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The effect of HNSCC-derived soluble factors on the proliferation and function of immune cells

Smith, Joanne D.


Joanne D. Smith


Victoria L. Green

John (Professor of tumour immunology) Greenman


Malignant epithelium and associated stromal cells secrete soluble factors which may influence tumour evasion of host immunity. The effect of these factors on the proliferation and function of individual immune cell populations has been investigated along with the role of hypoxia.

The conditioned medium (CM) from four HNSCC cell lines, primary-derived fibroblasts, cultured in both normoxic and hypoxic conditions, and overnight-dispersed tumour CM was collected. Cytokine profiles were determined using a Quantibody cytokine array™ and ELISA. The CM was added to Tregulatory cells (CD4⁺CD25⁺CD127ˡᵒ), Teffector cells (CD4⁺CD25⁻) and cytotoxic T cells (CD8⁺) isolated from healthy donors and HNSCC patients. MTS assays and flow cytometry were used to assess changes in proliferation and percentage of immune cells. CFSE suppression assays, ELISA and flow cytometry were undertaken to measure changes in function of Tregulatory cells, Teffectors cells and CD8⁺ T cells obtained from healthy PBMC.

A significant increase in the proliferation of whole PBMC from patient and healthy donors was observed upon the addition all HNSCC-derived CM, with healthy PBMC proliferating to an even greater extent compared with the patient samples. Tregulatory and Teffector cell percentages within healthy PBMC increased while CD8⁺ T cell percentage decreased in many cases. Also, HNSCC-derived CM was able to increase the suppressive activity of Tregs in 40% of samples. The CM caused an increase in Th1 type cytokines IFN-γ and IL-2 in at least 50% of samples with little change to Th2 cytokine levels and was also able to significantly reduce the function of CD8⁺ T cells in at least 50% of samples.

In conclusion, the secretome of HNSCC epithelial cells, primary-derived fibroblasts and overnight dispersed tumour has the ability to alter the proliferation and function of individual sets of immune cells, potentially to a greater extent in the presence of other cell types.


Smith, J. D. (2016). The effect of HNSCC-derived soluble factors on the proliferation and function of immune cells. (Thesis). University of Hull. Retrieved from

Thesis Type Thesis
Deposit Date Jun 20, 2017
Publicly Available Date Feb 23, 2023
Keywords Biological sciences
Public URL
Additional Information Department of Biological, Biomedical and Environmental Sciences, The University of Hull
Award Date Feb 1, 2016


Thesis (5.5 Mb)

Copyright Statement
© 2016 Smith, Joanne D. All rights reserved. No part of this publication may be reproduced without the written permission of the copyright holder.

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