Top

Published in:

Open Access 01-12-2015 | Research

Changes in peripheral blood lymphocytes in polycythemia vera and essential thrombocythemia patients treated with pegylated-interferon alpha and correlation with JAK2 ^V617F allelic burden

Authors: Magdalena Kovacsovics-Bankowski, Todd W. Kelley, Olga Efimova, Soo Jin Kim, Andrew Wilson, Sabina Swierczek, Josef Prchal

Published in: Experimental Hematology & Oncology | Issue 1/2015

Abstract

Background

Pegylated-interferon alpha (PegINFα) treatment of patients with polycythemia vera (PV) and essential thrombocythemia (ET) has resulted in long-term clinical response, decreased JAK2 ^V617F allelic burden and restoration of polyclonal hematopoiesis. The mechanisms of the beneficial effects of PegINFα are not clear, but available evidence suggests direct suppression of JAK2-mutated clone, induction of dormant stem cells to proliferation, and augmentation of an immune effect against PV and ET clones.

Methods

We analyzed the phenotype and frequency of peripheral blood lymphocytes (PBL) from PegINFα treated patients and compared them to patients treated with hydroxyurea (HU). Samples collected at various time points before and during treatment were analyzed using multicolor flow cytometry.

Results

We found that PegINFα increased the frequency of peripheral blood CD4⁺ Foxp3⁺ regulatory T cells (Treg). Highly suppressive Treg, characterized by co-expression of CD39 and HLA-DR, were also increased in PBL from PegINFα treated patients. We observed an augmentation of cycling CD8⁺ T cells, NK cells, and of poorly activated CD38⁺CD8⁺ T cells. Our results also suggest that PegINFα increased the frequency of PD-1⁺ CD4⁺ helper cells and PD-1⁺ CD4⁺ Foxp3⁺ Treg cells. None of these changes were present in HU treated patients. We analyzed the correlation between changes in different T cell populations in the peripheral blood with the changes in JAK2 ^V617F allelic burden in clonal granulocytes. Augmentation of Ki-67⁺ Treg, HLA-DR⁺ CD39⁺ Treg, Helios⁺ Treg and HLA-DR⁺ CD38⁺ CD8⁺ T cells correlated with an increase in JAK2 ^V617F allelic burden. We also found a positive correlation between PD-1⁺ Treg and JAK2 ^V617F allelic burden; however, the number of available patients was small (n = 7).

Conclusions

We report marked changes in frequencies of PBL subsets after PegINFα treatment, suggesting an immunomodulatory effect by PegINFα. Generation of a more suppressive immune response, as measured by an increase in highly suppressive Treg and poorly activated CD8⁺ T cells, correlated with a poor molecular response. In this study, we have not identified changes in the PBL that would indicate the presence of an effective anti-tumor response.Trial registration NCT01259856, December 7. 2010 and NCT01259817, December 6. 2010, Grant #1P01CA108671-O1A2, July 17. 2006, Sponsor: MPDRC/NIH, NCI-2012-00269, January 12. 2011 and NCI-2012-00268, January 12. 2011

Available only for authorised users

Baxter EJ, Scott LM, Campbell PJ, East C, Fourouclas N, Swanton S, Vassiliou GS, Bench AJ, Boyd EM, Curtin N, Scott MA, Erber WN, Green AR, Cancer genome project. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet. 2005;365(9464):1054–61.PubMedCrossRef

James C, Ugo V, Le Couédic JP, Staerk J, Delhommeau F, Lacout C, Garçon L, Raslova H, Berger R, Bennaceur-Griscelli A, Villeval JL. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434(7037):1144–8.PubMedCrossRef

Kralovics R, Passamonti F, Buser AS, Teo SS, Tiedt R, Passweg JR, Tichelli A, Cazzola M, Skoda RC. A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med. 2005;352(17):1779–90.PubMedCrossRef

Levine RL, Wadleigh M, Cools J, Ebert BL, Wernig G, Huntly BJ, Boggon TJ, Wlodarska I, Clark JJ, Moore S, Adelsperger J. Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell. 2005;7(4):387–97.PubMedCrossRef

Liu E, Jelinek J, Pastore YD, Guan Y, Prchal JF, Prchal JT. Discrimination of polycythemias and thrombocytoses by novel, simple, accurate clonality assays and comparison with PRV-1 expression and BFU-E response to erythropoietin. Blood. 2003;101(8):3294–301.PubMedCrossRef

Kiladjian JJ, Cassinat B, Chevret S, Turlure P, Cambier N, Roussel M, Bellucci S, Grandchamp B, Chomienne C, Fenaux P. Pegylated interferon-alfa-2a induces complete hematologic and molecular responses with low toxicity in polycythemia vera. Blood. 2008;112(8):3065–72.PubMedCrossRef

Quintás-Cardama A, Kantarjian H, Manshouri T, Luthra R, Estrov Z, Pierce S, Richie MA, Borthakur G, Konopleva M, Cortes J, Verstovsek S. Pegylated interferon alfa-2a yields high rates of hematologic and molecular response in patients with advanced essential thrombocythemia and polycythemia vera. J Clin Oncol. 2009;27(32):5418–24.PubMedPubMedCentralCrossRef

Kiladjian JJ, Mesa RA, Hoffman R. The renaissance of interferon therapy for the treatment of myeloid malignancies. Blood. 2011;117(18):4706–15.PubMedCrossRef

King KY, Matatall KA, Shen CC, Goodell MA, Swierczek SI, Prchal JT. Comparative long-term effects of interferon α and hydroxyurea on human hematopoietic progenitor cells. Exp Hematol. 2015;43(10):912–8.PubMedCrossRef

10.

Drake CG. Combined immune checkpoint blockade. Semin Oncol. 2015;42(4):656–62.PubMedCrossRef

11.

Atkins MB, Sznol M. Cancer immunotherapy: past progress and future directions. Semin Oncol. 2015;42(4):518–22.PubMedCrossRef

12.

Fattovich G, Betterle C, Brollo L, Pedini B, Giustina G, Realdi G, Alberti A, Ruol A. Autoantibodies during α-interferon therapy for chronic hepatitis B. J Med Virol. 1991;34(2):132–5.PubMedCrossRef

13.

Dumoulin FL, Leifeld L, Sauerbruch T, Spengler U. Autoimmunity induced by interferon-α therapy for chronic viral hepatitis. Biomed Pharmacother. 1999;53(5):242–54.PubMedCrossRef

14.

Huber JP, Farrar JD. Regulation of effector and memory T-cell functions by type I interferon. Immunology. 2011;132(4):466–74.PubMedPubMedCentralCrossRef

15.

Golding A, Rosen A, Petri M, Akhter E, Andrade F. Interferon-alpha regulates the dynamic balance between human activated regulatory and effector T cells: implications for antiviral and autoimmune responses. Immunology. 2010;131(1):107–17.PubMedPubMedCentral

16.

Sprengers D, Stoop JN, Binda RS, Kusters JG, Haagmans BL, Carotenuto P, Artsen A, van der Molen RG, Janssen HL. Induction of regulatory T-cells and interleukin-10-producing cells in non-responders to pegylated interferon-alpha therapy for chronic hepatitis B. Antiviral therapy. 2007;12(7):1087.PubMed

17.

Flaherty LE, Othus M, Atkins MB, Tuthill RJ, Thompson JA, Vetto JT, Haluska FG, Pappo AS, Sosman JA, Redman BG, Moon J. Southwest Oncology Group S0008: a phase III trial of high-dose interferon Alfa-2b versus cisplatin, vinblastine, and dacarbazine, plus interleukin-2 and interferon in patients with high-risk melanoma—an intergroup study of cancer and leukemia Group B, Children’s Oncology Group, Eastern Cooperative Oncology Group, and Southwest Oncology Group. J Clin Oncol. 2014;32(33):3771–8.PubMedPubMedCentralCrossRef

18.

Chevolet I, Schreuer M, Speeckaert R, Neyns B, Hoorens I, van Geel N, Krüse V, Hennart B, Allorge D, Van Gele M, Brochez L. Systemic immune changes associated with adjuvant interferon-α2b-therapy in stage III melanoma patients: failure at the effector phase? Melanoma Res. 2015;25(4):357–61.PubMedCrossRef

19.

Borsellino G, Kleinewietfeld M, Di Mitri D, Sternjak A, Diamantini A, Giometto R, Höpner S, Centonze D, Bernardi G, Dell’Acqua ML, Rossini PM. Expression of ectonucleotidase CD39 by Foxp3 + Treg cells: hydrolysis of extracellular ATP and immune suppression. Blood. 2007;110(4):1225–32.PubMedCrossRef

20.

Getnet D, Grosso JF, Goldberg MV, Harris TJ, Yen HR, Bruno TC, Durham NM, Hipkiss EL, Pyle KJ, Wada S, Pan F. A role for the transcription factor Helios in human CD4+, CD25+ regulatory T cells. Mol Immunol. 2010;47(7):1595–600.PubMedPubMedCentralCrossRef

21.

Kisielewicz A, Schaier M, Schmitt E, Hug F, Haensch GM, Meuer S, Zeier M, Sohn C, Steinborn A. A distinct subset of HLA-DR+-regulatory T cells is involved in the induction of preterm labor during pregnancy and in the induction of organ rejection after transplantation. Clin Immunol. 2010;137(2):209–20.PubMedCrossRef

22.

Schober L, Radnai D, Schmitt E, Mahnke K, Sohn C, Steinborn A. Term and preterm labor: decreased suppressive activity and changes in composition of the regulatory T-cell pool. Immunol Cell Biol. 2012;90(10):935–44.PubMedCrossRef

23.

Zabransky DJ, Nirschl CJ, Durham NM, Park BV, Ceccato CM, Bruno TC, Tam AJ, Getnet D, Drake CG. Phenotypic and functional properties of Helios + regulatory T cells. PLoS ONE. 2012;7(3):e34547.PubMedPubMedCentralCrossRef

24.

Croft M. Co-stimulatory members of the TNFR family: keys to effective T-cell immunity? Nat Rev Immunol. 2003;3(8):609–20.PubMedCrossRef

25.

Redmond WL, Ruby CE, Weinberg AD. The role of OX40-mediated co-stimulation in T-cell activation and survival. Crit Rev Immunol. 2009;29(3):187–201.PubMedPubMedCentralCrossRef

26.

Bastidas S, Graw F, Smith MZ, Kuster H, Gunthard HF, Oxenius A. CD8+ T cells are activated in an antigen-independent manner in HIV-infected individuals. J Immunol. 2014;192(4):1732–44.PubMedCrossRef

27.

Kestens L, Vanham G, Gigase P, Young G, Hannet I, Vanlangendonck F, Hulstaert F, Bach BA. Expression of activation antigens, HLA-DR and CD38, on CD8 lymphocytes during HIV-1 infection. AIDS. 1992;6(8):793–7.PubMedCrossRef

28.

Riley CH, Brimnes MK, Hansen M, Jensen MK, Hasselbalch HC, Kjaer L, Svane IM. Interferon-alpha induces marked alterations in circulating regulatory T cells, NK cell subsets and dendritic cells in patients with JAK2 -positive essential thrombocythemia and polycythemia vera. Eur J Haematol. 2015.

29.

Riley CH, Hansen M, Brimnes MK, Hasselbalch HC, Bjerrum OW, Svane IM, Jensen MK. Expansion of circulating CD56bright natural killer cells in patients with JAK2-positive chronic myeloproliferative neoplasms during treatment with interferon-α. Eur J Haematol. 2015;94(3):227–34.PubMedCrossRef

30.

Riley CH, Jensen MK, Brimnes MK, Hasselbalch HC, Bjerrum OW, Straten PT, Svane IM. Increase in circulating CD4(+) CD25(+) Foxp3(+) T cells in patients with Philadelphia-negative chronic myeloproliferative neoplasms during treatment with IFN-alpha. Blood. 2011;118(8):2170–3.PubMedCrossRef

31.

Prchal JT, Throckmorton DW, Carroll AJ, Fuson EW, Gams RA, Prchal JF. A common progenitor for human myeloid and lymphoid cells. Nature. 1978;274(5671):590–1.PubMedCrossRef

32.

Jelinek J, Prchal JT. Oxygen-dependent regulation of erythropoiesis. Methods Enzymol. 2004;381:201–10.PubMedCrossRef

33.

Roederer M. Spectral compensation for flow cytometry: visualization artifacts, limitations, and caveats. Cytometry. 2001;45(3):194–205.PubMedCrossRef

34.

Nussenzveig RH, Swierczek SI, Jelinek J, Gaikwad A, Liu E, Verstovsek S, Prchal JF, Prchal JT. Polycythemia vera is not initiated by JAK2V617F mutation. Exp Hematol. 2007;35(1):32–8.PubMedCrossRef

35.

Tefferi A, Thiele J, Vannucchi AM, Barbui T. An overview on CALR and CSF3R mutations and a proposal for revision of WHO diagnostic criteria for myeloproliferative neoplasms. Leukemia. 2014;28(7):1407–13.PubMedCrossRef

36.

Hervas-Stubbs S, Perez-Gracia JL, Rouzaut A, Sanmamed MF, Le Bon A, Melero I. Direct effects of type I interferons on cells of the immune system. Clin Cancer Res. 2011;17(9):2619–27.PubMedCrossRef

37.

Tough DF. Modulation of T-cell function by type I interferon. Immunol Cell Biol. 2012;90(5):492–7.PubMedCrossRef

38.

Kelley TW, Swierczek S, Efimova O, Wilson AR, Kim SJ, Prchal J. The number of peripheral blood CD4+, CD25+, FOXP3+ regulatory T-cells is a marker of response in patients with JAK2 V617F positive myeloproliferative neoplasms treated with pegylated-interferon-α. ASH abstract, 2013.

39.

Suuu SS, He H, Kong LB, Zhang YG, Zhao SX, Wang RQ, Zheng HW, Sun DX, Nan YM, Yu J. Regulatory phenotype, PD-1 and TLR3 expression in T cells and monocytes from HCV patients undergoing antiviral therapy: a randomized clinical trial. PLoS ONE. 2014;9(4):e93620.CrossRef

40.

Kovacsovics-Bankowski M, Chisholm L, Vercellini J, Tucker CG, Montler R, Haley D, Newell P, Ma J, Tseng P, Wolf R, Vetto JT. Detailed characterization of tumor infiltrating lymphocytes in two distinct human solid malignancies show phenotypic similarities. J Immunother Cancer. 2014;2(1):1.CrossRef

Title: Changes in peripheral blood lymphocytes in polycythemia vera and essential thrombocythemia patients treated with pegylated-interferon alpha and correlation with JAK2 V617F allelic burden
Authors: Magdalena Kovacsovics-Bankowski
Todd W. Kelley
Olga Efimova
Soo Jin Kim
Andrew Wilson
Sabina Swierczek
Josef Prchal
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: Experimental Hematology & Oncology / Issue 1/2015
Electronic ISSN: 2162-3619
DOI: https://doi.org/10.1186/s40164-016-0057-y

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2015

Complete remission after single agent blinatumomab in a patient with pre-B acute lymphoid leukemia relapsed and refractory to three prior regimens: hyperCVAD, high dose cytarabine mitoxantrone and CLAG

Cell therapy must be regulated as medicine

Novel melanoma therapy

A systematic classification of megakaryocytic dysplasia and its impact on prognosis for patients with myelodysplastic syndromes

New CD20 alternative splice variants: molecular identification and differential expression within hematological B cell malignancies

p27KIP1 and PTEN cooperate in myeloproliferative neoplasm tumor suppression in mice

Keynote webinar | Spotlight on antibody–drug conjugates in cancer