CTLA-4 in mesothelioma patients: tissue expression, body fluid levels and possible relevance as a prognostic factor

Cancer Immunology, Immunotherapy volume 65, pages909–917(2016)Cite this article

Abstract

CTLA-4 function as a negative regulator of T cell-mediated immune response is well established, whereas much less is known about the immunoregulatory role of its soluble isoform (sCTLA-4). No data are available on CTLA-4 expression and prognostic impact in malignant pleural mesothelioma (MPM). We investigated, by immunohistochemistry, CTLA-4 expression in tumor tissues and, by ELISA, sCTLA-4 levels in sera and matched pleural effusions from 45 MPM patients. Prognostic effect of CTLA-4 expression on overall survival (OS) was assessed through Cox regression and prognostic significance expressed as death rate ratio (HR). We found that 56.0 % of MPM tissues expressed CTLA-4 with variable intensity and percentage of positive cells estimated by the immunoreactive score. sCTLA-4 levels were significantly higher in sera (S-sCTLA-4) than in pleural effusions (PE-sCTLA-4) (geometric mean ratio = 2.70, P value = 0.020). CTLA-4 expression at the tissue level was higher in the epithelioid histological subtype than in the sarcomatoid, whereas at the serum level, it was higher in the sarcomatoid subtype. A homogeneous favorable prognostic effect was found for CTLA-4 overexpression in tissue, serum and pleural effusion. Interestingly, only the PE-sCTLA-4 was found to be a statistically significant positive prognostic factor (HR = 0.37, 95 % CI = 0.18–0.77, P value = 0.007). Indeed, PE-sCTLA-4 correlated with CTLA-4 expression in tissues, whereas this latter expression showed a weak association with OS. To confirm our findings, further experimental evidences obtained from a larger cohort of MPM patients are required. However, our results would indicate a positive correlation of PE-sCTLA-4 levels and OS in MPM patients.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

US$ 39.95

Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3

Abbreviations

APC:

Antigen-presenting cells

BNG:

Pulmonary benign disease

CI:

Confidence intervals

CTLA-4:

Cytotoxic T lymphocyte antigen-4

FFPE:

Formalin-fixed paraffin embedded

GM:

Geometric mean

HR:

Hazard ratio

IHC:

Immunohistochemistry

IRS:

Immunoreactive score

mCTLA-4:

Membrane CTLA-4

MPM:

Malignant pleural mesothelioma

OS:

Overall survival

PE:

Pleural effusion

PE-sCTLA-4:

Soluble CTLA-4 in PE

sCTLA-4:

Soluble CTLA-4

S-sCTLA-4:

Soluble CTLA-4 in serum

Teffs:

Effector T cells

Tregs:

Regulatory T cells

References

  1. 1.

    Robinson BW, Lake RA (2005) Advances in malignant mesothelioma. N Engl J Med 353:1591–1603

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Porpodis K, Zarogoulidis P, Boutsikou E, Papaioannou A, Machairiotis N, Tsakiridis K, Katsikogiannis N, Zaric B, Perin B, Huang H et al (2013) Malignant pleural mesothelioma: current and future perspectives. J Thorac Dis 5(Suppl 4):S397–S406

    PubMed  PubMed Central  Google Scholar 

  3. 3.

    Ismail-Khan R, Robinson LA, Williams CC Jr, Garrett CR, Bepler G, Simon GR (2006) Malignant pleural mesothelioma: a comprehensive review. Cancer Control 13:255–263

    PubMed  Google Scholar 

  4. 4.

    Remon J, Reguart N, Corral J, Lianes P (2015) Malignant pleural mesothelioma: new hope in the horizon with novel therapeutic strategies. Cancer Treat Rev 41:27–34

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Maker AV, Attia P, Rosenberg SA (2005) Analysis of the cellular mechanism of antitumor responses and autoimmunity in patients treated with CTLA-4 blockade. J Immunol 175:7746–7754

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Calabrò L, Morra A, Fonsatti E, Cutaia O, Amato G, Giannarelli D, Di Giacomo AM, Danielli R, Altomonte M, Mutti L, Maio M (2013) Tremelimumab for patients with chemotherapy-resistant advanced malignant mesothelioma: an open-label, single-arm, phase 2 trial. Lancet Oncol 14:1104–1111

    Article  PubMed  Google Scholar 

  7. 7.

    Calabrò L, Morra A, Fonsatti E, Cutaia O, Fazio C, Annesi D, Lenoci M, Amato G, Danielli R, Altomonte M et al (2015) Efficacy and safety of an intensified schedule of tremelimumab for chemotherapy-resistant malignant mesothelioma: an open-label, single-arm, phase 2 study. Lancet Respir Med 3:301–309

    Article  PubMed  Google Scholar 

  8. 8.

    Walker LS, Sansom DM (2011) The emerging role of CTLA-4 as a cell-extrinsic regulator of T cell responses. Nat Rev Immunol 11:852–863

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Krummel MF, Allison JP (1996) CTLA-4 engagement inhibits IL-2 accumulation and cell cycle progression upon activation of resting T cells. J Exp Med 183:2533–2540

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Brunner MC, Chambers CA, Chan FK, Hanke J, Winoto A, Allison JP (1999) CTLA-4-mediated inhibition of early events of T cell proliferation. J Immunol 162:5813–5820

    CAS  PubMed  Google Scholar 

  11. 11.

    Engelhardt JJ, Sullivan TJ, Allison JP (2006) CTLA-4 overexpression inhibits T cell responses through a CD28-B7-dependent mechanism. J Immunol 177:1052–1061

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Zheng Y, Manzotti CN, Burke F, Dussably L, Qureshi O, Walker LS, Sansom DM (2008) Acquisition of suppressive function by activated human CD4 + CD25- T cells is associated with the expression of CTLA-4 not FoxP3. J Immunol 181:1683–1691

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Oaks MK, Hallett KM (2000) Cutting edge: a soluble form of CTLA-4 in patients with autoimmune thyroid disease. J Immunol 164:5015–5018

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Ward FJ, Dahal LN, Wijesekera SK, Abdul-Jawad SK, Kaewarpai T, Xu H, Vickers MA, Barker RN (2013) The soluble isoform of CTLA-4 as a regulator of T-cell responses. Eur J Immunol 43:1274–1285

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Saverino D, Brizzolara R, Simone R, Chiappori A, Milintenda-Floriani F, Pesce G, Bagnasco M (2007) Soluble CTLA-4 in autoimmune thyroid diseases: relationship with clinical status and possible role in the immune response dysregulation. Clin Immunol 123:190–198

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Simone R, Brizzolara R, Chiappori A, Milintenda-Floriani F, Natale C, Greco L, Schiavo M, Bagnasco M, Pesce G, Saverino D (2009) A functional soluble form of CTLA-4 is present in the serum of celiac patients and correlates with mucosal injury. Int Immunol 21:1037–1045

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Ueda H, Howson JM, Esposito L, Heward J, Snook H, Chamberlain G, Rainbow DB, Hunter KM, Smith AN, Di Genova G et al (2003) Association of the T-cell regulatory gene CTLA-4 with susceptibility to autoimmune disease. Nature 423:506–511

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Simone R, Tenca C, Fais F, Luciani M, De Rossi G, Pesce G, Bagnasco M, Saverino D (2012) A soluble form of CTLA-4 is present in paediatric patients with acute lymphoblastic leukaemia and correlates with CD1d + expression. PLoS One 7(9):e44654

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Erfani N, Razmkhah M, Ghaderi A (2010) Circulating soluble CTLA-4 (sCTLA-4) is elevated in patients with breast cancer. Cancer Invest 28:828–832

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Contardi E, Palmisano GL, Tazzari PL, Martelli AM, Falà F, Fabbi M, Kato T, Lucarelli E, Donati D, Polito L et al (2005) CTLA-4 is constitutively expressed on tumor cells and can trigger apoptosis upon ligand interaction. Int J Cancer 117:538–550

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Laurent S, Queirolo P, Boero S, Salvi S, Piccioli P, Boccardo S, Minghelli S, Morabito A, Fontana V, Pietra G et al (2013) The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production. J Transl Med 11:108

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Salvi S, Fontana V, Boccardo S, Merlo DF, Margallo E, Laurent S, Morabito A, Rijavec E, Dal Bello MG, Mora M, Ratto GB, Grossi F, Truini M, Pistillo MP (2012) Evaluation of CTLA-4 expression and relevance as a novel prognostic factor in patients with non-small cell lung cancer. Cancer Immunol Immunother 61:1463–7214

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Shah KV, Chien AJ, Yee C, Moon RT (2008) CTLA-4 is a direct target of Wnt/beta-catenin signalling and is expressed in human melanoma tumors. J Invest Dermatol 128:2870–2879

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Mao H, Zhang L, Yang Y, Zuo W, Bi Y, Gao W, Deng B, Sun J, Shao Q, Qu X (2010) New insights of CTLA-4 into its biological function in breast cancer. Curr Cancer Drug Targets 10:728–736

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Antczak A, Pastuszak-Lewandoska D, Górski P, Domańska D, Migdalska-Sęk M, Czarnecka K, Nawrot E, Kordiak J, Brzeziańska E (2013) CTLA-4 expression and polymorphisms in lung tissue of patients with diagnosed non-small-cell lung cancer. Biomed Res Int 2013:576486

    Article  PubMed  PubMed Central  Google Scholar 

  26. 26.

    Marubini E, Valsecchi MG (2004) Analysing survival data from clinical trials and observational studies. Wiley & Sons, New York

    Google Scholar 

  27. 27.

    Liang H, Zou G (2008) Improved AIC selection strategy for survival analysis. Comput Stat Data Anal 52:2538–2548

    Article  PubMed  PubMed Central  Google Scholar 

  28. 28.

    DiCiccio TJ, Efron B (1996) Bootstrap confidence intervals. Stat Sci 11:189–228

    Article  Google Scholar 

  29. 29.

    Leung HT, Bradshaw J, Cleaveland JS, Linsley PS (1995) Cytotoxic T lymphocyte-associated molecule-4, a high-avidity receptor for CD80 and CD86, contains an intracellular localization motif in its cytoplasmic tail. J Biol Chem 270:25107–25114

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Tai X, Van Laethem F, Pobezinsky L, Guinter T, Sharrow SO, Adams A, Granger L, Kruhlak M, Lindsten T, Thompson CB et al (2012) Basis of CTLA-4 function in regulatory and conventional CD4(+) T cells. Blood 119:5155–5163

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Valdés L, San-José E, Estévez JC, González-Barcala FJ, Alvarez-Dobaño JM, Golpe A, Valle JM, Penela P, Vizcaíno L, Pose A (2010) Cholesterol in pleural exudates depends mainly on increased capillary permeability. Transl Res 155:178–184

    Article  PubMed  Google Scholar 

  32. 32.

    Chen YQ, Shi HZ, Qin XJ, Mo WN, Liang XD, Huang ZX, Yang HB, Wu C (2005) CD4 + CD25 + regulatory T lymphocytes in malignant pleural effusion. Am J Respir Crit Care Med 172:1434–1439

    Article  PubMed  Google Scholar 

  33. 33.

    Esposito L, Hunter KM, Clark J, Rainbow DB, Stevens H, Denesha J, Duley S, Dawson S, Coleman G, Nutland S et al (2014) Investigation of soluble and transmembrane CTLA-4 isoforms in serum and microvesicles. J Immunol 193:889–900

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  34. 34.

    Cao J, Zhang L, Huang S, Chen P, Zou L, Chen H, Xiang Y, Lai X, Ren G (2011) Aberrant production of soluble co-stimulatory molecules CTLA-4 and CD28 in patients with chronic hepatitis B. Microb Pathog 51:262–267

    CAS  Article  PubMed  Google Scholar 

  35. 35.

    Steiner K, Moosig F, Csernok E, Selleng K, Gross WL, Fleischer B, Bröker BM (2001) Increased expression of CTLA-4 (CD152) by T and B lymphocytes in Wegener’s granulomatosis. Clin Exp Immunol 126:143–150

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Shen Y, Yang T, Wang T, Chen L, Wen F (2013) Elevated circulating cytotoxic T lymphocyte-associated antigen-4 levels are related to lung function and inflammation in chronic obstructive pulmonary disease. Respirology 18:885–886

    Article  PubMed  Google Scholar 

  37. 37.

    Musk AW, Olsen N, Alfonso H, Reid A, Mina R, Franklin P, Sleith J, Hammond N, Threlfall T, Shilkin KB, de Klerk NH (2011) Predicting survival in malignant mesothelioma. Eur Respir J 38:1420–1424

    CAS  Article  PubMed  Google Scholar 

  38. 38.

    Guazzelli A, Hussain M, Krstic-Demonacos M, Mutti L (2015) Tremelimumab for the treatment of malignant mesothelioma. Expert Opin Biol Ther 15:1819–1829

    CAS  Article  PubMed  Google Scholar 

  39. 39.

    Ward FJ, Dahal LN, Khanolkar RC, Shankar SP, Barker RN (2014) Targeting the alternatively spliced soluble isoform of CTLA-4: prospects for immunotherapy? Immunotherapy 6:1073–1084

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgments

Supported in part by Grants from the Italian Ministry of Health (5 × 1000 funds 2011), AIL Sezione La Spezia and Fondazione Umberto Veronesi (Grant 2013), Milan, Italy. The authors would like to thank Dr. Roberto Benelli for image analysis and graphical support.

Author information

Author notes

    Affiliations

    1. Division of Histopathology and Cytopathology, ASL5, La Spezia, Italy

      Silvio Roncella, Paola Ferro, Maria Cristiana Franceschini, Alessandro Valentino & Franco Fedeli

    2. Department of Internal Medicine (DIMI), University of Genoa, Genoa, Italy

      Stefania Laurent

    3. Unit of Clinical Epidemiology, IRCCS AOU San Martino-IST, Genoa, Italy

      Vincenzo Fontana, Domenico Franco Merlo & Marcello Ceppi

    4. Division of Histopathology and Cytopathology, IRCCS AOU San Martino-IST, Genoa, Italy

      Sandra Salvi, Serena Varesano & Simona Boccardo

    5. Unit of Clinical Oncology, ASL5, La Spezia, Italy

      Antonella Vigani

    6. Unit of Tumor Epigenetics, IRCCS AOU San Martino-IST, Genoa, Italy

      Anna Morabito, Massimo Romani & Maria Pia Pistillo

    7. Division of Pneumology, ASL5, La Spezia, Italy

      Pier Aldo Canessa

    8. Unit of Radiodiagnostic, ASL5, La Spezia, Italy

      Ugo Giannoni & Ilan Rosenberg

    9. Blood Transfusion Centre, IRCCS AOU-San Martino-IST, Genoa, Italy

      Salvatore Riggio

    10. Department of Experimental Medicine, University of Genoa, Genoa, Italy

      Daniele Saverino

    11. Unit of Molecular Oncology and Angiogenesis, IRCCS AOU San Martino-IST, Genoa, Italy

      Alessandro Poggi

    Authors
    1. Silvio Roncella
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Stefania Laurent
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Vincenzo Fontana
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Paola Ferro
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Maria Cristiana Franceschini
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Sandra Salvi
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Serena Varesano
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Simona Boccardo
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Antonella Vigani
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Anna Morabito
      View author publications

      You can also search for this author in PubMed Google Scholar

    11. Pier Aldo Canessa
      View author publications

      You can also search for this author in PubMed Google Scholar

    12. Ugo Giannoni
      View author publications

      You can also search for this author in PubMed Google Scholar

    13. Ilan Rosenberg
      View author publications

      You can also search for this author in PubMed Google Scholar

    14. Alessandro Valentino
      View author publications

      You can also search for this author in PubMed Google Scholar

    15. Franco Fedeli
      View author publications

      You can also search for this author in PubMed Google Scholar

    16. Domenico Franco Merlo
      View author publications

      You can also search for this author in PubMed Google Scholar

    17. Marcello Ceppi
      View author publications

      You can also search for this author in PubMed Google Scholar

    18. Salvatore Riggio
      View author publications

      You can also search for this author in PubMed Google Scholar

    19. Massimo Romani
      View author publications

      You can also search for this author in PubMed Google Scholar

    20. Daniele Saverino
      View author publications

      You can also search for this author in PubMed Google Scholar

    21. Alessandro Poggi
      View author publications

      You can also search for this author in PubMed Google Scholar

    22. Maria Pia Pistillo
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Maria Pia Pistillo.

    Ethics declarations

    Conflict of interest

    The authors declare that they have no conflict of interest.

    Additional information

    S. Roncella and S. Laurent contributed equally to this work.

    Rights and permissions

    Reprints and Permissions

    About this article

    Verify currency and authenticity via CrossMark

    Cite this article

    Roncella, S., Laurent, S., Fontana, V. et al. CTLA-4 in mesothelioma patients: tissue expression, body fluid levels and possible relevance as a prognostic factor. Cancer Immunol Immunother 65, 909–917 (2016). https://doi.org/10.1007/s00262-016-1844-3

    Download citation

    Keywords

    • CTLA-4
    • Mesothelioma
    • Pleural effusion
    • Prognostic factor
    • Overall survival