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CTL- vs T reg lymphocyte-attracting chemokines, CCL4 and CCL20, are strong reciprocal predictive markers for survival of patients with oesophageal squamous cell carcinoma

View Article: PubMed Central - PubMed

ABSTRACT

Background:: Tumoural infiltration of T lymphocytes is determined by local patterns of specific chemokine expression. In this report, we examined the roles of CCL4 and CCL20 in the accumulation of CD8+ cytotoxic T lymphocytes (CTLs) and regulatory T (Treg) lymphocytes in oesophageal squamous cell carcinoma (ESCC), and determined the correlations between chemokine expressions and ESCC patients' survival.

Methods:: Reverse transcriptase–PCR and immunohistochemistry (IHC) staining were performed to examine the expressions of interested genes. Flow cytometry was adopted to check the expressions of CCL4- and CCL6-specific receptors, CCR5 and CCR6, on CTLs and Treg cells. In addition, transwell assay was carried on.

Results:: The CCL4 expression was significantly correlated with the expression of CTL markers (CD8 and Granzyme B), whereas CCL20 was positively correlated with Treg markers (FoxP3 and IL-10). Consistently, CCR5 was found to be mainly expressed on CD8+ T lymphocytes, while CCR6 showed prevalence on Treg lymphocytes and the frequencies of CCR5+CD8+ CTLs and CCR6+ Treg cells were higher in TIL compared with PBMC. Respectively, CCL4 and CCL20 recruited CD8+ and regulatory T cells in vitro. Importantly, high levels of CCL4 in the lesions of ESCC patients predicted prolonged survival. Furthermore, CCL4high/CCL20low group demonstrated better overall survival, whereas CCL4low/CCL20low and CCL4low/CCL20high groups showed the worst overall survival.

Conclusions:: Our data showed that CCL4 and CCL20 recruit functionally different T lymphocyte subsets into oesophageal carcinoma, indicating CCL4 and CCL20 are potential predictors of ESCC patients' survival.

No MeSH data available.


Related in: MedlinePlus

Expressions of CCL4 and CCL20 are elevated in tumour tissues. Both marginal and tumour tissues were checked by immunohistochemistry for the expressions of CCL4 and CCL20. Then, the immunohistochemical scores were calculated as described in the Materials and Methods. (A) Representative photographs of marginal and tumour tissue sections stained with specific primary antibodies or non-immune rabbit IgG, × 400. (B) Scores of CCL4 and CCL20 were significantly higher in malignant tissues (n=128) than that in marginal tissues (n=75).
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fig4: Expressions of CCL4 and CCL20 are elevated in tumour tissues. Both marginal and tumour tissues were checked by immunohistochemistry for the expressions of CCL4 and CCL20. Then, the immunohistochemical scores were calculated as described in the Materials and Methods. (A) Representative photographs of marginal and tumour tissue sections stained with specific primary antibodies or non-immune rabbit IgG, × 400. (B) Scores of CCL4 and CCL20 were significantly higher in malignant tissues (n=128) than that in marginal tissues (n=75).

Mentions: Next, we checked the expressions of CCL4 and CCL20 in tumour and marginal tissues. The IHC analysis showed that only 68.0% of marginal tissues (51 out of 75) were stained positively for CCL4, compared with 93.75% of tumour tissues (120 out of 128) (Figure 4A). The average scores of CCL4 were different between marginal and tumour tissues (1.26±1.50 vs 3.22±2.61; t=5.914, P<0.0001; Figure 4B). For CCL20 staining, 89.84% of tumour samples (115 out of 128) were CCL20 positive in contrast with 54.67% of marginal tissues (41 out of 75) (Figure 4A). The average IHC scores of tumour tissues were obviously higher than marginal tissues (2.28±1.89 vs 0.89±1.25; t=5.648, P<0.0001; Figure 4B). This result was also reflected in the RT–PCR analysis. In paired samples, mRNA intensities of CCL4 and CCL20 were significantly increased in tumour tissues (tCCL4=2.268, P<0.05; tCCL20=2765, P<0.01; Supplementary Figure 3).


CTL- vs T reg lymphocyte-attracting chemokines, CCL4 and CCL20, are strong reciprocal predictive markers for survival of patients with oesophageal squamous cell carcinoma
Expressions of CCL4 and CCL20 are elevated in tumour tissues. Both marginal and tumour tissues were checked by immunohistochemistry for the expressions of CCL4 and CCL20. Then, the immunohistochemical scores were calculated as described in the Materials and Methods. (A) Representative photographs of marginal and tumour tissue sections stained with specific primary antibodies or non-immune rabbit IgG, × 400. (B) Scores of CCL4 and CCL20 were significantly higher in malignant tissues (n=128) than that in marginal tissues (n=75).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4559838&req=5

fig4: Expressions of CCL4 and CCL20 are elevated in tumour tissues. Both marginal and tumour tissues were checked by immunohistochemistry for the expressions of CCL4 and CCL20. Then, the immunohistochemical scores were calculated as described in the Materials and Methods. (A) Representative photographs of marginal and tumour tissue sections stained with specific primary antibodies or non-immune rabbit IgG, × 400. (B) Scores of CCL4 and CCL20 were significantly higher in malignant tissues (n=128) than that in marginal tissues (n=75).
Mentions: Next, we checked the expressions of CCL4 and CCL20 in tumour and marginal tissues. The IHC analysis showed that only 68.0% of marginal tissues (51 out of 75) were stained positively for CCL4, compared with 93.75% of tumour tissues (120 out of 128) (Figure 4A). The average scores of CCL4 were different between marginal and tumour tissues (1.26±1.50 vs 3.22±2.61; t=5.914, P<0.0001; Figure 4B). For CCL20 staining, 89.84% of tumour samples (115 out of 128) were CCL20 positive in contrast with 54.67% of marginal tissues (41 out of 75) (Figure 4A). The average IHC scores of tumour tissues were obviously higher than marginal tissues (2.28±1.89 vs 0.89±1.25; t=5.648, P<0.0001; Figure 4B). This result was also reflected in the RT–PCR analysis. In paired samples, mRNA intensities of CCL4 and CCL20 were significantly increased in tumour tissues (tCCL4=2.268, P<0.05; tCCL20=2765, P<0.01; Supplementary Figure 3).

View Article: PubMed Central - PubMed

ABSTRACT

Background:: Tumoural infiltration of T lymphocytes is determined by local patterns of specific chemokine expression. In this report, we examined the roles of CCL4 and CCL20 in the accumulation of CD8+ cytotoxic T lymphocytes (CTLs) and regulatory T (Treg) lymphocytes in oesophageal squamous cell carcinoma (ESCC), and determined the correlations between chemokine expressions and ESCC patients' survival.

Methods:: Reverse transcriptase&ndash;PCR and immunohistochemistry (IHC) staining were performed to examine the expressions of interested genes. Flow cytometry was adopted to check the expressions of CCL4- and CCL6-specific receptors, CCR5 and CCR6, on CTLs and Treg cells. In addition, transwell assay was carried on.

Results:: The CCL4 expression was significantly correlated with the expression of CTL markers (CD8 and Granzyme B), whereas CCL20 was positively correlated with Treg markers (FoxP3 and IL-10). Consistently, CCR5 was found to be mainly expressed on CD8+ T lymphocytes, while CCR6 showed prevalence on Treg lymphocytes and the frequencies of CCR5+CD8+ CTLs and CCR6+ Treg cells were higher in TIL compared with PBMC. Respectively, CCL4 and CCL20 recruited CD8+ and regulatory T cells in vitro. Importantly, high levels of CCL4 in the lesions of ESCC patients predicted prolonged survival. Furthermore, CCL4high/CCL20low group demonstrated better overall survival, whereas CCL4low/CCL20low and CCL4low/CCL20high groups showed the worst overall survival.

Conclusions:: Our data showed that CCL4 and CCL20 recruit functionally different T lymphocyte subsets into oesophageal carcinoma, indicating CCL4 and CCL20 are potential predictors of ESCC patients' survival.

No MeSH data available.


Related in: MedlinePlus