neutrophil ALP (NAP) score demonstrated cytochemically (r = 0.832). mNAP levels in various hematological disorders were evaluated by % mNAP+ cells and MFI. The levels in aplastic anemia and polycythemia vera were significantly higher, and in chronic myelocytic leukemia and paroxysmal nocturnal hemoglobinuria (PNH), the levels were significantly lower compared with the levels in healthy volunteers. Two-color immunofluorescence with anti-ALP and anti-CD16 showed that the PNH clone was essentially negative for mNAP, whereas residual normal neutrophils (CD16+) had levels slightly higher than those in normal individuals. Highly reproducible results were obtained in the blood samples which were stored at 4 degrees C for at least 24 hr without any treatment prior to immunofluorescent staining. No degradation of fluorescent intensity was seen 4 days after staining and fixation. The mNAP assay is simple, without subjective evaluation for quantification, and is useful for differential diagnosis of hematological disorders.

journal_name

Am J Hematol

authors

Shibano M,Machii T,Nishimori Y,Nakamoto I,Ueda E,Masuhara K,Kitani T

doi

10.1002/(sici)1096-8652(199901)60:1<12::aid-ajh3>3

subject

Has Abstract

pub_date

1999-01-01 00:00:00

pages

12-8

issue

1

eissn

0361-8609

issn

1096-8652

pii

10.1002/(SICI)1096-8652(199901)60:1<12::AID-AJH3>3

journal_volume

60

pub_type

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