Increased tumor-associated mast cells facilitate thyroid cancer progression by inhibiting CD8<sup>+</sup> T cell function through galectin-9

As an important component of solid tumors, mast cells show specific phenotypes in various tumor microenvironments. However, the precise mechanism of mast cell accumulation and the phenotypic features of thyroid cancer (TC) remain largely unknown. Here, we found that mast cells were obviously recruited to tumor tissue by TC-derived stem cell factor (SCF). With tumor progression, mast cell levels increased gradually. In addition, intratumoral mast cells expressed higher levels of the immunosuppressive molecule galectin-9, which effectively suppresses CD8⁺ T-cell antitumor immunity in vitro. Blocking galectin-9 on tumor-infiltrating mast cells reversed the immunosuppression of CD8⁺ T cells. In conclusion, our data elucidated novel protumorigenic and immunosuppressive roles of mast cells in TC. In addition, our results indicated that blocking mast cells may impede tumor progression and ameliorate the prognosis of TC patients.

Key words
Mast cells; Thyroid cancer; Stem cell factor; Galectin-9; CD8⁺ T cells

Authorship

Yanli Hou

Department of Oncology and Hematology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, ChinaChongqing Hospital of Traditional Chinese MedicineChinaChongqing, ChinaDepartment of Oncology and Hematology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China

* These authors contributed equally to this work.

http://orcid.org/0000-0002-6897-6515

Qiang Wang

Department of Orthodontics, Deyang Stomatological Hospital, Deyang, Sichuan, ChinaDeyang Stomatological HospitalChinaDeyang, Sichuan, ChinaDepartment of Orthodontics, Deyang Stomatological Hospital, Deyang, Sichuan, China

* These authors contributed equally to this work.

http://orcid.org/0000-0002-2145-8542

Li Su

http://orcid.org/0000-0001-9590-3402

Ying Zhu

http://orcid.org/0000-0002-5416-4065

Yun Xiao Correspondence: Fei Feng: <ff451836670@163.com> | Yun Xiao: <xiaoyunsss@hotmail.com>

http://orcid.org/0000-0002-5565-391X

Fei Feng Correspondence: Fei Feng: <ff451836670@163.com> | Yun Xiao: <xiaoyunsss@hotmail.com>

http://orcid.org/0000-0003-4886-5487

Correspondence: Fei Feng: <ff451836670@163.com> | Yun Xiao: <xiaoyunsss@hotmail.com>

*
These authors contributed equally to this work.

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (4)
Tables (1)

Thumbnail

Figure 1
The mast cell percentage was increased in thyroid cancer (TC), and this increase was correlated with disease progression. A, Dot plots of molecule labelling for mast cell (CD45⁺ CD117⁺ FcεRI⁺ cells) gating on CD45⁺ cells in tumor and non-tumor tissue of TC patients. B, Mast cell percentage in CD45⁺ cells among TNM stages (I+II vs III+IV) in tumor and non-tumor tissues of TC patients. The cumulative results of 113 patients with TC are shown. C, Intratumoral mast cell percentage among TNM stages is shown. The horizontal bars represent means±SE, and each ring represents 1 patient in Panels B and C. *P<0.05; **P<0.01; (ANOVA). ns: P>0.05.

Thumbnail

Figure 2
Thyroid cancer (TC)-derived stem cell factor (SCF) mediated mast cell recruitment in TC tissues. A, The expression of Ki-67 on mast cells in tumor and non-tumor tissues. Red and blue histograms represent staining of Ki-67 in tumor and non-tumor tissues, respectively; black, isotype control. B, The correlations between mast cell percentage in CD45⁺ cell and SCF concentration in TC were detected (n=109). C, Differences in SCF levels between tumor tissues and autologous non-tumor tissues and between tumor or non-tumor tissue culture supernatants (TTCS and autologous NTCS) are shown. D, Migration of tumor-associated mast cells was detected by transwell assay (n=3). Each ring or dot in Panels A-C represents 1 patient. Data are reported as means±SE. **P<0.01 (ANOVA). ns: P>0.05.

Thumbnail

Figure 3
The expression of galectin-9 was upregulated on mast cells in thyroid cancer (TC) tissues. A, The expression of galectin-9 on mast cells in TC tumor and non-tumor tissues (n=8). Red and blue histograms represent labelling of galectin-9 in tumor and non-tumor tissues, respectively; black, isotype control. B, Expression of galectin-9 on HMC-1 cells (human mast cell line) stimulated by 50% tumor or non-tumor tissue culture supernatants (TTCS and autologous NTCS) for 24 h was detected by immunofluorescence staining. Red, galectin-9; blue, DAPI-stained nuclei. Scale bar: 50 microns. C, Expression of galectin-9 on hCBMCs stimulated by 50% TTCS and autologous NTCS for 24 h (n=3). Red and blue histograms represent staining of galectin-9 in tumor and non-tumor tissues, respectively; black, isotype control. Each dot in Panel A represents 1 patient. Data are reported as means±SE. **P<0.01 (t-test).

Thumbnail

Figure 4
Tumor-associated mast cells suppressed CD8⁺ T-cell antitumor immunity by galectin-9. A, The relationship between mast cells and CD8⁺ T cells in thyroid cancer (TC) is shown. B, The expression of TIM-3 on CD8⁺ T cells in TC tumor and non-tumor tissues (n=8). Red and blue histograms represent staining of TIM-3 in tumor and non-tumor tissues, respectively; black, isotype control. C, Mast cells sorted from tumor or non-tumor tissues were co-cultured with CFSE-labelled autologous peripheral CD8⁺ T cells with or without anti-galectin-9 antibody for 5 days. Representative data and statistical analysis of CD8⁺ T-cell proliferation and granzyme-B secretion are shown as means and SE (n=5). Each dot in Panels A and B represents 1 patient. Data are reported as means±SE. *P<0.05; **P<0.01 (t-test or ANOVA). CFSE: carboxyfluorescein succinimidyl ester.

Thumbnail

Table 1
Clinical features of 113 patients with thyroid cancer.

Figure 1 The mast cell percentage was increased in thyroid cancer (TC), and this increase was correlated with disease progression. A, Dot plots of molecule labelling for mast cell (CD45⁺ CD117⁺ FcεRI⁺ cells) gating on CD45⁺ cells in tumor and non-tumor tissue of TC patients. B, Mast cell percentage in CD45⁺ cells among TNM stages (I+II vs III+IV) in tumor and non-tumor tissues of TC patients. The cumulative results of 113 patients with TC are shown. C, Intratumoral mast cell percentage among TNM stages is shown. The horizontal bars represent means±SE, and each ring represents 1 patient in Panels B and C. *P<0.05; **P<0.01; (ANOVA). ns: P>0.05.

Figure 2 Thyroid cancer (TC)-derived stem cell factor (SCF) mediated mast cell recruitment in TC tissues. A, The expression of Ki-67 on mast cells in tumor and non-tumor tissues. Red and blue histograms represent staining of Ki-67 in tumor and non-tumor tissues, respectively; black, isotype control. B, The correlations between mast cell percentage in CD45⁺ cell and SCF concentration in TC were detected (n=109). C, Differences in SCF levels between tumor tissues and autologous non-tumor tissues and between tumor or non-tumor tissue culture supernatants (TTCS and autologous NTCS) are shown. D, Migration of tumor-associated mast cells was detected by transwell assay (n=3). Each ring or dot in Panels A-C represents 1 patient. Data are reported as means±SE. **P<0.01 (ANOVA). ns: P>0.05.

Figure 3 The expression of galectin-9 was upregulated on mast cells in thyroid cancer (TC) tissues. A, The expression of galectin-9 on mast cells in TC tumor and non-tumor tissues (n=8). Red and blue histograms represent labelling of galectin-9 in tumor and non-tumor tissues, respectively; black, isotype control. B, Expression of galectin-9 on HMC-1 cells (human mast cell line) stimulated by 50% tumor or non-tumor tissue culture supernatants (TTCS and autologous NTCS) for 24 h was detected by immunofluorescence staining. Red, galectin-9; blue, DAPI-stained nuclei. Scale bar: 50 microns. C, Expression of galectin-9 on hCBMCs stimulated by 50% TTCS and autologous NTCS for 24 h (n=3). Red and blue histograms represent staining of galectin-9 in tumor and non-tumor tissues, respectively; black, isotype control. Each dot in Panel A represents 1 patient. Data are reported as means±SE. **P<0.01 (t-test).

Figure 4 Tumor-associated mast cells suppressed CD8⁺ T-cell antitumor immunity by galectin-9. A, The relationship between mast cells and CD8⁺ T cells in thyroid cancer (TC) is shown. B, The expression of TIM-3 on CD8⁺ T cells in TC tumor and non-tumor tissues (n=8). Red and blue histograms represent staining of TIM-3 in tumor and non-tumor tissues, respectively; black, isotype control. C, Mast cells sorted from tumor or non-tumor tissues were co-cultured with CFSE-labelled autologous peripheral CD8⁺ T cells with or without anti-galectin-9 antibody for 5 days. Representative data and statistical analysis of CD8⁺ T-cell proliferation and granzyme-B secretion are shown as means and SE (n=5). Each dot in Panels A and B represents 1 patient. Data are reported as means±SE. *P<0.05; **P<0.01 (t-test or ANOVA). CFSE: carboxyfluorescein succinimidyl ester.

Table 1 Clinical features of 113 patients with thyroid cancer.

Variables	No. of patients
Gender
Male	29
Female	84
Age (years)
<55	64
≥55	49
Tumor size (cm)
<2	52
≥2	61
Tumor (T) invasion
T1+T2	79
T3+T4	34
Lymphoid nodal (N) status
N0+N1	72
N2+N3	41
Distant metastasis (M) status
M0	105
M1	8
TNM stage
I+II	79
III+IV	34

How to cite

copy

Associação Brasileira de Divulgação Científica Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto SP Brazil, Tel. / Fax: +55 16 3315-9120 - Ribeirão Preto - SP - Brazil
E-mail: bjournal@terra.com.br

Acompanhe os números deste periódico no seu leitor de RSS

Versão para download de PDF

PDF

Inglês

Versões e tradução automática

Versão original do texto

English

Tradução automática

Google Translator
Microsoft Translator

Como citar

RIS

BIBTEX

Outros formatos de citação e exportação:

SciELO - Scientific Electronic Library Online
Rua Dr. Diogo de Faria, 1087 – 9º andar – Vila Clementino 04037-003 São Paulo/SP - Brasil
E-mail: scielo@scielo.org