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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

Expression levels of CCL4 and CCL20 correlate with, respectively, local expressions of (A) CTL markers (CD8 and Granzyme B) and (B) regulatory T lymphocytes markers (FoxP3 and IL-10). A total of 123 fresh tumour tissues from ESCC patients were collected and processed. RNA was extracted. Reverse transcriptase–PCR assay of interested genes was performed. Spearman's test was performed to determine the correlations among gene expressions.
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fig1: Expression levels of CCL4 and CCL20 correlate with, respectively, local expressions of (A) CTL markers (CD8 and Granzyme B) and (B) regulatory T lymphocytes markers (FoxP3 and IL-10). A total of 123 fresh tumour tissues from ESCC patients were collected and processed. RNA was extracted. Reverse transcriptase–PCR assay of interested genes was performed. Spearman's test was performed to determine the correlations among gene expressions.

Mentions: As reported, CCL4 is involved in the migration of not only CD8+ T lymphocytes but also Treg lymphocytes (Harlin et al, 2009; Schlecker et al, 2012). CCL20 mainly dictates the movement of Treg lymphocytes (Chen et al, 2011; Cook et al, 2014). However, the impacts of CCL4 and CCL20 on T lymphocytes in ESCC have not been reported. To elucidate how the tumoural accumulation of T lymphocytes was affected by CCL4 and CCL20 in ESCC, we performed RT–PCR assay to detect the associations between chemokines and T lymphocyte markers in malignant sites. As shown in Figure 1A, the expressions of CCL4 were positively related with cytotoxic T lymphocyte (CTL) markers, CD8 and Granzyme B (rCD8=0.257, P<0.005; rGranB=0.273, P<0.01). Meanwhile, the intensity of CCL20 was clearly correlated with Treg lymphocyte markers: FoxP3 and IL-10 (rFoxP3=0.240, P<0.01; rIL-10=0.387, P<0.005; Figure 1B). Furthermore, the mRNA levels of CTL markers were not statistically associated with that of CCL20 (rCD8=−0.024, P>0.05; rGranB=0.107, P>0.05; Supplementary Figure 1B), whereas the expressions of FoxP3 and IL-10 were independent on CCL4 levels (rFoxP3=0.103, P>0.05; rIL-10=0.159, P>0.05; Figure 1B). These data suggest that CCL4 and CCL20 are probably implicated in the migration and retention of CD8+ CTLs and Treg lymphocytes, respectively, into ESCC lesions.


CTL- vs T reg lymphocyte-attracting chemokines, CCL4 and CCL20, are strong reciprocal predictive markers for survival of patients with oesophageal squamous cell carcinoma
Expression levels of CCL4 and CCL20 correlate with, respectively, local expressions of (A) CTL markers (CD8 and Granzyme B) and (B) regulatory T lymphocytes markers (FoxP3 and IL-10). A total of 123 fresh tumour tissues from ESCC patients were collected and processed. RNA was extracted. Reverse transcriptase–PCR assay of interested genes was performed. Spearman's test was performed to determine the correlations among gene expressions.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Expression levels of CCL4 and CCL20 correlate with, respectively, local expressions of (A) CTL markers (CD8 and Granzyme B) and (B) regulatory T lymphocytes markers (FoxP3 and IL-10). A total of 123 fresh tumour tissues from ESCC patients were collected and processed. RNA was extracted. Reverse transcriptase–PCR assay of interested genes was performed. Spearman's test was performed to determine the correlations among gene expressions.
Mentions: As reported, CCL4 is involved in the migration of not only CD8+ T lymphocytes but also Treg lymphocytes (Harlin et al, 2009; Schlecker et al, 2012). CCL20 mainly dictates the movement of Treg lymphocytes (Chen et al, 2011; Cook et al, 2014). However, the impacts of CCL4 and CCL20 on T lymphocytes in ESCC have not been reported. To elucidate how the tumoural accumulation of T lymphocytes was affected by CCL4 and CCL20 in ESCC, we performed RT–PCR assay to detect the associations between chemokines and T lymphocyte markers in malignant sites. As shown in Figure 1A, the expressions of CCL4 were positively related with cytotoxic T lymphocyte (CTL) markers, CD8 and Granzyme B (rCD8=0.257, P<0.005; rGranB=0.273, P<0.01). Meanwhile, the intensity of CCL20 was clearly correlated with Treg lymphocyte markers: FoxP3 and IL-10 (rFoxP3=0.240, P<0.01; rIL-10=0.387, P<0.005; Figure 1B). Furthermore, the mRNA levels of CTL markers were not statistically associated with that of CCL20 (rCD8=−0.024, P>0.05; rGranB=0.107, P>0.05; Supplementary Figure 1B), whereas the expressions of FoxP3 and IL-10 were independent on CCL4 levels (rFoxP3=0.103, P>0.05; rIL-10=0.159, P>0.05; Figure 1B). These data suggest that CCL4 and CCL20 are probably implicated in the migration and retention of CD8+ CTLs and Treg lymphocytes, respectively, into ESCC lesions.

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