ISAC XX
WORKSHOP
Carleton C. Stewart
Sigrid J. Stewart
RPCI
LFC
IMMUNOPHENOTYPING
ACUTE LEUKEMIAS
RPCI
LFC
CD3
CD14
HLADr
CD45
This antibody combination is designed to give a differential. CD45 vs SSC is used
to define lymphocytes, monocytes and granulocytes. CD14 further defines
monocytes while CD3 gives T-cells and HLADr identifies B-cells and NK cells. In
bone marrow, progenitor cells are CD4 dim HLADr+ and erythroid cells or
platelets are CD45 negative.
CD7
CD13
CD2
CD19
This combination is used for three reasons. 1) Normal T-cells express CD2 and
CD7, which are often expressed at abnormal levels by malignant T-cells. NK cell
malignancy is usually CD7+CD2-. 2) CD2+CD7+CD13+ cells represent aberrant
co expression of the myeloid antigen, CD13, on acute T lineage lymphocytic
leukemias and lymphoproliferative disorders. 3) Co expression of CD2 or CD7 or
CD13 with CD19 defines aberrant expression of these markers on B lineage
malignancy. Finally, CD19 is often co-expressed on CD13 AML-FAB/M2 with a
t(8,21) translocation.
CD5
This combination is designed to resolve B-lineage lymphoproliferative disorders.
Lambda Clonal excess of kappa or lambda on CD19 positive cells or CD19CD5 positive
CD19
cells are explicitly defined.
Kappa
CD20
CD11c
CD22
CD25
This combination is used to further characterize B-lymphoproliferative disorders
and to define the degree of maturity of acute B-lineage leukemias. In addition,
hairy cell leukemia can be classified by its unique high expression of CD11c.
Aberrant expression of the T-cell marker CD25 on B-cells is diagnostic for
lymphoproliferative diseases when it is expressed.
CD5
CD19
CD10
CD34
This antibody combination is designed to resolve the cells within the maturation of
both T and B-cell lineages. The earliest T-cells are CD5+CD10+CD34+, which
differentiate into CD5+CD10+ by losing CD34 and finally into mature T-cells that
express only CD5. This combination can be used to evaluate the maturity of a T
lineage acute leukemia. In a like manner, the maturation of the two distinct B-cell
lineages: CD19+CD5- and CD19+CD5+ can be defined. The earliest B-cells co
express both CD34 and CD10. As maturation occurs, they lose CD34, then CD10
to become mature B-cells.
CD11b
CD13
CD33
CD34
This combination resolves both normal and malignant myeloid cells. The most
immature myeloid cells express only CD34 and CD13. FAB M0 and M1 will
generally be positive for only these two markers. As these cells differentiate, they
acquire low CD33 and low CD11b expression. A dramatic increase in CD11b
expression with gradual loss of CD34 expression is characteristic of differentiation
into the granulocytic lineage and the malignant counterpart is FAB M2. In contrast,
an increase in CD33 expression and reduction of CD34 expression is characteristic
of differentiation into the monocytic lineage (FAB M4). APL has characteristics
similar to both the early granulocytic and monocytic lineage leukemias suggesting
they may be derived from precursors at the branch point.
CD15
CD56
CD19
CD34
This combination is used to define aberrant antibody expression on hematopoietic
malignancies. CD56 is expressed early on progenitor cells (CD34+CD56+) that
may also co-express CD15. We have shown that in acute leukemia, coexpression of CD15, CD56 and CD34 is associated with the t(8,21) translocation,
which results in a very bad prognosis. CD15 is also expressed on granulocytes
and some B-cells.
CD41
CD71
CD45
CD34
This combination is used to define erythroleukemia FAB/M6 and
megakaryoblastic leukemia FAB/M7. Activated HPCs co- express CD71, the
transferrin receptor, as well as CD45 and CD34. Erythroid progenitor cells can
be distinguished by their very high levels of CD71 and CD34 expression. As
they differentiate into precursor normoblasts they lose expression of both CD45
and CD34 and express only CD71. When HPCs differentiate into the
megakaryocytic lineage they acquire CD41 and lose CD45 before losing CD34.
CD38
CD138
CD45
CD56
Myeloma cells are the only hematopoietic cells that express CD138. In addition,
CD38 is expressed at very high levels and CD56 is often (~80% of the time)
expressed. CD45 is expressed at low levels on immature plasma cells, but is
completely absent on mature plasma cells.
Types of Antigen Expression
Ab1
Ab1
Ab1
F
Ab2
E
Ab2
Ab2
D
Ab1
C
Ab2
B
Ab2
Ab2
A
Ab1
Ab1
INTERPRETING COEXPRESSION
4
10
C
D
1
9
A2
1
10
C
D
1
9
3
10
2
10
CD19 -->
3
10
2
10
1
1
10
10
4
10
4
10
B 19
18
CD19 -->
3
1
10
2
10
3
CD7 -->
CD7
5
10
20
4
4
10
C
D
1
9
10
2
10
CD7 -->
21
10
C6
3
C
D
7
4
1
10
2
10
3
CD2 -->
CD2
1
10
4
D2
3
10
2
10
CD19 -->
R1
1
1
10
10
7
8
10
1
10
2
CD2 -->
CD2
10
3
10
4
3
4
10
1
10
2
CD7
-->
CD7
10
3
10
4
T LINEAGE ACUTE LEUKEMIA
C
D
1
4
C
D
4
5
C
D
1
4
CD45
HLADr
4
10
4
10
4
10
1
HLADr
2
5
2
FL1 -->
CD45
10
3
10
1
10
10
1
2
4
10
C
D
4
5
10
C
D
1
4
3
10
1
FL1 -->
10
1
2
10
3
10
2
FL2 -->
10
FL2 -->
C
D
1
4
3
10
3
10
R1
6
R5
7
4
10
1
10
2
FL3 -->
HLADr
10
3
10
4
8
10
1
10
2
HLADr
FL3 -->
10
3
10
4
3
10
10
2
10
3
10
10
1
10
2
10
3
10
4
10
CD8
1
10
10
CD3
10
2
10
FL1-Height -->
10
1
FL1-Height -->
10
1
2
10
10
2
FL2-Height -->
10
FL2-Height -->
C
D
4
10
4
10
1
10
2
FL3-Height -->
CD8
10
3
10
4
10
4
6
R5
C
D
3
7
4
3
10
3
10
3
10
3
10
R1
3
CD8
FL3 -->
3
5
2
C
D
4
2
4
10
4
10
4
10
1
10
8
1
FL3 -->
CD3
2
FL1 -->
3
7
4
FL1 -->
1
10
10
1
4
1
2
3
10
2
10
C
D
3
1
10
2
10
C
D
4
1
C
D
4
10
5
10
2
3
10
R1
4
10
4
1
10
4
10
T LINEAGE ACUTE LEUKEMIA
8
10
1
10
2
FL3-Height -->
CD8
10
3
10
4
1
10
2
10
3
10
4
10
4
10
3
10
1
CD7
10
2
10
3
10
4
10
1
10
10
CD7
10
2
10
FL1 -->
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
10
3
10
1
10
2
FL3 -->
CD2
4
6
R5
C
D
7
7
4
10
3
10
3
10
3
10
FL2 -->
C
D
1
9
4
3
10
4
10
4
10
4
10
R1
3
2
5
2
C
D
1
9
10
FL3 -->
CD2
FL3 -->
1
2
1
CD2
FL1 -->
1
10
10
1
1
10
2
2
10
C
D
7
10
10
2
10
C
D
1
9
1
C
D
1
9
3
3
10
10
4
4
10
10
T LINEAGE ACUTE LEUKEMIA
10
3
10
4
8
10
1
10
2
CD2
FL3 -->
10
3
10
4
4
10
10
1
10
2
10
3
10
4
1
10
1
10
CD5
2
10
3
10
10
4
1
10
10
3
2
FL1 -->
CD5
10
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
C
D
1
9
10
4
10
1
10
2
FL3 -->
CD10
4
6
R5
C
D
5
7
4
2
3
3
10
3
10
3
10
3
10
4
10
4
10
4
10
FL2 -->
C
D
1
9
2
5
R1
10
10
CD10
2
1
1
CD10
1
10
10
1
10
10
2
10
C
D
5
2
3
3
10
10
4
4
10
3
2
10
C
D
1
9
1
C
D
1
9
10
T LINEAGE ACUTE LEUKEMIA
10
3
10
4
8
10
1
10
2
FL3 -->
CD10
10
3
10
4
1
10
2
10
3
10
4
10
10
1
1
10
2
10
3
10
4
10
10
1
CD5
10
2
FL1 -->
10
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
10
3
10
1
10
2
FL3 -->
CD19
4
6
R5
C
D
5
7
4
10
3
10
3
10
3
10
C
D
3
4
4
3
10
4
10
4
10
4
10
FL2 -->
3
2
5
R1
C
D
3
4
10
CD19
2
10
1
CD19
1
2
2
10
2
10
CD5
1
10
3
10
4
10
4
10
3
10
C
D
5
10
10
2
10
C
D
3
4
1
C
D
3
4
1
3
10
4
10
T LINEAGE ACUTE LEUKEMIA
10
3
10
4
8
10
1
10
2
FL3 -->
CD19
10
3
10
4
1
10
2
10
3
10
4
3
10
2
1
10
10
10
10
10
FL1 -->
10
3
FITC-CD7
10
10
FL1 -->
2
FITC-CD3
1
10
4
10
3
10
2
10
1
FL2 -->
PE-CD4
4
4
T-Lineage
Acute Lymphocytic Leukemia
10
1
10
2
10
3
10
4
10
1
10
FL3 -->
FL1 -->
10
3
10
4
TC-CD8
TC-CD2
1
10
2
FL1 -->
10
3
FITC-CD5
10
4
10
10
3
10
FL1 -->
1
10
1
10
10
2
FITC-CD45
10
3
10
2
FL2 -->
10
PE-CD34
3
10
2
FL2 -->
10
1
10
PE-CD19
10
4
4
4
FITC-CD3
2
FL3 -->
10
1
10
2
FL1 -->
10
3
FITC-CD5
10
4
10
1
10
2
10
3
10
4
FL3 -->
PP-HLADr
08076102
MARKER EXPRESSION ON T-CELL ALL
MARKER
CD1
CD2
CD3
CD4
CD5
CD7
CD8
CD10
CD11c
CD13
CD19
0
PERCENT
63
71
41
61
84
93
55
16
2
25
0
MARKER
CD20
CD22
CD33
CD34
CD38
CD45b
CD45d
CD45n
HLADr
Kappa
Lambda
n = 54
> 10% of leukemia cells express the marker
RPCI
LFC
PERCENT
0
6
2
27
78
15
85
4
13
0
0
B LINEAGE ACUTE LEUKEMIA
C
D
1
4
C
D
1
4
C
D
4
5
CD45
HLADr
4
10
4
10
4
10
1
HLADr
2
5
FL1 -->
CD45
10
3
10
1
2
10
10
2
1
10
1
4
10
6
R5
3
10
10
C
D
1
4
FL1 -->
2
1
10
10
3
3
2
FL2 -->
10
C
D
1
4
10
3
10
R1
C
D
4
5
7
4
10
1
10
2
HLADr
FL3 -->
10
3
10
4
8
10
1
10
2
10
FL3 -->
HLADr
3
10
4
10
3
10
4
10
10
1
1
10
2
10
3
10
4
10
10
1
10
2
CD5
10
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
10
FL1 -->
3
10
1
10
2
FL3 -->
CD19
4
6
R5
C
D
5
7
4
10
3
10
3
10
3
10
C
D
3
4
4
3
10
4
10
4
10
4
10
3
2
5
R1
C
D
3
4
10
CD19
2
10
1
CD19
1
2
2
10
10
CD5
1
10
3
10
4
10
4
10
3
2
10
2
2
10
1
1
10
C
D
5
10
C
D
3
4
10
C
D
3
4
1
3
10
4
10
B LINEAGE ACUTE LEUKEMIA
10
3
10
4
8
10
1
10
2
FL3 -->
CD19
10
3
10
4
4
10
10
1
10
2
10
3
10
4
1
10
1
CD5
10
2
10
3
10
10
4
1
10
10
CD5
10
2
10
FL1 -->
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
10
3
10
1
10
2
FL3 -->
CD10
4
6
R5
C
D
5
7
4
10
3
10
3
10
3
10
C
D
1
9
4
3
10
4
10
4
10
4
10
3
2
5
R1
C
D
1
9
10
CD10
2
2
1
CD10
1
1
10
1
10
10
2
10
C
D
5
2
3
3
10
10
4
4
10
3
2
10
C
D
1
9
1
C
D
1
9
10
B LINEAGE ACUTE LEUKEMIA
10
3
10
4
8
10
1
10
2
FL3 -->
CD10
10
3
10
4
10
1
10
2
10
3
10
10
4
10
1
CD20
10
2
10
3
10
2
10
1
CD22
10
10
FL1 -->
3
10
3
10
1
10
2
FL3 -->
CD22
10
4
6
R5
C
D
2
0
7
4
10
3
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
C
D
1
1
c
4
10
2
4
10
3
10
3
10
3
10
CD22
4
10
4
10
C
D
1
1
c
2
1
5
R1
10
2
10
2
1
10
4
CD22
1
10
10
3
10
4
1
2
10
C
D
2
0
10
10
2
10
C
D
1
1
c
1
C
D
1
1
c
1
3
3
10
10
4
4
10
10
B LINEAGE ACUTE LEUKEMIA
3
10
4
8
10
1
10
2
FL3 -->
CD22
10
3
10
4
4
4
4
10
10
10
B LINEAGE ACUTE LEUKEMIA
5
2
10
1
10
2
10
1
2
10
1
C
D
3
3
10
3
10
3
10
3
C
D
1
3
10
C
D
1
3
10
1
7
3
1
10
2
10
10
3
10
1
10
4
CD33
2
10
10
3
10
10
4
CD19
10
1
CD33
3
10
FL1 -->
10
2
2
10
10
6
R5
C
D
3
3
7
4
10
1
10
2
FL3 -->
CD19
4
3
10
1
FL1 -->
10
1
2
10
10
2
FL2 -->
10
C
D
1
3
4
1
10
4
10
3
10
3
10
R1
3
10
3
5
2
C
D
1
3
10
2
CD19
4
10
4
10
1
10
8
1
10
3
10
4
8
10
1
10
2
FL3 -->
CD19
10
3
10
4
MARKER EXPRESSION ON
B-CELL ALL
MARKER
CD1
CD2
CD3
CD4
CD5
CD7
CD8
CD10
CD11c
CD13
CD19
PERCENT
0
6
0
0
1
0
0
72
1
32
98
MARKER
CD20
CD22
CD33
CD34
CD38
CD45b
CD45d
CD45n
HLADr
Kappa
Lambda
n =284
RPCI
LFC
PERCENT
33
88
3
81
85
3
83
33
96
2
4
4
10
10
4
PRE-B-CELLS
3
10
1
10
CD19 PE -->
2
10
10
1
2
C
D
1
9
C
D
1
9
FL2 -->
BONE MARROW
10
10
3
BLOOD
10
1
10
2
CD10
FL3 -->
10
3
10
4
10
1
10
2
CD10
CD10
TC -->
10
3
10
4
RPCI
LFC
11065003
02178081.004
C
D
1
9
C
D
2
0
CD10
C
D
3
4
CD10
C
D
4
5
CD19
CD19
10
1
10
2
10
3
10
10
4
10
1
CD45
10
2
10
3
10
2
10
1
CD45
10
10
1
FL1-Hei ght -->
3
10
3
10
1
10
2
FL3-Hei ght -->
HLADr
4
6
R5
C
D
4
5
7
4
10
3
10
10
FL1-Height -->
2
FL2-Height -->
1
10
2
C
D
1
4
4
10
2
4
10
3
10
3
3
10
HLADr
4
10
4
C
D
1
4
2
1
5
R1
10
2
10
2
1
10
4
HLADr
1
10
10
3
10
4
1
2
10
C
D
4
5
10
10
2
10
C
D
1
4
1
C
D
1
4
1
3
3
10
10
4
4
10
10
ACUTE MYELOID LEUKEMIA
10
3
10
4
8
10
1
10
2
10
HLADr
FL3-Hei ght -->
3
10
4
10
3
10
10
10
1
CD33
2
10
4
10
1
10
1
HLADr
3
10
FL3-Height -->
10
2
2
10
4
6
R5
C
D
3
3
10
10
3
3
8
1
10
5
10
C
D
1
3
10
2
4
10
6
R5
7
4
10
HLADr
FL1-Height -->
1
10
10
1
CD33
3
2
10
4
10
10
2
FL1-Height -->
10
3
FL1-Height -->
10
3
10
3
10
FL2-Height -->
C
D
1
3
2
3
5
R1
10
10
4
10
4
10
2
1
2
HLADr
1
10
10
1
10
4
10
3
10
4
10
4
10
3
2
10
2
2
10
1
1
10
C
D
3
3
10
C
D
1
3
10
C
D
1
3
1
3
10
4
10
ACUTE MYELOID LEUKEMIA
7
4
8
10
1
10
2
FL3-Height -->
HLADr
10
3
10
4
1
10
2
10
3
10
4
10
10
2
10
3
10
CD11b
10
10
10
2
10
3
10
CD33
6
R5
C
D
1
1
b
7
4
10
1
10
2
CD33
FL3-Hei ght -->
10
3
10
4
4
1
10
10
FL1-Hei ght -->
3
2
2
10
1
FL1-Height -->
10
1
2
10
10
2
FL2-Height -->
10
10
2
10
3
10
3
10
3
10
C
D
1
3
4
1
1
4
10
CD33
10
5
R1
3
10
4
2
C
D
1
3
1
10
1
4
1
2
10
10
4
10
CD11b
3
10
4
10
4
10
3
10
10
10
2
10
C
D
1
3
1
C
D
1
3
C
D
1
1
b
1
3
10
4
10
ACUTE MYELOID LEUKEMIA
8
10
1
10
2
CD33
FL3-Hei ght -->
10
3
10
4
10
1
10
2
10
3
10
4
10
4
10
3
2
10
C
D
3
8
10
10
2
10
C
D
3
4
1
C
D
3
4
1
3
10
4
10
ACUTE MYELOID LEUKEMIA
8
10
1
10
2
10
3
10
4
10
3
10
4
10
1
10
10
CD38
10
2
10
FL1-Height -->
3
10
1
FL1-Height -->
10
1
2
10
10
2
FL2-Height -->
10
10
2
6
R5
C
D
3
8
7
4
10
1
10
2
FL3-Height -->
HLADr
10
3
10
4
4
10
3
10
3
10
3
10
FL2-Height -->
C
D
3
4
4
1
10
3
5
R1
3
10
2
HLADr
2
C
D
3
4
1
4
10
1
10
2
HLADr
4
10
4
10
CD38
10
1
8
10
1
10
2
FL3-Height -->
HLADr
10
3
10
4
1
10
2
10
3
10
4
10
10
2
10
3
10
4
2
10
1
CD7
10
10
FL1-Hei ght -->
3
10
1
10
2
FL3-Hei ght -->
CD2
4
6
R5
C
D
7
7
4
10
3
10
1
FL1-Height -->
10
1
2
10
10
2
FL2-Height -->
10
C
D
1
3
4
10
3
4
10
3
10
3
10
3
10
CD2
4
10
4
10
C
D
1
3
2
10
5
R1
2
1
10
2
10
2
1
1
CD2
1
1
10
10
2
10
CD7
10
10
3
10
4
10
4
10
3
10
C
D
7
10
10
2
10
C
D
1
3
1
C
D
1
3
1
3
10
4
10
ACUTE MYELOID LEUKEMIA
10
3
10
4
8
10
1
10
2
FL3-Height -->
CD2
10
3
10
4
4
10
1
4
10
4
10
ACUTE MYELOID LEUKEMIA
2
5
3
10
2
10
1
1
10
10
C
D
3
4
FL1 -->
2
1
10
10
3
3
2
FL2 -->
10
C
D
3
4
10
3
10
R1
C
D
1
5
4
10
1
10
2
10
3
10
7
10
4
1
10
2
10
3
10
4
CD15
10
10
1
10
2
CD15
3
10
1
FL1-Height -->
10
1
2
10
10
2
FL2-Height -->
10
FL1-Hei ght -->
10
6
R5
C
D
1
5
7
4
10
1
10
2
FL3-Hei ght -->
CD56
10
3
10
3
10
3
10
2
3
4
10
4
10
4
10
C
D
3
4
4
10
10
5
R1
3
10
2
CD56
2
C
D
3
4
1
FL3 -->
CD56
1
1
8
FL3 -->
FL1 -->
10
6
R5
10
3
10
4
8
10
1
10
2
FL3-Hei ght -->
CD56
10
3
10
4
4
10
2
1
10
CD45 PerCP -->
CD45
10
10
2
CD34 PE -->
0
10
20
10
10
30
1
10
CD34
90
80
40
50
60
70
SSC
Side Scatter -->
3
3
100
110
120
10
4
4
FEATURES OF
ERYTHROLEUKEMIA (FAB M6)
10
20
30
40
50
60
70
80
Forward Scatter -->
FSC
90
100
110
120
10
1
10
2
CD45 PerCP -->
CD45
10
3
10
4
10
20
30
40
50
60
70
80
90
100
110
120
Side Scatter -->
SSC
12017021016
Phenotype of Myeloid Component of
Erythroleukemia
marker
CD45
CD13
CD4
CD11c
CD34
CD38
HLADr
% of
leukemias
100
100
87
87
87
87
87
marker
CD33
CD32
CD11b
CD15
CD16
CD41
CD7
% of
leukemias
80
47
40
33
20
15
7
3/18 erythroleukemias did not exhibit a myeloid component
4/16 erythroleukemias exhibit a megakaryocytic component
MARKER EXPRESSION ON
AML CELLS
MARKER
CD2
CD3
CD4
CD5
CD7
CD8
CD11b
CD13
CD14
CD15
CD16
PERCENT
15
<1
73
<1
19
2
50
95
15
79
6
MARKER
CD19
CD32
CD33
CD34
CD38
CD45b
CD45d
CD56
CD64
HLADr
PERCENT
5
73
76
62
61
7
97
14
51
73
n = 957
> 10% of leukemia cells express the marker
4% of specimens diagnosed by hematopathology as AML were found
to have ALL or were normal by Flow Cytometry.
RPCI
LFC
IMMUNOPHENOTYPING
LYMPHOPROLIFERATIVE
DISEASE
multiple myeloma
T-cell lymphomas
RPCI
LFC
1
10
2
10
3
10
3
10
2
10
1
FL2-Height -->
10
10
PE-CD56
3
10
2
FL2-Height -->
10
1
10
PE-CD56
10
4
10
4
MULTIPLE MYELOMA
4
10
1
10
3
10
4
FITC-CD45
10
2
FL1-Height -->
10
3
FITC-CD5
10
4
10
2
10
1
FL1-Height -->
1
10
FITC-CD20
10
3
10
4
10
3
10
2
10
1
10
FL2-Height -->
2
4
FITC-CD38
PC-CD19
10
FL3-Height -->
FL1-Height -->
10
1
10
2
10
3
10
4
FL3-Height -->
TC-CD22
11296291
BONE MARROW FROM MULTIPLE
MYELOMA PATIENT STAINED WITH CD38
CD56 AND CD45
1
2
3
4
P
E
C
D
5
6
10
1
10
2
FITC-CD38
10
3
10
4
BONE MARROW FROM MULTIPLE
MYELOMA PATIENT STAINED FOR CD38
CD56 CD45
AND DNA CONTENT
GATED ON
MONONUCLEAR CELLS
GATED ON
CD38+CD56+CD45- CELLS
700
120 100 90 80 70 60 50 40 30 20 10
800
500
Number
600
400
300
200
100
10 30 50 70 90 110
140
170
200
10 30 50 70 90 110 140 170 200 230
230
DNA
4
10
C
D
4
B
4
A2
1
10
4
10
3
3
3
10
10
10
C
D
4
2
10
LAMBDA -->
C
D
7
2
10
2
10
KAPPA -->
LAMBDA -->
R1
1
1
1
10
10
10
3
4
10 1
10 2
10 3
6
C
5
R5
7
10 4
10 1
KAPPA -->
10 2
10 3
10 4
8
10 1
10 3
10 4
CD19 CD2
-->
CD19
-->
CD8
CD3
10 2
Lymphoma
4
10
C
D
4
D
2
1
4
C
D
4
3
10
4
E
10
10
C
D
7
3
10
10
2
2
10
10
10
KAPPA -->
LAM B DA -->
R1
1
1
1
10
10
10
3
4
10
1
10
2
KAPPA -->
CD3
10
3
10
6
F
3
2
LAM B DA -->
5
R5
7
4
10
1
10
2
CD19
-->
CD8
10
3
10
4
8
10
1
10
2
CD19
-->
CD2
Re active Lymph Node
Typical T-cell Lymphoma or Reactive Lymph Node:
10
3
10
4
4
10
1
10
2
10
3
10
4
10
CD3 FITC -->
2
1
10
10
1
10
2
10
3
10
4
10
FITC-CD3
10
10
4
3
FITC-CD5
10
CD5 FITC -->
2
10
3
10
2
1
10
CD7 FITC -->
10
1
FITC-CD7
10
10
1
10
2
CD2 TC -->
10
3
TC-CD2
10
4
2
10
3
TC-CD8
4
FITC-CD3
1
CD8 PerCP -->
CD8 PerCP -->
10
10
10
CD4 PE -->
2
10
1
1
10
2
CD3 FITC -->
10
10
3
3
PE-CD4
TC-CD8
10
3
PE-CD4
10
CD4 PE -->
10
4
10
4
NON MALIGNANT
LYMPH NODE -T CELLS
10
1
10
2
CD10 TC -->
10
3
TC-CD10
10
4
10
4
10
2
10
10
1
10
FL2 -->
2
10
1
FL2 -->
10
PE-CD3
3
3
TC-CD4
10
10
4
4
SEZARY CELL LEUKEMIA
10
1
10
2
10
3
10
4
10
FL1 -->
1
10
2
10
3
10
4
FL3 -->
PE-CD3
10
2
1
10
FL1 -->
2
10
1
10
10
FL2 -->
10
3
3
PE-CD25
FITC-CD45
10
10
4
4
FITC-CD71
10
1
10
2
FL3 -->
10
3
FITC-CD3
10
4
10
1
10
2
10
3
10
4
FL3 -->
PP-HLADr
11126101
4
2
10
3
10
10
3
TC-CD3
3
10
TC-CD8
4
10
2
10
3
10
4
FITC-CD57
4
4
10
1
FL3-Height -->
10
FL1-Height -->
10
1
10
2
FL3-Height -->
10
3
TC-CD2
10
4
3
FITC-CD45
3
10
2
10
1
FL1-Height -->
2
10
4
10
FITC-CD7
10
FL3-Height -->
2
FL1-Height -->
10
10
4
10
3
10
2
FL2-Height -->
10
1
PE-CD56
1
FL3-Height -->
10
1
10
TC-CD3
10
10
FITC-CD3
10
1
3
10
2
10
4
10
10
3
2
10
10
2
1
10
FL1-Height -->
10
1
1
FL1-Height -->
10
10
FITC-CD3
3
10
2
10
FL2-Height -->
10
1
PE-CD4
10
10
4
10
4
T-CELL
Large Granular Lymphocytic Leukemia
10
1
10
2
10
3
10
4
FL3-Height -->
PP-HLADr
08226202
IMMUNOPHENOTYPING
LYMPHOPROLIFERATIVE
DISEASE
B-cell/CLL
RPCI
LFC
10
4
FITC-CD20
10
2
1
10
CD20 FITC -->
2
10
CD19 PE -->
10
3
10
3
PC-CD19
3
10
2
10
10
10
10
1
1
CD19 PE -->
PC-CD19
10
4
4
NON-MALIGNANT
LYMPH NODE-B CELLS
10
1
10
2
10
3
10
4
10
1
CD5 FITC -->
10
2
10
3
10
10
4
10
2
10
3
10
4
TC-CD10
TC-CD22
4
10
PE-CD79
1
10
2
KAPPA FITC -->
10
3
FITC-kappa
10
4
3
2
1
10
10
10
10
CD79 PE -->
10
10
2
1
10
IgM PE -->
2
10
1
10
LAMBDA PE -->
10
PE-IgM
3
3
PE-lambda
10
10
4
4
FITC-CD5
1
CD22 TC -->
CD10 TC -->
10
1
10
2
IgD FITC -->
10
3
FITC-IgD
10
4
10
1
10
2
10
3
10
4
CD23 FITC -->
FITC-CD23
01148261
4
4
10
A
10
C
D
1
9FL2-Height -->
1
B
C
D
1
9
3
1
10
10
R1
3
1
10
2
10
3
10
4
10
FL3-Height -->
10
4
10
C
D
1
9
C
D
1
9
3
10
2
10
3
10
10
1
1
10
10
4
10
1
Fluorescence Two Height -->
1
F
3
10
4
3
4
10
G
10
10 2
3
CD7
2
10
3
10
H
10
2
10
1
1
10
10
Fluorescence Two Height -->
4
10
10
3
10
R1
1
C
D
1
3CD13 -->
2
1
10
4
10
10
4
CD19
3
Fluorescence One Height -->
10
CD19 -->
C
D
1
9
3
10
10
CD10
2
Fluorescence Two Height -->
Fluorescence Thr ee Height -->
10 2
FL3-Height -->
10
10
CD2
2
4
10
2
1
2
2
10
10
10
10
3
1
1
10
CD7
E
10
3
Fluorescence One Height -->
CD2
4
1
D
C
D
1
3CD13 -->
10
Fluorescence Two Height -->
10
10
2
10
1
C
10
2
10
4
3
10
2
10
C
D
1
9FL2-Height -->
3
10
4
2
10 4
10
1
10
2
10
3
Fluorescence Thr ee Height -->
CD10
10 4
10 1
10
2
CD19 -->
CD19
Aberrant Antigen Coexpression in B-cell lymphoma
10 3
10
4
4
A
L
a
m
b
d
a
L
a
m
b
d
a
Kappa
C
D
1
3
D
B
K
a
p
p
a
CD19
CD19
E
C
D
1
3
CD33
C
C
D
3
3
CD19
Problem Case
CD19
F
1
10
2
10
3
10
4
4
10
1
10
2
10
3
10
3
10
2
10
1
CD20 FITC -->
10
CD5 FITC -->
10
2
10
1
CD19 PE -->
10
3
FITC-CD20
10
10
10
PE-CD19
2
10
10
1
CD19 PE -->
10
3
PE-CD19
10
4
4
CHRONIC LYMPHOCYTIC LEUKEMIA
4
10
1
10
2
10
3
10
4
10
4
CD22 TC -->
CD10 TC -->
1
10
2
KAPPA FITC -->
10
3
FITC-Kappa
10
4
10
1
10
2
IgD FITC -->
10
3
FITC-IgD
10
4
3
10
2
10
1
10
10
10
CD79 PE -->
PE-CD79
3
10
2
10
1
IgM PE -->
PE-IgM
10
10
4
10
3
10
2
10
1
10
LAMBDA PE -->
PE-Lambda
4
4
FITC-CD5
TC-CD10
TC-CD22
GATED ON CD19 POSITIVE B-CELLS FOR CLARITY
10
1
10
2
10
3
CD23 FITC -->
FITC-CD23
01078121
F L2 -He ig ht -->
10
3
1
1
10
10
2
2
10
10
3
10
F L2 -->
4
4
10
10
ABERRANT ANTIGEN EXPRESSION
ON FOLLICULAR LYMPHOMA
1 01
1 02
1 03
1 04
1 01
F L3 -->
1 02
1 03
1 04
F L3 -H eigh t -->
4
4
10
10
TC-CD2
TC-CD10
CD19 EXPRESSION ON LYMPH NODE T-CELLS
FL2 -He ight -->
10
3
1
1
10
10
2
2
10
10
3
10
F L2 -He ig ht -->
(B-CELLS REMOVED FOR CLARITY)
05124063
1 01
1 02
FL1 -H eight -->
1 03
FITC-CD7
1 04
1 01
1 02
FL3 -H eight -->
1 03
TC-CD2
1 04
11215024
05245403
10
10
3
10
3
10
10
4
2
10
3
FITC-Kappa
10
4
1
10
2
IgD FITC -->
10
3
FITC-IgD
10
3
10
4
10
4
4
4
2
10
3
10
10
10
10
10
2
B-cells
2
IgM PE -->
10
KAPPA FITC -->
10
TC-CD22
10
PE-IgM
TC-CD10
CD19+
1
CD22 TC -->
10
1
2
10
2
1
3
10
2
10
1
10
10
CD20 FITC -->
1
10
3
10
4
4
gated on
10
1
CD10 TC -->
10
LAMBDA PE -->
10
10
FITC-CD5
PE-Lambda
FITC-CD20
3
10
2
4
CD79 PE -->
3
1
10
10
2
CD5 FITC -->
PE-CD79
10
10
CD19 PE -->
1
1
PE-CD19
10
10
3
10
2
CD19 PE -->
10
1
10
PE-CD19
10
4
10
4
4
Small Non-Cleaved
(Burkitt’s) Lymphoma
10
1
10
2
10
3
CD23 FITC -->
FITC-CD23
12197181
4
10
FITC-CD19
1
10
2
10
3
10
4
3
10
2
1
10
10
10
10
10
FL1-Height -->
2
1
10
FL2-Height -->
2
10
1
FL2-Height -->
10
10
3
3
PC-CD19
PC-CD19
10
10
4
4
MANTLE CELL
10
1
10
2
10
3
10
4
FL3-Height -->
FL1-Height -->
FITC-CD5
10
1
10
2
10
3
FL3-Height -->
TC-CD10
TC-CD22
10
2
1
10
FL2-Height -->
PE-IgM
2
10
1
10
10
FL2-Height -->
10
3
3
PE-lambda
10
10
4
4
gated on CD19+ B-cells
10
1
10
2
FL1-Height -->
10
3
FITC-kappa
10
4
10
1
10
2
10
3
10
4
FL1-Height -->
FITC-IgD
07266341
10
4
2
1
10
FL2-Height -->
2
10
10
1
10
FL2 -->
10
10
3
3
PE-CD19
PE-CD19
10
10
4
4
ABERRANT ANTIGEN EXPRESSION
ON FOLLICULAR LYMPHOMA
10
1
10
2
FL3 -->
10
3
TC-CD2
10
4
10
1
10
2
FL3-Height -->
10
3
TC-CD10
10
4
10
PE-CD19
3
3
2
1
10
FL2 -->
10
10
2
10
FL2 -->
10
1
10
PE-CD19
10
4
4
ABERRANT ANTIGEN EXPRESSION
ON CLL/SLL
10
1
10
2
10
3
10
4
10
FL3 -->
2
10
3
10
10
2
FL3 -->
10
3
PC-CD19
10
4
3
10
2
10
FL2 -->
1
1
PE-CD19
3
10
2
10
1
10
10
10
10
4
4
FITC-CD7
10
FL2 -->
10
FL1 -->
TC-CD2
PE-CD13
1
10
1
10
2
FL3 -->
10
3
TC-CD10
10
4
4
4
10
1
2
A
10
3
C
D
1
9
4
B
5
6
C
D
2
0
10
2
10
R1
FL2-Height -->
1
10
2
10
R5
FL1-Height -->
4
10
1
10
2
10
3
7
1
10
D
2
4
10
3
2
10
3
2
10
Fluorescence Two Height -- >
3
10
2
4
4
10
1
10
10
3
Fluorescence
One Height -->
CD5
10
E
5
6
2
10
4
10
L
A
M
B
D
A
10
2
8
10
3
Fluorescence Three Height - ->
CD20
10
F
1
2
10
R1
10
10
4
2
1
1
10
10
10
10
3
3
1
7
4
10
KAPPA
Fluorescence One Height -->
4
2
FL1-Height -->
R5
R1
10
1
3
10
3
C
D
2
2
10
10
R1
CD22
1
1
10
FL3-Height -->
CD5
C
D
1
9
2
8
10
FL1-Height -->
10
10
10
4
10
3
1
10
3
C2
1
FL2-Height -->
1
10
4
10
L
A
M
B
D
A
3
4
Fluorescence Two Height -- >
3
4
10
1
10
2
4
10
3
Fluorescence One Height -->
Typical B-cell Lymphoma
KAPPA
10
4