Cancer Immunology, Immunotherapy

, Volume 55, Issue 10, pp 1247–1257 | Cite as

The immunologically active site of prothymosin α is located at the carboxy-terminus of the polypeptide. Evaluation of its in vitro effects in cancer patients

  • Margarita Skopeliti
  • Ioannis F. Voutsas
  • Persefoni Klimentzou
  • Marinos L. Tsiatas
  • Alexander Beck
  • Aristotelis Bamias
  • Maria Moraki
  • Evangelia Livaniou
  • Monica Neagu
  • Wolfgang Voelter
  • Ourania E. Tsitsilonis
Original Article

Abstract

Prothymosin α (proTα) is a 109 amino acid long polypeptide presenting distinct immunoenhancing activity in vitro and in vivo. Recent reports suggest that in apoptotic cells, proTα is cleaved by caspases at its carboxy(C)-terminus generating potentially bioactive fragments. In this study, we identified the peptide segment of proTα presenting maximum immunomodulatory activity. Calf thymus proTα was trypsinised, and the five fragments produced (spanning residues 1–14, 21–30, 31–87, 89–102 and 103–109) were tested for their ability to stimulate healthy donor- and cancer patient-derived peripheral blood mononuclear cell (PBMC) proliferation in autologous mixed lymphocyte reaction (AMLR), natural killer and lymphokine-activated killer cell activity, intracellular production of perforin, upregulation of adhesion molecules and CD25 expression. ProTα(89–102) and proTα(103–109) significantly fortified healthy donor-lymphocytes’ immune responses to levels comparable to those induced by intact proTα. These effects were more pronounced in cancer patients, where peptides proTα(89–102) and proTα(103–109) partly, however significantly, restored the depressed AMLR and cytolytic ability of PBMC, by simulating the biological activity exerted by intact proTα. ProTα(1–14), proTα(21–30) and proTα(31–87) marginally upregulated lymphocyte activation. This is the first report showing that proTα’s immunomodulating activity can be substituted by its C-terminal peptide(s). Whether generation and externalization of such immunoactive proTα fragments occurs in vivo, needs further investigation. However, if these peptides can trigger immune responses, they may eventually be used therapeutically to improve some PBMC functions of cancer patients.

Keywords

Prothymosin α fragmentation Immune responses Cell proliferation Cytotoxicity Anticancer activity 

Abbreviations

AMLR

Autologous mixed lymphocyte reaction

BRM

Biologic response modifiers

IFN

Interferon

IL-2

Interleukin-2

IL-2R

Interleukin-2 receptor

LAK cells

Lymphokine-activated killer cells

NK cells

Natural killer cells

NLS

Nuclear localization signal

PBMC

Peripheral blood mononuclear cells

PMN

Polymorphonuclear cells

proTα

Prothymosin α

Τα1

Thymosin α1

Notes

Acknowledgements

Supported in part by Grants 7285/02 from the GSRT, Greece (to OET and MN) and IKYDA 61/2003 from the Hellenic State Scholarships Foundation and the Deutscher Akademischer Austausch Dienst (to OET and WV). We thank Dr. N. Cacoullos for critically reviewing the manuscript and Dr. C. N. Baxevanis for help with FACS analysis. OET and WV wish to dedicate this report to the memory of Athanassios (Nassos) A. Haritos on his eleventh death anniversary.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Margarita Skopeliti
    • 1
  • Ioannis F. Voutsas
    • 1
  • Persefoni Klimentzou
    • 2
  • Marinos L. Tsiatas
    • 3
  • Alexander Beck
    • 4
  • Aristotelis Bamias
    • 3
  • Maria Moraki
    • 5
  • Evangelia Livaniou
    • 2
  • Monica Neagu
    • 6
  • Wolfgang Voelter
    • 7
  • Ourania E. Tsitsilonis
    • 1
  1. 1.Department of Animal and Human Physiology, Faculty of BiologyUniversity of AthensAthensGreece
  2. 2.Institute of RRPNCSR “Demokritos”AthensGreece
  3. 3.Department of Clinical Therapeutics, “Alexandra” Hospital, School of MedicineUniversity of AthensAthensGreece
  4. 4.Central Laboratory, Department of Internal Medicine IVUniversity Clinic of TuebingenTuebingenGermany
  5. 5.Blood Transfusion Unit“St. Savas” Cancer HospitalAthensGreece
  6. 6.Immunology LaboratoryVictor Babes National Research InstituteBucharestRomania
  7. 7.Biochemistry InstituteUniversity of TuebingenTuebingenGermany

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