2019 IRTA1 MNDA in MZL
2019 IRTA1 MNDA in MZL
2019 IRTA1 MNDA in MZL
From the Department of Laboratory Medicine, Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, OH.
DOI: 10.1093/AJCP/AQY144
© American Society for Clinical Pathology, 2018. All rights reserved. Am J Clin Pathol 2019;151:337-343 337
For permissions, please e-mail: journals.permissions@oup.com DOI: 10.1093/ajcp/aqy144
Wang and Cook / IRTA1 and MNDA in MZL
lymphoma (FL).21,22 MNDA therefore may be useful in for RNA integrity. The bacterial gene DapB (cat 312039;
the differential diagnosis of MZL vs FL. The relation- Advanced Cell Diagnostics) served as a negative con-
ship, if any, between MNDA and IRTA1 expression and trol in each run. Cases with RNA in situ hybridization
correlations of MNDA and IRTA1 expression with mor- (RISH) signal for IRTA1 associated with more than 20%
phologic features have not been previously characterized. of neoplastic cells were classified as positive.
In this study, we examine the expression of MNDA
and IRTA1 in 127 lymphomas, including 80 MZLs and 47 IHC
other small B-cell neoplasms. MNDA was examined by
IHX was performed using antibodies for MNDA
IHC, while IRTA1, due to a lack of commercially avail-
(anti-MNDA, clone ab188566, 1:40; Abcam) and CD21
able antibodies for IHC, was studied using a novel RNA
(anti-CD21, mouse monoclone 1F8; Abcam) on a
in situ hybridization assay.23 In lymph node samples in-
Ventana BenchMark XT with an OptiView Amplification
volved by MZL, results of MNDA and IRTA1 studies
Kit and an OptiView DAB IHC Detection Kit (Roche
were correlated with the architectural growth pattern and
Ventana Medical Systems). Nuclear staining for MNDA
❚Table 1❚
Demographics of MZL Cases
Diagnosis Age, Median (Range), y Sex, F/M, No. (%)
NMZL, n = 23
Conventional (adult) type, n = 22 66 (52-89) 10 (45)/12 (55)
Pediatric type, n = 1 15 1F
MALT, n = 31 68 (42-85) 16 (52)/15 (48)
SMZL, n = 26 64 (43-84) 18 (69)/8 (31)
MALT, mucosa-associated lymphoid tissue; NMZL, nodal marginal zone lymphoma; SMZL, splenic marginal zone lymphoma.
examined by IRTA1 ISH, but insufficient tissue remained expression was similar in NMZL (12/22, 54%), MALT
for evaluation of MNDA staining. (21/31, 68%), and SMZL (18/26, 69%) (P = .51, χ2 test).
Results of IRTA1 and MNDA studies are detailed Expression of either IRTA1 or MNDA was found in 64
in ❚Table 2❚ and illustrated in ❚Image 2❚ and ❚Image 3❚. (84%) of 76 MZLs, while coexpression of both markers
Overall, IRTA1 expression was identified in 31 (42%) of was less frequently seen (22/73, 30%).
74 MZLs vs one (2%) of 43 non-MZL cases (P < .001, In 36 lymph node samples involved by MZL (four
Fisher exact test). Within MZL, IRTA1 expression was SMZLs, 10 MALTs, and 22 NMZLs), the expression of
similar in NMZL (10/23, 43%) and MALT (16/31, 52%) MNDA and IRTA1 was assessed in relationship to the
and slightly lower in SMZL (5/20, 25%) (P = .17, χ2 test). morphologic features. IRTA1 expression showed no cor-
The single non-MZL case positive for IRTA1 ISH was a relation with predominantly monocytoid cytology (6/15
grade 1 to 2 FL with staining in approximately 20% of [40%] cases with >50% monocytoid cells vs 12/21 [57%]
cells, located predominantly within the neoplastic folli- with <50% monocytoid cells; P = .50, Fisher exact test).
cles. This case of FL contained CD10-positive monotypic IRTA1 expression was observed less commonly in cases
B cells by flow cytometry, coexpressed CD10 and BCL2 with a diffuse pattern (4/16, 25%) vs cases with a nondif-
by IHC, and was morphologically typical for a low-grade fuse growth pattern (14/20, 70%) (P = .018, Fisher exact
FL. MNDA expression was observed in 51 (64%) of 79 test). CD21 stains, available in 15 of the 16 cases with a
MZLs vs 21 (45%) of 46 non-MZL cases (P = .06, Fisher diffuse growth pattern on routine H&E stains, showed
exact test). Notably, however, MNDA expression was sig- background residual dendritic cell meshworks in three
nificantly less common in FL (3/14, 21%) compared with (75%) of four cases positive for IRTA1 and six (54%) of
MZL (P = .003, Fisher exact test).Within MZL, MNDA 11 cases negative for IRTA1 (P = .60, Fisher exact test).
❚Table 2❚
IRTA1 and MNDA Expression in MZL and Other Small B-Cell Neoplasmsa
No./Total No. (%)
Characteristic IRTA1 MNDA Either Both
All MZLs (n = 80) 31/74 (42) 51/79 (64) 64/76 (84) 22/73 (30)
NMZL (n = 23) 10/23 (43) 12/22 (54) 18/22 (82) 7/22 (32)
MALT (n = 31) 16/31 (52) 21/31 (68) 26/31 (84) 12/31 (39)
SMZL (n = 26) 5/20 (25) 18/26 (69) 20/23 (87) 3/20 (15)
LPL (n = 9) P vs all MZLs 0/7 (0) .040 3/8 (37) .25 3/7(43) .02 0/8 (0) .10
FL (n = 14) P vs all MZLs 1/14 (7) .015 3/14 (21) .003 3/14 (21) <.001 1/14 (7) .10
CLL (n = 15) P vs all MZLs 0/13 (0) .003 8/15 (53) .56 8/13 (61) .12 0/14 (0) .017
MCL (n = 9) P vs all MZLs 0/9 (0) .023 7/9 (78) .72 7/9 (78) .64 0/9 (0) .10
CLL, chronic lymphocytic leukemia; FL, follicular lymphoma; IRTA1, immune receptor translocation-associated protein 1; LPL, lymphoplasmacytic lymphoma;
MALT, mucosa-associated lymphoid tissue; MCL, mantle cell lymphoma; MNDA, myeloid nuclear differentiation antigen; MZL, marginal zone lymphoma; NMZL,
nodal marginal zone lymphoma; SMZL, splenic marginal zone lymphoma.
❚Image 2❚ Examples of myeloid nuclear differentiation antigen (MNDA) and immune receptor translocation-associated pro-
tein 1 (IRTA1) staining in selected marginal zone lymphoma (MZL) cases. A case of nodal MZL (NMZL) is negative for MNDA
but shows staining for IRTA1. A case of splenic MZL (SMZL) involving a hilar lymph node shows staining for both MNDA and
IRTA1. A case of gastric mucosa-associated lymphoid tissue (MALT) lymphoma shows staining for both MNDA and IRTA1. (All
images ×400.)
LMO2) and are associated with BCL2 or BCL6 transloca- marginal zones are reportedly negative.11,12,14,15,17 Previous
tions. MCL expresses CD5 and cyclin D1 and/or SOX11, studies have reported IRTA1 expression in 67% to 73% of
while CLL/small lymphocytic lymphoma expresses CD5 NMZLs and 93% of extranodal MALT lymphomas,17,18
and LEF1. LPL lacks specific phenotypic markers but is but antibodies used in these prior studies are not commer-
highly associated with the MYD88 L265P point mutation cially available, limiting the utility of this potential marker.
detectable by molecular studies.26-29 MZLs, in contrast, In this study, we designed a novel RNA ISH method23 to
often remain a diagnosis of exclusion. Translocations evaluate IRTA1 expression. Using RISH, IRTA1 stain-
involving the MALT1 gene are highly specific for extran- ing was found in 43% of NMZLs, 52% of extranodal
odal MALT lymphomas, but these rearrangements are MALT lymphomas, and 25% of SMZLs. The cause of
found in only a subset of cases, are predominantly re- the lower incidence of IRTA1 staining noted by RISH in
stricted to cases arising at specific anatomic sites, and do this study compared with prior IHC reports is uncertain,
not occur in cases of nodal MZL or SMZL.5,30 For these but this might reflect differential stability of IRTA1 RNA
reasons, there remains a pressing need for biomarkers that vs protein or perhaps staining by IHC that is not com-
❚Image 3❚ Examples of myeloid nuclear differentiation antigen (MNDA) and immune receptor translocation-associated protein
1 (IRTA1) staining in selected non–marginal zone lymphoma cases. Cases of follicular lymphoma (FL) and lymphoplasmacytic
lymphoma (LPL) are negative for both MNDA and IRTA1. A case of mantle cell lymphoma (MCL) is positive for MNDA and
negative for IRTA1. (Top left panel ×100, all other images ×400.)
comparing FL and MZL identified MNDA as strongly elucidate the factors promoting IRTA1 expression in
differentiating between these two entities.22 In this study, MZL with or without a diffuse growth pattern.
we confirmed that MNDA expression is seen in normal Overall, these results demonstrate frequent expres-
splenic marginal zone B cells and is absent in normal sion of IRTA1 and MNDA in a variety of MZLs. MNDA
monocytoid B cells. Within small B-cell neoplasms, expression is particularly useful in cases with a differential
MNDA positivity was frequent in MZL and uncommon diagnosis with FL. IRTA1 offers particular advantages in
in FL (51/79 [64%] vs 3/14 [21%], respectively; P = .003). differential diagnosis as it appears to be highly specific
MNDA expression was frequent in other small B-cell for MZL, being rarely expressed in other lymphomas
neoplasms, such that the utility of MNDA IHC staining examined. The RISH assay employed in this study can
appears limited to the specific differential diagnosis of FL be adopted for routine clinical use on commercially avail-
vs MZL. This result is consistent with prior reports show- able IHC platforms and provides a practical alternative to
ing MNDA expression in most cases of small B-cell lym- IHC given that suitable IRTA1 antibodies remain com-
phomas other than FL.21,22 MNDA expression was seen mercially unavailable. These additional markers there-
12. Hatzivassiliou G, Miller I, Takizawa J, et al. IRTA1 and IRTA2, 24. Wotherspoon AC. Extranodal and splenic small B-cell lym-
novel immunoglobulin superfamily receptors expressed in B phoma. Mod Pathol. 2013;26(suppl 1):S29-S41.
cells and involved in chromosome 1q21 abnormalities in B 25. Bogusz AM, Bagg A. Genetic aberrations in small B-cell lym-
cell malignancy. Immunity. 2001;14:277-289. phomas and leukemias: molecular pathology, clinical relevance
13. Miller I, Hatzivassiliou G, Cattoretti G, et al. IRTAs: a and therapeutic targets. Leuk Lymphoma. 2016;57:1991-2013.
new family of immunoglobulinlike receptors differentially 26. Ondrejka SL, Lin JJ, Warden DW, et al. MYD88 L265P so-
expressed in B cells. Blood. 2002;99:2662-2669. matic mutation: its usefulness in the differential diagnosis of
14. Jöhrens K, Shimizu Y, Anagnostopoulos I, et al. T-bet-positive and bone marrow involvement by B-cell lymphoproliferative disor-
IRTA1-positive monocytoid B cells differ from marginal zone B ders. Am J Clin Pathol. 2013;140:387-394.
cells and epithelial-associated B cells in their antigen profile and 27. Swerdlow SH, Kuzu I, Dogan A, et al. The many faces of
topographical distribution. Haematologica. 2005;90:1070-1077. small B cell lymphomas with plasmacytic differentiation
15. Falini B, Tiacci E, Pucciarini A, et al. Expression of the IRTA1 and the contribution of MYD88 testing. Virchows Arch.
receptor identifies intraepithelial and subepithelial marginal 2016;468:259-275.
zone B cells of the mucosa-associated lymphoid tissue (MALT). 28. Hamadeh F, MacNamara SP, Aguilera NS, et al. MYD88
Blood. 2003;102:3684-3692. L265P mutation analysis helps define nodal lymphoplasma-
16. Ikeda JI, Kohara M, Tsuruta Y, et al. Immunohistochemical cytic lymphoma. Mod Pathol. 2015;28:564-574.