Flow cytometric quantification of intraperitoneal free tumor cells in patients with peritoneal metastasis

Kitayama, Joji; Emoto, Shigenobu; Yamaguchi, Hironori; Ishigami, Hironori; Kamei, Takao; Yamashita, Hiroharu; Seto, Yasuyuki; Matsuzaki, Kei; Watanabe, Toshiaki

doi:10.1002/cyto.b.21126

Cited by 13 publications

(15 citation statements)

References 30 publications

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“…Flow cytometry had very good sensitivity and specificity (>95%). This study extended the work of Kitiyama et al on ascites and peritoneal lavages where tumor cells and leukocytes were distinguished in a similar way.…”

Section: Detection Of Solid Tumor Cells In Body Fluids (N=1)supporting

confidence: 84%

Issue highlights – May 2016

Gratama¹

2016

Cytometry Part B Clinical

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Section: Detection Of Solid Tumor Cells In Body Fluids (N=1)supporting

confidence: 84%

Issue highlights – May 2016

Gratama¹

2016

Cytometry Part B Clinical

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“…7-Amino-actinomycin D (AAD) Staining Solution (BD Biosciences) was added 10 min before analysis to gate out nonviable cells. EpCAM-positive and 7-AAD-negative gated cells were analysed as tumour cells, as reported previously ( Kitayama et al , 2014 ).…”

Section: Methodsmentioning

confidence: 99%

IFN-γ from lymphocytes induces PD-L1 expression and promotes progression of ovarian cancer

et al. 2015

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Background:PD-L1 (programmed cell death 1 ligand 1) on tumour cells suppresses host immunity through binding to its receptor PD-1 on lymphocytes, and promotes peritoneal dissemination in mouse models of ovarian cancer. However, how PD-L1 expression is regulated in ovarian cancer microenvironment remains unclear.Methods:The number of CD8-positive lymphocytes and PD-L1 expression in tumour cells was assessed in ovarian cancer clinical samples. PD-L1 expression and tumour progression in mouse models under conditions of altering IFN-γ signals was assessed.Results:The number of CD8-positive cells in cancer stroma was very high in peritoneally disseminated tumours, and was strongly correlated to PD-L1 expression on the tumour cells (P<0.001). In mouse models, depleting IFNGR1 (interferon-γ receptor 1) resulted in lower level of PD-L1 expression in tumour cells, increased the number of tumour-infiltrating CD8-positive lymphocytes, inhibition of peritoneal disseminated tumour growth and longer survival (P=0.02). The injection of IFN-γ into subcutaneous tumours induced PD-L1 expression and promoted tumour growth, and PD-L1 depletion completely abrogated tumour growth caused by IFN-γ injection (P=0.01).Conclusions:Interferon-γ secreted by CD8-positive lymphocytes upregulates PD-L1 on ovarian cancer cells and promotes tumour growth. The lymphocyte infiltration and the IFN-γ status may be the key to effective anti-PD-1 or anti-PD-L1 therapy in ovarian cancer.

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“…28,29 Flow cytometry has been shown to be a reliable and rapid method that combines the high specificity of morphological methods and the high sensitivity of molecular methods. 28,29 Flow cytometry has been shown to be a reliable and rapid method that combines the high specificity of morphological methods and the high sensitivity of molecular methods.…”

Section: Discussionmentioning

confidence: 99%

Flow cytometry as the new ‘gold standard’ for detection of free tumour cells in abdominal lavage fluid in gastric cancer patients: A comparative study of molecular and conventional methods

et al. 2016

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Aim:The ideal method for detection of free tumour cells in abdominal lavage fluid should be rapid, reliable and widely available. Flow cytometry potentially covers these properties, but there are only a few studies directly comparing flow cytometry for detecting gastric cancer cells to other methods. Therefore, we compared free tumour-cell detection in abdominal lavage fluids using cytology with immunocytochemistry, RT-qPCR and flow cytometry. Methods: Peritoneal lavage fluid samples were collected from 10 patients. Detection of free tumour cells was performed using cytological and immunocytochemical analysis, and using RT-qPCR and flow cytometry. Tumour cells were detected according to CEA and CK20 mRNA expression levels using RT-qPCR, and with the epithelial markers EpCAM/CD326 and CEA using flow cytometry. Results: The sensitivity and specificity of cytology were 40% and 100%, respectively, while the RT-qPCR had a sensitivity of 80% and specificity of 100%. Flow cytometry showed no false-negative results and only one false-positive result, giving a sensitivity and specificity of 100% and 80%, respectively. The RT-qPCR and flow cytometry could detect free tumour cells in each of the three false-negative cases with cytology. In the samples with detected tumour cells, tumour cell clusters were observed with imaging flow cytometry, providing additional morphological confirmation. Conclusion: The combination of quantitative and qualitative data from flow cytometry and the images of tumour cells in borderline cases provide a rapid, reliable and reproducible method for detection of free tumour cells in abdominal lavage fluid, which is necessary for intraoperative selection of patients for intraperitoneal chemotherapy.

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Flow cytometric quantification of intraperitoneal free tumor cells in patients with peritoneal metastasis

Abstract: TLR measured with FACS is an excellent reflection of the tumor spread in the peritoneal cavity and could be a reliable diagnostic biomarker to determine the severity of PM as well as effectiveness of IP chemotherapy.

Cited by 13 publications

References 30 publications

Issue highlights – May 2016

Issue highlights – May 2016

IFN-γ from lymphocytes induces PD-L1 expression and promotes progression of ovarian cancer

Flow cytometry as the new ‘gold standard’ for detection of free tumour cells in abdominal lavage fluid in gastric cancer patients: A comparative study of molecular and conventional methods

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