Bruce Greig

2006 Bethesda International Consensus recommendations on the immunophenotypic analysis of hematolymphoid neoplasia by flow cytometry: Optimal reagents and reporting for the flow cytometric diagnosis of hematopoietic neoplasia†

Immunophenotyping by flow cytometry has become standard practice in the evaluation and monitoring of patients with hematopoietic neoplasia. However, despite its widespread use, considerable variability continues to exist in the reagents used for evaluation and the format in which results are reported. As part of the 2006 Bethesda Consensus conference, a committee was formed to attempt to define a consensus set of reagents suitable for general use in the diagnosis and monitoring of hematopoietic neoplasms. The committee included laboratory professionals from private, public, and university hospitals as well as large reference laboratories that routinely operate clinical flow cytometry laboratories with an emphasis on lymphoma and leukemia immunophenotyping. A survey of participants successfully identified the cell lineage(s) to be evaluated for each of a variety of specific medical indications and defined a set of consensus reagents suitable for the initial evaluation of each cell lineage. Elements to be included in the reporting of clinical flow cytometric results for leukemia and lymphoma evaluation were also refined and are comprehensively listed. The 2006 Bethesda Consensus conference represents the first successful attempt to define a set of consensus reagents suitable for the initial evaluation of hematopoietic neoplasia.

2006 Bethesda International Consensus recommendations on the immunophenotypic analysis of hematolymphoid neoplasia by flow cytometry: Recommendations for training and education to perform clinical flow cytometry†

As clinical flow cytometry practices continue to expand and immunophenotyping for leukemia and lymphoma becomes more widespread, the need for defined guidelines for training of medical professionals is imperative. Standards of expected knowledge and skills are necessary to ensure reliable test results as well as provide direction to those who are considering adding flow cytometry to their clinical laboratory practice. Before now, no clear guidelines have been established for defining the areas of responsibility, education and training standards, and credentials that would be required to perform clinical flow cytometry for leukemia and lymphoma.

Subcutaneous panniculitic T‐cell lymphoma in a cardiac allograft recipient

Background:  Post‐transplant lymphoproliferative disorder (PTLD) is the third leading cause of death in heart transplant patients beyond the immediate peri‐operative period (Ouseph R, Denny DM, Erbeck KM. J Am Soc Echocardiogr 1998; 11: 758; Armitage JM, Kormos RL, Stuart RS, et al. J Heart Lung Transplant 1991; 10: 877; Swinnen LJ, Mullen M, Carr TJ, et al. Blood 1995; 86: 3333; Ying AJ, Myerowitz D, Marsh WL. Ann Thorac Surg 1997; 64: 1822). The majority of PTLD cases are of B‐cell origin whereas T‐cell neoplasms have been reported as rare, aggressive, and late complications of solid‐organ transplantation (Fatio R, Sütsch G, Mayer K, et al. Transplant Proc 1998; 30: 1118).

Stabilization media increases recovery in paucicellular cerebrospinal fluid specimens submitted for flow cytometry testing.

Flow cytometric immunophenotpying (FCI) of cerebrospinal fluid (CSF) and other paucicellular fluids has been demonstrated to have increased sensitivity in detection of lymphoma and leukemia when compared to cytomorphology [(1) de Graaf et al., Cytometry Part B 2011, 80B:271–281; (2) Szamosi et al., CLSI Document H56‐A—Body Fluid Analysis for Cellular Composition; Approved Guideline, Wayne, PA: Clinical and Laboratory Standards Institute, 2006; (3) Kraan et al., Flow Cytometric Immunophenotyping of Cerebrospinal Fluid. Current Protocols in Cytometry, Hoboken, NJ: Wiley, 2008]. However, low cellularity has been an historical problem with these samples. Several studies indicate that immediate addition of a stabilization media (e.g., RPMI with fetal calf serum (FCS)) to CSF improves the cell yield for FCI [(1) de Graaf et al.]. Such stabilization medias can, however, significantly increase cost.

B-ALL minimal residual disease flow cytometry: an application of a novel method for optimization of a single-tube model.

OBJECTIVES Optimizing a clinical flow cytometry panel can be a subjective process dependent on experience. We develop a quantitative method to make this process more rigorous and apply it to B lymphoblastic leukemia/lymphoma (B-ALL) minimal residual disease (MRD) testing. METHODS We retrospectively analyzed our existing three-tube, seven-color B-ALL MRD panel and used our novel method to develop an optimized one-tube, eight-color panel, which was tested prospectively. RESULTS The optimized one-tube, eight-color panel resulted in greater efficiency of time and resources with no loss in diagnostic power. CONCLUSIONS Constructing a flow cytometry panel using a rigorous, objective, quantitative method permits optimization and avoids problems of interdependence and redundancy in a large, multiantigen panel.

A QA program for MRD testing demonstrates that systematic education can reduce discordance among experienced interpreters

Minimal residual disease (MRD) in B lymphoblastic leukemia (B‐ALL) by flow cytometry is an established prognostic factor used to adjust treatment in most pediatric therapeutic protocols. MRD in B‐ALL has been standardized by the Childrens Oncology Group (COG) in North America, but not routine clinical labs. The Foundation for National Institutes of Health sought to harmonize MRD measurement among COG, oncology groups, academic, community and government, laboratories.