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Updated: Oct 16, 2026
Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018
A prospective study of CD38/45 flow cytometry and immunofluorescence microscopy to detect blood plasma cells in
T E Witzig1, C Meyers, T Therneau
1Department of Internal Medicine, Mayo Clinic and Mayo Foundation, Rochester, MN 55905, USA. witzig@mayo.edu
Abstract:
Malignant plasma cells can be detected in the blood of patients with multiple myeloma (MM) using flow cytometry (FC), immunofluorescence microscopy (IM), or a variety of molecular techniques. Increased numbers of light chain-restricted blood plasma cells as detected by IM is associated with a diagnosis of overt MM and a decreased overall survival. The IM technique is time consuming; therefore, a prospective study was designed to test whether CD38 CD45 FC could simplify the procedure. Blood samples from 769 patients with plasma cell proliferative disorders were studied prospectively by FC and IM over a one-year period. The FC technique was performed on 1 ml of whole blood after ammonium chloride red blood cell lysis and utilized anti-CD38PE and anti-CD45PerCP. The number of CD38+ 45- events were enumerated and compared to the number of light chain-restricted plasma cells detected by the standard IM technique. In 46% (353/769) of cases > or = 1 CD38+ CD45- events were detected by FC whereas IM was positive for light chain restricted plasma cells in 33%; there was concordance between FC and IM in 73% of cases. In 20% of cases FC was positive and IM was negative; however, in 7% of cases FC was negative yet light chain-restricted plasma cells were detected by IM. FC was positive in 88% (134/153) of cases where the IM technique showed a high number of circulating plasma cells. This study demonstrates that two-color CD38/45 FC identifies most cases with a high IM result and reduces the workload in the clinical laboratory. The prognostic implications of a positive FC screen but a negative IM will require long-term patient follow-up.
Insights
Flow cytometry (FC) using CD38/45 markers can efficiently detect malignant plasma cells in multiple myeloma (MM) patients. This method simplifies diagnosis and reduces laboratory workload compared to immunofluorescence microscopy (IM).
Area of Science:
- Hematology
- Clinical Pathology
- Immunology
Background:
- Malignant plasma cells in multiple myeloma (MM) are detectable via flow cytometry (FC), immunofluorescence microscopy (IM), and molecular methods.
- Increased light chain-restricted plasma cells in blood, identified by IM, correlate with overt MM and reduced survival.
- The IM technique is labor-intensive, necessitating a simpler diagnostic approach.
Purpose of the Study:
- To evaluate the efficacy of CD38/45 two-color flow cytometry (FC) as a simplified method for detecting malignant plasma cells.
- To compare the diagnostic performance of FC with the standard immunofluorescence microscopy (IM) technique in plasma cell proliferative disorders.
Main Methods:
- A prospective study involved 769 patients with plasma cell proliferative disorders over one year.
- Blood samples were analyzed using CD38/45 two-color FC and standard IM.
- FC involved ammonium chloride lysis of red blood cells followed by staining with anti-CD38PE and anti-CD45PerCP.
Main Results:
- FC detected CD38+ CD45- events in 46% of cases, while IM identified light chain-restricted plasma cells in 33%.
- Concordance between FC and IM was observed in 73% of cases.
- FC showed higher sensitivity, being positive in 88% of cases with high IM results, and identified additional cases not detected by IM.
Conclusions:
- Two-color CD38/45 FC effectively identifies most cases with high IM results, significantly reducing clinical laboratory workload.
- FC offers a more streamlined approach for detecting circulating plasma cells in MM patients.
- Further long-term follow-up is needed to clarify the prognostic significance of FC-positive, IM-negative results.
