IMPORTANCIA DE PARAMETROS CLINICO-BIOLOGICOS en la ESTRATIFICACION del TRATAMIENTO en LMA

IMMUNOPHENOTYPE OF MALIGNANT CLONE
Implications of management
Department of Haematology
Cancer Research Centre
University Hospital
J.F. San Miguel
University of Salamanca, Spain

BACKGROUND
IMMUNOPHENOTYPING
- Acute Leukemias & Lymphoproliferative disorders:
· Mandatory for diagnosis & monitoring
- Multiple Myeloma:
· Restricted to research
· Differential diagnosis of unusual cases

IMMUNOPHENOTYPING of PLASMA CELL in
MULTIPLE MYELOMA
Characterization of antigenic profile of PC
- Prognostic influence of specific antigens
- Correlation protein expression & gene markers
- MRD investigation in MM

Prognostic influence of antigenic profile in Myeloma
Controversial results (CD10, CD20, Myeloid...)
- Technical pitfalls
. Differences in the Clone of MoAb & Flurochromes
. Single vs multiparametric labelling
. Criteria for definition of positivity
- Treatment heterogeneity*
* Uniformely treated with GEM2000 protocol
protocol (VBCMP/VBAD
(VBCMP/VBAD (x6)
(x6) followed
followed by ASCT)
627 patients;
patients; median OS: 62 months; RFS: 36 months

Antigenic profile of plasma cells
Multiparametric flow-cytometric approach
Panel of MoAbs
(double acquisition)
(quadruple combinations)
1st
1st STEP
2nd
2nd STEP
Total cellularity
Specific selection of PCs*
FITC
PE
PE/Cy5
APC
CD38
CD56
CD19
CD45
C CS
CD138
CD28
CD33
CD38
S
C C
S S
S SSS
ED ED
D
M
D
M
E E
CD20
CD117
CD138
CD38
R R
M
O
M
O
R ROOFFSSNN
TRANSF
A
10
100
0
10
101
1
10
102
2
10
103
3
10
104
4
TRANSF
ARR
10
100
0
10
101
1
10
102
2
10
103
3
10
104
4
T
F
F IIT
T C
C :: flu
flu o
o r
r escein
escein iis
s o
o th
th iocy
iocy a
a n
n a
a tte
e ;; PE
PE :: p
p h
h y
y c
c o
o eritrin
eritrin ;;
T
CD3
CD3 8
8 FITC
FITC ->
->
P
P E
E /C
/C y
y 5
5 :: P
P E
E //c
c y
y anin
anin e-
e- 5;
5; AP
AP C:
C: aloph
aloph y
y c
c o
o c
c y
y a
a nin
nin e
e
G
G a
a tte
e CD
CD 3
3 8
8 FI
FI TC
TC ->
->
*
* 3
3 ,,000
000 CP
CP o
o b
b tta
a iined
ned fro
fro m
m 10
10 6
106 acqu
acqu iired
red leuk
leuk ocytes
ocytes

Patterns of antigenic expression in PC of MM
n= 627 patients
Patterns of expression
n= 627 patients
-ve
-
+weak
+
++bright
++
-/+het
-/+
CD38
-
80%
20%
-
CD138
2%*
25%
63%
-
CD19
94%
-
1%
5%
CD20
83%
6%
7%
4%
CD28
60%
11%
20%
9%
CD33
80%
4%
7%
9%
CD45
70%
10%
10%
10%
CD117
65%
14%
16%
5%
CD56
22%
10%
58%
10%
* Usually
Usually associ
a
ated
ssociated with apoptotic
poptotic cells
cells

Prognostic influence of PC antigenic expression (I)
CD56
CD117
-- CD117 + (n=195)
-- CD56 +
CD56
(n=272)
1,0
-- CD117 +
1,0
1,0
-- CD117 -ve (n=388)
-- CD56 ve
CD56 ve (n=355)
,9
-- CD117 -ve (n=388
,8
,8
,8
,7
38 m
,6
,6
,6
PFS
41 m
,5
,4
,4
33 m
,4
31 m
,2
,2
,3
,2
0,0
0,0
p=0.05
,1
p=0.01
0,0
48
48
42
42
36
36
30
30
24
24
18
18
12
12
6
6
0
0
60
60
54
54
0
6
12
18
24
30
36
42
48
54
60
66
72
-- CD117 +(n=195)
-- CD56 +
CD56
(n=272)
-- CD117 +(n=195
1,0
1,0
1.0
-- CD117 -ve (n=388)
-- CD56 ve
CD56 ve (n=355)
-- CD117 -ve (n=388
,8
,8
.8
OS
NR
OS ,6
,6
NR
.6
,4
,4
59 m
58 m
.4
,2
,2
.2
0,0
0,0
p=0.03
p=0.1
0.0
48
48
42
42
36
36
30
30
24
24
18
18
12
12
6
6
0
0
60
60
54
54
0
6
12
18
24
30
36
42
48
54
60
66
72
Months from diagnosis
Months from diagnosis

Prognostic influence of PC antigenic expression (II)
CD19
CD28
-- CD28 + (n=235)
1,0
-- CD19 +
CD19
(n=47)
1,0
1,0
-- CD28 + (n=235
,9
-- CD19 -ve
CD19 -ve (n=580)
-- CD28 -ve (n=368)
,9
,9
-- CD28 -ve
,8
,8
,8
,7
,7
,7
,6
,6
,6
36m
37 m
,5
,5
,5
PFS ,4
28 m
,4
,4
31 m
,3
,3
,3
,2
,2
,2
,1
p=0.2
,1
,1
p=0.05
0,0
p=0.0
0
6
12
18
24
30
36
42
48
54
60
0,0
0,0
6
12
18
24
30
36
42
48
54
60
0
0
1,0
-- CD28 +
CD28
(n=235)
-- CD19 +
CD19
(n=47)
1,0
1,0
,9
-- CD28 -ve (n=368)
,9
,9
-- CD28 -ve
-- CD19 -ve
CD19 -ve (n=580)
,8
,8
,8
,7
62 m
,7
,7
66m
,6
,6
,6
OS ,5
,5
,5
54 m
,4
54 m
,4
,4
,3
,3
,3
,2
,2
,2
,1
p=0.06
,1
,1
p=0.1
0,0
0,0
0,0
0
6
12
18
24
30
36
42
48
54
60
0
6
12
18
24
30
36
42
48
54
60
Months from diagnosis
Months from diagnosis

Prognostic influence of PC antigenic expression (III)
PFS (median)
p
OS (median)
p
CD20
-
n=
n= 473
37 m
0.5
65 m
0.7
+
n= 100
33 m
62 m
CD33
-
n=
n= 463
36 m
0.2
62 m
0.3
+
n= 118
32 m
58 m
CD45
-
n=
n= 514
36 m
0.8
59 m
0.7
+
n=100
37 m
62 m

Prognostic influence of PC antigenic expression (V)
PFS (median)
p
OS (median)
p
CD20
-
n= 473
37 m
0.5
65 m
0.7
+
n= 100
33 m
62 m
CD33
-
n= 463
36 m
0.2
62 m
0.3
+
n= 118
32 m
58 m
CD45
-
n= 444
36 m
0.8
59 m
0.7
+
n=141
37 m
62 m
PFS
OS
-ve
37 m
1.0
1.0
1.0
1.0
+weak
35 m
.9
.9
+weak
35
.9
.9
.8
.8
++bright 45 m
.8
.8
.7
.7
-/+ het
36 m
.7
.7
CD45
.6
.6
.6
.6
.5
.5
.5
.5
-ve
58 m
.4
.4
.4
.4
+weak
NR
.3
.3
+weak
.3
.3
.2
.2
++bright
NR
.2
.2
p=0.6
.1
.1
-/+ het
50 m
p=0.5
.1
.1
-/+ het
50 m
0.0
0.0
0.0
0.0
66
66
60
60
54
54
48
48
42
42
36
36
30
30
24
24
18
18
12
12
6
6
0
0
66
66
60
60
54
54
48
48
42
42
36
36
30
30
24
24
18
18
12
12
6
6
0
0

Prognostic influence of phenotypic
phenotypic
phenotypic
y
phenotypic profiles
CD56 & CD117
CD56 & CD28
CD117 & CD28
+ve
-- CD117 +veCD28-ve (n=277)
1,0
-- C
--C D56
D56 ve
ve
D56
56
CD
CD
CD117-ve
117-ve
117-v
117-ve (n
( =
n 303)
=
--CD56 ve CD28+ve (n=87)
1,0
-- C
--C D56 ve
D56 ve
D56
CD28+ve
8+ve
28+v (n
(n =87
=
)
87)
1,0
-- CD117 +ve
CD117
CD28-ve
CD28-ve(n
1,0
-- C
--C D56
D56 ve
ve CD
CD 117-ve
117-ve (n
( =
n 303)
1,0
-- C
--C D56 ve
D56 ve CD28+ve
8+ve (n
(n =87
=
)
87)
1,0
(n=303
=
)
303
1,0
-- CD117 +veCD28-ve (n
,9
,9
-- ot
o h
t er
h s
er (n=
(
372)
n=
,9
-- oth
t ers
er (n=140)
14
n= 0)
14
,9
-- others (n=37
n=
2)
140)
37
-- oth
th ers (n=87)
=87)
=87)
8
,8
,8
,8
,7
,7
,7
,6
,6
31
31 m
m
,6
m
37
37 m
m
NR
NR
,5
,5
,5
,4
,4
PFS
,4
PFS
41
41 m
m
29
29 m
m
33
33 m
m
,3
,3
,3
,2
,2
,2
p=0.00
0.0 9
0
p=0.00
0.0 5
0
p=0.001
,1
p=0.
p=0 0
. 0
0 9
0
p=0.
p=0 0
. 0
0 5
,1
0
,1
p=0.00
=
1
0.00
,1
0,0
0,0
0,0
0
6
12
18
24
30
36
42
48
54
60
66
72
0
6
12
18
24
30
36
42
48
54
60
66
72
0
6
12
18
24
30
36
42
48
54
60
66
72
Months from
fr
di
d a
i g
a no
n si
s s
i
Months
Months
Months
Mo
fr
fr om
om
from
o
dia
dia g
dia no
n s
gnos i
s s
is
Months
Months
Months
Month
Mo
Month
fro
fr m
om
fro
dia
dia
di
dia gnos
gnos
ag
a
is
is
i
Mont
Mon hs from
fr
diagnos
dia
is
i
gnosis
gnos
gnosis
gnos
1,0
1.0
1,0
,9
.9
,9
,8
.8
,8
,7
NR
NR
.7
NR
NR
,7
NR
NR
OS ,6
.6
,6
OS
,5
.5
,5
NR
NR
42
42 m
m
NR
NR
,4
.4
,4
p=0.002
p=0.002
0.002
p=0.00
0.002
p=0.0
0.002
p=0.
,3
.3
,3
,2
.2
,2
p=0
p= .
p=0 0
. 1
0.01
0
0.01
p=0.00
0.0 2
0
p=0
p= .
p=0 0
. 4
0.04
0
0.04
,1
.1
,1
0,0
0.0
0,0
0
6
12
18
24
30
36
42
48
54
60
66
72
0
6
12
18
24
30
36
42
48
54
60
66
72
0
6
12
18
24
30
36
42
48
54
60
66
72
Months from
fr
diag
dia nos
g
i
nos s
Mont
Mon hs from
fr
diagnos
dia
is
i
gnos
Months
Months
Months
Mo
from
from
from
fro
fr
fro
dia
dia
di g
dia nos
gnos
g
gnos is
is
nos
Months
Months
Months
Mo
from
from
from
fro
fr
fro
dia
dia
di g
dia no
n s
gnos i
s s
is
Months from diagnosis
i
is

Prognostic influence of phenotypic profiles
CD56 & CD117
CD56 & CD28
CD56 & CD117
1.0
1.0
CD56+CD117+ n= 130
45 m
.9
CD5
CD5 6+CD
6+CD 28
28 -- n
n =
= 1116
1116
41
41 m
m
CD56+CD117+ n= 130
45 m
.9
+/- or +/-
n=267
36 m
+/+
+/+ or
or -/
-/ --
n
n =266
=266
36
36 m
m
+/- or +/-
n=267
36 m
.8
.8
CD56-CD117-
n=186
31 m
CD5
CD5 6-CD
6-CD 28
28 +
+
n
n =116
=116
29
29 m
m
CD56-CD117-
n=186
31 m
.7
.7
.6
.6
.5
PFS .5
.4
.4
.3
.3
.2
.2
p=0.001
p=0.
p=0. 0
0 1
1
p=0.001
.1
.1
0.0
Months
Months fr
fr om
om di
di agn
agn o
o s
s iis
s
0.0
0
6
12
18
24
30
36
42
48
54
60
66
72
0
6
12
18
24
30
36
42
48
54
60
66
72
Months
Months fr
fr om
om di
di agn
agn o
o s
s iis
s
Months
Months fr
fr om
om di
di agn
agn o
o s
s iis
s
CD28 & CD117
CD2
CD2 8-CD
8-CD 117
117 +
+
n
n =
= 142
142
45
45 m
m
1.0
+/
+/ -- o
o r
r +
+ //--
n
n =
= 327
327
37
37 m
m
.9
CD2
CD2 8+CD
8+CD 117
117 --
n
n =114
=114
29
29 m
m
.8
.7
.6
.5
PFS .4
.3
.2
.1
p
p =
= 0
0 ..000
000 5
5
0.0
0
6
12
18
24
30
36
42
48
54
60
66
72

Multivariate analysis for RFS and OS
RFS
OS
P
P
% BMPC by FCM >20%
0.01
NS
% BMPC in S-phase 2.5%
0.03
0.004
Non-hyperdiploidy DNA
0.05
NS
ISS
0.003
0.0001
Calcium 11 mg/dl
0.005
NS
Platelets 130 · 109
0 /L
0.017
0.04
CD56- CD117-
0.004
0.02

IMMUNOPHENOTYPING of PLASMA CELL in
MULTIPLE MYELOMA
Characterization of antigenic profile of PC
- Prognostic influence of specific antigens
- Correlation
Correlation protein expression
expression & genomic changes
- MRD investigation in MM

Correlation between chromosomal abnormalities &
DNA content
Hyperdiploid
Non-hyperdiploid
P
Del(13q)
19% (32/166)
45% (87/194)
.001
IGH translocations:
Total
10% (15/142)
49% (87/175)
.001
t(11;14)
4% (6/142)
20% (36/175)
.05
no t(11;14)
6% (9/142)
29% (51/175)
Mateo et al; Clin. Can. Res, 2005

Correlation between antigen expression & DNA content
Hyperdiploid (414)
Non-hyperdiploid (455)
P
CD20
+
28% (40/142)
72% (102/142)
.0001
CD28
+
39% (123/319)
61% (196/319)
.0001
CD33
+
63% (103/164)
37% (61/164)
.0001
CD56
-
35% (118/340)
65% (134/201)
.0001
CD117 -
36% (192/526)
64% (334/526)
.0001
Number of cases: 869
Mateo et al; Clin. Can. Res, 2005

Correlation between
antigenic expression & chromosomal abnormalities
Del 13
p
t(11;14)
p
Other IGH
CD20
-
33% (36/73)
NS
8% (19/244)
.0001
20% (49/244)
+
33% (92/277)
32% (21/66)
14% (9/66)
CD56
-
35% (51/144)
NS
24% (31/129)
.001
20% (26/129)
+
30% (69/266)
6% (11/197)
19% (36/197)
CD117 -
37%
37% (85/232)
.002
16% (34/211)
22% (46/211)
+
26% (31/121)
6% (6/101)
.001
13% (13/101)
FISH analysis (n=410)
Mateo et al; Clin. Can. Res, 2005

Incidence of Aberrant Phenotypes in PC from MM
N= 627 patients
80%
80
57%
60
92%
50
40
25%
30%
30
20%
21%
20
13%
10
0
CD38
CD56
CD28 CD33
CD117
sIg
CD20
Infra-
Over- expression
expression
Asynchronous expression
expression

II. Antigen over-expression
Normal BM
Multiple Myeloma
Antigenic
Antigenic pattern
pattern
% cases
Antigenic pattern
% cases
CD56
CD56
heterogeneous
heterogeneous
homogeneous
100%
57%
( 10% +weak)
(+strong)
012
34
10
10
01
102
103
104
10
10
10
10
10
Gate CD 38 ->
CD38 ->
CD28
CD28
homogeneous
70%/30%
25%
(-ve/w
(-ve/w eak)
eak)
(+strong)
10
10
01
102
103
104
10
10
01
102
103
104
Gate CD38 ->
CD38 ->
CD33
CD33
homogeneous
80%/20%
20%
(-ve/w
(-ve/w eak)
eak)
(+strong)
01
2
3
4
10
10
01
102
103
104
10
10
10
10
10
Gate CD38 ->
CD38 ->

Additional useful markers
Normal BM
Multiple Myeloma
Antigenic
Antigenic pattern
pattern
% cases
Antigenic
Antigenic pattern
pattern
% cases
CD19
CD19
heterogeneous
heterogeneous
homogeneous
100%
93%
( 80% +ve cells)
(-ve)
01
2
3
4
10
10
01
10 2
10 3
10 4
10
10
10
10
10
Gate CD 38 ->
CD38 ->
CD45
CD45
homogeneous
homogeneous
(+strong)
94%
(-ve)
83%
10
10
01
10
10
23
10 4
10
10
01
10 2
10 3
10 4
CD45 ->
CD45 ->

Distinction between myelomatous & normal PC
Selected
Select
e ed
Selected CD38high
CD38hi
CD38
plas
pl
ma
asma
plas
ce
c lls
cells
lls
n-PC
n-PC
MM-PC
CD19
MM-P
CD19
CD45
CD56

PROTOCOL for MRD
Double Acquisition
Panel of MoAb
(quadruple combinations)
SSC
FITC
PE
PE/Cy5
APC
1er
1
STEP
Total BM cellularity
CD38
CD56
CD19
CD45
Total BM cellularity
CD138
CD28
CD33
CD38
TRANSFORMED
10
100
0
10
101
1
10
102
2
10
103
3
10
104
4
TRANSFORMED
CD20
CD117
CD138
CD38
CD38 FITC ->
F
F IIT
T C
C :: flu
flu o
o r
r escein
escein iis
s o
o tioc
tioc iana
iana te
te ;; PE
PE :: p
p h
h y
y c
c o
o e
e r
r ytrine;
ytrine; P
P E
E /C
/C y
y 5
5 ::
PE
PE /cy
/cy a
a n
n iine5
ne5 ;; A
A P
P C
C :: a
a llo
llo -p
-p hy
hy c
c o
o -cy
-cy a
a nine
nine
CSS
D
2nd
2
STEP
MER
Identify the phenotypic aberrancy
O
Live Gate on selected
F
characteristic of malignant PC
S
fraction*
on*
NAR
0
0
1
1
2
2
3
3
4
4
PATIENT-SPECIFIC PROBE
10
10
10
10
10
10
10
10
10
10
T
Gate CD38 FITC ->
* 3,000 gated PC from 106
106 acquired leukocytes

MRD: Sensitivity of Immunophenotyping
Dilutional Experiments
1:1
1:10
1:100
15%
1.4%
0.13%
MM- PC
MM- PC
MM- PC
->
->
->
6
6
6
5
5
5
D
D
D
C
C
C
0
12
3
4
0
1
234
10
10
10
012
103
104
10
10
10
10
10
10
10
10
10
10
CD38 ->
Gate CD38 ->
Gate CD38 ->
1:1000
1:10000
0.015%
0.0014%
MM- PC
. .
MM- PC
. ...
.. ...... .
.
. .. .
.... .. . .
. .
. ... .
. ....
.. .
. . .
.. .. . .
.
.. .
...
. .... . .......... ..
.
->
.
. ...
.
->
.. .
. ..
6
56
.
. . . .
.
.
..
.
.
. .. .. .
.. . .
D5
. . .
. .... .
Sensitivity
... .
. ..
Sensitiv
C
.
. .. ..
. ..
.
CD
. .... . ..
.
..
..
. . .. .
. .
. ..
. ... .. .
. .. .
.
. ..
. .
. ..
.
..
level: 10-4
10
10
10
01
102
103
104
10
10
01
102
10 3
104
Gate CD38 ->
Gate CD38 ->

MRD: parameters analyzed
.
.
Total cellularity
.
Total cellularity
PC compartment
..
.
. .... .
.
.
..
..
.
.
% myelomatous PC %MM-PC
% normal PC %N-PC
Proportion of N-PC referred to the
total-PC
%N-PC/ total-PC

Spanish multi-centre protocol GEM (2000-2004)
Diagnosis
VBCMP/VBAD (x4)
Non-responding
Stem cell collection
VBCMP/VBAD (x2)
Double-Trx
MRD
investigation
ASCT (1º)
Complete remission
Partial response
3m post-ASCT
(negative electrophoresis)*
(positive electrophoresis)
ASCT (2º)
Mini-ALO-Trx
Maintenance
Relapse
* Immunofixation either ve or +ve

Correlation between immunophenotyping & electrophoretic responses
at three months post-ASCT (n=200)
Partial response
Complete remission
EF-positive
IFx-positive
IFx-negative
p
MRD evaluation
n=74 cases
n=27 cases (21%)
n=99 cases (79%)
MM-PC
0.76 ± 0.9 #
0.76 ± 0.9
0.28 ± 0.4
0.1 ± 0.3
0.006
MRDnegative cases
8% #
8%
MRDnegative cases
33%
64%
0.003
% n-PC/TPC
42 ± 33 #
42 ± 33
69 ± 34
86 ± 25
0.008
Results expressed as mean ± SD
# p significance between EF-positive (PR) and CR IFE-positive cases: p=0.005; p= 0.0001; p=0.0001
MRD-ve: <0.01% residual MM PC

Changes in PC compartment following ASCT
20
14
10
1
0.8
0,3
0,25
0,21
0,2
n-PC
%
0,15
0,14
0,1
0,05
0,035
MM-PC
0
0,03
0,007
0
100
80
80
77
60
%
40
% n-PC / TPC
20
0
4
0
VBCMP/
Diagnosis
(x6)
3m post-Trx
12m post-Trx
VBAD
Results expressed as mean

Changes in PC distribution following ASCT
Diagnosis
Post-Trx
N-PC
MM-PC
MM-PC
CD19
CD19
CD45
CD45
CD56
CD56
Gate CD38

Impact on RFS of MRD (by immunophenotyping) in BM
obtained 3 months post-ASCT in 200 patients
%MM-PC
1,0
p=0.0001
,9
,8
,7
NR
-- <0.01% MM-PC
,6
-- 0.01% to 1% MM-PC
,5
aliv
40m
-- 1% MM-PC
v
,4
sure
,3
e
23m
e-fr
,2
psa
,1
Rel
0,0
0
6
12
18
24
30
36
42
48
54
60
66
72
78
Months from immunophenotypical analysis (3 months post-ASCT)

Impact on RFS of MRD (by immunophenotyping) in BM
obtained 3 months post-ASCT in 99 patients in CR (IFx-)
%MM-PC
%N-PC / total PC
1,1
1,0
p=0,02
p=0,01
1,0
,9
,9
,8
,8
NR
,7
NR
,7
,6
,6
al
,5
36m
iv
,5
ivv
32m
,4
,4
suree ,3
,3
e-fr
ps
,2
,2
a
Rel
,1
,1
0,0
0,0
0
6
12
18
24
30
36
42
48
54
60
66
0
6
12
18
24
30
36
42
48
54
60
66
Months from immunophenotypical analysis (3 months post-ASCT)
-- <0.01% MM-PC (MRD ve)
-- 75 % N-PC/total PC
-- 0.01% MM-PC
-- < 75 % N-PC/total PC

Impact on RFS of immunophenotyping
at 3 months post-ASCT in 200 patients
%MM-PC
1,0
p=0.0001
,9
,8
,7
NR
,6
-- <0.01% MM-PC (n=82)
,5
alivv
40m
-- 0.01% to 1% MM-PC (n=100)
v
-- 0.01% to 1% MM-PC
,4
sure
-- 1% MM-PC (n=18)
e
-- 1% MM-PC
,3
e
23m
e-fr
,2
psa
,1
Rel
0,0
0
6
12
18
24
30
36
42
48
54
60
66
72
78
Months from immunophenotypical analysis (3 months post-ASCT)

Impact on RFS of immunophenotyping
at 3 months post-ASCT in 99 CR (IF-) patients
%MM-PC
%N-PC / total PC
1,1
1,0
p=0,02
p=0,01
1,0
,9
,9
,8
,8
NR
,7
NR
,7
,6
,6
al
,5
36m
iv
,5
ivv
32m
,4
,4
suree ,3
,3
e-fr
ps
,2
,2
a
Rel
,1
,1
0,0
0,0
0
6
12
18
24
30
36
42
48
54
60
66
0
6
12
18
24
30
36
42
48
54
60
66
Months from immunophenotypical analysis (3 months post-ASCT)
-- <0.01% MM-PC (n=66)
-- 75 % N-PC/total PC (n=80)
-- 0.01% MM-PC (n=33)
-- < 75 % N-PC/total PC (n=19)

CONCLUSIONS
Inmunophenotypic studies have clinical value:
- Prognostic influence of antigen expression
- Correlation genetic changes/protein profile
- Investigation of MRD (immunophenotypic remission)

Grupo Español de Mieloma (GEM/Pethema)
Hospitales
Hospitales
Clínico de Barcelona
General de Segovia
12 Octubre (Madrid)
Cruces (Bilbao)
Clínico de Salamanca
St. Coloma de Gramanet
Clínico de San Carlos (Madrid)
(Barcelona)
Hospital de Badalona
Gregorio Marañon (Madrid)
Clínico de Asturias
Carlos Haya (Málaga)
Fr. Peset (Valencia)
H. Tauli (Gerona)
Universitario de Canarias
Huesca
Rio Ortega (Valladolid)
Palencia
Cínico de Zaragoza
Alcira (Valencia)
Hospital General de Jerez
H. Del Mar (Barcelona)
Ramón y Cajal (Madrid)
Mahón (Baleares)
Morales Meseguer (Murcia)
Clínico de Málaga
La Fe (Valencia)
Xeral Cies (Vigo)
C.U. de Navarra
Plasencia
Galdakao (Vizcaya)
Cáceres
Clínico de Valladolid
Algeciras
Sant Pau (Barcelona)
Ávila
Arnau Vilanova (Lérida)
Jaén
Universitario de Santiago
S. Pau i Sta Tecla (Tarragona)
General Universitario de Valencia
General de Guadalajara
Universitario de Getafe (Madrid)
Sagunto (Valencia)
Insular de las Palmas
Son Dureta (Mallorca)
H. de La Princesa (Madrid)
Cuenca
Severo Ochoa (Madrid)
Alicante SUS
Juan XIII (Tarragona)
M. Valdecilla (Santander)
Toledo
Albacete
Gandía (Valencia)
H. Del Bierzo
Vall D´Hebrón (Barcelona)
Fundación Jiménez Díaz (Madrid)
San Jorge (Huesca)
Elda (Alicante)
Verge de la Cinta (Tortosa)
V. Del Rosel (Cartagena)
Alarcos (Ciudad Real)
Castellón
Salamanca´Group:
Mataró (Madrid)
Mutua Tarrasa
Juán Canalejo (Coruña)
Consorcio Tarrasa
G.Mateo, N.Gutierrez, R.Lopez, M.Mateos,
Ferrol
C. Corachán (Barcelona)
R.Garcia-Sanz, A.Orfao