Fig. 1Detecting immunoglobulin (IG) and T cell receptor (TCR) gene rearrangement by 4% Metaphor agarose gel (A) and 8% non-denaturing polyacylamide gel (B) using the BIOMED-2 protocols. Results of IG/TCR gene clonality detection are indicated by the presence or absence of a PCR product of the expected size. All reactions on non-denaturing polyacrylamide gels and Metaphor agarose gels revealed the same results. The case of the photos showed positivity for TCRBA and TCRBC on both gels. The expected size of the PCR products were as follows: IGHA 310-360 bp; IGHB 250-295 bp; IGHC 100-170 bp; IGHD 110-290, 390-420 bp; IGHE 100-130 bp; IGKA 120-160, 190-210, 260-300 bp; IGKB 201-250, 270-300, 350-390 bp; TCRBA 240-285 bp; TCRBB 240-285 bp; TCRBC 170-210, 285-325 bp; TCRGA 145-255; TCRGB 80-220 bp. M, DNA size marker; PC, positive control; NC, negative control.
Table 1.Characteristics of the non-Hodgkin lymphomas and reactive lymphoproliferative diseases
Characteristic |
NHL |
RLD |
No. |
158 (100) |
61 (100) |
Age (yr) |
|
|
< 30 |
21 (13.3) |
12 (19.7) |
30-49 |
27 (17.1) |
19 (31.1) |
50-64 |
49 (31.0) |
18 (29.5) |
> 65 |
61 (38.6) |
12 (19.7) |
Mean (range) |
57.8 (7-88) |
50.8 (3-78) |
Sex |
|
|
Male |
94 |
36 |
Female |
64 |
25 |
Subtype |
|
|
MCL |
4 |
NA |
FL |
6 |
NA |
MZL |
3 |
NA |
MALTL |
8 |
NA |
DLBCL |
49 |
NA |
BLL/L |
8 |
NA |
PTCL-U |
41 |
NA |
ALCL |
7 |
NA |
N-NK/T |
12 |
NA |
AITL |
12 |
NA |
ATL/L |
2 |
NA |
TLL/L |
6 |
NA |
Table 2.Clonality of IG and TCR targets in B cell lymphoma, T cell lymphoma and reactive lymphoproliferative disease
|
BCL (n = 78)
|
TCL (n = 80)
|
RLD (n = 61) |
|
Mature BCL (n = 70) |
BLL/L (n = 8) |
Mature TCL (n = 74) |
TLL/L (n = 6) |
IGH+IGK |
67 (95.7) |
4 (50.0) |
6 (8.1) |
0 (0) |
2 (3.3) |
TCRB+TCRG |
2 (2.9) |
5 (62.5) |
62 (83.8) |
4 (66.7) |
2 (3.3) |
Co-existing clonality |
2 (2.9) |
2 (25.0) |
6 (8.1) |
0 (0) |
0 (0) |
Polyclonality |
3 (4.3) |
1 (12.5) |
12 (16.2) |
2 (33.3) |
57 (93.4) |
Table 3.Genotypic profiles of B cell lymphoma (BCL)
|
Mature BCL
|
Precursor BCL
|
Total (n = 78) |
|
MCL (n = 4) |
FL (n = 6) |
MZL (n = 3) |
MALTL (n = 8) |
DLBCL (n = 49) |
Subtotal (n = 70) |
BLL/L (n = 8) |
IGH genes |
|
|
|
|
|
|
|
|
No rearrangement |
1 (25.0) |
1 (16.7) |
0 (0) |
0 (0) |
8 (16.3) |
10 (4.3) |
5 (62.5) |
15 (23.1) |
VDJ cases |
2 (50.0) |
5 (83.3) |
3 (100) |
8 (100) |
38 (77.6) |
56 (64.0) |
3 (37.5) |
59 (75.6) |
DH-JH cases |
1 (25.0) |
0 (0) |
1 (33.3) |
4 (50.0) |
29 (59.2) |
35 (50.0) |
1 (12.5) |
36 (46.2) |
All IGH VDJ+DH |
3 (75.0) |
5 (83.3) |
3 (100) |
8 (100) |
41 (83.7) |
60 (85.7) |
3 (37.5) |
63 (80.8) |
IGK genes |
|
|
|
|
|
|
|
|
No rearrangement |
0 (0) |
1 (16.7) |
1 (33.3) |
4 (50.0) |
31 (63.3) |
37 (52.9) |
7 (87.5) |
44 (56.4) |
Total IGK |
4 (100) |
5 (83.3) |
2 (66.7) |
4 (50.0) |
18 (36.7) |
33 (47.1) |
1 (12.5) |
34 (43.6) |
Combined IG genes |
|
|
|
|
|
|
|
|
All IGH+IGK |
4 (100) |
6 (100) |
3 (100) |
8 (100) |
46 (93.8) |
67 (95.7) |
4 (50.0) |
71 (91.0) |
TCRB genes |
|
|
|
|
|
|
|
|
Vβ-Jβ genes |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
1 (2.0) |
1 (1.4) |
1 (12.5) |
2 (2.6) |
Dβ-Jβ genes |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
1 (12.5) |
1 (1.3) |
Total TCRB |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
1 (2.0) |
1 (1.4) |
2 (25.0) |
3 (3.8) |
TCRG genes |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
1 (2.0) |
1 (1.4) |
5 (62.5) |
6 (7.7) |
Combined TCR genes |
|
|
|
|
|
|
|
|
All TCRB+TCRG |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
2 (4.1) |
2 (2.9) |
5 (62.5) |
7 (9.0) |
Polyclonality |
0 (0) |
0 (0) |
0 (0) |
0 (0) |
3 (6.1) |
3 (6.1) |
1 (12.5) |
4 (5.1) |
Table 4.Genotypic profiles of T cell lymphoma (TCL)
|
Mature TCL
|
Precursor TCL
|
Total (n = 80) |
|
PTCL-U (n = 41) |
ACLC (n = 7) |
N-NK/T (n = 12) |
AITL (n = 12) |
ATL/L (n = 2) |
Subtotal (n = 74) |
TLL/L (n = 6) |
TCRB genes |
|
|
|
|
|
|
|
|
No rearrangement |
22 (53.7) |
4 (57.1) |
5 (41.7) |
5 (41.7) |
0 (0) |
36 (48.6) |
4 (75.0) |
40 (50.0) |
Vβ-Jβ cases |
12 (29.3) |
2 (28.6) |
2 (16.7) |
5 (41.7) |
2 (100) |
23 (31.1) |
1 (16.7) |
24 (30.0) |
Dβ-Jβ cases |
13 (31.7) |
4 (57.1) |
5 (41.7) |
3 (25.0) |
1 (50.0) |
26 (35.1) |
1 (16.7) |
27 (33.8) |
Total TCRB |
19 (46.3) |
4 (57.1) |
7 (58.3) |
6 (50.0) |
2 (100) |
38 (51.4) |
2 (33.3) |
40 (50.0) |
TCRG genes |
|
|
|
|
|
|
|
|
No rearrangement |
19 (46.3) |
4 (57.1) |
8 (66.7) |
4 (33.3) |
0 (0) |
35 (47.3) |
4 (66.7) |
39 (48.8) |
Total TCRG |
21 (51.2) |
3 (42.9) |
4 (33.3) |
8 (66.7) |
2 (100) |
38 (51.4) |
2 (33.3) |
40 (50.0) |
Total TCR |
37 (90.2) |
5 (71.4) |
8 (66.7) |
10 (83.3) |
2 (100) |
62 (83.8) |
4 (66.7) |
66 (82.5) |
IGH genes |
|
|
|
|
|
|
|
|
VDJ cases |
1 (2.4) |
0 (0) |
0 (0) |
2 (16.7) |
0 (0) |
3 (4.0) |
0 (0) |
3 (3.8) |
DH-JH cases |
3 (7.3) |
0 (0) |
0 (0) |
1 (8.3) |
0 (0) |
4 (5.4) |
0 (0) |
4 (5.0) |
All IGH VDJ+DH |
3 (7.3) |
0 (0) |
0 (0) |
2 (16.7) |
0 (0) |
5 (6.8) |
0 (0) |
5 (6.3) |
IGK genes |
1 (2.4) |
0 (0) |
0 (0) |
1 (8.3) |
0 (0) |
2 (2.7) |
0 (0) |
2 (2.5) |
Combined IG genes |
|
|
|
|
|
|
|
|
All IGH+IGK |
3 (7.3) |
0 (0) |
0 (0) |
2 (16.7) |
0 (0) |
5 (6.8) |
0 (0) |
5 (6.3) |
Polyclonality |
4 (9.8) |
2 (28.6) |
4 (33.3) |
2 (16.7) |
0 (0) |
12 (16.2) |
2 (33.3) |
14 (17.5) |