cytology of cerebrospinal and serous body cavity · pdf filemy objectives • review normal...
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Cytology of Cerebrospinaland Serous Body Cavity
Fluids
Alex Thurman
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My Objectives
• Review normal cytology of cerebrospinal
fluid (CSF) and serous cavity fluids
• Discuss abnormal CSF and serous cavity
fluid cytology with a focus on neoplasia
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Your Objectives
• Be able to identify normal components of
CSF and serous cavity fluids
• Be able to distinguish benign from
potentially malignant cells which may be
present in these fluids
• Learn about the clinical context in which
CSF and serous cavity fluid studies are
performed
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Outline
• Normal CSF
• Normal serous fluids
• Abnormal fluids (case-based)
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Cerebrospinal Fluid
• CSF is a fluid which surrounds and
cushions the brain and spinal cord
• Produced by cells of the choroid plexus,
which line the ventricular system of the
brain
• Total volume in an adult is ~150 mL
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www.wikipedia.org
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Cerebrospinal Fluid
• CSF is generally obtained via lumbar
puncture (LP), introduced in 1891
• Malignant cells in CSF first described by a
French neurologist in 1904
• Procedure involves passing a needle
through the intervertebral space of the
lower lumbar vertebrae
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www.wikipedia.org
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Cerebrospinal Fluid
• Sensitivity of a single CSF analysis for detection of
malignancy is 54%, increases to 84% with a
second sample
• Sensitivity also depends on sample volume (more
is better!) and extent of disease
• Specificity is high, and false-positive diagnoses
occur at a rate of 2-3%
• Typically due to overcalling reactive lymphocytes
or peripheral blood contamination as
leukemia/lymphoma
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Normal Cells of CSF
• Keep in mind, CSF is a low-cellularity fluid,
typically <5 cells/mm3 (equivalent to <5000
cells/mL)
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Normal Cells of CSF
1. Lymphocytes
2. Monocytes
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Cibas and Ducatman, 2009
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Ali and Cibas, 2012
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Normal Cells of CSF
3. Ependymal cells/choroid plexus cells
– taken together, make up the ventricular lining
cells
– round to slightly oval, paracentral to eccentric
nuclei
– dispersed chromatin texture
– moderately abundant cytoplasm
– may be seen singly or in small clusters
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Ali and Cibas, 2012
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Normal Cells of CSF
4. Germinal matrix cells
– lie beneath ependymal cells
– exfoliate in the setting of hemorrhage (so-
called germinal matrix hemorrhage)
– more common in neonates (especially
preterm infants)
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Normal Cells of CSF
4. Germinal matrix cells
– small cells with fine chromatin, scant
cytoplasm, and high nucleus-to-cytoplasm
ratio
– often clustered (→ nuclear molding)
– may mimic a small cell malignancy
– typically accompanied by hemosiderin-laden
macrophages
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Cibas and Ducatman, 2009
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Normal Cells of CSF
5. Brain tissue
– seen only in samples taken directly from the
ventricles (i.e., means of sampling has to
traverse brain tissue to reach CSF)
– therefore, should not be seen in LP samples
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Cibas and Ducatman, 2009
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Normal Cells of CSF
5. Brain tissue
– fragments of brain tissue have a fibrillary
texture and may contain glial cells, neurons,
and/or capillaries
– may rarely see isolated neurons (large,
angular cells with round to ovoid nucleus and
prominent central nucleolus)
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Sheaff and Singh, 2013
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Ali and Cibas, 2012
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Normal Cells of CSF
6. Bone marrow
– if LP needle is inserted too far anteriorly, CSF
can be contaminated with bone marrow from
vertebral body
– sample will contain benign marrow elements
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Cibas and Ducatman, 2009
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Ali and Cibas, 2012
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Normal Cells of CSF
1. Lymphocytes
2. Monocytes
3. Ependymal cells/choroid
plexus cells
4. Germinal matrix cells
5. Brain tissue
6. Bone marrow
common
uncommon
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Normal Cells of CSF
• So… should you see mesothelial cells in a
CSF obtained by lumbar puncture?
• Why not?
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Serous Body Cavities
• Pleural, pericardial, and peritoneal cavities
are lined by a single layer of mesothelial
cells (“serosa”)
• These are potential spaces (contain a
small amount of fluid, enough to lubricate
surfaces for breathing, heartbeat, etc.)
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Serous Body Cavities
• In disease, fluid can accumulate, causing
an effusion
• Effusions may be transudative (low LDH,
low protein) or exudative (high LDH, high
protein) depending on causative factors
• Specimens are obtained by inserting a
needle into the cavity of interest and
removing fluid
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Serous Body Cavities
• Fluid analysis is more sensitive than blind biopsy
in detecting serosal malignancy (71% vs. 45%),
likely because fluid is more representative of the
entire cavity
• Sensitivity can be increased further with analysis
of serial samples
• Specificity is high, and false-positives occur in
<1% of cases
• When they do occur, typically due to overcalling
mesothelial atypia
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Normal Cells of Serous Fluids
1. Mesothelial cells
– often numerous
– round cells with dense, two-toned cytoplasm
(perinuclear zone lighter, peripheral zone
darker) and outer “lacy skirt”
– round, central to paracentral nucleus with
(typically) single nucleolus
– generally isolated, although occasional
clusters show intercellular windows
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Normal Cells of Serous Fluids
1. Mesothelial cells
– if reactive, may be binucleated,
multinucleated, or vacuolated
– if atypia is present, look for a spectrum
(spectrum = reassuring, two separate
populations = suspicious)
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Davidson et al., 2012
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Davidson et al., 2012
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Davidson et al., 2012
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Davidson et al., 2012; www.wikipedia.org
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Davidson et al., 2012
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Davidson et al., 2012
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Davidson et al., 2012
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Normal Cells of Serous Fluids
2. Histiocytes/macrophages
– smaller nucleus than mesothelial cells
– nucleus oval to reniform (bean-like, folded or
clefted)
– fine chromatin, inconspicuous nucleoli
– pale, ill-defined, vacuolated cytoplasm
– may contain phagocytized debris (RBCs = recent
hemorrhage, hemosiderin = remote hemorrhage)
– no intercellular windows
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Davidson Davidson et al., 2012
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Davidson et al., 2012
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Sheaff and Singh, 2013
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Normal Cells of Serous Fluids
3. Inflammatory cells (lymphocytes,
neutrophils, eosinophils)
– minor component of most effusions
– larger proportion may be present in inflammatory
conditions
– neutrophils considered significant if ≥25% of total
cells
– eosinophilic effusions (≥10% eosinophils) may be
associated with allergic, parasitic, neoplastic, or
drug-related processes, among others
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Davidson et al., 2012
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Sheaff and Singh, 2013
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Davidson et al., 2012
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Davidson et al., 2012
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Normal Cells of Serous Fluids
1. Mesothelial cells
2. Histiocytes/macrophages
3. Inflammatory cells (lymphocytes,
neutrophils, eosinophils)
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General Features of Malignant Cells in CSF and Serous Fluids
• Sometimes, you can make the diagnosis
without a microscope…
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Bibbo and Wilbur, 2008
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General Features of Malignant Cells in CSF and Serous Fluids
1. Atypical nuclear chromatin (hyperchromasia
or immature/blast-like chromatin)
2. Macronucleoli
3. Nuclear pleomorphism/nuclear membrane
irregularity
4. High nucleus-to-cytoplasm ratio (N/C ratio)
5. Clusters (especially of 12 or more cells)
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General Features of Malignant Cells in CSF and Serous Fluids
• Keeping these in mind…
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Case 1
• 63-year-old male smoker presents to
Emergency Department with exacerbation
of chronic cough
• Cough typically productive of pale green
sputum, but patient has recently seen
admixed blood
• Has also noted a 40 pound weight loss
over the last 6 months
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Case 1
• Afebrile, somewhat rapid breathing
• Decreased lung sounds over the left
hemithorax
• A chest x-ray is ordered…
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www.wikipedia.org
“white out”
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Case 1
• Afebrile, somewhat rapid breathing
• Decreased lung sounds over the left
hemithorax
• A chest x-ray is ordered…
…and diagnostic thoracentesis is
subsequently performed
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Sheaff and Singh, 2013
>12
cells??
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Sheaff and Singh, 2013
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Sheaff and Singh, 2013
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Sheaff and Singh, 2013
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Case 1
• Final diagnosis:
“Metastatic adenocarcinoma.”
• Confirmed as lung primary on
immunohistochemical analysis of cytologic
specimen
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Lung Cancer
• Likely the most common source of new
cancer worldwide
• Estimated 222,000 new cases and
157,300 deaths from lung cancer in U.S. in
2010
• Types include adenocarcinoma (most
common in U.S.), squamous cell
carcinoma, and small cell carcinoma
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Lung Cancer
• Risk factors include smoking and asbestos
exposure (asbestos is a risk factor for both
lung cancer and mesothelioma)
• The presence of a malignant effusion
significantly increases lung cancer stage
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Krueger and Buja, 2013
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Case 2
• 54-year-old female tanning enthusiast is
brought to the Emergency Department by
her daughter for neck stiffness
• Daughter says her mother has been
complaining of headaches for several
weeks, but became concerned when her
mom developed a painful, stiff neck
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Case 2
• Afebrile, mildly tachycardic
• Exam notable for neck stiffness and a
concerning skin lesion on the left posterior
calf
![Page 69: Cytology of Cerebrospinal and Serous Body Cavity · PDF fileMy Objectives • Review normal cytology of cerebrospinal fluid (CSF) and serous cavity fluids • Discuss abnormal CSF](https://reader034.vdocuments.mx/reader034/viewer/2022051305/5a7dae1e7f8b9a49588dc2fd/html5/thumbnails/69.jpg)
www.wikipedia.org
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Case 2
• CT scan shows mild meningeal
enhancement, no intracranial masses
• A lumbar puncture is performed
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Sheaff and Singh, 2013; www.wikipedia.org
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Sheaff and Singh, 2013
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Cibas and Ducatman, 2009
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Bibbo and Wilbur, 2008; www.wikipedia.org
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Case 2
• Final diagnosis:
“Malignant.”
• Biopsy of suspicious skin lesion showed
malignant melanoma
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Melanoma
• Most significant predisposing factor is exposure to
ultraviolet radiation (sunlight, tanning beds)
• Intense, intermittent exposure is particularly
harmful (“indoor job with outdoor hobbies”)
• Incidence has increased dramatically in recent
decades, likely due to a combination of increased
sun exposure and higher detection rates
• Metastases may appear many years after excision
of the primary lesion
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Krueger and Buja, 2013
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Krueger and Buja, 2013
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Case 3
• 56-year-old male presents with 2 weeks of
bloating and abdominal distension
• Has also noticed a decreased appetite
• Upon further questioning, decreased
appetite is due to an early feeling of
“fullness” when eating meals
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Case 3
• Afebrile, somewhat shallow breathing
• Abdomen distended, tympanitic to
percussion, positive fluid wave
• Diagnostic paracentesis is performed
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www.wikipedia.org
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Davidson et al., 2012
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Sheaff and Singh, 2013
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Sheaff and Singh, 2013
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Case 3
• Final diagnosis:
“Metastatic adenocarcinoma.”
• Confirmed as gastric adenocarcinoma
(diffuse type) on gastrectomy specimen
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Gastric Cancer
• One of the most commonly diagnosed cancers
worldwide
• Risk factors include Helicobacter pylori infection
and a diet rich in cured and/or smoked foods
• Incidence has declined in Western Europe and
North America but remains high in Asia
• Screening programs in Japan (similar to regular
colonoscopies in the U.S.) have improved survival
• The presence of malignant ascites is a poor
prognostic marker
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Krueger and Buja, 2013
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Krueger and Buja, 2013
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Case 4
• 58-year-old male presents with a 10-day
history of shortness of breath
• Also reports a recent history of cough and
left-sided chest pain, especially with deep
breathing
• Nonsmoker, although reports that he has
worked as a shipbuilder for almost 40
years
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Case 4
• Afebrile, but breathing somewhat rapidly
• Decreased breath sounds and dullness to
percussion over left hemithorax,
approximately to level of fourth rib
• A chest x-ray is performed…
![Page 91: Cytology of Cerebrospinal and Serous Body Cavity · PDF fileMy Objectives • Review normal cytology of cerebrospinal fluid (CSF) and serous cavity fluids • Discuss abnormal CSF](https://reader034.vdocuments.mx/reader034/viewer/2022051305/5a7dae1e7f8b9a49588dc2fd/html5/thumbnails/91.jpg)
www.wikipedia.org
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Case 4
• Afebrile, but breathing somewhat rapidly
• Decreased breath sounds and dullness to
percussion over left hemithorax,
approximately to level of fourth rib
• A chest x-ray is performed…
…followed by diagnostic thoracentesis
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Ali and Cibas, 2012
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Ali and Cibas, 2012
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Davidson et al., 2012
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Sheaff and Singh, 2013
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Ali and Cibas, 2012
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Davidson et al., 2012
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Case 4
• Final diagnosis:
“Atypical, highly concerning for
malignancy.”
• Confirmed as malignant mesothelioma on
immunohistochemical analysis of cytologic
specimen
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Mesothelioma
• One of the only malignancies directly
associated with exposure to a natural
substance (asbestos)
• Asbestos exposure can be documented in
>80% of cases—even spouses and
children of asbestos workers are at risk
• High-risk occupations including
construction, shipbuilding, pipefitting
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Mesothelioma
• Mechanism of carcinogenesis involves
chronic irritation of serosal surface by
asbestos fibers, ultimately leading to
malignant transformation
• This process may take 15-60 years (long
latency period)
• Typically presents as a unilateral pleural
effusion (detected on initial chest x-ray in 80-
95% of patients)
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Krueger and Buja, 2013
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Case 5
• 30-year-old male presents with low-grade
headache, fatigue, and inability to smile
• Recently vacationed at Martha’s Vineyard
about 2 weeks ago
• Recalls multiple tick bites during his trip
• Upon further questioning, he also reports
mild joint pain and a rash
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Case 5
• Temperature 100.2 degrees F, other vital
signs normal
• Right facial nerve paralysis and mild neck
stiffness noted, neurologic exam otherwise
normal
• Ring-like rash with central clearing on right
arm
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www.wikipedia.org
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Krueger and Buja, 2013
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Case 5
• Head CT scan negative
• Lumbar puncture is performed
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Sheaff and Singh, 2013
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Cibas and Ducatman, 2009
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Sheaff and Singh, 2013
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Cibas and Ducatman, 2009
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Case 5
• Final diagnosis:
“Lymphocytic pleocytosis with plasma
cells, favor reactive, correlate with
clinical findings and pending
serologies.”
• Serologic testing positive for Lyme disease
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Lyme Disease
• Causative agent in U.S. is spirochete Borrelia
burgdorferi
• Rodents are primary reservoir, spread to humans
via bite of infected Ixodes tick (“deer tick”)
• Highest risk areas in U.S. are Northeast region
and western Wisconsin
• Symptoms occur within weeks to months of
exposure
• Treatment is with oral or intravenous antibiotics
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www.wikipedia.org
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www.wikipedia.org
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Krueger and Buja, 2013
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www.wikipedia.org
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www.wikipedia.org
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Lyme Disease
• Plasma cells in CSF may be associated
with other conditions, including viral
meningitis, tuberculosis, syphilis, multiple
sclerosis
• May need flow cytometry to distinguish
florid B-cell pleocytosis in Lyme disease
from leukemia/lymphoma
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Summary
• Knowing what things are benign helps you to
identify what things are malignant
• General features of malignant cells are:
1. Atypical chromatin
2. Macronucleoli
3. Nuclear pleomorphism/nuclear membrane
irregularity
4. High N/C ratio
5. Clustering of cells
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Quiz
1. Which of the following is NOT a general
feature of malignant cells?
A. Atypical nuclear chromatin
B. Nuclear pleomorphism/nuclear membrane
irregularity
C. Low nucleus-to-cytoplasm ratio
D. High nucleus-to-cytoplasm ratio
E. Clusters (especially of 12 or more cells)
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Quiz
2. Which of the following is NOT a normal cell
which may be found in a cerebrospinal fluid
sample?
A. Lymphocytes
B. Monocytes
C. Ependymal cells/choroid plexus cells
D. Germinal matrix cells
E. Ciliated bronchial epithelial cells
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Quiz
3. TRUE or FALSE? Analysis of serial
samples DECREASES the sensitivity of
cerebrospinal and serous body cavity
fluid analyses.
A. TRUE
B. FALSE
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Quiz
4. TRUE or FALSE? Asbestos exposure is
a risk factor for both lung cancer AND
mesothelioma.
A. TRUE
B. FALSE
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Quiz
5. TRUE or FALSE? It is important to
identify malignant cells in cerebrospinal
and serous body cavity fluids because
malignant involvement has a significant
impact on patient prognosis.
A. TRUE
B. FALSE
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References
1. Ali SZ, Cibas ES. Serous Cavity Fluid and Cerebrospinal Fluid
Cytopathology. Essentials in Cytopathology, vol. 11. 2012.
2. Bibbo M, Wilbur D. Comprehensive Cytopathology. 3rd ed. 2008.
3. Cibas ES, Ducatman BS. Cytology: Diagnostic Principles and
Clinical Correlates. 3rd ed. 2009.
4. Davidson B, Firat P, Michael CW. Serous Effusions: Etiology,
Diagnosis, Prognosis, and Therapy. 2012.
5. Krueger GRF, Buja LM. Atlas of Anatomic Pathology with Imaging:
A Correlative Diagnostic Companion. 2013.
6. Sheaff MT, Singh N. Cytopathology: An Introduction. 2013.
7. www.wikipedia.org