Cytoplasmic IgM in leukemic B cells by flow cytometry

Karin C. Loftin, James M. Reuben, Evan M Hersh, Daniel Sujansky

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The presence of intracellular cytoplasmic immunoglobulin M (IgM) in leukemic cells from patients with acute lymphocytic leukemia (ALL) and chronic lymphocytic leukemia (CLL) was investigated by flow cytometry. The objective of the study was to develop a reproducible flow cytometric method. A Burkitt's lymphoma-derived B-cell line, Daudi, and a pre-B ALL, Nalm-6, served as prototypes. Normal B cells and cells from patients with chronic myelogenous leukemia (CML) were used as negative controls. Cytoplasmic μ was expressed in 77.3 ± 7.5% (n = 10) of Nalm-6 cells. CALLA+ ALL and CML cells lacked cytoplasmic μ. The surface-membrane immunoglobulin on the viable B cells was blocked with purified goat anti-human IgM. Subsequently, the B cells were fixed in cold absolute methanol and stained with a fluorescein-conjugated goat anti-human IgM to demonstrate cytoplasmic IgM. After the surface-membrane IgM was blocked, normal B cells had no cytoplasmic IgM (0.3-0.5% positive cells) detectable by flow cytometry. However, the peripheral blood lymphocytes from five patients with CLL and the Daudi cells contained cytoplasmic IgM, ranging from 7.8 to 76.7% and 45.7 to 89.3%, respectively. We conclude that cytoplasmic μ in Nalm-6, CLL, and Daudi cells can be easily and rapidly demonstrated by flow cytometry.

Original languageEnglish (US)
Pages (from-to)1379-1387
Number of pages9
JournalLeukemia Research
Volume9
Issue number11
DOIs
StatePublished - 1985
Externally publishedYes

Fingerprint

Immunoglobulin M
Flow Cytometry
B-Lymphocytes
B-Cell Chronic Lymphocytic Leukemia
Precursor Cell Lymphoblastic Leukemia-Lymphoma
B-Cell Antigen Receptors
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Goats
B-Cell Leukemia
Membranes
Burkitt Lymphoma
Fluorescein
Methanol
Lymphocytes
Cell Line

Keywords

  • B cells
  • CLL
  • cytoplasmic μ
  • Daudi
  • flow cytometry
  • Nalm-6

ASJC Scopus subject areas

  • Cancer Research
  • Hematology
  • Oncology

Cite this

Cytoplasmic IgM in leukemic B cells by flow cytometry. / Loftin, Karin C.; Reuben, James M.; Hersh, Evan M; Sujansky, Daniel.

In: Leukemia Research, Vol. 9, No. 11, 1985, p. 1379-1387.

Research output: Contribution to journalArticle

Loftin, Karin C. ; Reuben, James M. ; Hersh, Evan M ; Sujansky, Daniel. / Cytoplasmic IgM in leukemic B cells by flow cytometry. In: Leukemia Research. 1985 ; Vol. 9, No. 11. pp. 1379-1387.
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