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Characteristics of immunosuppressive regulatory T cells in cutaneous squamous cell carcinomas and role in metastasis

Characteristics of immunosuppressive regulatory T cells in cutaneous squamous cell carcinomas and role in metastasis
Characteristics of immunosuppressive regulatory T cells in cutaneous squamous cell carcinomas and role in metastasis
BACKGROUND: Non-melanoma skin cancer is the most common cancer worldwide, and cutaneous squamous cell carcinomas (SCCs) account for substantial morbidity and mortality because of their potential for metastasis. SCCs are surrounded by an immune cell infiltrate containing regulatory T cells (Tregs). The aim of this study was to characterise Tregs in SCCs and investigate whether increased Treg numbers in primary skin SCCs are associated with subsequent metastasis.

METHODS: Lymphocytes were extracted from freshly excised skin SCC tumours and corresponding peripheral blood and normal skin. Flow cytometry was used for T-cell analysis and cell sorting. Tritiated thymidine based lymphocyte proliferation assays and interferon ? (IFN?) ELISPOT assays were used to assess peritumoral lymphocyte function in vitro. Immunohistochemistry was performed on primary cutaneous SCC sections from tumours that subsequently metastasised and from those that did not after 5-year follow-up.

FINDINGS: Increased frequencies of CD3+CD4+CD25hiCD127loFOXP3+ Tregs were found in SCCs (21·5% of CD4+ immune infiltrate, n=60 tumours) compared with corresponding peripheral blood (5·4%) and normal skin (7·6%). SCC Tregs expressed significantly higher levels of the co-stimulatory molecules OX40 (37·2% of FOXP3+ cell population, n=10 tumours) and 4-1BB (12·6%, n=9) than peritumoral non-regulatory T cells and Tregs from peripheral blood and normal skin (p=0·0005). The inhibitory receptor CTLA4 and the transcription factor Helios were expressed at high levels in peritumoral Tregs. SCC Tregs significantly suppressed phytohaemagglutinin-stimulated peritumoral CD4+ T-cell proliferation (p=0·005, n=10), peritumoral CD8+ T-cell proliferation (p=0·015, n=9), and IFN? secretion by CD4+ effector T cells (p=0·026, n=10). Increased in-vitro proliferation of phytohaemagglutinin-stimulated peritumoral CD4+ T cells was shown after the addition of anti-OX40 antibodies (p=0·0078, n=9 tumours) and anti-4-1BB antibodies (p=0·0039, n=9). Immunohistochemistry showed fewer CD8+ T cells in SCCs that metastasised (n=29) than in non-metastatic SCCs (n=26) (28·5% of immune infiltrate vs 44·6%%, p<0·0001) and more FOX3+ Tregs (28·5% of immune infiltrate vs 49·3%, p<0·0001).

INTERPRETATION: Our study shows that immunosuppressive Tregs are present in the immune infiltrate of cutaneous SCCs, and contribute to ineffective anti-tumour immune responses, thereby permitting SCC development and promoting metastasis.

FUNDING: Wellcome Trust, National Institute for Health Research
0140-6736
S59
Lai, Chester
29ba48ea-2d38-497f-8cf9-400237f6a3a0
August, Suzannah
1fc940e8-45d7-46b0-8da7-efdfcfcbebbb
Behar, Ramnik
cae7deb5-66d5-4f1b-9b21-751a614345a3
Polak, Marta
e0ac5e1a-7074-4776-ba23-490bd4da612d
Ardern-Jones, Mike
7ac43c24-94ab-4d19-ba69-afaa546bec90
Theaker, Jeff
2e0133cc-197b-479a-9465-458e534deb63
Al-Shamkhani, Aymen
0a40b3ce-9d71-4d41-9369-7212f0a84504
Healy, Eugene
400fc04d-f81a-474a-ae25-7ff894be0ebd
Lai, Chester
29ba48ea-2d38-497f-8cf9-400237f6a3a0
August, Suzannah
1fc940e8-45d7-46b0-8da7-efdfcfcbebbb
Behar, Ramnik
cae7deb5-66d5-4f1b-9b21-751a614345a3
Polak, Marta
e0ac5e1a-7074-4776-ba23-490bd4da612d
Ardern-Jones, Mike
7ac43c24-94ab-4d19-ba69-afaa546bec90
Theaker, Jeff
2e0133cc-197b-479a-9465-458e534deb63
Al-Shamkhani, Aymen
0a40b3ce-9d71-4d41-9369-7212f0a84504
Healy, Eugene
400fc04d-f81a-474a-ae25-7ff894be0ebd

Lai, Chester, August, Suzannah, Behar, Ramnik, Polak, Marta, Ardern-Jones, Mike, Theaker, Jeff, Al-Shamkhani, Aymen and Healy, Eugene (2015) Characteristics of immunosuppressive regulatory T cells in cutaneous squamous cell carcinomas and role in metastasis. [in special issue: Spring Meeting for Clinician Scientists in Training 2015] The Lancet, 385, supplement 1, S59. (doi:10.1016/S0140-6736(15)60374-9). (PMID:26312881)

Record type: Article

Abstract

BACKGROUND: Non-melanoma skin cancer is the most common cancer worldwide, and cutaneous squamous cell carcinomas (SCCs) account for substantial morbidity and mortality because of their potential for metastasis. SCCs are surrounded by an immune cell infiltrate containing regulatory T cells (Tregs). The aim of this study was to characterise Tregs in SCCs and investigate whether increased Treg numbers in primary skin SCCs are associated with subsequent metastasis.

METHODS: Lymphocytes were extracted from freshly excised skin SCC tumours and corresponding peripheral blood and normal skin. Flow cytometry was used for T-cell analysis and cell sorting. Tritiated thymidine based lymphocyte proliferation assays and interferon ? (IFN?) ELISPOT assays were used to assess peritumoral lymphocyte function in vitro. Immunohistochemistry was performed on primary cutaneous SCC sections from tumours that subsequently metastasised and from those that did not after 5-year follow-up.

FINDINGS: Increased frequencies of CD3+CD4+CD25hiCD127loFOXP3+ Tregs were found in SCCs (21·5% of CD4+ immune infiltrate, n=60 tumours) compared with corresponding peripheral blood (5·4%) and normal skin (7·6%). SCC Tregs expressed significantly higher levels of the co-stimulatory molecules OX40 (37·2% of FOXP3+ cell population, n=10 tumours) and 4-1BB (12·6%, n=9) than peritumoral non-regulatory T cells and Tregs from peripheral blood and normal skin (p=0·0005). The inhibitory receptor CTLA4 and the transcription factor Helios were expressed at high levels in peritumoral Tregs. SCC Tregs significantly suppressed phytohaemagglutinin-stimulated peritumoral CD4+ T-cell proliferation (p=0·005, n=10), peritumoral CD8+ T-cell proliferation (p=0·015, n=9), and IFN? secretion by CD4+ effector T cells (p=0·026, n=10). Increased in-vitro proliferation of phytohaemagglutinin-stimulated peritumoral CD4+ T cells was shown after the addition of anti-OX40 antibodies (p=0·0078, n=9 tumours) and anti-4-1BB antibodies (p=0·0039, n=9). Immunohistochemistry showed fewer CD8+ T cells in SCCs that metastasised (n=29) than in non-metastatic SCCs (n=26) (28·5% of immune infiltrate vs 44·6%%, p<0·0001) and more FOX3+ Tregs (28·5% of immune infiltrate vs 49·3%, p<0·0001).

INTERPRETATION: Our study shows that immunosuppressive Tregs are present in the immune infiltrate of cutaneous SCCs, and contribute to ineffective anti-tumour immune responses, thereby permitting SCC development and promoting metastasis.

FUNDING: Wellcome Trust, National Institute for Health Research

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Published date: 26 February 2015
Organisations: Clinical & Experimental Sciences

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Local EPrints ID: 390062
URI: https://eprints.soton.ac.uk/id/eprint/390062
ISSN: 0140-6736
PURE UUID: fdc2b721-1261-40a8-8f1d-fb3fdec4c59f
ORCID for Aymen Al-Shamkhani: ORCID iD orcid.org/0000-0003-0727-4189

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Date deposited: 17 Mar 2016 16:08
Last modified: 31 Jan 2019 01:37

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