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SUPPLEMENTARY MATERIAL miR expression in MYC-negative DLBCL/BL with partial trisomy 11 is similar to classical Burkitt lymphoma and different from diffuse large B-cell lymphoma
Michalina Zajdel*1, Grzegorz Rymkiewicz*2, Magdalena Chechlinska1, Katarzyna Blachnio2, Barbara Pienkowska-Grela2, Beata Grygalewicz2, Krzysztof Goryca3, Maria Cieslikowska1, Zbigniew Bystydzienski2, Pawel Swoboda1, Jan Walewski4 and Jan K. Siwicki1
1Department of Immunology, 2Department of Pathology and Laboratory Diagnostics, 3Department of Genetics, 4Department of Lymphoid Malignancies, Maria Sklodowska-Curie Memorial Centre and Institute of Oncology, Warsaw, Poland
*These authors contributed equally to this work
Correspondence should be addressed to Jan Konrad Siwicki e-mail: [email protected]
1 Pathological review and immunohistochemical analysis All lymphomas were investigated by hematopathologist and a final consensus diagnosis was made based on histopathological and immunohistochemical features. When multiple biopsies were available, the primary biopsy was selected. In 8 secondary DLBCL,NOS cases the material analyzed was taken from relapsing/transformed disease. Almost all cases were examined for expression of the antigens listed in Table S1. Staining was done in an autostainer (Dako), where tissue sections were incubated for 20 minutes with antibodies diluted by the Dako manufacturer.
TABLE S1 The antibodies used for IHC analyses
MoAb CD3 CD5 CD10 CD20 CD43 CD44 CD56 BCL2 BCL6 MUM1 Ki-67 Cyclin D1 TdT MYC Clone F7.2.38 SP19 56C6 L26 DF-T1 DF1485 123C3 124 PG-B6p MUM1p MIB-1 SP4/EP-12 B6 NCL-cMYC Dilution 1:50 FLEX FLEX FLEX FLEX 1:50 FLEX FLEX FLEX FLEX FLEX FLEX 1:20 1:50 Source D N MoAb, monoclonal antibodies; D, Dako (Glostrum, Denmark); N, Novocastra Leica Microsystems (Berlin, Germany)
Flow cytometry immunophenotyping
FNAB samples were collected into K2 - EDTA tube with PBS. 50 µl (<1x106 nucleated cells) of cellular suspension obtained by a hematopathologist from FNAB was mixed and incubated with 20µl of the relevant mouse (or rat) anti-human surface or cytoplasmic monoclonal antibody (MoAb) (15 min. at room temperature in the dark environment), immediately after delivery to the laboratory. FNAB samples were prepared by a standard red blood cell lyse/wash technique with FACSlysing Solution (BD, San Jose) and were stained for 3 or 4- color immunophenotyping. Samples were washed once in PBS solution without Ca/Mg ions and then suspended in 0,5ml of 1% paraformaldehyde in PBS. B-NHLs cells were stained with MoAb combinations with the following fluorochromes: FITC/ PE or RPE/ PerCP or PerCP-Cy-5.5 or PE-Cy7/ APC1. All samples were stained with isotype controls, corresponding to the MoAb type used in the experiment to establish negative fluorescence signal threshold for each fluorochrome, and to evaluate non-specific binding. An optimum panel of MoAbs, and their clones used for FCM analysis recommended to evaluate BL,
DLBCL/BL, and DLBCL,NOS are listed in the Table S2. The intracytoplasmic staining of BCL2 (BD) was performed after permeabilization with PermeaFix (BD). Photomultiplier tube
1 *FITC, PE, RPE, PerCP, PerCP-Cy5.5, PE-Cy7, APC represent respectively: fluorescein isothiocyanate, phycoerythrin, R-phycoerythrin, peridinin-chlorophyll protein, PerCP with a cyanine dye (Cy5.5), PE with a cyanine dye (Cy7) and allophycocyanin
2 and compensation settings on the FACSCalibur cytometer were adjusted daily, with initial setup using Calibrate beads and FACSComp software (BD Biosciences). Final settings were based on staining patterns of peripheral blood lymphocytes stained with CD3FITC, CD19PE, CD45PerCP-CY-5.5 and CD5 APC.
TABLE S2 The antibodies used for FCM M g 6 I a , 5 2 R G D ^ c * 0 8 β L b 7 G 2 9 6 3 5 0 2 5 0 1 1 2 4 g 1 1 D 3 1 9 4 5 8 0 3 3 8 2 C I g 1 5 4 4 2 2 2 1 7 8 5 4 - C λ L A
2 1 7 3 2 1 6 I / G G , D D D D / & o D D D D D D D D D D D D C A κ g g D M D D D D D D 1 6 D C C C C I I L C C C C C C C C C C C C B M F 1 C C g C C C C C G I H
g D , I C A g I ^ 3 2 3 E - 4 2 1 P 5 2 0 / 3 L . 1 1 0 5 5 1 1 3 9 5 - . 6 2 2 2 - - - C 0 1 Y 1 8 3 8 7 a e - - 1 1 4 1 1 - 1 3 - - 1 . L L T M 8 3 7 1 7 3 1 3 X 7 0 n / 1 3 C 8 A X 1 I C C 4 F M 1 T / / / / 2 1 - / o C C M 2 N K K K G A M D A O 2 7 P P F l I 0 K H - 5 F 0 2 -
1 M S L 2 4 2 N l S S S S - 1 H H E A , - L S J 4 L C O T O , H - F - C c D B 3 8 S L C 7 S S X M Y 7 R 2 B M M C N B B B M V H T B E
S BD Serotec Invitrogen Caltag MoAb, monoclonal antibodies; S, source; ^two different clones of CD23 antibody, *CD38, HB7 clone conjugated with FITC (fluorescein isothiocyanate) and PE (phycoerythrin); BCL2, an intracytoplasmic antigen was assessed following permeabilization of cells; κ/λ, (kappa/lambda) antibodies against light chains; IgD, IgM, IgG and IgA, antibodies against heavy chains; IgG1/IgG2a and IgG1, antibodies of isotype control conjugated with FITC/PE and APC (allophycocyanin), respectively.
TABLE S3 Clinical and immunophenotypic characteristics of patients with Burkitt lymphoma ) e (
) l ) ) n ) ) a o M i M M d M M t ) o C C C a ] ] C C x C n z F F e F i F F / / / a s l / / H % %
r [ . [ I C / a C C
t C C ( o c y
1 7 x H
H H o , H H 6 e I N 7 6 l I I
I I e (
- ( ( / L ( ( i (
D r g )
0 8 2 0 o C n 3 K C A 1 3 ( 2 L 4 B
m l D D D C D u a C C C B d T C o N
1* 40/F ov e +h − − + +h + 100 100
2* 52/F ov e +h − − + +h − 100 100
3* 46/M st e +h − + + +h nd 100 90
4* 22/M c n +h − + + +h + 95 100
5* 75/F br e +h − + + +h + 100 100
6* 53/M ing e +h − + + +h − 100 94
3 7* 33/M c n +h − nd + +h + nd 100
8* 61/F c n +h − + + +h + 99 100
9* 53/F c n +h − + + +h + 100 100
10* 44/F ax n +h − nd + +h + 100 100
11* 32/M s e +h − + + +h + 100 100
12* 18/M c n +h − + + +h + 95 100
13* 25/M c n +h − nd + +h + 100 100
14* 27/M c n +h − + + +h + 95 100
15* 60/F ing/br e +h − + + +h +/− 100 100
16* 25/M bm e +h − nd + +h + nd 100
17* 39/M liv e +h − + + +h + 100 100
18* 21/M st e +h − + + +h nd 95 100
19* 41/M ab e +h − nd + +h nd 95 100
20* 62/F c n +h −/+* + + +h + 95 100
21* 28/M ab e +h − + + +h +/− 95 100
22* 19/F ab e +h −/+* + + +h +/− 100 100
23* 37/M ab e +h −/+* nd + +h + 95 100
24* 20/M c e +h − nd + +h +/− 98 100
25* 58/M ab e +h −/+* − + +h + 95 100
26* 37/M s e +h − + + +h + 100 100
27^ 29/M ing n +h −/+* + + +h + 100 100
28^ 37/M ax n +h − + + +h + 100 100
29^ 36/M ax n +h − nd + +h + nd 100
30^ 29/M ax n +h − nd + +h +/− nd 100
31^ 30/M c n +h − + + +h +/− 100 90
32^ 36/M c n +h − nd + +h + nd 100
33^ 33/M ab n +h − + + +h + 100 100
34^ 40/M c n +h − nd + +h + nd 100
4 BL cases expressed CD20(+)higher, BCL2(–) (29 out of 34 cases), showed median Ki67 and CD71 values of 100%, and were CD10(+), CD38(+)higher, CD3(–), CD5(–), TdT(–) and CD34(–), as evaluated by IHC and FCM. No., case number; *, sporadic variant of BL; ^, human immunodeficiency virus-associated BL; tumor localization: ov – ovary, st – soft tissue tumor, c – cervical lymph node, br – breast gland tumor, ing – inguinal lymph node, ax – axillary lymph node, s – stomach tumor, bm - bone marrow, liv – liver tumor, ab – abdominal tumor (excluding ovary and stomach involvement); IHC/FCM – immunohistochemical and flow cytometry evaluation of antigens (CD20, BCL2, BCL6, CD10, CD38, CD43, Ki-67 and CD71) expression; antigen expression in neoplastic cells: +, in 100% of cells, +/–, in more than 20% to <100% of cells, –/+,* very weak BCL2 expression in less than 20% of cells, –, no expression (<20% cells); +h, a given antigen has higher expression in B-NHL cells compared to normal B/T lymphocytes; Ki-67 [%] and CD71[%], percentage of BL cells expressing Ki-67 and/or CD71; nd, not determined.
TABLE S4 Clinical and immunophenotypic characteristics of patients with DLBCL/BL including D/THLs ) e (
l ) ) ) ) n a ) o i d M M M M C ) t ) o
C C C C a ] x H ] n C M z F e I F F F i a / / / / ( s l H % C
% r [ . I / a [ t C C C C
1 F (
o c y
( x 1
7 H H H H D o e , 6 N 7 l I
I I I 3 6 e
/ ( ( ( ( - L n
4
D i r g i ) l 2 0 0 8 o C D C n A K c 2 1 3 L ( B y C
m l D D D C u C a C C C B T d o N
1 18/M ab e +d −/+* − + +h + − 95 100
2 64/M ing n +d − + + + − − 100 100
3* 31/F ab e +d − + + +h + − 100 100
4 56/M ing n +d + + + +h +/− − 100 100
5* 22/F c n +d +h + + + +/− − 30 100
6* 38/M c n +b − + + +h + + 100 100
7^ 50/M st e +d − + + + + − 90 100 DLBCL/BL cases expressed CD20, but the expression was (+)dim/lower compared with the expression of CD20 in BL and BNHLs[11q] (6 out of 7 cases), showed median Ki67 and CD71 values of 100%, and were CD10(+), and CD3(–), CD5(–), TdT(–) and CD34(–), as evaluated by IHC and FCM. No., case number; *, double/triple hit lymphoma of the DLBCL/BL; ^, human immunodeficiency virus-associated case; tumor localization: ab – abdominal tumor, ing - inguinal lymph node, c – cervical lymph node, st – soft tissue tumor; IHC/FCM – immunohistochemical and flow cytometry evaluation of antigens (CD20, BCL2, BCL6, CD10, CD38, CD43, cyclin D1, Ki-67 and CD71) expression; antigen expression in
5 neoplastic cells: +, in 100% of cells, +/–, in more than 20% to <100% of cells, –/+*, very weak BCL2 expression in less than 20% of cells, –, no expression (<20% cells); +h, a given antigen has higher expression in B-NHL cells compared to normal B/T lymphocytes; +d, a given antigen has dim/low expression in B-NHL cells compared to normal B/T lymphocytes; Ki-67 [%] and CD71[%], percentage of DLBCL/BL cells expressing Ki-67 and CD71; nd, not determined.
TABLE S5 Clinical and immunophenotypic characteristics of patients with B- NHLs[11q] ) e (
l ) ) n ) ) a o i d M M M M ) t ) o C C a ] ] C C x n C M z F e F i F F a / / s l / / % H % C r
[ [ . I a / t C C
F C C ( o c y ( x 1 7
H H o e , H H 6 N 7 6 l I
I 3 I I e
- / ( ( L ( (
4 i
D r g ) 0 8 2 0 o C D K C n A 1 3 2 L ( B C m D D l D C u a C C C B T d o N
1 18/M c n +h − + + + +/− 100 100
2 25/M ab n +h − + + + + 100 95
3 38/F t e +h − + + + +/− 100 100
4 23/M c n +h − + + + + 100 100
5 22/M ing n +h − + + +h + 100 100
6 35/M c/t e +h − + + +h +/− 100 100
7 40/M ab n +h − + + +h +/− 100 97
8 29/M c n +h − + + +h + 100 100
9 20/M ab n +h − + + + − 100 100
10 32/M c/t e +h − + − + +/− 100 100
11 22/M c n +h − + + +h + 100 96
12 62/M ab n +h − + + +h +/− 100 100 B-NHL[11q] cases expressed CD20(+)higher,BCL2(–),BCL6(+), CD10(+) (11 out of 12 cases), showed median Ki67 and CD71 values of 100%, and were CD38(+)higher or bright, CD3(–), CD5(–) TdT(–) and CD34(–), as evaluated by FCM and IHC.
No., case number; tumor localization: c – cervical lymph node, ab – abdominal tumor (including stomach involvement), t – tonsil tumor, ing - inguinal lymph node; IHC/FCM – immunohistochemical and flow cytometry evaluation of antigens (CD20, BCL2, BCL6, CD10, CD38, CD43, Ki-67 and CD71) expression; antigen expression in neoplastic cells: +,
6 in 100% of cells, +/–, in more than 20% to <100% of cells; -, no expression (<20% cells); +h, a given antigen has higher expression in B-NHL cells compared to normal B/T lymphocytes; Ki-67 [%] and CD71[%], percentage of B-NHLs[11q] cells expressing Ki-67 and CD71.
TABLE S6 Clinical and immunophenotypic characteristics of patients with primary DLBCL,NOS ) e (
l ) ) ) ) ) n ) a o
i d M M M M M ) t M ) o C C C C C a ] ] C x C n C z F e F F F F i a / F / / / / H s l H % % / r
I [ . [ I / a t C C C C C (
( C
o c y
x 1 7
H H 1 H H H o e , 6 N H 7 6 l I
I I I I e
- / I ( ( ( ( ( L
i
M
( D r g )
2 0 0 8 3 C o U K C 5 n A 2 1 3 4 L ( B
m D M l D D D D C u a C C C C C B T d o N
1* 67/F t e +d +/− + + +/− − +h + 100 100
2* 35/M t e +h − + nd nd − +b + 100 100
3* 36/M c n +h +b + + +/− − +b + 40 100
4* 72/F thy e +h − + nd nd − +d + nd 100
5* 55/F ab n +h − + nd +/− − − − nd 100
6* 86/F c n +h +b + − + + +/− − nd 100
7* 54/M t e +h +b + + +/− + +b +/− 85 100
8* 79/F ax n +b − nd +/− − nd nd +/− 60 nd
9* 49/M c n +d − + + + − +/− − 80 100
10* 78/F ab n +h +h + nd +/− + +b + nd 100
11* 18/M c n +h − + nd − − +b +/− 50 88
12* 68/F c e +h +b + + +/− + +b + 80 100
13* 56/M t e +h +h + + + − +b + 90 82
14* 52/F thy e +b − + nd nd − +h +/− nd 100
15* 43/F ab e +h +b + nd nd − +/− + nd 100
16* 66/M ab e +h +b + nd nd − +/− +/− 60 84
17* 48/M c n +d +h + nd − + − +/− nd 88
18* 57/F st e +/- +b + + +/− − +/− +/− nd 100
7 19* 66/F sp e +d − + + nd − +/− − 100 100
20*1 47/M c n +d +b + + nd + +b +/− nd 100 21*1 39/M ax n +h − + + nd − +b − nd 100
22^ 54/F br e +h +b − nd + − +/− + nd 100
23^ 58/M c n +b +h − nd + − +h + nd 100
24^ 77/F thy e +d +b − nd + − +/− + nd 100
25^ 66/M c n +b +b − + + − +h + 100 100
26^ 60/F s e +/− − − − + − +/− − nd 100
27^ 56/M ab e +/− +b − + + − − − 80 92
28^ 72/M ing n +b +b − + + − nd +/− 90 100
29^ 54/F s e +b +/− − + + − nd − 95 nd
30^ 56/M c n +d +b − nd + − − + nd 75
31^ 50/M ax n +b +b − + + − nd + 50 nd
32^ 70/M c n +b +h − +/− + − +d +/− 90 100
33^ 38/M ab n +h +b − +/− + − +/− + 90 94
34^ 80/F br e +h +b − +/− + − +/− +/− 90 100
35^ 69/M c n +b +b − nd + − − + nd 100
36^ 31/F c e +d +b − + + − − − nd 100
37^ 31/M ad e +h +b − + + − +b + 100 100
38^ 90/F st e +h − − nd +/− − +b − 100 100
39^ 73/F ing n +h +b − nd + − +/− +/− nd 75
40^ 68/F ab n +d nd − nd + − +b nd nd 100
41# 60/M ab e +b +b − − + + +b +/− 80 80 *All germinal center B-cell-like DLBCL,NOS cases (cases: 1-21). showed heterogeneous higher bright dim/lower CD20 expression [(+) , (+) , (+) or (+/–)], with median Ki67 and CD71 values of 90% and 100%, respectively, and were CD10(+) and CD3(–), as evaluated by IHC and FCM. ^All non-germinal center B-cell-like DLBCL,NOS cases (cases: 22-40) showed heteroge- neous CD20 expression [(+)higher, (+)bright, (+)dim/lower or (+/–)], with median Ki67 and CD71 val- ues of 90% and 100%, respectively, and were MUM1(+), CD10(–) and CD3(–), as evaluated by FCM and IHC.
8 No., case number; *germinal center B-cell-like-, ^non-germinal center B-cell-like-, #CD5(+)- DLBCL,NOS immunophenotypic subgroups.*1germinal center B-cell-like, human immunodeficiency virus-associated DLBCL,NOS; tumor localization: t – tonsil tumor, c – cervical lymph node, thy – thyroid tumor, ab – abdominal tumor (excluding stomach, adrenal gland, and spleen involvement), ax – axillary lymph node, st – soft tissue tumor, sp – spleen tumor, br – breast gland tumor, s – stomach tumor, ing - inguinal lymph node, ad - adrenal gland tumor; IHC/FCM – immunohistochemical and flow cytometry evaluation of antigens (CD20, BCL2, CD10, BCL6, MUM1, CD5, CD38, CD43, Ki-67 and CD71) expression; antigen expression in neoplastic cells: +, in 100% of cells, +/-, in more than 20% to <100% of cells, -/+,* very weak BCL2 expression in less than 20% of cells, −, no expression (<20% cells); +h, a given antigen has higher expression in B-NHL cells compared to normal B/T lymphocytes; +b, a given antigen has same expression in B-NHL cells compared to normal B/T lymphocytes, +d a given antigen has dim expression in B-NHL cells compared to normal B/T lymphocytes; Ki-67 [%] and CD71[%], percentage of primary DLBCL,NOS cells expressing Ki-67 and/or CD71; nd, not determined.
TABLE S7 Clinical and immunophenotypic characteristics of patients with secondary DLBCL,NOS w o ) l e
(
n ) l ) ) ) ) n o ) a o M ) s i d d M M M M ) t M C e o C L C C C C a C s ] ] x F C n z / H a F e F H F F i a H / F / / / I s l % % b / I
r C ( N
[ [ . / a ( t C C C C
-
C o n c y H # x 1 7
# B H H H H I o o e , 1
H N 7 6 i l I (
I I 6 I e
- / I e t ( ( ( (
i
( D r g L M # a d )
2 0 8 3 o K C a 5 A n C U 0 2 3 4 m L ( r 1 m r B D l g D D D M C u o D a C f C C C B T s d C o n a N r T
1* 61/F ab e FL +d +b + + nd − − − nd 90
2* 61/F ing n FL +h +b + + − − +h +b 60 100
3* 72/F ing n FL +h +h + + nd − +/− − 50 80
MALT 4^ 72/F st e +h +/− − − nd − − − 95 100 /MZL MALT 5^ 67/F br e +h +/− − − nd − − +b 75 100 /MZL CLL/ 6# 56/M st e +d +h − − nd + +h +b nd 57 SLL CLL/ 7# 28/M c n +d +h +/− − +/− + +/− +b 55 57 SLL CLL/ 8# 62/F c n + d +h +/− − + + + h +b 80 100 SLL All secondary DLBCL,NOS cases showed heterogeneous CD20 expression [(+)higher, (+)bright or(+/–)], with median Ki67 and CD71 values of 68% and 95%, respectively, and were CD3 negative, as evaluated by IHC and FCM.
9 No., case number; *germinal center B-cell-like-, ^ non-germinal center B-cell-like-, #CD5(+)- secondary DLBCL,NOS immunophenotypic subgroups; tumor localization: ab – abdominal tumor, ing - inguinal lymph node, st – soft tissue tumor, br – breast gland tumor, c – cervical lymph node; FL, transformation of follicular lymphoma to DLBCL, NOS; MALT/MZL, transformation of MALT/marginal zone lymphoma to DLBCL, NOS, CLL/SLL, transformation of chronic lymphocytic leukemia /small lymphocytic lymphoma to DLBCL, NOS (Richter syndrome); IHC/FCM – immunohistochemical and flow cytometry evaluation of antigens (CD20, BCL2, BCL6, CD5, CD10, CD38, CD43, MUM1, Ki-67 and CD71) expression; antigen expression on neoplastic cells: +, in 100% of cells, +/-, in more than 20% to <100% of cells, −, no expression (<20% cells); +h, a given antigen has higher expression in B-NHL cells compared to normal B/T lymphocytes; +d, a given antigen has dim expression in B-NHL cells compared to normal B/T lymphocytes; Ki-67 [%] and CD71[%], percentage of secondary DLBCL,NOS cells expressing Ki-67 and/or CD71; nd, not determined.
Flow cytometry algorithm of aggressive B-NHL’s diagnosis On the basis of the published data and our own experience, we employed a practical FCM approach for diagnosis of aggressive B-NHLs [1-16], including the following criteria: 1) BL: CD45(+)weaker/CD20(+)higher>CD19(+)bright>CD22(+)/CD10(+)/BCL2(–)/CD43(+)/ FMC7(+)/ CD38(+)higher/CD81(+)higher/CD138(+/–)/CD16&CD56(–/+)/CD56(–/ +)/CD44(–)/CD79β(+)/ κ(+) or λ(+)/IgM(+)/CD5(–)/CD11c(–)/CD23/ (–)/CD25(–)/CD200(–)/CD52(+) and 100% of cells positive for CD71(+++). 2) B- NHLs[11q]:CD45(+)weaker/CD20(+)higher>CD19(+)bright>CD22(+)/CD10(+)/BCL2(–)/CD43(+)/ FMC7(+)/CD38(+)/CD81(+)higher/CD138(+/–)/CD16&CD56(+/–)/CD56(+/-)/CD44(–) or CD44(+)weaker /CD79β(+)/κ(+) or λ(+)/IgM(+)/IgD(+) or IgM(+) or IgM(+)/IgG(+)/CD5(–) /CD11c(–)/CD23(–)/CD25(–)/CD200(–)/CD52(+) and 100% of cells positive for CD71(+++). 3) D/THLs (double/triple hit lymphomas): CD45(+)/CD20(+)weaker
10 CD23(–)/ often CD25(+)/CD52(+) or loss/decreased of CD52 expression and 60-100% of cells positive for CD71(+++/++). 5) DLBCL, non-GCB (non-germinal centre B-cell-like): CD45(+)/CD20(+) or CD20(+)weaker or CD20(–) < CD19(+)bright/CD22(+/–)/CD10(–)/BCL2(+)/CD43(+)/ FMC7(+/–)/ CD38(+/–) or CD38(–)/CD138(+/–) or CD138(–)/CD16&CD56(–)/CD56(–)/CD44(+)higher/CD79β(+)/clonality of surface light and heavy chain immunoglobulins can be very diverse/CD5(–)/often CD11c(+)/CD23(–)/often CD25(+)higher/CD52(+) or loss/decreased of CD52/CD81/CD200 expression and 60-100% of cells positive for CD71(+++/++). 6) Secondary DLBCL: represents the transformation of chronic lymphocytic leukemia or small lymphocytic lymphoma (CLL/SLL)/MALT lymphoma/marginal zone lymphoma (MZL) and follicular lymphoma (FL) to high-grade lymphoma. Comparison of the FCM immunophenotypes of the previous or coexistent CLL/SLL/MALT/MZL/FL has revealed minor modulations of antigen expression without major alterations in most secondary DLBCL cases. The majority of secondary DLBCL based on SLL/CLL are characterized by a loss/decrease of CD52, CD62L and increased CD71 expression [2].
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