From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
Correspondence To the editor: The PRDX2 gene is transcriptionally silenced and de novo methylated in Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma In a recent issue of Blood, Agrawal-Singh et al reported the identification of PRDX2 (peroxiredoxin 2) as a novel tumor suppressor gene in acute myeloid leukemia (AML).1 The authors observed recurrent histone H3 deacetylation and DNA hypermethylation at
the PRDX2 promoter region in primary leukemia samples. Epigenetic alterations correlated with low expression of PRDX2 protein and were associated with poor prognosis in AML patients. Moreover, the authors demonstrated that this gene acts as a growth inhibitor
Figure 1. Promoter hypermethylation and lack of PRDX2 expression in Hodgkin and Reed-Sternberg (HRS) cells. (A) Localization of the 27K BeadArray (Illumina) array tags cg03860466 and cg00155609 (indicated by arrows) with respect to the PRDX2 TSS. (B) PRDX2 mRNA expression (Affymetrix U95 microarray, tag 39729_at) in cHL cell lines, normal B-cell populations (10 3 GCB cells, 5 3 naive B cells, 5 3 memory B cells), 15 diffuse large B-cell lymphomas (DLBCLs), 11 Burkitt lymphomas (BLs), 6 follicular lymphomas (FLs), 11 B-cell chronic lymphocytic leukemias (CLLs), and 5 large cell lymphomas (LCLs). Mean PRDX2 promoter methylation level based on 27K BeadArray (Illumina) array (tags: cg03860466, cg00155609) in the cHL cell lines is given in parentheses. Methylation and expression data were taken from Ammerpohl et al2 and Kuppers ¨ et al,3 respectively. (C) Bisulfite pyrosequencing of PRDX2 promoter region in cHL cell lines. Bars represent the cytosine guanine dinucleotide (CpG) sites analyzed within the sequenced region (genomic position of the CpG sites: black bar, chr19:12912528; gray bar, chr19:12912542). No bar indicates a methylation level of 0%. (D) Bisulfite pyrosequencing of PRDX2 promoter region in microdissected HRS cells (nodular sclerosis, cases 1, 2, 4, 5; mixed cellularity, 3) and nontumoral bystander cells (Lymph) (in duplicate) (genomic position of the CpG sites: black bar, chr19:12912528; gray bar, chr19:12912542). No bar indicates a methylation level of 0%. M, methylated DNA control (Millipore); POOL, whole blood DNAs, pooled; U, unmethylated DNA control (whole genome amplified DNA).
3672
BLOOD, 5 JUNE 2014 x VOLUME 123, NUMBER 23
From www.bloodjournal.org by guest on September 11, 2016. For personal use only. BLOOD, 5 JUNE 2014 x VOLUME 123, NUMBER 23
in myeloid cells. Here, we provide evidence that PRDX2 methylation and transcriptional silencing is recurrent not only in AML but also in classical Hodgkin lymphoma (cHL), further emphasizing the importance of loss of this tumor suppressor in hematologic malignancies. By analyzing methylation profiles of 5 cHL cell lines (27K BeadArray; Illumina),2 we observed hypermethylation of the PRDX2 gene compared with normal B-cell populations, including germinal center B (GCB) cells and a lymphoblastoid B-cell line. Elevated PRDX2 methylation was present in cHL cell lines L428, KMH2, L1236, and UHO1, but not HDLM2, and was indicated by tags flanking the PRDX2 transcription starting site (TSS) (cg03860466, –217 bp from TSS; cg00155609, 1196 bp from TSS) (Figure 1A). As observed in AML, elevated methylation correlated with lack of PRDX2 messenger RNA (mRNA) in the cHL cell lines (Figure 1B). In contrast, expression was present in germinal center, naive, and memory B cells and other lymphoma and leukemia specimens (except 3/11 Burkitt lymphomas) showing consistent expression of PRDX2 in normal B-cell populations and the majority of other hematologic malignancies (Figure 1B).3 This is consistent with the observation that the PRDX2 promoter region is rich in motifs recognized by transcription factors widely expressed in B cells, including PAX5, ELF1, TCF4, OCT-1, and nuclear factor kB.4 Together, these findings suggest epigenetic silencing as the mechanism responsible for the downregulation of PRDX2 in cHL (Figure 1B). Encouraged by these results, we designed a bisulfite pyrosequencing assay to measure DNA methylation at 2 CpG sites (chr19: 12912528 corresponding to the cg00155609 array tag and chr19: 12912542) in the 59 region of PRDX2. The sequenced region encompassed the position chr19:12 912 503-12 912 544 (hg19) (forward primer: 59-GGTTTAGGGTTTATTTGG-39; reverse primer: biotin-59-CCACCAACACTAAAATCAA-39; sequencing primer: 59-TGTTTAAGTAGGTATGAATA-39). The assay was validated on cHL cell lines showing similar methylation levels of PRDX2 as measured by the microarray (Figure 1C). We applied the assay to primary HRS cells from cHL patients. Because HRS cells constitute usually ;1% of the cellular infiltrate in an affected lymph node, we used laser microdissection to obtain a pure population of HRS cells. For each patient, 200 HRS cells, 200 nontumoral bystander cells, and a membrane control were microdissected. DNA extracted from these samples was bisulfite converted and pyrosequenced, including methylated and nonmethylated technical controls. Samples from 5 patients were analyzed in duplicate. In 2 patients (#2 and #4), we indeed observed hypermethylation of the PRDX2 promoter region in microdissected HRS cells compared with corresponding bystander cells (Figure 1D). This demonstrates that PRDX2 is recurrently hypermethylated in primary HRS cells and provides a rationale for our observation based on U133 plus 2.0 expression profiles that primary microdissected HRS cell populations (n 5 12) show a 0.58-fold downregulation of PRDX2 expression compared with GCB cells (n 5 10).5 Under physiological conditions, PRDX2 scavenges reactive oxygen species modulating their intracellular levels.6 Interestingly, we recently reported that genetic as well as epigenetic alterations in cHL frequently target genes related to reactive oxygen species homeostasis, putatively contributing to loss of B-cell identity and survival of HRS cells.7 Here we propose PRDX2 as a new player among these genes. Moreover, our results suggest epigenetic silencing of this gene in cHL extending the findings by Agrawal-Singh et al.1
CORRESPONDENCE
3673
Markus Schneider University of Duisburg-Essen, Essen, Germany Marcin Szaumkessel Polish Academy of Sciences, Poznan, ´ Poland Julia Richter Christian-Albrechts-University Kiel, Kiel, Germany University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany Ole Ammerpohl Christian-Albrechts-University Kiel, Kiel, Germany University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany Martin-Leo Hansmann Goethe University, Frankfurt am Main, Germany Ralf Kuppers ¨ University of Duisburg-Essen, Essen, Germany Reiner Siebert Christian-Albrechts-University Kiel, Kiel, Germany University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany Maciej Giefing Polish Academy of Sciences, Poznan, ´ Poland M. Schneider and M. Szaumkessel contributed equally to this study. Acknowledgments: This work was supported by a grant from the Polish National Science Centre (2011/01/D/NZ2/00095); the Deutsche Forschungsgemeinschaft (grant GRK1431); the Kinderkrebsinitiative Buchholz/Holm-Seppensen, infrastructure (R.S.); the Federation of Biochemical Societies, long-term fellowship (M.G.) and short-term fellowship “Collaborative Experimental Scholarships for Central and Eastern Europe” (M. Szaumkessel); the Polish Ministry of Science and Higher Education, “Support for International Mobility of Scientists” fellowship (M.G.); and a European Molecular Biology Organization, short-term fellowship (M. Szaumkessel). Contribution: M. Schneider and M. Szaumkessel performed research; J.R. and O.A. designed experiments; M.-L.H. contributed analytical tools and samples; R.K. provided gene expression data and revised the manuscript; R.S. analyzed data and wrote parts of the manuscript; and M.G. designed and performed research, analyzed data, and wrote parts of the manuscript. Conflict-of-interest disclosure: The authors declare no competing financial interests. Correspondence: Maciej Giefing, Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska ´ 32, 60-479 Poznan, ´ Poland; e-mail: [email protected].
References 1. Agrawal-Singh S, Isken F, Agelopoulos K, et al. Genome-wide analysis of histone H3 acetylation patterns in AML identifies PRDX2 as an epigenetically silenced tumor suppressor gene. Blood. 2012;119(10): 2346-2357. 2. Ammerpohl O, Haake A, Pellissery S, et al. Array-based DNA methylation analysis in classical Hodgkin lymphoma reveals new insights into the mechanisms underlying silencing of B cell-specific genes. Leukemia. 2012;26(1): 185-188. 3. Kuppers ¨ R, Klein U, Schwering I, et al. Identification of Hodgkin and ReedSternberg cell-specific genes by gene expression profiling. J Clin Invest. 2003; 111(4):529-537.
From www.bloodjournal.org by guest on September 11, 2016. For personal use only. 3674
BLOOD, 5 JUNE 2014 x VOLUME 123, NUMBER 23
CORRESPONDENCE
4. ENCODE Project Consortium. A user’s guide to the Encyclopedia of DNA Elements (ENCODE). PLoS Biol. 2011;9(4):e1001046. 5. Brune V, Tiacci E, Pfeil I, et al. Origin and pathogenesis of nodular lymphocytepredominant Hodgkin lymphoma as revealed by global gene expression analysis. J Exp Med. 2008;205(10):2251-2268. 6. Hall A, Karplus PA, Poole LB. Typical 2-Cys peroxiredoxins—structures, mechanisms and functions. FEBS J. 2009;276(9):2469-2477.
7. Giefing M, Winoto-Morbach S, Sosna J, et al. Hodgkin-Reed-Sternberg cells in classical Hodgkin lymphoma show alterations of genes encoding the NADPH oxidase complex and impaired reactive oxygen species synthesis capacity. PLoS ONE. 2013;8(12): e84928.
© 2014 by The American Society of Hematology
To the editor: Phenotypically aberrant clonal T cells in the lungs of patients with type II refractory celiac disease Type II refractory celiac disease (RCD II) is a rare condition characterized by a massive accumulation of intraepithelial lymphocytes with a natural killer/T phenotype and containing clonal T-cell rearrangements.1 Its severe prognosis is largely due to the development of an aggressive enteropathy-type–associated T-cell lymphoma (EATL). RCD II is a diffuse digestive but also extradigestive disease.2 Various pulmonary manifestations have been reported in celiac patients3-6 but little, if anything, is known about their mechanisms, even though the presence of a T lymphoid alveolitis had been described. One study has shown the presence of aberrant T cells in the lungs of some celiac patients.7 We confirm and extend these data by reporting here 7 patients with RCD II in whom the presence of specific aberrant T lymphocytes and clonal T-cell receptor gamma (TCR-g) rearrangements were detected in bronchoalveolar lavage (BAL), and/or in bronchial biopsies, or in mediastinal lymph nodes, all of which argue for the lung as an overt extradigestive site of the disease. All patients exhibited a massive digestive intraepithelial infiltration by cCD31, CD3–, CD4–, CD8–, and CD1031 T cells containing clonal TCR-g rearrangements associated in some way with the presence of similar T cells in skin and/or bone marrow (Table 1). Five of the 7 also exhibited the same phenotypically aberrant cells in BAL and/or bronchial biopsy T cells, accounting by fluorescenceactivated cell sorter analysis for 0.5% to 74% of total BAL lymphocytes. Lung cells also contained rearranged TCR-g T-cell clones, which were not always identical to those detected in the digestive mucosa and/or in blood. EATL developed in 4 of 7 patients. Two of them died of cutaneous lymphoma and 2 from digestive lymphoma.
Various clinical respiratory manifestations (chronic cough, hemoptysis, organized pneumonia) can occur in celiac patients, including the intrapulmonary location of RCD II–specific T cells in clinically asymptomatic patients.7 We confirm and extend the latter observation by showing the presence of clonal T cells in the lungs of 7 patients with RCD II in alveolar and/or bronchial epithelium or mediastinal lymph nodes. Interestingly, these clonal T-cells in some instances harbored different TCR-g rearrangements from those detected in the digestive mucosa or in circulating blood, ruling out their detection in the lung as mere blood contamination. Taken together, these data strongly argue for airway epithelium as an additional specific target of RCD II with its potentially most severe complication— aggressive local EATL. This should alert the clinicians in charge of such patients about the importance of including pulmonary evaluation in their routine workups. Indeed, these patients can be clinically and/or radiologically asymptomatic and yet exhibit abnormal T cells, with a potential for transformation into aggressive malignant cells. Jean Pastre´ Service de Pneumologie, Hopital ˆ Europeen ´ Georges Pompidou, Universite´ Paris Descartes, Paris, France Karine Juvin Service de Pneumologie, Hopital ˆ Europeen ´ Georges Pompidou, Universite´ Paris Descartes, Paris, France Georgia Malamut Service de Gastro-enterologie, ´ Hopital ˆ Europeen ´ Georges Pompidou, Universite´ Paris Descartes, Paris, France
Table 1. Characteristics of patients with refractory celiac disease type II and specific pulmonary involvement Age Patient Sex (years)
Aberrant T cells in digestive sample (%)*
Extradigestive localization of aberrant T cells
Aberrant T cells in BAL (%)*
Clonal TCR-g rearrangement in the lung
Site
Follow-up (months)
Outcome Died of digestive
1
F
36
60
Skin, bone marrow, lung
NA
1
BAL, BB
60
2
M
39
65
Bone marrow, lung
0.5
1
BAL
96
3
F
60
85
Bone marrow, lung
1
–
4
F
68
60
Skin, bone marrow, lung
0.5
1
5
F
51
80
Bone marrow, lung
1
6
F
57
70
Skin, bone marrow, blood,
7
F
31
40
Bone marrow, lung
lymphoma Alive
72
Alive
BAL
36
Died of skin
1
BAL, BB
36
Alive
74
1
BAL, BB
24
Died of skin
NA
1
Mediastinal
28
Died of digestive
lymphoma
lung
lymphoma lymph nodes
BB, bronchial biopsy; F, female; M, male; NA, not available. *Percentage of total sample lymphocytes.
lymphoma
From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
2014 123: 3672-3674 doi:10.1182/blood-2014-02-553263
The PRDX2 gene is transcriptionally silenced and de novo methylated in Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma Markus Schneider, Marcin Szaumkessel, Julia Richter, Ole Ammerpohl, Martin-Leo Hansmann, Ralf Küppers, Reiner Siebert and Maciej Giefing
Updated information and services can be found at: http://www.bloodjournal.org/content/123/23/3672.full.html Articles on similar topics can be found in the following Blood collections Lymphoid Neoplasia (2370 articles) Information about reproducing this article in parts or in its entirety may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests Information about ordering reprints may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#reprints Information about subscriptions and ASH membership may be found online at: http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American Society of Hematology, 2021 L St, NW, Suite 900, Washington DC 20036. Copyright 2011 by The American Society of Hematology; all rights reserved.
Correspondence To the editor: The PRDX2 gene is transcriptionally silenced and de novo methylated in Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma In a recent issue of Blood, Agrawal-Singh et al reported the identification of PRDX2 (peroxiredoxin 2) as a novel tumor suppressor gene in acute myeloid leukemia (AML).1 The authors observed recurrent histone H3 deacetylation and DNA hypermethylation at
the PRDX2 promoter region in primary leukemia samples. Epigenetic alterations correlated with low expression of PRDX2 protein and were associated with poor prognosis in AML patients. Moreover, the authors demonstrated that this gene acts as a growth inhibitor
Figure 1. Promoter hypermethylation and lack of PRDX2 expression in Hodgkin and Reed-Sternberg (HRS) cells. (A) Localization of the 27K BeadArray (Illumina) array tags cg03860466 and cg00155609 (indicated by arrows) with respect to the PRDX2 TSS. (B) PRDX2 mRNA expression (Affymetrix U95 microarray, tag 39729_at) in cHL cell lines, normal B-cell populations (10 3 GCB cells, 5 3 naive B cells, 5 3 memory B cells), 15 diffuse large B-cell lymphomas (DLBCLs), 11 Burkitt lymphomas (BLs), 6 follicular lymphomas (FLs), 11 B-cell chronic lymphocytic leukemias (CLLs), and 5 large cell lymphomas (LCLs). Mean PRDX2 promoter methylation level based on 27K BeadArray (Illumina) array (tags: cg03860466, cg00155609) in the cHL cell lines is given in parentheses. Methylation and expression data were taken from Ammerpohl et al2 and Kuppers ¨ et al,3 respectively. (C) Bisulfite pyrosequencing of PRDX2 promoter region in cHL cell lines. Bars represent the cytosine guanine dinucleotide (CpG) sites analyzed within the sequenced region (genomic position of the CpG sites: black bar, chr19:12912528; gray bar, chr19:12912542). No bar indicates a methylation level of 0%. (D) Bisulfite pyrosequencing of PRDX2 promoter region in microdissected HRS cells (nodular sclerosis, cases 1, 2, 4, 5; mixed cellularity, 3) and nontumoral bystander cells (Lymph) (in duplicate) (genomic position of the CpG sites: black bar, chr19:12912528; gray bar, chr19:12912542). No bar indicates a methylation level of 0%. M, methylated DNA control (Millipore); POOL, whole blood DNAs, pooled; U, unmethylated DNA control (whole genome amplified DNA).
3672
BLOOD, 5 JUNE 2014 x VOLUME 123, NUMBER 23
From www.bloodjournal.org by guest on September 11, 2016. For personal use only. BLOOD, 5 JUNE 2014 x VOLUME 123, NUMBER 23
in myeloid cells. Here, we provide evidence that PRDX2 methylation and transcriptional silencing is recurrent not only in AML but also in classical Hodgkin lymphoma (cHL), further emphasizing the importance of loss of this tumor suppressor in hematologic malignancies. By analyzing methylation profiles of 5 cHL cell lines (27K BeadArray; Illumina),2 we observed hypermethylation of the PRDX2 gene compared with normal B-cell populations, including germinal center B (GCB) cells and a lymphoblastoid B-cell line. Elevated PRDX2 methylation was present in cHL cell lines L428, KMH2, L1236, and UHO1, but not HDLM2, and was indicated by tags flanking the PRDX2 transcription starting site (TSS) (cg03860466, –217 bp from TSS; cg00155609, 1196 bp from TSS) (Figure 1A). As observed in AML, elevated methylation correlated with lack of PRDX2 messenger RNA (mRNA) in the cHL cell lines (Figure 1B). In contrast, expression was present in germinal center, naive, and memory B cells and other lymphoma and leukemia specimens (except 3/11 Burkitt lymphomas) showing consistent expression of PRDX2 in normal B-cell populations and the majority of other hematologic malignancies (Figure 1B).3 This is consistent with the observation that the PRDX2 promoter region is rich in motifs recognized by transcription factors widely expressed in B cells, including PAX5, ELF1, TCF4, OCT-1, and nuclear factor kB.4 Together, these findings suggest epigenetic silencing as the mechanism responsible for the downregulation of PRDX2 in cHL (Figure 1B). Encouraged by these results, we designed a bisulfite pyrosequencing assay to measure DNA methylation at 2 CpG sites (chr19: 12912528 corresponding to the cg00155609 array tag and chr19: 12912542) in the 59 region of PRDX2. The sequenced region encompassed the position chr19:12 912 503-12 912 544 (hg19) (forward primer: 59-GGTTTAGGGTTTATTTGG-39; reverse primer: biotin-59-CCACCAACACTAAAATCAA-39; sequencing primer: 59-TGTTTAAGTAGGTATGAATA-39). The assay was validated on cHL cell lines showing similar methylation levels of PRDX2 as measured by the microarray (Figure 1C). We applied the assay to primary HRS cells from cHL patients. Because HRS cells constitute usually ;1% of the cellular infiltrate in an affected lymph node, we used laser microdissection to obtain a pure population of HRS cells. For each patient, 200 HRS cells, 200 nontumoral bystander cells, and a membrane control were microdissected. DNA extracted from these samples was bisulfite converted and pyrosequenced, including methylated and nonmethylated technical controls. Samples from 5 patients were analyzed in duplicate. In 2 patients (#2 and #4), we indeed observed hypermethylation of the PRDX2 promoter region in microdissected HRS cells compared with corresponding bystander cells (Figure 1D). This demonstrates that PRDX2 is recurrently hypermethylated in primary HRS cells and provides a rationale for our observation based on U133 plus 2.0 expression profiles that primary microdissected HRS cell populations (n 5 12) show a 0.58-fold downregulation of PRDX2 expression compared with GCB cells (n 5 10).5 Under physiological conditions, PRDX2 scavenges reactive oxygen species modulating their intracellular levels.6 Interestingly, we recently reported that genetic as well as epigenetic alterations in cHL frequently target genes related to reactive oxygen species homeostasis, putatively contributing to loss of B-cell identity and survival of HRS cells.7 Here we propose PRDX2 as a new player among these genes. Moreover, our results suggest epigenetic silencing of this gene in cHL extending the findings by Agrawal-Singh et al.1
CORRESPONDENCE
3673
Markus Schneider University of Duisburg-Essen, Essen, Germany Marcin Szaumkessel Polish Academy of Sciences, Poznan, ´ Poland Julia Richter Christian-Albrechts-University Kiel, Kiel, Germany University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany Ole Ammerpohl Christian-Albrechts-University Kiel, Kiel, Germany University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany Martin-Leo Hansmann Goethe University, Frankfurt am Main, Germany Ralf Kuppers ¨ University of Duisburg-Essen, Essen, Germany Reiner Siebert Christian-Albrechts-University Kiel, Kiel, Germany University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany Maciej Giefing Polish Academy of Sciences, Poznan, ´ Poland M. Schneider and M. Szaumkessel contributed equally to this study. Acknowledgments: This work was supported by a grant from the Polish National Science Centre (2011/01/D/NZ2/00095); the Deutsche Forschungsgemeinschaft (grant GRK1431); the Kinderkrebsinitiative Buchholz/Holm-Seppensen, infrastructure (R.S.); the Federation of Biochemical Societies, long-term fellowship (M.G.) and short-term fellowship “Collaborative Experimental Scholarships for Central and Eastern Europe” (M. Szaumkessel); the Polish Ministry of Science and Higher Education, “Support for International Mobility of Scientists” fellowship (M.G.); and a European Molecular Biology Organization, short-term fellowship (M. Szaumkessel). Contribution: M. Schneider and M. Szaumkessel performed research; J.R. and O.A. designed experiments; M.-L.H. contributed analytical tools and samples; R.K. provided gene expression data and revised the manuscript; R.S. analyzed data and wrote parts of the manuscript; and M.G. designed and performed research, analyzed data, and wrote parts of the manuscript. Conflict-of-interest disclosure: The authors declare no competing financial interests. Correspondence: Maciej Giefing, Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska ´ 32, 60-479 Poznan, ´ Poland; e-mail: [email protected].
References 1. Agrawal-Singh S, Isken F, Agelopoulos K, et al. Genome-wide analysis of histone H3 acetylation patterns in AML identifies PRDX2 as an epigenetically silenced tumor suppressor gene. Blood. 2012;119(10): 2346-2357. 2. Ammerpohl O, Haake A, Pellissery S, et al. Array-based DNA methylation analysis in classical Hodgkin lymphoma reveals new insights into the mechanisms underlying silencing of B cell-specific genes. Leukemia. 2012;26(1): 185-188. 3. Kuppers ¨ R, Klein U, Schwering I, et al. Identification of Hodgkin and ReedSternberg cell-specific genes by gene expression profiling. J Clin Invest. 2003; 111(4):529-537.
From www.bloodjournal.org by guest on September 11, 2016. For personal use only. 3674
BLOOD, 5 JUNE 2014 x VOLUME 123, NUMBER 23
CORRESPONDENCE
4. ENCODE Project Consortium. A user’s guide to the Encyclopedia of DNA Elements (ENCODE). PLoS Biol. 2011;9(4):e1001046. 5. Brune V, Tiacci E, Pfeil I, et al. Origin and pathogenesis of nodular lymphocytepredominant Hodgkin lymphoma as revealed by global gene expression analysis. J Exp Med. 2008;205(10):2251-2268. 6. Hall A, Karplus PA, Poole LB. Typical 2-Cys peroxiredoxins—structures, mechanisms and functions. FEBS J. 2009;276(9):2469-2477.
7. Giefing M, Winoto-Morbach S, Sosna J, et al. Hodgkin-Reed-Sternberg cells in classical Hodgkin lymphoma show alterations of genes encoding the NADPH oxidase complex and impaired reactive oxygen species synthesis capacity. PLoS ONE. 2013;8(12): e84928.
© 2014 by The American Society of Hematology
To the editor: Phenotypically aberrant clonal T cells in the lungs of patients with type II refractory celiac disease Type II refractory celiac disease (RCD II) is a rare condition characterized by a massive accumulation of intraepithelial lymphocytes with a natural killer/T phenotype and containing clonal T-cell rearrangements.1 Its severe prognosis is largely due to the development of an aggressive enteropathy-type–associated T-cell lymphoma (EATL). RCD II is a diffuse digestive but also extradigestive disease.2 Various pulmonary manifestations have been reported in celiac patients3-6 but little, if anything, is known about their mechanisms, even though the presence of a T lymphoid alveolitis had been described. One study has shown the presence of aberrant T cells in the lungs of some celiac patients.7 We confirm and extend these data by reporting here 7 patients with RCD II in whom the presence of specific aberrant T lymphocytes and clonal T-cell receptor gamma (TCR-g) rearrangements were detected in bronchoalveolar lavage (BAL), and/or in bronchial biopsies, or in mediastinal lymph nodes, all of which argue for the lung as an overt extradigestive site of the disease. All patients exhibited a massive digestive intraepithelial infiltration by cCD31, CD3–, CD4–, CD8–, and CD1031 T cells containing clonal TCR-g rearrangements associated in some way with the presence of similar T cells in skin and/or bone marrow (Table 1). Five of the 7 also exhibited the same phenotypically aberrant cells in BAL and/or bronchial biopsy T cells, accounting by fluorescenceactivated cell sorter analysis for 0.5% to 74% of total BAL lymphocytes. Lung cells also contained rearranged TCR-g T-cell clones, which were not always identical to those detected in the digestive mucosa and/or in blood. EATL developed in 4 of 7 patients. Two of them died of cutaneous lymphoma and 2 from digestive lymphoma.
Various clinical respiratory manifestations (chronic cough, hemoptysis, organized pneumonia) can occur in celiac patients, including the intrapulmonary location of RCD II–specific T cells in clinically asymptomatic patients.7 We confirm and extend the latter observation by showing the presence of clonal T cells in the lungs of 7 patients with RCD II in alveolar and/or bronchial epithelium or mediastinal lymph nodes. Interestingly, these clonal T-cells in some instances harbored different TCR-g rearrangements from those detected in the digestive mucosa or in circulating blood, ruling out their detection in the lung as mere blood contamination. Taken together, these data strongly argue for airway epithelium as an additional specific target of RCD II with its potentially most severe complication— aggressive local EATL. This should alert the clinicians in charge of such patients about the importance of including pulmonary evaluation in their routine workups. Indeed, these patients can be clinically and/or radiologically asymptomatic and yet exhibit abnormal T cells, with a potential for transformation into aggressive malignant cells. Jean Pastre´ Service de Pneumologie, Hopital ˆ Europeen ´ Georges Pompidou, Universite´ Paris Descartes, Paris, France Karine Juvin Service de Pneumologie, Hopital ˆ Europeen ´ Georges Pompidou, Universite´ Paris Descartes, Paris, France Georgia Malamut Service de Gastro-enterologie, ´ Hopital ˆ Europeen ´ Georges Pompidou, Universite´ Paris Descartes, Paris, France
Table 1. Characteristics of patients with refractory celiac disease type II and specific pulmonary involvement Age Patient Sex (years)
Aberrant T cells in digestive sample (%)*
Extradigestive localization of aberrant T cells
Aberrant T cells in BAL (%)*
Clonal TCR-g rearrangement in the lung
Site
Follow-up (months)
Outcome Died of digestive
1
F
36
60
Skin, bone marrow, lung
NA
1
BAL, BB
60
2
M
39
65
Bone marrow, lung
0.5
1
BAL
96
3
F
60
85
Bone marrow, lung
1
–
4
F
68
60
Skin, bone marrow, lung
0.5
1
5
F
51
80
Bone marrow, lung
1
6
F
57
70
Skin, bone marrow, blood,
7
F
31
40
Bone marrow, lung
lymphoma Alive
72
Alive
BAL
36
Died of skin
1
BAL, BB
36
Alive
74
1
BAL, BB
24
Died of skin
NA
1
Mediastinal
28
Died of digestive
lymphoma
lung
lymphoma lymph nodes
BB, bronchial biopsy; F, female; M, male; NA, not available. *Percentage of total sample lymphocytes.
lymphoma
From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
2014 123: 3672-3674 doi:10.1182/blood-2014-02-553263
The PRDX2 gene is transcriptionally silenced and de novo methylated in Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma Markus Schneider, Marcin Szaumkessel, Julia Richter, Ole Ammerpohl, Martin-Leo Hansmann, Ralf Küppers, Reiner Siebert and Maciej Giefing
Updated information and services can be found at: http://www.bloodjournal.org/content/123/23/3672.full.html Articles on similar topics can be found in the following Blood collections Lymphoid Neoplasia (2370 articles) Information about reproducing this article in parts or in its entirety may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests Information about ordering reprints may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#reprints Information about subscriptions and ASH membership may be found online at: http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American Society of Hematology, 2021 L St, NW, Suite 900, Washington DC 20036. Copyright 2011 by The American Society of Hematology; all rights reserved.
