Correlations between pruritus and CC chemokine ligand 17 in patients with mycosis fungoides / 中华皮肤科杂志

Objective:To investigate molecules involved in the occurrence of pruritus in patients with mycosis fungoides (MF) .Methods:Totally, 522 patients with MF were enrolled from Peking University First Hospital from October 2009 to August 2021, and the incidence of pruritus was calculated. The patients were grouped according to whether they suffered from pruritus or not. RNA sequencing data on biopsied skin lesions of 49 patients were analyzed to identify differentially expressed genes between patients with pruritus and those without; enzyme-linked immunosorbent assay and immunohistochemical techniques were performed to determine the protein expression of CC chemokine ligand 17 (CCL17) in serum samples from 88 MF patients, and in tissue samples from 81 MF patients, respectively; flow cytometry was conducted to detect markers for T lymphocyte activation and differentiation in peripheral blood samples from 46 MF patients to identify peripheral blood lymphocyte subsets associated with pruritus. Statistical analysis was carried out by using chi-square test, Mann-Whitney U test, and Spearman correlation analysis. Results:Among the 522 patients with MF, 305 were males and 217 were females; 347 were diagnosed with early-stage MF, and 175 with advanced MF. The incidence of pruritus was 67.2% (351/522) in the patients with MF, and significantly higher in the patients with advanced MF (81.7%, 143/175) than in those with early-stage MF (59.9%, 208/347; χ2 = 25.03, P < 0.001) . RNA sequencing showed that CCL17 mRNA expression was significantly higher in the MF patients with pruritus than in those without (fold change = 10.09, P < 0.001) . The serum CCL17 concentration was significantly elevated in the patients with pruritus (1 017.05[377.12, 4 831.80] pg/ml) compared with those without (361.66 [180.47, 500.08] pg/ml; Z = -4.57, P < 0.001) , and correlated with pruritus scores ( r = 0.57, P = 0.010) . In both early and advanced stages of MF, the serum CCL17 concentration was significantly higher in the patients with pruritus than in those without ( Z = -3.68, P < 0.001; Z = -2.54, P = 0.011, respectively) . Immunohistochemical staining revealed that there was no significant difference in the relative quantification value of CCL17 between the patients with pruritus and those without ( Z = -1.84, P = 0.066) . The percentage of CD3 +CD4 +CD26 -CCR4 + malignant T cells significantly increased in the MF patients with pruritus than in those without ( Z = -2.03, P = 0.043) , and was positively correlated with serum CCL17 concentrations ( r = 0.49, P < 0.001) . Conclusions:Both CCL17 mRNA expression in lesional tissues and serum CCL17 concentrations increased in MF patients with pruritus, and CCL17 was associated with the occurrence of pruritus. CCL17 may be involved in the occurrence of pruritus through the recruitment of CD3 +CD4 +CD26 -CCR4 + malignant T cells.

Mechanism of Macrophage-Derived Chemokine/CCL22 Production by HaCaT Keratinocytes

BACKGROUND: CC chemokine ligand 17 (CCL17) and CCL22 are the functional ligands for CCR4. We previously reported that inhibitors of nuclear factor-kappa B and p38 mitogen-activated protein kinase (p38 MAPK), but not of extracellular signal-related kinase (ERK), inhibited tumor necrosis factor (TNF)-alpha- and interferon (IFN)-gamma-induced production of CCL17 by the human keratinocyte cell line, HaCaT. Further, an inhibitor of epidermal growth factor receptor (EGFR) enhanced the CCL17 production by these keratinocytes. OBJECTIVE: To identify the mechanism underlying CCL22 production by HaCaT cells. METHODS: We investigated the signal transduction pathways by which TNF-alpha and IFN-gamma stimulate HaCaT cells to produce CCL22 by adding various inhibitors. RESULTS: TNF-alpha- and IFN-gamma-induced CCL22 production was inhibited by PD98059, PD153035, Bay 11-7085, SB202190, c-Jun N-terminal kinase (JNK) inhibitor II, and Janus kinase (JAK) inhibitor 1. CONCLUSION: Our results indicate that CCL22 production in HaCaT cells is dependent on ERK, EGFR, p38 MAPK, JNK, and JAK and is mediated by different signal pathways from those regulating CCL17 production. Altogether, our previous and present results suggest that EGFR activation represses CCL17 but enhances CCL22 production by these cells.

Expression and function of chemokine TARC/CCR4 at fetal-maternal interface in first trimester / 中华妇产科杂志

Objective To investigate the expression and function of thymus and activation regulated chemokine (TARC) and its special receptor CCR4 at placenta villous in the first trimester placenta villous.Methods Placenta villous was collected from healthy women undergoing artificial abortion at 6 to 8 weeks of gestation,mRNA levels of TARC,CCR4 were analyzed using semi-quantitative reverse transcription (RT)-PCR methods.Immunohistochemistry assay was used to assess the protein localization and expression of TARC,CCR4.Additionally,extravillous cytotrophoblasts were isolated and cultured.Expression of TARC and CCR4 was measured by immunofluorescence assay.Invasion of cell line HTR8/SVneo was analyzed by transwell assay at concentration of 10,25,50 and 100 ng/ml of TARC matched with RPMI 1640 fetal bovine serum free eulture medium as control group.In the mean time,blocking experiment was also added to detect TARC regulating cell invasion,which were classified into four groups:control,100 ng/ml rhTARC,20 μg/ml anti-TARC + 100 ng/ml rhTARC,100 ng/ml rhTARC + 20 μg/ml IgG.The influence of 100 ng/ml TARC on expression level of integrin-α5 and integrin-β1 were measured by using western-blot assay.Results (1)In vivo assay:expression of TARC and CCR4 mRNA were detectable in first trimester placenta villous,TARC protein was localized in cytotrophoblasts,syncytiotrophoblasts and cell column especially on the distal portion,while CCR4 protein was localized on invading interstitial cytotrophobalsts.(2)In vitro assay:①TARC,CCR4 was also expressed in primary isolated extravillous cytotrophoblasts by immunofluorescence assay; ②Matrigel invasion assay demonstrated that TARC had specific dose dependent stimulatory effects on the cells invading through the matrigel precoated filter,the number of cells migration into the lower chamber were:142 ± 31 at 10 ng/ml group,161 ±46 at 25 ng/ml group,201 ±30 at 50 ng/ml group,312 ±48 at 100 ng/ml group,117 ± 33 at control group,the significant response observed from 25 ng/ml (P ＜ 0.05)and reached a peak effect at 100 ng/ml (P ＜ 0.01); ③Blocking experiment demonstrated that when trophoblast invasion was monitored in response to TARC neutralizing antibody (15 μg/ml) together with rhTARC 100 ng/ml.The stimulatory activity of rhTARC was completely overcome,with the cells invasion into the lower chambers were 100 ng/ml rhTARC,20 μg/ml anti-TARC + 100 ng/ml rhTARC,100 ng/ml rhTARC +20 μg/ml IgG,control:313 ±47,113 ±41,287 ±75 and 128 ±23,respectively;④Western-blot assay demonstrated that if cells were treated with 100 ng/ml rhTARC,the expression of integrin-α5 were significantly increased(P ＜0.01),integrin-β1 level also increased when compared with control(P ＜0.05).Conclusion TARC was expressed specifically at human fetal-maternal interface.Trophoblast invasion and migration mainly was regulated by up-regulation integrin-α5 and integrin-β1,which plays an role in trophoblasts differentiation and placentation.

The Relationship between Atopic Dermatitis, Thymus and Activation-Regulated Chemokine/CCL17, Quality of Life, and Attention Deficit-Hyperactivity Disorder in Preschool Children / 소아알레르기및호흡기학회지

PURPOSE: Atopic dermatitis may impair quality of life and lead to attention deficit-hyperactivity disorder (ADHD). Thymus and activation-regulated chemokine (TARC)/CCL17 may serve as a new biomarker for atopic dermatitis. We investigated the relationship between TARC and the severity of atopic dermatitis, quality of life, and ADHD. METHODS: A total of 249 preschool children who had atopic dermatitis were enrolled. Parents of the patients filled out a questionnaire on the past history of allergic diseases, quality of life, and ADHD. In each patient, total immunoglobulin (Ig) E and specific IgE to nine foods and inhalant allergens, total eosinophil counts, and TARC levels were measured. We evaluated the severity of atopic dermatitis by using the scoring atopic dermatitis (SCORAD) score and divided the patients into three groups; mild (40). RESULTS: In a total of 249 children, 222 children (89.2%) had a history of atopic dermatitis. Children with allergic sensitization had a higher SCORAD score, total IgE levels, and total eosinophil counts, but not TARC levels. Three groups by the SCORAD score showed significant differences in quality of life index and TARC levels but not in ADHD index. TARC level was correlated with the SCORAD score, but not with the quality of life index and ADHD index. The SCORAD score was correlated with the quality of life index but not with the ADHD index. CONCLUSION: Serum TARC levels may be associated with the severity of atopic dermatitis but not with the degree of quality of life and ADHD. Disease severity of atopic dermatitis in preschool children may be associated with the degree of quality of life but not with the level of ADHD.