Transcript Group Five - Angelfire

Immune
System Neoplasms
Contents
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What are Neoplasms?
Classification
Neoplasm Differentiation
Immunophenotypes
Neoplastic Diseases
Detection of Neoplasms - FACS
- Morphology
What are Neoplasms?
Any new and
abnormal growth;
specifically a new
growth of tissue in
which the growth is
uncontrolled and
progressive.
Occurrence of Neoplasms
Common in patients with:
 primary immunodeficiency diseases,
 AIDS
 immunosuppressed transplant
patients.
Updated REAL/WHO Classification -- B-cell neoplasms
I. Precursor B-cell neoplasm: precursor B-acute lymphoblastic
leukemia/lymphoblastic lymphoma (B-ALL, LBL)
II. Peripheral B-cell neoplasms
A. B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma
B. B-cell prolymphocytic leukemia
C. Lymphoplasmacytic lymphoma (incl. WM)/immunocytoma
D. Mantle cell lymphoma
E. Follicular lymphoma
F. Extranodal marginal zone B-cell lymphoma of MALT type
G. Nodal marginal zone B-cell lymphoma (+/- monocytoid B-cells)
H. Splenic marginal zone lymphoma (+/- villous lymphocytes)
I. Hairy cell leukemia
J. Plasmacytoma/plasma cell myeloma
K. Diffuse large B-cell lymphoma
L. Burkitt's lymphoma
T-cell and putative NK-cell neoplasms
I. Precursor T-cell neoplasm: precursor T-acute lymphoblastic
leukemia/lymphoblastic lymphoma (T-ALL, LBL)
II. Peripheral T-cell and NK-cell neoplasms
A. T-cell chronic lymphocytic leukemia/prolymphocytic leukemia
B. T-cell granular lymphocytic leukemia
C. Mycosis fungoides/Sezary's syndrome
D. Peripheral T-cell lymphoma, not otherwise characterized
E. Hepatosplenic gamma/delta T-cell lymphoma
F. Subcutaneous panniculitis-like T-cell lymphoma
G. Angioimmunoblastic T-cell lymphoma
H. Extranodal T-/NK-cell lymphoma, nasal type
I. Enteropathy-type intestinal T-cell lymphoma
J. Adult T-cell lymphoma/leukemia (HTLV 1+)
K. Anaplastic large cell lymphoma, primary systemic type
L. Anaplastic large cell lymphoma, primary cutaneous type
M. Aggressive NK-cell leukemia
Hodgkin's lymphoma (Hodgkin's disease)
I. Nodular lymphocyte-predominant Hodgkin's lymphoma
II. Classical Hodgkin's lymphoma
A. Nodular sclerosis Hodgkin's lymphoma
B. Lymphocyte-rich classical Hodgkin's lymphoma
C. Mixed cellularity Hodgkin's lymphoma
D. Lymphocyte depletion Hodgkin's lymphoma
Neoplasm Categories
Immunophenotypes
Different stages of neoplasm express different cell
surface receptors
2 sets:
B-Cells
T-cells
Antigenic characteristics of
T cell lymphoblastic lymphoma/leukemia
3 Examples of Neoplastic
Diseases
Burkitt’s Lymphoma
T-cell Lymphoblastic Leukemia
Hodgkin’s Disease
T-cell Lymphoblastic Leukemia
Characteristics
Patients tend to be young adults/adolescents
50-80% present with mediastinal mass
Peripheral blood involvement > 30% at
presentation
Flow cytometry – excellent – since immature
T cell phenotype (while a normal cellular
phenotype) is not expected in the sample
most commonly submitted (LN, PBL, CSF,
pleural fluid).
T-cell Lymphoblastic Leukemia
Neoplasm of immature T cells.
Double positive for CD4 and CD8 and little or no CD3
on the surface.
T cell receptor rearrangement is not completed yet but
still express TdT (terminal deoxynucleotidyl transferase).
These neoplams present as leukemia.
Detection of Neoplasms
FACS
Morphology
Cells labeled with monoclonal
antibody conjugated to
fluorescent dye
Directed in
thin stream
Stream broken
up into droplets
Laser beam
at stream
Fluoresced by photocell
FACS detect neoplasm by using fluorescence
dye as markers to mark the the ligands and
receptors on the affected cells.
Different stages of neoplasm express different
cell surface receptors
FACS dyes different receptors in different
colour.
FACS is very efficient as it can sort up to
300,000 individual cells in one second.
Case Study
History – 33 year old male with pleural
effusion
Cytology –
Diagnosis: Pleural fluid: Lymphocytic effusion
composed of small lymphocytes, some
macrophages and mesothelial cells.
Comment: Patient has lymphoma on a recent
biopsy. A specimen was sent to immunology for
flow studies.
Supplemental report: Pleural fluid – flow analysis
of the specimen shows cells consistent with
lymphoblastic lymphoma.
FACS profile
Marker
Description
Total T Cells (Pan T)
33
CD5
Pan T & Most Ig+CLL
99
CD20
B Cells
0
CD19
B Cells
0
Kappa
Surface Kappa light chain
Lambda
Surface Lambda light chain
CD38
Thymocytes, NK, Activated T, B Subsets
HLA-DR
HLA-DR
3
CD10
Calla
96
CD34
Human Progenitor Cell Antigen
4
Glycophorin AErythroid
CD14
Monocytes/promonocytes & 40 % AML
CD45
Leukocytes
TDT
Terminal Deoxynucleotidyl transferase
CD4
Helper/Inducer T Cells
CD8
Cytotoxic/Suppressor T Cells
CD2
Total T Cells
CD7
Pan T, T-ALL, NK cells
CD25
Anti-IL2 receptor
1
CD1a
Thymocytes
CD4+CD8+ Dual Marker
97
CD3+CD1a+ Dual Marker
32
TDT+CD10+ Dual Marker
94
CD7+CD38+ Dual Marker
99
Data
CD3
0
0
100
3
1
100
96
98
99
100
99
99
The patient’s pleural fluid
demonstrates the presence
of small to large cells by
light scattering properties.
These cells phenotype as T
cells expressing CD2,
CD1a, CD38, CD7, CD5,
CD10, dual express CD4 &
CD8, and TdT. Some CD3
was expressed. There was
no significant expression of
DR, CD34, myeloid and B
cell markers. These results
are consistant with the
diagnosis of a
lymphoblastic lymphoma of
T cell common thymocyte
origin.
IMMUNOPHENOTYPIC CHARACTERISTICS OF CHRONIC T-CELL
LEUKEMIAS
TdT
CD1
CD2
CD3
TcR-
TcR-
CD4+/CD8CD4+/CD8+
CD4-/CD8+
CD4-/CD8CD5
CD7
CD16
CD56
CD57
CD25
HLA-DR
- = <10% are positive
+/- = 10 to 25% are positive
+ = 25 to 75% are positive
+ + = >75% are positive
T-PLL
++
++
++
+
+/+/++
+ +S
+/-
T-CLL
++
++
++
+
+/+/++
++
+/-
ATLL
++
++
++
++
++
+/+ +S
+/-
CTLL
++
++
++
++
++
++/+/+/-
LGL Leukemias
T-LGL (CD3+)
NK-LGL (CD3-)
++
++
++
++
+/+/++
+/++
++
++
++
++/++
+/++
++
+
+/+/+
+
LGL = large granular lymphocytic s = strong antigen expression
T-PLL/T-CLL = T-cell prolvmphocytic leukemia/ T chronic LL
ATLL = adult T cell leukemia/lymphoma
CTLL = cutaneous T-cell leukemia/lymphoma
Morphology
This method involves staining the
chromosomes and other components of
the neoplasms and examine under a
microscope to view the cells. Different
neoplasms will stain differently, so they
can be distinguished and classified.
Conclusions
Flow Cytometry is an important stand
alone technology and a secondary
consultative resource to other standard
technologies
Flow Cytometry has proven to be a
widely used diagnostic technology that
will continue to expand even in today’s
health care environment.
References
Seventh Edition Basic and Clinical Immunology
Daniel P.Stites, Abba I. Terr
Fourth Edition Immunology A Short Course
Eli Benjamini, Richard Coico, Geoffrey Sunshine
Clinical Immunology
John Bradley, James McCluskey
Immunophenotyping
Carleton C. Stewart, Janet K. A. Nicholson