NON HODGKIN LYMPHOMA (NHL)

European Federation of Cytology Societies 4tu
Annual Tutorial in Cytopathology
Trieste, June 66-10, 2011
NON HODGKIN LYMPHOMA (NHL)
NHL Classification
The World Health Organization Classification of Lymphomas (2001)
B-CELL
B
CELL NEOPLASMS
T-CELL
T
CELL AND NK
NK-CELL
CELL NEOPLASMS
Precursor B cell neoplasm
•Precursor B-lymphoblastic leukemia/lymphoma
•Precursor
Precursor B-cell
B cell acute lymphoblastic leukemia
Precursor T-cell neoplasm
•Precursor T-lymphoblastic lymphoma/leukemia
•Precursor
Precursor T-cell
T cell acute lymphoblastic leukemia
Mature (peripheral) B-cell neoplasms
•B-cell chronic lymphocytic leukemia/
small lymphocytic lymphoma (SLL/CLL)
•B-cell
B ll prolymphocytic
l
h
ti lleukemia
k i
Lymphoplasmacytic lymphoma (LPCL)
•Splenic marginal zone B-cell lymphoma
(with/without villous lymphocytes)
•Hairy cell leukemia
•Plasma cell myeloma/plasmacytoma
•Extranodal marginal zone B-cell lymphoma
of MALT type (MALT)
•Nodal marginal zone B-cell lymphoma
(with/without monocytoid Bcells) (MZL)
•Follicular lymphoma (FL)
•Mantle-cell lymphoma (MCL)
•Diffuse large B-cell lymphoma (DLBCL)
•Burkitt lymphoma/Burkitt cell leukemia (BL)
Mature (peripheral) T-cell neoplasms
•T-cell prolymphocytic leukemia
•T-cell granular lymphocytic leukemia
•NK-cell
NK ll llymphomaleukemia
h
l k i (NKL)
•Adult T-cell lymphoma/leukemia
(HTLV-1 positive)
•Extranodal NK/T-cell lymphoma, nasal type
•Enteropathy-type
E t
th t
T-cell
T ll llymphoma
h
•Hepatosplenic gamma-delta T-cell lymphoma
•Subcutaneous panniculities-like T-cell lymphoma
•Mycosis fungoides/Sezarysyndrome
•Anaplastic
A
l ti llarge-cellll llymphoma,
h
T/
T/nullll cell,
ll
primary cutaneous type
•Peripheral T-cell lymphoma,
not otherwise characterized (PTL)
•Angioimmunoblastic
A i i
bl ti T
T-cellll llymphoma
h
•Anaplastic large-cell lymphoma, T/null cell,
primary systemic type
Cytology, phenotype (FC), IGH quantitative assessment (PCR), l
h
( )
(
)
IGH integrity evaluation (FISH) of lymphoid cells
IGH probe, split signal
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
•
IGH (14q32) is the most frequently involved in different translocations of B‐cell NHL
t (14;18)(q32;q21) in FL and a subset of DLBCL
t (14;18)(q32;q21) in FL
and a subset of DLBCL
t (11;14)(q13;q32) in MCL
t (q14;q32) in 60 % in MM with different chromosomal partners
t (9;14)(p13;q32) in 50% of LPCL and some cases of MZL
t (8;14)(q24;q32) in 100% of BL
• The The IGH FISH DNA probe, split signal, is a mixture of two fluorochrome:
IGH FISH DNA probe, split signal, is a mixture of two fluorochrome:
the green labelled DNA probe that binds to a 612 kb telomeric segment, and
the red labelled that binds to a 460 kb centromeric segment, to the IGH breakpoint
•
Therefore IGH FISH DNA probe
probe, split signal should detect any breakage involving the IGH
locus at chromosome 14q32.
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Indirect FC markers of malignancy
g
y
CD5/CD19 co‐expression
/
p
g
CD19/CD10 co‐expression in >50% of the gated cells
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
ICC-FC sub-classification of small/medium cell size NHL
phenotype
LPCL
SLL
CLL
MCL
FL
MZL/
MALT
SNCL
PTL
NKL
k:λ>4:1 or <1:3
+
+
+
+
+
+/
+/-
-
-
CD19/CD5+
-
+
+
-
-
-
-
-
C
CD19/CD10+
/C
-
-
-
+
-
+
-
-
CD19/CD23+
+/-
+
-
-
-
-
-
-
CD19/FMC7+
+/-
-
+
+/-
-
-
-
-
CD19/CD38+
+
-
-
-
-
-
-
-
bcl-2+/CD10
-
-
-
+
-
-
-
-
CD4+/CD8- or CD4-/CD8+
-
-
-
-
-
-
+
-
CD5+ CD19-
-
-
-
-
-
-
+
-
CD2/CD3+CD7- or
CD2/CD7+CD3-
-
-
-
-
-
-
+
-
CD3+/- CD56+ CD19-
-
-
-
-
-
-
-
+
Chronic lymphatic leukaemia/small lymphocytic
lymphoma (CLL/SLL)
•
•
•
•
•
•
Incidence: 7-8%
7 8% of all NHL.
NHL
Cytology: Smears show monotonous, dispersed cell populations
composed of small lymphocytes with clumped chromatin and
occasional small,
occasional,
small nucleoli; mitoses are absent or rare.
rare Accelerate
phases of CLL/SLL show larger, nucleolated,
cells
(paraimmunoblasts) and mitoses, and should not be confused with
Richter’s syndrome, which is a large-cell “histiocytic” lymphoma
arising
i i on a CLL/SLL background.
b k
d
ICC: light chain restriction, CD5+, CD20+, CD23+; Ki 67 is positive
in a small percentage of the cells.
FC CD5/CD19+,
FC:
CD5/CD19 CD19/CD23+,
CD19/CD23 CD19+/CD10-,
CD19 /CD10 κ or λ light
li ht chain
h i
restriction.
FISH: trisomy 12, 11q22 (ATM), 12, 13q14.3, 13q34.3 (LAMP1), and
17p13 1 (p53) as prognostic markers.
17p13.1
markers
Differential diagnosis: Non-specific hyperplasia ML, FL, MZL.
Chronic lymphatic leukemia/small lymphocytic lymphoma
(CLL/SLL)
CLL/SLL: A monotonous monomorphous cell population of small lymphocytes with clumped
chromatin and occasional, small, nucleoli.
Chronic lymphatic leukemia/small lymphocytic lymphoma (SLL/CLL)
SLL/CLL cy: a monomorphic, dispersed cell population composed by small lymphocytes and
occasional larger nuclei
nuclei. “glandular
glandular bodies”)are
bodies )are present in the background
background.
ICC of SLL/CLL on cytospins
y
showing negativity for
lambda and positivity for
kappa light chains
(peroxidase/anti peroxidase
immunostain)
lambda
kappa
Chronic lymphatic leukemia/small lymphocytic lymphoma (CLL/SLL)
CLL/SLL: FNA smear and corresponding FC showing CD19/CD5 and CD19/CD23 coexpression and kappa light chain restriction.
Chronic lymphatic leukemia/small lymphocytic lymphoma (CLL/SLL)
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Lymphoplasmacytoid Lymphoma, Immunocytoma (LPCL)
•
•
•
•
•
Incidence: 1-2%
1 2% of all NHL.
NHL
Cytology: dispersed cell population composed of small lymphocytes
with plasmacytoid differentiation; mature plasma cells are also
present.
ICC: CD20+, CD5-,CD10-, CD23-, Ki67+ in a variable percentage
(10-20%) of cells.
cells
FC: CD5-/CD19+, CD19+/CD23+, CD19+/CD10-, CD19+/CD38+, κ
or λ light chain restriction.
Differential diagnosis: Non-specific hyperplasia, SLL/CLL.
Lymphoplasmacytoid Lymphoma, Immunocytoma (LPCL)
LPCL dispersed small lymphocytes with clumped chromatin and plasmacytoid differentiation
Lymphoplasmacytoid Lymphoma (LPCL)
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Mantle cell lymphoma (MCL)
•
•
•
•
•
•
Incidence: 7-10% of all NHL.
C t l
Cytology:
di
dispersed
d cellll population
l ti composed
d off small/medium-size
ll/ di
i
lymphocytes with irregular nuclei and a thin rim of pale cytoplasm.
Nuclei may have slightly irregular shape, clumped chromatin and
small nucleoli,
nucleoli or may be cleaved.
cleaved Mitoses are scanty whereas they
may be numerous in the “blastoid” variant
ICC. CD20+, CD5+,CD10-, CD23-, Ki67+ in a variable percentage
((10-20%)) of the cells.
FC: CD5+/CD19+, CD19/FMC7+, CD19+/CD23-, CD19+/CD10-, κ
or λ light chain restriction.
(
)(q q )
FISH: t(11;14)(q13;q32).
Differential diagnosis: Non-specific hyperplasia, SLL/CLL,FL, MZL.
Mantle cell lymphoma (MCL)
MCL: dispersed cell population composed by medium-size lymphocytes. Nuclei may be
slightly irregular in shape or cleaved and have clumped chromatin and small nucleoli
nucleoli.
Mantle cell lymphoma (MCL)
Mantle cell lymphoma (MCL)
Cyclin D1 - bcl-1
MCL: dispersed cell population composed
by medium-size lymphocytes with irregular
nuclei. Nuclei are slightly irregular in
shape or cleaved and have clumped
chromatin and small nucleoli
nucleoli.
MCL: nuclear positivity for cyclin D1.
Mantle cell lymphoma (MCL)
Ki67
MCL: Medium-size lymphocytes with
irregular nuclei and clumped chromatin.
Frequent mitoses may be seen.
MCL: Ki67 immunostain may shows nuclear
positivity in a high percentage e of cells.
Mantle cell lymphoma (MCL)
MCL: Medium-size lymphocytes
with irregular nuclei and clumped
chromatin. Nuclei may show a
kind of moulded pattern.
.
MCL: FISH demonstration of
t(11;14)(q13;q32): locus 14q32
(IGH) is labelled in green and
locus 11q13 ((CNDD1/PRAD1))
is labelled in orange. Note
fusion signals (yellow) in which
the green signals overlap the
orange signals.
Mantle cell lymphoma (MCL)
FC of MCL showing CD19/CD5 and CD19/FMC7 co-expression and lambda light chain
restriction.
Follicular lymphoma (FL)
•
•
•
•
•
•
Incidence: 22-30% of all NHL.
Cytology: dispersed cell population of medium- to large-size lymphocytes,
with irregular nuclei and a thin rim of pale cytoplasm. Nuclei may have
irregular shape,clumped chromatin and inconspicuous nucleoli in grade I,
or large nucleiand two or more marginal nucleoli in grades II and III.
III
ICC: CD20+,CD5-,CD10+,CD23-,Ki67+ in a variable % of the cells.
FC: CD5-/CD19+, CD19+/FMC7+, CD19+/CD23-, CD19+/CD10+,
CD10/bcl 2+ κ or λ light chain restriction.
CD10/bcl-2+,
restriction
FISH: t(14;18)(q32;q21).
Differential diagnosis: Non-specific hyperplasia, SLL/CLL,MCL.
Phenotype
Follicular lymphoma (FL)
Low-grade FL: a monotonous population of small lymphocytes with clumped chromatin
and inconspicuous nucleoli. Scattered large cells are present in the background.
Low-grade FL: Ki-67 show positivity in a relative small percentage of the cells.
Follicular lymphoma (FL)
Low-grade FL: a monotonous population of small
lymphocytes with clumped chromatin and
inconspicuous nucleoli. Scattered large cells are
present in the background.
Low-grade FL: Ki-67 show positivity in a relative small
percentage of the cells.
Follicular lymphoma (FL)
Intermediate-grade
Intermediate
grade FL: a relative monomorphous population of medium
medium-sized
sized lymphocytes
with clumped chromatin, nuclear abnormalities and one or more nucleoli.
intermediate-grade FL: Ki-67 show positivity in a large percentage of the cells.
Follicular lymphoma (FL)
High-grade FL: a relative monomorphous population of large lymphoid cells with dispersed
chromatin, nuclear abnormalities and one or more large nucleoli.
High-grade FL: Ki-67 show positivity in almost all the large cells present in the smear.
Follicular lymphoma (FL)
High-grade
Hi
h
d FL
FL: a relative
l ti polymprphous
l
h
population
l ti off llarge llymphoid
h id cells
ll with
ith di
dispersed
d
chromatin, nuclear abnormalities and one or more large nucleoli. Scattered small
lymphocytes are present in the background.
Follicular lymphoma (FL)
Low--grade FL
Low
High--grade FL
High
FC of FL showing CD19/CD10 co-expression, bcl-2 overexpression and kappa light chain restriction.
Follicular lymphoma (FL)
FL: a relative polymorphous population of medium
medium-sized
sized lymphocytes with dispersed
chromatin, nuclear abnormalities, and one or more small nucleoli. FC shows CD10/bcl-2 coexpression that reflects the follicles bcl-2 histological positivity.
Follicular lymphoma (FL) with dim or no light chains expression
t(14;18)(q32;q21)
IIn cases off FL (CD10/19 co-expression)
i ) with
ith di
dim or no lilight
ht chains
h i expression,
i
FISH may b
be
helpful in diagnosis by the demonstration of t14-18. Bcl2 locus (18q21,labelled red), overlaps
IGH locus (14q32, labelled in green) giving a signal of fusion (yellow).
IGH gene rearrangement in lymphoma with dim or no light chains expression
FC in a lymphoma without light
chains expression
Detection of clonal IGH gene rearrangements: In PCR, rearranged DNA is amplified with a series
of consensus primers that are complementary to sequences of variable regions framework 1,
framework 2
2, and framework 3 and to joining regions (JH) of the IGH gene
gene. Monoclonal and
polyclonal PCR products can be discerned by their size distributions. There are several
methodologies for evaluating PCR amplifications; here conventional electrophoresis in ethidium
bromide–stained agarose gels (AGE). If a significant population of cells contains the same unique
IGH rearrangement, it appears as a well-defined band on each of the AGE here reported. A clonal
population of cells is indicated by the presence of a distinct non–germline band, whereas normal
or reactive populations have only germline bands and should produce multiple bands (smears).
Marginal zone B-cell lymphoma (MZL)
•
•
•
•
•
Incidence: 2-3% of all NHL.
Cytology: dispersed cell population composed of small lymphocytes with
irregular nuclei. Nuclei may have slightly irregular shape, dense chromatin
pale cytoplasm
y p
observed in
and indistinct nucleoli,, or be cleaved. The rim of p
histological sections is rare or undetectable on the smears. Monocytoid
cells and plasma cells may be intermingled, mitoses are scanty.
ICC: CD20
CD20+,, CD5
CD5-,CD10-,
,CD10 , CD23
CD23-,, Ki67
Ki67+/-.
/.
FC: CD5-/CD19+, CD19+/FMC7-, CD19+/CD23-, CD19+/CD10-, κ or λ light
chain restriction.
Differential diagnosis: Non-specific
Non specific hyperplasia,
hyperplasia SLL/CLL,MCL,
SLL/CLL MCL FL.
FL
Marginal zone B-cell lymphoma (MZL)
MZL: ultrasound guided FNC of the spleen with diffuse splenomegaly (note the tip of the
needle).
) FNC
C shows a relative monomorphous population off small lymphocytes with clumped
chromatin. FC shows CD5-, CD10-,kappa light chain restriction with CD19/kappa coexpression.
Extranodal B-cell lymphoma of mucosa associated lymphoid
tissue (MALT lymphoma)
MALT lymphoma: ultrasound guided FNC of an extremely thick bowel wall with former
inconclusive endoscopic biopsies. FNC shows atypical lymphoid cells in a necrotic background.
FC shows CD5- and kappa light chain restriction.
Diffuse large B-cell lymphoma (DLBCL)
•
•
•
•
•
Incidence: 30-40% of all NHL.
C t l
Cytology:
di
dispersed
d large
l
l
lymphoid
h id cells
ll with
ith irregular
i
l nuclei,
l i one or
more large nucleoli and a wider rim of pale cytoplasm. Variants of
DLBCL are centroblastic, immunoblastic T-cell/histiocyte-rich and
anaplastic Nuclei may be slightly irregular in shape or cleaved and
anaplastic.
have dense chromatin and indistinct nucleoli. Lymphocyte and
plasma cells may be intermingled, Mitoses are scanty.
percentage
g
ICC: CD19+,, CD20+,, 22+,, CD79a+,, Ki67+ in a variable p
(10-20%) of the cells.
FC: CD5-/CD19+, CD19+/FMC7-, CD19+/CD23-, CD10-, κ or λ light
chains restriction.
DD: HL, metastases.
Diffuse large B-cell lymphoma (DLBCL)
DLBCL : dispersed cell population composed by large lymphoid cells with irregular nuclei, one
or more large nucleoli. Scatterd small lymphocytes are present in the background.
DLBCL : ICC:CD20+.
Diffuse large B-cell lymphoma (DLBCL)
DLBCL : dispersed cell population composed by large lymphoid cells with irregular nuclei
nuclei, one or
more large nucleoli, immature chromatin and nuclear crushes. Cytoplasm may be present or not.
Plasma cell myeloma/plasmacytoma
•
•
•
•
•
Incidence: 15% of haematological malignancies.
Cytology: in typical cases show dispersed cell population composed
of small/medium-size plasma cells. Cells show one or two nuclei
with
ith clumped
l
d ““cartwheel”
t h l” chromatin
h
ti and
d smallll nucleoli.
l li C
Cytoplasm,
t l
when present are well defined with evident perinuclear Golgi
apparatus. In poorly differentiated cases nuclei are atypical and may
show marked anisonucleosis.
ICC. CD20-/+, CD5-, CD10-, CD23-, CD38, CD79a, EMA, Ki67+ in a
variable percentage.
FC: CD38
CD38+,, CD38/56 +/-,
/ , CD10
CD10-,, κ or λ light chain restriction.
Differential diagnosis: non-haematological tumours in poorly
differentiated cases.
Plasma cell myeloma/plasmacytoma
y
p
y
Extramedullary plasmacytoma shows a dispersed cell population of small and medium
size plasma cells. Cells show one or two nuclei with clumped “cartwheel” chromatin and
small nucleoli.
nucleoli Cytoplasm,
Cytoplasm when present are well defined with evident perinuclear Golgi
apparatus. Note binucleated cells.
Plasma cell myeloma/plasmacytoma
FC of Plasma cell myeloma/plasmacytoma : CD19-, CD38+, CD38/56 co-expression in 70-80%
of the cases Lack of CD56 expression has been considered a negative prognostic factor.
Burkitt lymphoma (BL)
•
•
•
•
•
Incidence: 1-2% of all NHL.
Cytology: dispersed cell population composed of undifferentiated
medium sized lymphocytes with dispersed chromatin and one or
more small basophilic nucleoli; the cytoplasm is basophilic and
g , resulting
g in
sometimes vacuolated. Immature nuclei are fragile,
frequent nuclear crushes in the smear; a high mitotic index is
observed. Scattered macrophages with engulfed cytoplasm may
give a starry sky pattern to the smear.
ICC CD20+,
ICC:
CD20+ CD5-,
CD5 CD10+,
CD10+ Bcl-6+,
B l 6+ CD23-,
CD23 Ki67+ in
i a high
hi h
percentage of the cells.
FC: CD5-/CD19+, CD19+/FMC7-, CD19+/CD23-, CD19+/CD10+,
light chain often unexpressed.
unexpressed
FISH: t(8;14)(8q24;q32).
Burkitt lymphoma (BL)
BL: dispersed cell population composed by undifferentiated medium sized lymphocytes with
dispersed chromatin and one or more small basophilic nucleoli; Immature nuclei are fragile
resulting in frequent nuclear crushes in the smear; an high mitotic index is observed
observed.
Scattered macrophages with engulfed cytoplasm may give a starry sky pattern to the smear.
Burkitt lymphoma (BL)
BL: dispersed cell population composed by undifferentiated medium sized
lymphocytes with immature chromatin. Numerous mitoses may be observed. FC:
almost all the cells shows CD19/CD10 co
co-expression.
expression
anaplastic large cell lymphoma
•
•
•
•
•
•
•
•
Incidence: 3% of all NHL.
NHL
Cytology: dispersed and polymorphous large cells with very irregular
shape. Multilobated, horseshoe bi- and multinuclecleated cells are
present. The nuclei have coarse chromatin and larger nucleoli. The
background is generally polymorphous with mature lymphocytes,
granulocytes and eosinophils. Atypical mitoses and necrosis may be
seen.
ICC: CD20-, CD45+ CD15-, CD30++, Ki67+, EMA+, ALK-1+/-.
FC: CD5+/-, CD19+/-, aberrant phenotypes may be observed.
FISH: t(2;5)(p23;q35).
( ; )(p ;q )
PCR: clonal rearrangement of the T-cell receptor (90%).
DD: DLBCL, HL, metastases.
anaplastic large cell lymphoma
Ki67
Large anaplastic, nucleolated cells, Ki67+ with small lymphocytes and granulocytes in the
background.
anaplastic large cell lymphoma
ALK
Large anaplastic
anaplastic, nucleolated cells
cells, ALK+ interspersed between small lymphocytes
lymphocytes.
Peripheral T-cell lymphoma (PTL)
•
•
•
•
•
•
Incidence: 7-8% of all the NHL.
NHL
Cytology: dispersed and polymorphous medium- or large-sized cells
with irregular shape, vesicular nuclei and evident nucleoli. The
nuclei have irregular shape, coarse chromatin and one or more large
nucleoli. Binucleated cells are frequently observed. The background
is generally polymorphous; mature lymphocytes, granulocytes,
numerous eosinophils may be present.
ICC:
CC CD20-,
C 20 CD3+,
C 3
C
CD4+
C 10 CD23-,
CD10-,
C 23 Ki67+
6
in a variable
percentage of the cells.
FC: CD5+, CD19-, CD10-, CD2+ CD3+ CD7-.
PCR TC receptor rearranged
PCR:
d in
i most cases.
DD: B-cell NHL.
Peripheral T-cell lymphoma (PTL)
PTL shows a wide spectrum of cytological features: dispersed or loosely aggregated,
monomorphic or polymorphous small or medium
medium-sized
sized cells with irregular shape nuclei with
evident or inconspicuous nucleoli. The background may be polymorphous with mature
lymphocytes granulocytes and/or eosinophils.
Peripheral T-cell lymphoma (PTL)
CD 20
CD45 Ro
PTL shows p
polymorphous
y
p
medium-sized cells with irregular
g
shaped
p nuclei and small
nucleoli. FC: CD19-, CD10-, ICC shows CD20 negativity with few scattered positive cells
and diffuse positivity for CD45Ro.
Peripheral T-cell lymphoma (PTL)
PTL shows polymorphous medium-sized cells with irregular shaped nuclei and scattered small
lymphocytes in the background. FC: CD19-, CD5+, CD10-, CD4+, CD8-, CD2+, CD2/3+ CD7. Loss of CD7 expression is frequently observed in PTL.
Peripheral T-cell lymphoma (PTL)
CD45Ro
PTL shows medium-sized cells poorly differentiated lymphoid cells with irregular shaped nuclei
and small nucleoli. ICC shows diffuse positivity for CD45Ro. FC: CD19-, CD5+, CD2+,CD7-.
Precursor T-lymphoblastic
y p
lymphoma/leukemia
y p
((TLL))
•
•
•
•
•
IIncidence:
id
<1%
1% off allll the
h NHL.
NHL
Cytology:
dispersed
and
polymprphous
medium-sized
undifferentiated cells. Nuclei are round or convoluted with dispersed
p
chromathin and small nucleoli. High mitotic index and nuclear
fragility.
ICC: CD20-,
CD20 CD3+,
CD3+ CD10-,
CD10 TDT+,
TDT+ Ki67+ in a high percentage of the
cells.
FC: CD5 +/-, CD19-, CD10-, CD2+ CD3+ CD7-.
DD: BL.
Precursor T-lymphoblastic lymphoma/leukemia (TLL)
dispersed and polymprphous medium-sized poorly differentiated cells. Nuclei are irregular
with with clumped chromathin and small nucleoli. Numerpous mitosesmay be seen.
CD3
TLL: diffuse CD3+
NK-cell lymphoma/leukemia
•
•
•
•
•
Incidence: <1% of all the NHL.
NHL
Cytology: dispersed and monomorphous small- or medium-sized
undifferentiated cells. The nuclei are round or convoluted with
dispersed chromatin and small nucleoli. There is also a high mitotic
index and nuclear fragility.
ICC: CD20
CD20-, CD3+,
CD3+ CD10
CD10-, TDT+,
TDT+ CD56+ Ki67+ in a high
percentage of the cells.
FC: CD56+, CD5 -, CD19-, CD10-, CD2-CD3- CD7-.
DD: PTL, LLA, TLL.
NK ll l
NK‐cell lymphoma/leukemia
h
/l k i
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Blastic NK-cell lymphoma/leukemia
Blastic NK lymphoma: dispersed and monomorphous medium-size
medium size and poorly differentiated
or undifferentiated cells. The nuclei are irregular and fragile with dense chromatin and small
nucleoli. Nuclear crushes and atypical mitoses are present. FC is “mute” except for CD56+
Pitfalls and shortcomings !
Pitfalls and shortcomings !
Hodgkin Lymphoma
anaplastic large cell lymphoma
CD30
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
F, 57‐yrs,
F, 57
yrs, FNC of 50 mm nodule in the left thyroidal area
FNC of 50 mm nodule in the left thyroidal area
Cytological diagnosis:
y
g
g
T‐cell lymphoproliferative process
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Hi t l i l di
Histological diagnosis: benign type B1 ectopic thymoma
i b i t
B1 t i th
CD45Ro
CK-pan
CK
pan
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Thymic lymphocytes:
y
y p y
•mature “polyclonal” T‐cells profile: (CD2+,CD3+,CD5+,CD7+)
•CD10+ in an immature T‐cell subsets
•maturation: CD4‐ CD8‐ in the cortex, continues as CD4+ CD8+,
reaches maturation as CD4+ CD8‐ or CD4‐ CD8+ in the medulla.
CD4+/CD8+ is a specific feature of intrathymic T‐cells
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
• 40 yrs old female, HIV positive with right ‐ cervical swelling. The patient suffered from a FL, since 2 yrs, last chemotherapy 10 months before
• US: 2,5 cm large lymph node, oval with hilus preservation
US: 2 5 cm large lymph node oval with hilus preservation
• FC: CD5:10%, CD19:49%, CD23: 10%, FMC7: 0%, CD10:40%, CD10/19:40%, lambda light chains 40%, kappa light chains 0%.
Cytological FC diagnosis: FL
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Final histological diagnosis: reactive hyperplasia with Fi
l hi t l i l di
i
ti h
l i ith
florid follicular proliferation!
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Incidence, phenotypical and molecular data of non-lymphomatous,
clonal B-cell
B cell population: a review of the literature
Author/year
organ
sample
Berthold F, 1989
lymph node
histology
Matsubayashi S, 1990
thyroid
FNA
Cartagena N, 1992
lymph node
Jordan RC, 1996
n. of cases
1
clinical background
light chain restriction
FC‐ICC‐IHH ( pos./total)
IgH clonality PCR (pos./total)
follow‐up
Herpes virus 6
NP
+ (1/1)*
negative
15
Hashimoto thyroiditis
FC/ + (7/15)
NP
NR
FNA
86
NP
ICC/ + (1/86)
NP
NR
minor salivary glands
histology
50
Sjogren’s syndrome
NP
+ (4/50)
NR
Zhou XG, 1999
lymph node tonsils
histology
9
NR
NP
+ (9/9)
negative
Saxena A, 2000
stomach
histology
20
Helicobacter pylori gastritis
NP
+ (10/20)
negative
Kussik SJ, 2004
lymph node
histology and FNA
6
5 negative,1 HIV
FC/ + (6/6)*
+ (3/6)
negative
Chen H, 2006
thyroid
histology and FNA
21
Hashimoto thyroiditis
FC/ + (18/21)
‐
negative
Venkatraman L, 2006
lymph node
FNA
26
NHL, reactive, HIV (1case)
ICC+ (1‐HIV/26)
+ (1/26)
negative
Bangerter M, 2007
lymph node
FNA
131
NR
FC+ (3/131)
NP
negative
Nam‐Cha SH, 2008
lymph node
histology
8
Castleman’s, LES, EBV, HIV (1 case)
IHH/+ (8/8)*
+ (3/8)
6 negative, 2 developed NHL
Bhargava P, 2008
lymph node
FNA
1
HIV
FC/+ ‐
negative
Zeppa P, 2009
thyroid
FNA
34
Hashimoto thyroiditis
FC/+ (5/34)
‐
negative
Fan HB, 2009
liver
histology
40
HCV hepatitis
‐
+ (4/40)
negative
FUTURE PERSPECTIVES
FUTURE PERSPECTIVES
New antibodies for diagnosis
g
•
Schniederjan SD et al.: A novel flow cytometric antibody panel for distinguishing Burkitt lymphoma fromCD10+ diffuse large B‐cell lymphoma. Am J Clin Pathol. 2010 133 718
2010;133:718.
• ……expression of CD44 and CD54 differs significantly between BL and CD10+ DLBCL…..
between BL and CD10
DLBCL…..
• Leshchenko et al.: Genomewide DNA methylation analysis reveals novel targets for drug development in mantle cell lymphoma. Blood. 2010;116:1025.
…….CD37 surface expression in 93% MCL. CD37 f
i i 93% MCL
FUTURE PERSPECTIVES
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
Non Hodgkin lymphoma, monoclonal antibodies
Non
Hodgkin lymphoma monoclonal antibodies
and Flow Cytometry
• Monoclonal antibodies have a direct action against IgG constant regions and a cross‐fire effect (radio conjugated)
• Flow cytometry may quantify the target cells by the evaluation of antigen co‐
expression and their numerical evaluation
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
FUTURE PERSPECTIVES
FUTURE PERSPECTIVES
Högerkorp CM et al.: Identification of uniquely expressed transcription factors in highly Högerkorp
CM et al : Identification of uniquely expressed transcription factors in highly
purified B‐cell lymphoma samples. Am J Hematol. 2010;85:418.
Andréasson U et al.: Identification of molecular targets associated with transformed diffuse Andréasson
U et al : Identification of molecular targets associated with transformed diffuse
large B cell lymphoma using highly purified tumor cells. Am J Hematol. 2009;84:803.
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT
acknowledgments
•
•
•
•
Prof. L. Palombini
Prof. A. Vetrani
Prof. G. Troncone
Dr I Cozzolino
Dr. I. Cozzolino
•
•
•
•
•
•
•
Dr. A. Iaccarino
Dr C Frangella
Dr. C. Frangella
Dr. U. Malapelle
Dr. F. Plaitano
Dr. M. Russo
Dr. M. Salatiello
Dr. L.V. Sosa Fernandez
www.eurocytology.eu
y
gy
Dipartimento di Scienze Biomorfologiche e Funzionali, Università di Napoli “Federico II” IT

Download Report