Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping

McCoy, J. Philip; Overton, W. Roy

doi:10.1002/cyto.990180304

Cited by 47 publications

(49 citation statements)

References 54 publications

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“…The percentage of T lymphocytes in our group of control cases (see above) ranged from 9% to 83%, which is in agreement with published data (33,34). Therefore, the predominance of a T-cell population is not diagnostic per se of malignancy.…”

Section: Discussionsupporting

confidence: 92%

An approach to diagnosis of T‐cell lymphoproliferative disorders by flow cytometry

Gorczyca¹,

Weisberger²,

Liu³

et al. 2002

Cytometry

122

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T-cell lymphoproliferative disorders are among the most challenging diagnoses in hematopathology.Unlike the more common B-cell disorders, in which clonality is often readily discernible by surface immunoglobulin light chain restriction, there is no specific immunophenotypic signature that is diagnostic of a clonal T-cell population. Immunophenotypic criteria that are helpful in the diagnosis of T-cell neoplasms include T-cell subset antigen restriction, anomalous T-cell subset antigen expression, deletion or diminution of one of the pan T-cell antigens, a precursor T-cell phenotype, and expression of additional markers (e.g., CD30, CD20, major myeloid antigens, and TCR␥␦). Analysis of the inherent forward and orthogonal light scatter properties of the cell can also provide important diagnostic clues. None of these features is 100% specific, however, for aberrant expression of pan-T antigens may be seen in viral infections, B-cell malignancies, or in reactive changes following administration of certain medications. An increased CD4:CD8 ratio is often observed in Hodgkin's lymphoma. Based on the analysis of 87 neoplastic and 80 control cases, we conclude that flow cytometric features that are most suspicious for malignancy include the loss or markedly dim expression of CD45; complete loss of one or more pan-T antigens; diminished expression of more than two pan-T antigens in conjunction with altered light scatter properties; and CD4/CD8 dual-positive or dual-negative expression (except thymic lesions). Cytometry (Clin. Cytometry) 50:177-190, 2002.

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Section: Discussionsupporting

confidence: 92%

An approach to diagnosis of T‐cell lymphoproliferative disorders by flow cytometry

Gorczyca¹,

Weisberger²,

Liu³

et al. 2002

Cytometry

122

View full text Add to dashboard Cite

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“…coexpressed by nearly all CD19-positive B cells. 11,15 In contrast, we found that the expression of CD20 and FMC7 did not parallel that of CD19 and CD79a on splenic B cells. To exclude the possibility that the differences between the results of our study and those of previous reports are due to the use of different flow cytometry protocols, lymphocytes from normal peripheral blood and lymph nodes were analyzed using the same method and reagents as used for spleen cells.…”

Section: Discussioncontrasting

confidence: 70%

“…11 The frequency of g/d TCR-positive cells was 6% and 11% of all CD3 positive T cells in normal and reactive spleen, respectively. These values may appear much lower than those previously reported by studies which showed that g/d TCR-positive cells represent about 17% of all CD3 positive cells in the red pulp of the spleen.…”

Section: Discussionmentioning

confidence: 90%

“…values for percent positive cells were calculated as described. 11 Student's t-test was used to compare the frequency of lymphocyte subpopulations in normal (cadaveric) vs reactive (non-neoplastic) spleens. Matched t-test was used to assess the differences between paired results obtained for B-cell subsets.…”

Section: Discussionmentioning

confidence: 99%

“…[1][2][3][4] The abnormal profiles are compared to the pattern of cell marker expression normally seen in the organ/tissue being examined, and a diagnostic interpretation is made. While the normal reference ranges for differentiation antigens expressed by lymphoid cells from peripheral blood, bone marrow, and lymph nodes are well established, [11][12][13][14][15] there is little information concerning the immunophenotypic profile of lymphocytes found in normal or reactive spleen. The aim of our study was to provide a detailed immunophenotypic characterization of human spleen lymphocytes using three-and four-color flow cytometry.…”

mentioning

confidence: 99%

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Flow cytometric analysis of normal and reactive spleen

Colovai

Giatzikis

et al. 2004

Modern Pathology

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Spleen is surgically removed for both non-neoplastic and neoplastic pathologies. A significant proportion of splenectomy specimens require distinguishing between reactive and neoplastic conditions (eg lymphoma). To establish a 'normal' reference range for the spleen lymphocyte subsets, fresh samples of benign, reactive spleens obtained from adult patients (N ¼ 12) and samples of normal spleen obtained from cadaveric transplant donors (N ¼ 14) were analyzed using three-and four-color flow cytometry. Study of pan-B, -T, and -NK marker expression revealed that the frequency of T cells is higher and that of B cells is lower in reactive (nonneoplastic) compared to normal (cadaveric) spleen. Furthermore, our study established a frame of reference for cell markers commonly used for immunophenotyping of lymphoma, and identified discrete lymphocyte subsets, such as early plasma cells and T cells carrying the phenotype of the NK/T subset. These results will facilitate an accurate interpretation of the flow cytometric analysis of human spleen lymphocytes.

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Evaluation of a novel stable whole blood quality control material for lymphocyte subset analysis: Results from the UK NEQAS immune monitoring scheme

et al. 1996

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The UK NEQAS Immune Monitoring Scheme (UK NEQAS) evaluates the performance of laboratories routinely performing T-lymphocyte subset analysis on HIV-infected individuals. The scheme originally issued fresh whole blood, but a significant problem was that of analyte stability, especially 36 h postphlebotomy.To circumvent this problem, we have developed a novel stabilisation procedure that ensures retention of leucocyte light scatter and immunological staining characteristics for up to 300 days. In addition, the stabilised whole blood preparation is fully compatible with flow cytometer technology, incorporating either whole blood lysis or "no wash, no lyse" techniques. The ranges of interlaboratory coefficient of variation for the stabilised material are now tighter than those previously obtained with fresh whole blood. Development of this novel material has enabled overseas laboratories to participate in the UK NEQAS Immune Monitoring Scheme and could, in the future, lead to the production of reference and/or calibration reagents for leucocyte immunophenotyping. o 1996 Wiley-Liss. Inc.

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Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping

Cited by 47 publications

References 54 publications

An approach to diagnosis of T‐cell lymphoproliferative disorders by flow cytometry

An approach to diagnosis of T‐cell lymphoproliferative disorders by flow cytometry

Flow cytometric analysis of normal and reactive spleen

Evaluation of a novel stable whole blood quality control material for lymphocyte subset analysis: Results from the UK NEQAS immune monitoring scheme

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