Detection of melanoma cells in bone marrow using monoclonal antibodies( a comparison of fluorescence activated cell sorting (FACS) and conventional immunofluorescence (IF))

Dantas, Mary E.; Brown, Joseph P.; Thomas, Martin A.; Robinson, William A.; Glode, L. Michael

doi:10.1002/1097-0142(19830915)52:6<949::aid-cncr2820520602>3.0.co;2-2

Cited by 16 publications

(2 citation statements)

References 15 publications

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“…Detection of infrequent (< 1:10,000) cells in mixed cell populations is necessary for monitoring "minimal residual disease" in leukemia and lymphoma patients (1,8,10,12,17,18,20,23,25,30,36,40), detecting small numbers of metastatic cells in solid tumor patients (3,6,9,16,27,29,34), evaluating the efficacy of purging procedures designed to remove malignant or alloreactive cells from bone marrow harvested for transplantation (31)(32)(33)371, identifying fetal cells in maternal circulation (4,5,11,13), and assessing the frequency of mutational events (39,411. The high specificity of monoclonal antibodies has allowed the development of sensitive assays for such "rare-event analysis" using flow cytometry or manual microscopy, both of which are capable of detecting as few as one target cell per 100,000 background cells (27,361.…”

Section: Key Terms: Microcomputers Image Analysis Microscopymentioning

confidence: 99%

A digital image microscopy system for rare‐event detection using fluorescent probes

Lee¹,

Haseman

Reynolds³

1989

Cytometry

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Instrumentation for rare‐event analysis should be capable of reliably detecting infrequent cells (< 1:10,000) while both excluding false‐positive signals and including true positive cells found in multicell clumps. We have developed a digital image microscopy (DIM) system in which a cytospin of 2 million cells is scanned with an intensified video camera (ISIT) using an IBM PC AT microcomputer‐controlled microscope stage. PASCAL software controls the stage and analyzes video input, storing the location of positive cells to magnetic disk. The user can then “replay” each positive cell under computer control for either visual confirmation or analysis using other fluorescent probes. The computer requires 24 min to scan a cytoprep of 2 million cells, while playback for visual confirmation by the user averages 5 min. Using Hoechst–33342 premarked cells seeded into bone marrow as a model system, we found that the DIM system reliably detects one target cell per million marrow cells. With appropriate immunological markers, this system will aid in evaluating bone marrow purged of tumor cells prior to transplantation and should also be useful for detection of minimal residual disease in blood or bone marrow from patients with leukemia or solid tumors.

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Section: Key Terms: Microcomputers Image Analysis Microscopymentioning

confidence: 99%

A digital image microscopy system for rare‐event detection using fluorescent probes

Lee¹,

Haseman

Reynolds³

1989

Cytometry

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“…Such bone marrow involvement was correlated with an unfavorable prognosis in stage IV patients with advanced melanoma (20). The application of monoclonal antibodies specific for melanoma‐associated antigens, such as p97 for the detection of melanoma cells in bone marrow, has been tried unsuccessfully in a small panel of patients using immunofluorescence staining and fluorescence‐activated cell‐sorting techniques (22) This failure was probably due to the detection limit of 5% tumor cells per sample. In a recent report outlining a small series of patients undergoing lymphadenectomy, HMB45‐positive cells were detected in bone marrow in a subset of patients (23).…”

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confidence: 99%

Limitations of the immunocytochemical detection of isolated tumor cells in frozen samples of bone marrow obtained from melanoma patients

Schadendorf

Dorn-Beineke

Borelli

et al. 2003

Experimental Dermatology

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We report on a case of a 70-year-old woman with an ocular melanoma, which was diagnosed and treated 14 years ago. The patient was referred to the hospital with a suspected lymphoma. Cytological examination of bone marrow proved a marked infiltration with melanoma cells. Because detection of isolated tumor cells in the bone marrow of patients with various types of tumors was shown to be of prognostic significance and since current tumor-staging techniques are unable to detect single disseminated tumor cells or small aggregates of tumor cells, which might be the seed for subsequent metastatic relapse, we therefore evaluated the feasibility of immunocytochemical screening of bone marrow aspirates of 36 melanoma patients in different clinical stages using three monoclonal antibodies against melanoma-associated antigens in comparison with 43 non-melanoma control patients. Two of these antibodies (HMB45 and NKI-beteb) are directed against the melanoma antigen gp100/pmel17, whereas the third one (TA99) recognizes gp75/Tyrosinase-related protein 1 (TRP-1). None of the patients demonstrated a macroscopic bone marrow infiltration as was present in our patient with metastatic ocular melanoma. Seven (20.6%) of the 34 eligible melanoma patients presented with cells in the bone marrow positive for one or more of the above-mentioned melanosomal markers. Four of the positive patients were clinically free of tumors by the time of puncture, whereas the remaining 3 patients showed overt metastases in the subcutaneous fat (2 patients) and the brain (1 patient). On the other hand, 20 (66%) of the 29 patients with negative bone marrow findings also presented with clinical advanced disease with overt metastasis in the skin, lymph node, spleen, liver, lung, bone and brain. In conclusion, immunocytochemical screening of bone marrow samples is a feasible procedure that allows the detection of micrometastatic tumor cells in a subset of melanoma patients. Massive invasion of bone marrow with melanoma cells is a rare event even in far-advanced metastatic stages and no clear correlation between tumor load and bone marrow infiltration could be established.

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Prostate cancer: Flow cytometric methods for detection of bone marrow micrometastases

Hussain

KuKuruga²,

Biggar³

et al. 1996

Cytometry

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Up to 60% of patients with clinically localized prostate cancer will relapse despite potentially curative local treatment. Current staging tests have been limited in adequately identifying individual patients who are at a high risk for future relapse. Detection of bone marrow micrometastases may identify individuals destined to develop clinically detectable systemic metastases. Although immunohistochemistry and molecular approaches are being investigated, the most ideal test(s) are yet to be determined. In this report we describe methods for specific detection and isolation of prostate cancer micrometastases by multi‐parameter rare event flow cytometric analysis. A model was developed and validated using three human prostate cancer cell lines, healthy donor marrow, dual marker labeling for cytokeratin (epithelial‐specific marker) and CD45 (bone marrow‐specific marker). The detection sensitivity of this model was at the level of one prostate cancer cell in 100,000 nucleated bone marrow cells. As a part of an ongoing clinical study, bone marrow aspirates from 15 patients with newly diagnosed prostate cancer have been analyzed. Six patients were found to have cytokeratin positive/CD45 negative cells in their bone marrow aspirates. We conclude that flow cytometric rare event analysis provides a sensitive and specific assay for detection of bone marrow micrometastases in patients with clinically localized prostate cancer. © 1996 Wiley‐Liss, Inc.

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Detection of melanoma cells in bone marrow using monoclonal antibodies( a comparison of fluorescence activated cell sorting (FACS) and conventional immunofluorescence (IF))

Cited by 16 publications

References 15 publications

A digital image microscopy system for rare‐event detection using fluorescent probes

A digital image microscopy system for rare‐event detection using fluorescent probes

Limitations of the immunocytochemical detection of isolated tumor cells in frozen samples of bone marrow obtained from melanoma patients

Prostate cancer: Flow cytometric methods for detection of bone marrow micrometastases

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