Correlation of cytologic examination with ELISA assays for hyaluronan and soluble CD44v6 levels in evaluation of effusions

Alaa M Afify, Lin Chang Lynne, Lydia P Howell

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9 Citations (Scopus)

Abstract

Hyaluronan (HA) and its major cell surface receptor, CD44, play an important role in tumor growth, proliferation, neovascularization, and invasion. CD44 is an integral transmembrane protein and exists in standard form (CD44s), as well as a myriad of CD44 variants isoforms (CD44v1-v10). Functional fragments of the CD44 can be released from the cell membrane by proteolytic cleavage of extracellular domain producing soluble CD44. Although studies have proposed the use of serum HA and soluble CD44, specifically soluble CD44v6 (sCD44v6) levels, as a tumor markers, its diagnostic utility in body fluid samples has not been clearly established. The purpose of this study was to correlate HA and sCD44v6 levels in effusions with the cytology diagnosis and to assess their usefulness in differentiating between malignant and nonmalignant effusions. In this retrospective study we evaluated HA and sCD44v6 contents in 20 effusions from cytologically positive samples and 10 effusions from cytologically negative samples. Corresponding cytopathology slides were reviewed to confirm the diagnoses. Malignant effusions included 18 cases of metastatic adenocarcinomas (9 ovarian, 3 breast, 3 pulmonary, 3 adenocarcinoma of unknown primary) and 2 cases of lymphomas. The level of HA and sCD44v6 were measured using a sandwich enzyme-linked immunoadsorbent assay. For HA, we used hyaluronic acid quantitative test kit (Corgenix, Denver, CO) and for sCD44v6 we used Human sCD44v6 Instant ELISA (Bender MedSystems, Vienna, Austria). HA concentrations (μg/mL) and sCD44v6 concentrations (ng/mL) were calculated and correlated with clinical data as well as cytodiagnosis. The mean concentration of HA (22.42 ± 5 μg/mL) and sCD44v6 (70 ± 42 ng/mL) in the cytologically positive samples was significantly higher than those in the cytologically negative samples for HA (5.5 ± 5 μg/mL, P < 0.07) and sCD44V6 (17 ±1 0 ng/mL, P < 0.07). Using benign effusions as control and the upper limits of its mean levels for HA (10.5 μg/mL) as the positive boundary value, HA levels exceeded the boundary line in 17 out of 20 malignant effusions and 2 out of 10 benign effusions. Meanwhile, sCD44v6 exceeded the boundary line (27 ng/mL) in 18 out of 20 malignant effusions and 3 out of 70 benign effusions. The calculated sensitivity and specificity of this assay to the diagnosis of malignant effusions were 85 and 80% for HA and 90 and 70% for CD44v6, respectively. We conclude that the HA and sCD44v6 levels in body fluids correlate with the cytology diagnosis and could be used as an ancillary study in cytology to differentiate nonmalignant from malignant effusions.

Original languageEnglish (US)
Pages (from-to)105-110
Number of pages6
JournalDiagnostic Cytopathology
Volume35
Issue number2
DOIs
StatePublished - Feb 2007

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Hyaluronic Acid
Enzyme-Linked Immunosorbent Assay
Cell Biology
Body Fluids
Cytodiagnosis
Immunosorbents
Austria
Cell Surface Receptors
Carbon Monoxide
Tumor Biomarkers
Lymphoma
Protein Isoforms
Adenocarcinoma
Breast
Retrospective Studies

Keywords

  • Adenocarcinoma
  • CD44v6
  • Hyaluronan
  • Mesothelial cells
  • Serous fluid

ASJC Scopus subject areas

  • Anatomy

Cite this

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title = "Correlation of cytologic examination with ELISA assays for hyaluronan and soluble CD44v6 levels in evaluation of effusions",
abstract = "Hyaluronan (HA) and its major cell surface receptor, CD44, play an important role in tumor growth, proliferation, neovascularization, and invasion. CD44 is an integral transmembrane protein and exists in standard form (CD44s), as well as a myriad of CD44 variants isoforms (CD44v1-v10). Functional fragments of the CD44 can be released from the cell membrane by proteolytic cleavage of extracellular domain producing soluble CD44. Although studies have proposed the use of serum HA and soluble CD44, specifically soluble CD44v6 (sCD44v6) levels, as a tumor markers, its diagnostic utility in body fluid samples has not been clearly established. The purpose of this study was to correlate HA and sCD44v6 levels in effusions with the cytology diagnosis and to assess their usefulness in differentiating between malignant and nonmalignant effusions. In this retrospective study we evaluated HA and sCD44v6 contents in 20 effusions from cytologically positive samples and 10 effusions from cytologically negative samples. Corresponding cytopathology slides were reviewed to confirm the diagnoses. Malignant effusions included 18 cases of metastatic adenocarcinomas (9 ovarian, 3 breast, 3 pulmonary, 3 adenocarcinoma of unknown primary) and 2 cases of lymphomas. The level of HA and sCD44v6 were measured using a sandwich enzyme-linked immunoadsorbent assay. For HA, we used hyaluronic acid quantitative test kit (Corgenix, Denver, CO) and for sCD44v6 we used Human sCD44v6 Instant ELISA (Bender MedSystems, Vienna, Austria). HA concentrations (μg/mL) and sCD44v6 concentrations (ng/mL) were calculated and correlated with clinical data as well as cytodiagnosis. The mean concentration of HA (22.42 ± 5 μg/mL) and sCD44v6 (70 ± 42 ng/mL) in the cytologically positive samples was significantly higher than those in the cytologically negative samples for HA (5.5 ± 5 μg/mL, P < 0.07) and sCD44V6 (17 ±1 0 ng/mL, P < 0.07). Using benign effusions as control and the upper limits of its mean levels for HA (10.5 μg/mL) as the positive boundary value, HA levels exceeded the boundary line in 17 out of 20 malignant effusions and 2 out of 10 benign effusions. Meanwhile, sCD44v6 exceeded the boundary line (27 ng/mL) in 18 out of 20 malignant effusions and 3 out of 70 benign effusions. The calculated sensitivity and specificity of this assay to the diagnosis of malignant effusions were 85 and 80{\%} for HA and 90 and 70{\%} for CD44v6, respectively. We conclude that the HA and sCD44v6 levels in body fluids correlate with the cytology diagnosis and could be used as an ancillary study in cytology to differentiate nonmalignant from malignant effusions.",
keywords = "Adenocarcinoma, CD44v6, Hyaluronan, Mesothelial cells, Serous fluid",
author = "Afify, {Alaa M} and Lynne, {Lin Chang} and Howell, {Lydia P}",
year = "2007",
month = "2",
doi = "10.1002/dc.20585",
language = "English (US)",
volume = "35",
pages = "105--110",
journal = "Diagnostic Cytopathology",
issn = "8755-1039",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Correlation of cytologic examination with ELISA assays for hyaluronan and soluble CD44v6 levels in evaluation of effusions

AU - Afify, Alaa M

AU - Lynne, Lin Chang

AU - Howell, Lydia P

PY - 2007/2

Y1 - 2007/2

N2 - Hyaluronan (HA) and its major cell surface receptor, CD44, play an important role in tumor growth, proliferation, neovascularization, and invasion. CD44 is an integral transmembrane protein and exists in standard form (CD44s), as well as a myriad of CD44 variants isoforms (CD44v1-v10). Functional fragments of the CD44 can be released from the cell membrane by proteolytic cleavage of extracellular domain producing soluble CD44. Although studies have proposed the use of serum HA and soluble CD44, specifically soluble CD44v6 (sCD44v6) levels, as a tumor markers, its diagnostic utility in body fluid samples has not been clearly established. The purpose of this study was to correlate HA and sCD44v6 levels in effusions with the cytology diagnosis and to assess their usefulness in differentiating between malignant and nonmalignant effusions. In this retrospective study we evaluated HA and sCD44v6 contents in 20 effusions from cytologically positive samples and 10 effusions from cytologically negative samples. Corresponding cytopathology slides were reviewed to confirm the diagnoses. Malignant effusions included 18 cases of metastatic adenocarcinomas (9 ovarian, 3 breast, 3 pulmonary, 3 adenocarcinoma of unknown primary) and 2 cases of lymphomas. The level of HA and sCD44v6 were measured using a sandwich enzyme-linked immunoadsorbent assay. For HA, we used hyaluronic acid quantitative test kit (Corgenix, Denver, CO) and for sCD44v6 we used Human sCD44v6 Instant ELISA (Bender MedSystems, Vienna, Austria). HA concentrations (μg/mL) and sCD44v6 concentrations (ng/mL) were calculated and correlated with clinical data as well as cytodiagnosis. The mean concentration of HA (22.42 ± 5 μg/mL) and sCD44v6 (70 ± 42 ng/mL) in the cytologically positive samples was significantly higher than those in the cytologically negative samples for HA (5.5 ± 5 μg/mL, P < 0.07) and sCD44V6 (17 ±1 0 ng/mL, P < 0.07). Using benign effusions as control and the upper limits of its mean levels for HA (10.5 μg/mL) as the positive boundary value, HA levels exceeded the boundary line in 17 out of 20 malignant effusions and 2 out of 10 benign effusions. Meanwhile, sCD44v6 exceeded the boundary line (27 ng/mL) in 18 out of 20 malignant effusions and 3 out of 70 benign effusions. The calculated sensitivity and specificity of this assay to the diagnosis of malignant effusions were 85 and 80% for HA and 90 and 70% for CD44v6, respectively. We conclude that the HA and sCD44v6 levels in body fluids correlate with the cytology diagnosis and could be used as an ancillary study in cytology to differentiate nonmalignant from malignant effusions.

AB - Hyaluronan (HA) and its major cell surface receptor, CD44, play an important role in tumor growth, proliferation, neovascularization, and invasion. CD44 is an integral transmembrane protein and exists in standard form (CD44s), as well as a myriad of CD44 variants isoforms (CD44v1-v10). Functional fragments of the CD44 can be released from the cell membrane by proteolytic cleavage of extracellular domain producing soluble CD44. Although studies have proposed the use of serum HA and soluble CD44, specifically soluble CD44v6 (sCD44v6) levels, as a tumor markers, its diagnostic utility in body fluid samples has not been clearly established. The purpose of this study was to correlate HA and sCD44v6 levels in effusions with the cytology diagnosis and to assess their usefulness in differentiating between malignant and nonmalignant effusions. In this retrospective study we evaluated HA and sCD44v6 contents in 20 effusions from cytologically positive samples and 10 effusions from cytologically negative samples. Corresponding cytopathology slides were reviewed to confirm the diagnoses. Malignant effusions included 18 cases of metastatic adenocarcinomas (9 ovarian, 3 breast, 3 pulmonary, 3 adenocarcinoma of unknown primary) and 2 cases of lymphomas. The level of HA and sCD44v6 were measured using a sandwich enzyme-linked immunoadsorbent assay. For HA, we used hyaluronic acid quantitative test kit (Corgenix, Denver, CO) and for sCD44v6 we used Human sCD44v6 Instant ELISA (Bender MedSystems, Vienna, Austria). HA concentrations (μg/mL) and sCD44v6 concentrations (ng/mL) were calculated and correlated with clinical data as well as cytodiagnosis. The mean concentration of HA (22.42 ± 5 μg/mL) and sCD44v6 (70 ± 42 ng/mL) in the cytologically positive samples was significantly higher than those in the cytologically negative samples for HA (5.5 ± 5 μg/mL, P < 0.07) and sCD44V6 (17 ±1 0 ng/mL, P < 0.07). Using benign effusions as control and the upper limits of its mean levels for HA (10.5 μg/mL) as the positive boundary value, HA levels exceeded the boundary line in 17 out of 20 malignant effusions and 2 out of 10 benign effusions. Meanwhile, sCD44v6 exceeded the boundary line (27 ng/mL) in 18 out of 20 malignant effusions and 3 out of 70 benign effusions. The calculated sensitivity and specificity of this assay to the diagnosis of malignant effusions were 85 and 80% for HA and 90 and 70% for CD44v6, respectively. We conclude that the HA and sCD44v6 levels in body fluids correlate with the cytology diagnosis and could be used as an ancillary study in cytology to differentiate nonmalignant from malignant effusions.

KW - Adenocarcinoma

KW - CD44v6

KW - Hyaluronan

KW - Mesothelial cells

KW - Serous fluid

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