Human Cell

, Volume 32, Issue 2, pp 185–192 | Cite as

Diversity of cell phenotypes among MT-2 cell lines affects the growth of U937 cells and cytokine production

  • Hajime Nomura
  • Kunihiko UmekitaEmail author
  • Yuuki Hashikura
  • Kazumi Umeki
  • Ikuo Yamamoto
  • Yatsuki Aratake
  • Mineki Saito
  • Hiroo Hasegawa
  • Katsunori Yanagihara
  • Akihiko Okayama
Research Article


We previously reported the diversity of structure and integration sites of human T-cell leukemia virus type 1 (HTLV-1) provirus among different MT-2 cell lines. This raised the question as to whether cell phenotypes also differed among MT-2 cell lines. The influence of two different MT-2 cell lines (MT-2J and MT-2B) on the growth of the promonocytic leukemia cell line, U937, was investigated. Protein levels and mRNA expression of cytokines were also investigated. In addition, Western blot analysis of HTLV-1 regulatory proteins, Tax and HBZ, was also performed. Culture supernatant from MT-2B, but not MT-2J, cells showed marked suppressive effects on U937 cell growth. MT-2B showed high tumor necrosis factor (TNF)-α, TNF-β, and interferon (IFN)-γ both in protein levels of the culture supernatant and mRNA levels of the cells. Analysis using recombinant cytokines indicated that the suppressive effects of MT-2B were due, at least in part, to high levels of TNF-β and its synergic effects with IFN-γ in the culture supernatant. Protein levels of HTLV-1 Tax and HBZ were higher in MT-2B than those in MT-2J cells. These molecules have been reported to affect the cytokine production of HTLV-1 infected cells; therefore, the difference in these molecules may have accounted for the differences in cytokine production between MT-2J and MT-2B cells. Furthermore, because MT-2 cells showed a large variation of integrated HTLV-1 proviruses as well as cell phenotypes, it is important to exercise caution in the assessment and interpretation of experimental data from MT-2 cells.


HTLV-1 Cytokine production MT-2 cell lines Tumor necrosis factor Interferon gamma 



The authors would like to thank Ms Y. Kaseda (Miyazaki University) for her technical support and Dr Y. Ohkuwa (Miyazaki University) for his instruction concerning the statistical analysis. The authors would like to thank Enago ( for the English language review.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Not applicable.

Supplementary material

13577_2018_231_MOESM1_ESM.doc (92 kb)
Supplementary material 1 (DOC 92 KB)


  1. 1.
    Richardson JH, Höllsberg P, Windhagen A, et al. Variable immortalizing potential and frequent virus latency in blood- derived T-cell clones infected with human T-cell leukemia virus type I. Blood. 1997;89:3303–14.Google Scholar
  2. 2.
    Hinuma Y, Nagata K, Hanaoka M, et al. Adult T-cell leukemia: antigen in an ATL cell line and detection of antibodies to the antigen in human sera. Proc Natl Acad Sci USA. 1981;78:6476–80.CrossRefGoogle Scholar
  3. 3.
    Furukawa Y, Nakamura H, Sakamoto S, Miura Y. Cytomegalovirus gastritis as an initial manifestation of a patient with adult T-cell leukemia. Acta Haematol. 1988;80:216–8.CrossRefGoogle Scholar
  4. 4.
    Shahnaz S, Reich D, Arévalo-Valencia D, et al. HTLV-1-associated adult T cell leukemia lymphoma presenting as granulomatous Pneumocystis Jiroveci Pneumonia (PJP) and hypercalcemia. J Gen Intern Med. 2007;22:420–3.CrossRefGoogle Scholar
  5. 5.
    Satoh M, Toma H, Sato Y, et al. Reduced efficacy of treatment of strongyloidiasis in HTLV-I carriers related to enhanced expression of IFN-γ and TGF-β1. Clin Exp Immunol. 2002;127:354–9.CrossRefGoogle Scholar
  6. 6.
    Olière S, Douville R, Sze A, Belgnaoui SM, Hiscott J. Modulation of innate immune responses during human T-cell leukemia virus (HTLV-1) pathogenesis. Cytokine Growth Factor Rev. 2011;22:197–210.CrossRefGoogle Scholar
  7. 7.
    Datta A, Sinha-Datta U, Dhillon NK, Buch S, Nicot C. The HTLV-I p30 interferes with TLR4 signaling and modulates the release of pro- and anti-inflammatory cytokines from human macrophages. J Biol Chem. 2006;281:23414–24.CrossRefGoogle Scholar
  8. 8.
    Yasunaga JI, Sakai T, Nosaka K, et al. Impaired production of naive T lymphocytes in human T-cell leukemia virus type I-infected individuals: its implications in the immunodeficient state. Blood. 2001;97:3177–83.CrossRefGoogle Scholar
  9. 9.
    Sasaki H, Nishikata I, Shiraga T, et al. Overexpression of a cell adhesion molecule, TSLC1, as a possible molecular marker for acute-type adult T-cell leukemia. Blood. 2005;105:1204–13.CrossRefGoogle Scholar
  10. 10.
    Miyoshi I, Kubonishi I, Yoshimoto S, Shiraishi Y. A T-cell line derived from normal human cord leukocytes by co-culturing with human leukemic T-cells. Gan. 1981;72:978–81.Google Scholar
  11. 11.
    Miyoshi I, Kubonishi I, Yoshimoto S, et al. Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. Nature. 1981;294:770–1.CrossRefGoogle Scholar
  12. 12.
    Yamamoto N, Okada M, Koyanagi Y, Kannagi M, Hinuma Y. Transformation of human leukocytes by cocultivation with an adult T cell leukemia virus producer cell line. Science. 1982;217:737–9.CrossRefGoogle Scholar
  13. 13.
    Higashi T, Tsukada J, Yoshida Y, et al. Constitutive tyrosine and serine phosphorylation of STAT4 in T-cells transformed with HTLV-I. Genes Cells. 2005;10:1153–62.CrossRefGoogle Scholar
  14. 14.
    Yu F, Itoyama Y, Kira J, et al. TNF-beta produced by human T lymphotropic virus type I-infected cells influences the proliferation of human endothelial cells and fibroblasts. J Immunol. 1994;152:5930–8.Google Scholar
  15. 15.
    Hashikura Y, Umeki K, Umekita K, et al. The diversity of the structure and genomic integration sites of HTLV-1 provirus in MT-2 cell lines. Hum Cell. 2016;29:122–9.CrossRefGoogle Scholar
  16. 16.
    Harris P, Ralph P. Human leukemic models of myelomonocytic development: a review of the HL-60 and U937 cell lines. J Leukoc Biol. 1985;37:407–22.CrossRefGoogle Scholar
  17. 17.
    Onoda A, Takeda K, Umezawa M. Dysregulation of major functional genes in frontal cortex by maternal exposure to carbon black nanoparticle is not ameliorated by ascorbic acid pretreatment. Sci Total Environ. 2018;634:1126–35.CrossRefGoogle Scholar
  18. 18.
    Brazma A, Hingamp P, Quackenbush J, et al. Minimum information about a microarray experiment (MIAME)-toward standards for microarray data. Nat Genet. 2001;29:365–71.CrossRefGoogle Scholar
  19. 19.
    Giam CZ, Semmes OJ. HTLV-1 infection and adult T-cell leukemia/lymphoma-A tale of two proteins: Tax and HBZ. Viruses. 2016;8:161.CrossRefGoogle Scholar
  20. 20.
    Shiohama Y, Naito T, Matsuzaki T, et al. Absolute quantification of HTLV-1 basic leucine zipper factor (HBZ) protein and its plasma antibody in HTLV-1 infected individuals with different clinical status. Retrovirology. 2016;13:1–14.CrossRefGoogle Scholar
  21. 21.
    Takeuchi K, Kobayashi N, Hyun Nam S, Yamamoto N, Hatanaka M. Molecular cloning of cDNA encoding gp68 of adult T-cell leukaemia-associated antigen: evidence for expression of the pX IV region of human T-cell leukaemia virus. J Gen Virol. 1985;66:1825–9.CrossRefGoogle Scholar
  22. 22.
    Faulkner L, Patel M, Brickell PM, Katz DR. Regulation of c-fgr messenger RNA levels in U937 cells treated with different modulating agents. Immunology. 1992;76:65–71.Google Scholar
  23. 23.
    Joshi CV, Supriya P, Ajitkumar P. Growth inhibition of human promonocytic leukaemic U937 cells by interferon gamma is irreversible and not cell cycle phase-specific. Cytokine. 1999;11:673–8.CrossRefGoogle Scholar
  24. 24.
    Lee SH, Aggarwal BB, Rinderknecht E, Assisi F, Chiu H. The synergistic anti-proliferative effect of gamma-interferon and human lymphotoxin. J Immunol. 1984;133:1083–6.Google Scholar
  25. 25.
    Bakmiwewa SM, Weiser S, Grey M, et al. Synergistic induction of CXCL10 by interferon-gamma and lymphotoxin-alpha in astrocytes: possible role in cerebral malaria. Cytokine. 2016;78:79–86.CrossRefGoogle Scholar
  26. 26.
    Tschachler E, Bohnlein E, Felzmann S, Reitz MS. Human T-lymphotropic virus type I tax regulates the expression of the human lymphotoxin gene. Blood. 1993;81:95–100.Google Scholar
  27. 27.
    Sun SC, Ballard DW. Persistent activation of NF-kappa B by the Tax transforming protein of HTLV-1: hijacking cellular I kappa B kinases. Oncogene. 1999;18:6948–58.CrossRefGoogle Scholar
  28. 28.
    Hanon E, Goon P, Taylor GP, et al. High production of interferon gamma but not interleukin-2 by human T-lymphotropic virus type I-infected peripheral blood mononuclear cells. Blood. 2001;98:721–6.CrossRefGoogle Scholar
  29. 29.
    Ohsugi T, Kumasaka T. Low CD4/CD8 T-cell ratio associated with inflammatory arthropathy in human T-cell leukemia virus type I Tax transgenic mice. PLoS One. 2011;6:e18518.CrossRefGoogle Scholar
  30. 30.
    Paul NL, Lenardo MJ, Novak KD, et al. Lymphotoxin activation by human T-cell leukemia virus type I-infected cell lines: role for NF-KB. J Virol. 1990;64:5412–9.Google Scholar
  31. 31.
    Mitagami Y, Yasunaga J, Kinosada H, Ohshima K, Matsuoka M. Interferon-γ promotes inflammation and development of T-cell lymphoma in HTLV-1 bZIP factor transgenic mice. PLoS Pathog. 2015;11:e1005120.CrossRefGoogle Scholar
  32. 32.
    Etoh KI, Tamiya S, Yamaguchi K, et al. Persistent clonal proliferation of human T-lymphotropic virus type I-infected cells in vivo. Cancer Res. 1997;57:4862–7.Google Scholar
  33. 33.
    Araya N, Sato T, Ando H, et al. HTLV-1 induces a Th1-like state in CD4+ CCR4+ T cells. J Clin Investig. 2014;124:3431–42.CrossRefGoogle Scholar
  34. 34.
    Renu BL, Donna LR. Constitutive production of interleukin-6 and tumor necrosis factor-alpha from spontaneously proliferating T cells in patients with human T-cell lymphotropic virus type-I/II. Blood. 1991;3:571–4.Google Scholar

Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Hajime Nomura
    • 1
  • Kunihiko Umekita
    • 1
    Email author
  • Yuuki Hashikura
    • 1
  • Kazumi Umeki
    • 1
  • Ikuo Yamamoto
    • 1
  • Yatsuki Aratake
    • 2
  • Mineki Saito
    • 3
  • Hiroo Hasegawa
    • 4
  • Katsunori Yanagihara
    • 4
  • Akihiko Okayama
    • 1
  1. 1.Department of Rheumatology, Infectious Diseases and Laboratory Medicine, Faculty of MedicineUniversity of MiyazakiKiyotakeJapan
  2. 2.Department of Life MedicalKyushu University of Health and WelfareNobeokaJapan
  3. 3.Department of MicrobiologyKawasaki Medical SchoolKurashikiJapan
  4. 4.Department of Laboratory MedicineNagasaki University Graduate School of Biomedical SciencesNagasakiJapan

Personalised recommendations