Features of peripheral blood lymphocytes and tumor infiltrating lymphocytes subpopulations in patients with squamous cell head and neck cancer

• Tatiana N. Zabotina
• Antonina I. Chertkova
• Tsiklauri V.T.
• Korotkova O.V.
• Elena N. Zakharova
• Dmitry V. Tabakov
• Anna A. Borunova
• Panchuk I.O.
• Zaderenko I.A.
• Ali M. Mudunov
• Zaira G. Kadagidze

Abstract

The immune system plays an important role in the development, formation and progression of squamous cell carcinoma of the head and neck. Therefore, the study of the baseline state of systemic and local immunity and of the link of lymphocyte subpopulation composition with prevalence of the process and survival of patients is relevant.

Material and methods. Immunophenotyping of peripheral blood lymphocytes (PB) and lymphocytes infiltrating the tumor (TILs) by flow cytometry was carried out in 29 primary operable patients suffering squamous cell carcinoma of the head and neck prior to surgical treatment.

Results. In patients with early stages of disease there was an increase in the percentage of NK-cells, CD16⁺Perforin⁺- and CD11b⁺-lymphocytes, as well as cytotoxic potential of CD8⁺-cells. The number of activated CD25⁺-lymphocytes was increased in patients with stages III-IV and the ratio of the Teff/Treg was decreased versus group of comparison. The percentage of regulatory CD4⁺CD25⁺CD127^-/low-T-cells (Treg) was increased and the value of the Teff/Treg ratio was reduced in dead patients in contrast with survivors. Cytotoxic potential of CD8⁺-cells and CD8⁺Perforin^high-lymphocytes was elevated in living patients. The composition of TILs showed a significant increase of suppressor CD4⁺- and CD8⁺-T-cell populations compared with PB. There was also an increase in the percentage of CD8⁺- and CD4⁺-T-cells expressing the inhibitory receptor PD-1 compared with PB, and a high expression PD-L1 and PD-L2 on tumor cells.

Conclusion. Results indicate that simultaneous study of immune cells of peripheral blood and cells infiltrating tumor tissue is necessary.

Keywords:squamous cell head and neck cancer; peripheral blood lymphocytes; tumor infiltrating lymphocytes; effector lymphocytes; regulatory lymphocytes

References

1. Rivera C. Essentials of oral cancer. Int. J. Clin. Exp. Pathol. 2015; 8 (9): 11 884-94. URL: https://www.researchgate.net/publication/283455295_Essentials_Of_Oral_Cancer

2. Standardized indicators of the oncoepidemiological situation in 2016. Evraziyskiy onkologicheskiy zhurnal. 2018; 6 (2): 49-62. (in Russian)

3. Schoenfeld J.D. Immunity in head and neck cancer. Cancer Immunol. Res. 2015; 3 (1): 12-7. doi: 10.1158/2326-6066.CIR-14-0205.

4. Balermpas P., Michel Y., Wagenblast, J., Seitz O., et al. Tumor-infiltrating lymphocytes predict response to definitive chemoradiotherapy in head and neck cancer. Br. J. Cancer 2014; 110: 501-9. doi: 10.1038/bjc.2013.640.

5. Nasman A., Romanitan M., Nordfors C., Grun N., et al. Tumor infiltrating CD8+ and Foxp3+ lymphocytes correlate to clinical outcome and human papillomavirus (HPV) status in tonsillar cancer. PLoS One. 2012; 7: e38711. doi: 10.1371/journal.pone.0038711.

6. Economopoulou P., Agelaki S., Perisanidis C., Giotakis E.I., et al. The promise of immunotherapy in head and neck squamous cell carcinoma. Ann. Oncol. 2016; 27 (9): 1675-85. doi: 10.1093/annonc/mdw226.

7. Ferris R.L., Whiteside T.L., Ferrone S. Immune escape associated with functional defects in antigen-processing machinery in head and neck cancer. Clin. Cancer Res. 2006; 12 (13): 3890-5. doi: 10.1158/1078-0432.CCR-05-2750.

8. Jie H.B., Gildener-Leapman N., Li J., Srivastava R.M., et al. Intratumoral regulatory T cells upregulate immunosuppressive molecules in head and neck cancer patients. Br. J. Cancer. 2013; 109: 2629-35. doi: 10.1038/bjc.2013.645.

9. Fiorentini S., Licenziati S., Alessandri G., Castelli F., et al. CD11b expression identifies CD8+CD28+ T lymphocytes with phenotype and function of both naive/memory and effector cells. J. Immunol. 2001; 166 (2): 900-7.

10. McFarland H.I., Nahill S.R., Maciaszek J.W., Welsh R.M. CD11b (Mac-1): a marker for CD8+ cytotoxic T cell activation and memory in virus infection. J. Immunol. 1992; 149 (4): 1326-33. PMID:1500720.

11. Christensen J.E., Andreasen S.O., Christensen J.P., Thomsen A.R. CD11b expression as a marker to distinguish between recently activated effector CD8(+) T cells and memory cells. Int. Immunol. 2001; 13 (4): 593-600. PMID:11282998.

12. Fu B., Tian Z., Wei H. Subsets of human natural killer cells and their regulatory effects. Immunology. 2014; 141 (4): 483-9. doi: 10.1111/imm.12224.

13. Chikamatsu K., Sakakura K., Whiteside T.L., Furuya N. Relationships between regulatory T cells and CD8+ effector populations in patients with squamous cell carcinoma of the head and neck. Head Neck. 2007; 29 (2): 120-7. doi: 10.1002/hed.20490.

14. Schaefer C., Kim G.G., Albers A., Hoermann K., et al. Characteristics of CD4+CD25+ regulatory T cells in the peripheral circulation of patients with head and neck cancer. Br. J. Cancer. 2005; 92: 913-20. doi: 10.1038/sj.bjc.6602407.

15. Lau K.M., Cheng S.H., Lo K.W., Lee S.A., et al. Increase in circulating Foxp3+CD4+CD25high regulatory T cells in nasopharyngeal carcinoma patients. Br. J. Cancer. 2007; 96: 617-22. doi: 10.1038/sj.bjc.6603580.

16. Strauss L., Bergmann C., Szczepanski M., Gooding W., et al. A unique subset of CD4+CD25highFoxp3+ T cells secreting interleukin-10 and transforming growth factor-beta1 mediates suppression in the tumor microenvironment. Clin. Cancer Res. 2007; 13: 4345-54. doi: 10.1158/1078-0432.CCR-07-0472.

17. Yamada H., Martin P.J., Bean M.A., Braun M.P., et al. Monoclonal antibody 9.3 and anti-CD11 antibodies define reciprocal subsets of lymphocytes. Eur. J. Immunol. 1985, 15: 1164. doi: 10.1002/eji.1830151204.

18. Caruso A., Fiorentini S., Licenziati S., Alessandri G., et al. Expansion of rare CD8+ CD28- CD11b- T cells with impaired effector functions in HIV-1-infected patients. J. Acquir. Immune Defic. Syndr. 2000; 24 (5): 465-74. PMID:11035618.

19. Freedman M.S., Ruijs T.C., Blain M., Antel J.P. Phenotypic and functional characteristics of activated CD8+ cells: a CD11b-CD28-subset mediates noncytolytic functional suppression. Clin. Immunol. Immunopathol. 1991; 60 (2): 254-67.

20. Bron L., Jandus C., Andrejevic-Blant S., Speiser D.E., et al. Prognostic value of arginase-II expression and regulatory T-cell infiltration in head and neck squamous cell carcinoma. Int. J. Cancer. 2013; 132 (3): 85-93. doi: 10.1002/ijc.27728.

21. Tabachnyk M., Distel L.V., Buttner M., Grabenbauer G.G., et al. Radiochemotherapy induces a favourable tumour infiltrating inflammatory cell profile in head and neck cancer. Oral Oncol. 2012; 48 (7): 594-601. doi: 10.1016/j.oraloncology.2012.01.024.

22. Lechner A., Schlofier H., Rothschild S.I., Thelen M., et al. Characterization of tumor-associated T-lymphocyte subsets and immune checkpoint molecules in head and neck squamous cell carcinoma. Oncotarget. 2017; 8 (27): 44 418-33. doi: 10.18632/oncotarget.17901.

23. Jie H.B., Gildener-Leapman N., Li J., Srivastava R.M., et al. Intratumoral regulatory T cells upregulate immunosuppressive molecules in head and neck cancer patients. Br. J. Cancer. 2013; 109: 2629-35. doi: 10.1038/bjc.2013.645.

24. Zabotina T.N., Korotkova O.V., Borunova A.A., Tabakov D.V., et al. Multiparameter investigation of immunophenotype of tumor infiltrating lymphocytes in cancer patients. Rossiyskiy onkologicheskiy zhurnal. 2016; 21 (1-2): 51-4. (in Russian)

25. Chow L.Q.M., Haddad R., Gupta S., Mahipal A., et al. Antitumor activity of pembrolizumab in biomarker-unselected patients with recurrent and/or metastatic head and neck squamous cell carcinoma: results from the phase Ib KEYNOTE-012 expansion cohort. J. Clin. Oncol. 2016; 34 (32): 3838-45. doi: 10.1200/JCO.2016.68.1478.

All articles in our journal are distributed under the Creative Commons Attribution 4.0 International License (CC BY 4.0 license)