Expression Of Chemokine - Associated Cytokines In Chronic Lymphocytic Leukemia And Its Clinical Significance

ObjectiveChemokines are an8-10Kda protein which plays an important role in hematology and non-hematology cells chemotaxis. Chemokines assist lymphocyte assemblages in inflammatory tissue; at the same time in collaboration with chemokines receptor, regulate the migration of the leukemia cells within bone marrow (BM), lymphoid organs. Chemokines form a pro-survival circuitry by regulating lymphocyte trafficking, keeping extended lymphocyte survival. CXCR4/CXCL12, CXCR5/CXCL13, CCL3and CCL4perform significant function in homeostatic microenvironment and driving migration in B chronic lymphoproliferative disease (BCLPD). Therefore, we investigate the expression and significance of CXCR4, CXCR5, CCL3and CCL4levels by immunohistochemistry (IHC) in BCLPD, exploring its association with clinical or laboratory features and prognostic implication.MethodsCXCR4, CXCR5, CCL3and CCL4expression levels of87patients with BCLPD were detected by bone marrow pathology biopsy IHC. Eighty seven patients were composed of52cases of chronic lymphocytic leukemia (CLL),12cases of Waldenstrom’s macroglobulinemia (WM),4cases of follicular lymphoma (FL),4cases of splenic marginal zone lymphoma (SMZL),4cases of mantle cell lymphoma(MCL) and11cases of unclassification BCLPD (BCLPD-U). Statistical analysis was performed by software SPSS20.0and GraphPad Prism6.0. CXCR4, CXCR5, CCL3and CCL4expression level of different groups and relation between the two factors in CLL were performed by chi-square test. The Kaplan-Meier method was operated for drawing the time to first treatment (TTFT) curve and results were contrasted using the log-rank test. An effect was considered statistically significant at P<0.05.ResultsOf all87cases, positive expression of CXCR4in CLL and WM were51.9%,75.0%. SMZL were all showed weakly positive expression and weakly positive expression of MCL was75.0%. CXCR4expression in WM and SMZL, MCL were obvious statistics difference (P=0.003, P=0.014). BCLPD had no statistics difference in expression of CXCR5. Compared with other BCLPD, CCL3expression was the highest in FL and CLL, positive rate was75.0%and38.4%, respectively. Weakly positive expression of WM was91.6%and MCL50.0%. Obvious difference of CCL3expression was found between CLL and WM, CLL and MCL (P=0.022and P=0.012) and expression of CCL3in FL, WM were also distinct difference (P=0.009). CCL4positive expression in CLL, WM and FL were38.4%,66.7%and50.0%, respectively. Expression of CCL4in BCLPD had no statistics difference. We performed statistical analysis of CXCR4, CXCR5, CCL3and CCL4expression in various prognostic factors of CLL showing that CXCR4expression in del(17p13) positive group was higher than the negative group (P=0.019); CXCR4expression in non-response to chemotherapy group is higher than response to chemotherapy group (P=0.014); CCL3expression was related with expression of TK1(P=0.008). With a median follow-up of19months (range,3～138months), survival analysis showed that CXCR4, CXCR5, CCL3and CCL4were not markers of poor prognosis.ConclusionsCXCR4, CXCR5, CCL3and CCL4are showed different expression in BCLPD. CXCR4expression is related with clinical efficacy and has a certain prognostic significance. ObjectiveChemokines are composed of about50amino acids, and could regulate migration and mature of immune cells, lymphocytes. Chemokines form a prosurvival circuitry by regulating lymphocyte trafficking, keeping lymphocyte survival. CXCR4/CXCL12, CXCR5/CXCL13, CCL3and CCL4play an important role in chronic lymphocytic leukemia (CLL) microenvironment. Therefore, we investigate the expression of CXCR4, CXCR5, CCL3and CCL4in CLL expression and explore its association with clinical, laboratory features and prognostic implication.MethodsmRNA levels of CXCR4, CXCR5, CCL3and CCL4genes from CLL patients peripheral blood mononuclear cells (PBMC) and healthy donors purified CD19+B cells were quantified using qRT-PCR with SYBR Green. The expression level of relative mRNA was analyzed by2(-ΔCT) method. Statistical analysis was performed by SPSS (version20.0) and GraphPadPrism (version6.0). The difference of target gene mRNA expression in groups was described using the Mann-Whitney U test. The Kaplan-Meier method was performed for drawing TTFT (the time to first treatment) curves, and results were contrased using the log-rank test. An effect was considered statistically significant at P<0.05. ResultsOut of91CLL patients, CXCR4, CXCR5, CCL3and CCL4mRNA detection were found in all cases. Mean levels of CXCR4, CXCR5, CCL3and CCL4mRNA from CLL were0.4686(0.0220-2.5936),1.7548(0.0045-19.5115),0.1672(0.0063-13.0000) and0.2195(0.0010-13.1318), respectively. At the same time, mean levels of CXCR4, CXCR5, CCL3and CCL4mRNA from B cells of healthy donors were0.3275(0.1069-0.8100),2.5985(0.5700-3.7800),0.0272(0.0015-0.8700) and0.6050(0.3500-2.1700), respectively. There were significant difference expression levels of CCL3, CCL4between CLL patients and healthy donors (P=0.001and P=0.003). CXCR4expression in del(11q22.3) negative group was higher than the positive group (P=0.007), CCL3mRNA expression level was relation with β2-MG (P=0.039). Expression of CCL4mRNA was negative relation with absolute lymphocytes and TK1(P=0.035and P=0.035). Conclutions The CCL3and CCL4mRNA expression were aberrantly change in CLL patients. Expression of CCL3mRNA was related with β2-MG, indicating a significant prognosis CLL. ObjectiveWe study the expression of CXCL12in plasma, CXCL13, CCL3and CCL4in serum in chronic lymphocytic leukemia (CLL), exploring its association with clinical, laboratory features and prognostic implication.MethodsCXCL12in plasma, CXCL13, CCL3and CCL4in serum expression levels were detected by enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed by SPSS (version20.0) and GraphPadPrism (version6.0). The expression of CXCL12in plasma, CXCL13, CCL3and CCL4in serum in difference groups were described using the Mann-Whitney U test. The Kaplan-Meier method was performed for drawing TTFT (the time to first treatment) curves, and results were comtrasted by using the log-rank test. An effect was considered statistically significant at P<0.05.ResultsThe median expression levels of CXCL12in plasma, CXCL13, CCL3and CCL4in serum in50CLL patients were2103.21(12.10-3647.60) pg/ml,112.40(1.61-922.45) pg/ml,31.10(0.39-794.4) pg/ml and247.90(28.7-2327.3) pg/ml, respectively. In15healthy donors, median expression levels of CXCL12in plasma, CXCL13, CCL3and CCL4in serum were1768.30(1487.80-2457.80)pg/ml,16.73(1.39-105.50) pg/ml,19.90(3.80-68.40) pg/ml and28.20(7.10-53.10) pg/ml, respectively. Overall, the levels of CXCL12, CXCL13and CCL4in CLL patients were significantly higher than healthy controls (P=0.005, P=0.001and P<0.001). Statistics found that the expression of CXCL12in Binet A and B was obviously higher than Binet C (P=0.043); CXCL13was was related with age, β2-MG (P=0.042and P<0.001); CCL3expression was related with age,02-MG, IGVH, TK1(P=0.049, P=0.008, P=0.041and P=0.025); with a median follow-up of27months (range,1-138months), survival analysis showed that the patients with high serum CCL4levels had significantly shorter first time to treatment (TTFT) than the patients with low serum CCL4(P=0.009), indicating CCL4was a marker of poor prognosis.ConclutionsCLL patients showed increased CXCL12, CXCL13, CCL4levels compared to healthy controls. Survival analysis showed that the patients with high serum CCL4levels had significantly shorter TTFT.