Up-regulated expression of substance P in CD8+ T cells and NK1R on monocytes of atopic dermatitis.

BACKGROUND: Large numbers of CD8+ T cells were observed in atopic dermatitis (AD) skin, and monocytes from AD patients showed increased prostaglandin E2 production. However, little is known about the expression of substance P (SP) and its receptor NK1R in blood leukocytes of patients with AD. OBJECTIVE: To explore the expression of SP and NK1R in leukocytes of AD and the influence of allergens on SP and NK1R expression. METHODS: The expression levels of SP and NK1R in patients with AD were examined by flow cytometry, ELISA and a mouse AD model. RESULTS: The plasma SP level was 4.9-fold higher in patients with AD than in HC subjects. Both the percentage of SP expression in the population and mean fluorescence intensity (MFI) of SP expression were elevated in CD8+ T cells in the blood of AD patients. However, both the CD14+NK1R+ population and MFI of NK1R expression on CD14+ cells were enhanced in the blood of AD patients. Allergens ASWE, HDME and PPE failed to up-regulate SP expression in CD8+ T cells. However, allergens ASWE and HDME both enhanced NK1R expression on CD14+ blood leukocytes regardless of AD or HC subjects. OVA-sensitized AD mice showed an elevated proportion and MFI of SP-expressing CD8+ T cells in the blood, which agrees with the SP expression situation in human AD blood. Injection of SP into mouse skin did not up-regulate NK1R expression on monocytes. CONCLUSIONS: An elevated plasma SP level, up-regulated expression of SP and NK1R indicate that the SP/NK1R complex is important in the development of AD. Therefore, SP and NK1R antagonist or blocker agents may help to treat patients with AD. Trial registration Registration number: ChiCTR-BOC-16010279; Registration date: Dec., 28, 2016; retrospectively registered.

Upregulated expression of substance P (SP) and NK1R in eczema and SP-induced mast cell accumulation.

Substance P (SP) was reported to be associated with eczema and acts as a potent skin mast cell secretagogue. However, little is known of its expression in inflammatory cells in eczema and its ability in induction of mast cell accumulation. In the present study, we investigated expression of SP and neurokinin-1 receptor (NK1R) on peripheral blood leukocytes and mast cells from patients with eczema and influence of SP on mast cell accumulation by using flow cytometry analysis, trans-epithelial cell migration assay and mouse peritoneal model. The results showed that plasma SP and IL-17A levels in eczema patients were higher than that in healthy control subject. The percentages of SP+ and NK1R+ expression populations of monocytes, helper T cells, natural killer T cells and basophils in peripheral blood of eczema patients were markedly elevated. It was observed that not only absolute number of mast cells but also SP+ and NK1R+ mast cells are enhanced in the lesion skin of eczema. SP showed a potent chemoattractant action on mast cells as assessed by a mouse peritoneal model and a trans-endothelium cell migration assay. SP-induced mast cell accumulation appears a CD18/CD11a complex, L-selectin and ICAM-1-dependent event which can be blocked by a NK-1R antagonist RP67580. In conclusion, elevated expression of SP in patients with eczema and the ability of SP in induction of mast cell accumulation indicate strongly that SP is a potent proinflammatory mediator, which contributes to the pathogenesis of eczema. Inhibitors of SP and blockers of NK1R are likely useful agents for treatment of eczema.

Upregulated expression of CCR3 in rheumatoid arthritis and CCR3-dependent activation of fibroblast-like synoviocytes.

It is recognized that CC chemokine receptor 3 (CCR3) is associated with numerous inflammatory conditions and fibroblast-like synoviocyte (FLS) invasiveness correlates with articular damage in rheumatoid arthritis (RA). However, little is known of the expression and action of CCR3 on FLS in RA. In the present study, we investigated the expression of CCR3 on dispersed synovial tissue and peripheral blood cells in RA and influence of eotaxin-1 on FLS functions by using flow cytometry analysis, FLS challenge, and real-time PCR techniques. The results showed that approximately 7.0 % dispersed synovial cells are CCR3+ cells. Among those CCR3+ cells, 38.1, 23.8, and 20.6 % cells are CD90+CD14-CD3- (representing FLS), CD14+, and CD8+ cells, respectively, indicating that FLS is one of the major populations of CCR3+ cells in the synovial tissue of RA. In peripheral blood, CD14+ CCR3+ cells are elevated, but CD8+CCR3+ cells are reduced in RA. It was found that eotaxin-1 induced upregulated expression of CCR3 and matrix metalloproteinase (MMP)-9 messenger RNAs (mRNAs) in FLS. Since an antagonist of CCR3 suppressed the action of eotaxin-1, the event appeared CCR3 dependent. Moreover, we observed that interleukin (IL)-1ß induced markedly enhanced eotaxin-1 release from FLS, but TNF-α reduced eotaxin-1 release at 12 and 24 h following incubation. In conclusion, enhanced expression of CCR3 on synovial cells and increased levels of eotaxin-1 in plasma and synovial fluid (SF) of RA indicate that CCR3-mediated mechanisms may play an important role in RA. Blockage of eotaxin-1 provoked CCR3 and MMP-9 expression in FLS by antagonist of CCR3, implicating that anti-CCR3 agents may have therapeutic use for RA.

Upregulated expression of substance P in basophils of the patients with chronic spontaneous urticaria: induction of histamine release and basophil accumulation by substance P.

Human basophils have been implicated in the pathogenesis of chronic spontaneous urticaria (CSU), and substance P (SP) is a possible candidate as histamine-releasing factor in some patients with CSU. However, little is known of relationship between basophils and SP in CSU. In the present study, we investigated expression of SP and NK1R on basophils from patients with CSU, and influence of SP on basophil functions by using flow cytometry analysis, basophil challenge, and mouse sensitization model techniques. The results showed that plasma SP level and basophil numbers in CSU patients were higher than that in HC subject. The percentages of SP+ and NK1R+ basophils were markedly elevated in CSU blood in comparison with HC blood. Once added, SP induced up to 41.2 % net histamine release from basophils of CSU patients, which was comparable with that provoked by anti-IgE, and fMLP. It appeared that SP induced dramatic increase in blood basophil numbers of mice following peritoneal injection. Ovalbumin (OVA)-sensitized mice had much more SP+ and NK1R+ basophils in blood than non-sensitized mice. In conclusion, the elevated plasma concentration of SP, upregulated expression of SP and NK1R on basophils, and the ability of SP in induction of basophil degranulation and accumulation indicate strongly that SP is most likely a potent proinflammatory mediator, which contributes greatly to the pathogenesis of CSU through basophils. Inhibitors of SP and blockers of NK1R are likely useful agents for treatment of CSU.

Correlation of IL-18 with Tryptase in Atopic Asthma and Induction of Mast Cell Accumulation by IL-18.

Interleukin- (IL-) 18 and tryptase were previously reported to relate to asthma, but the correlation between these two potent proinflammatory molecules in asthma and their roles in mast cell accumulation remain uninvestigated. Using flow cytometric analysis technique and ovalbumin- (OVA-) sensitized mouse model, it was found that IL-18 and tryptase levels in the plasma of moderate and severe asthma were elevated, and they correlated well with each other. Tryptase and agonist peptides of protease activated receptor- (PAR-) 2 induced substantial quantity of IL-18 release. IL-18 and tryptase provoked mast cell accumulation in peritoneum of OVA-sensitized mice. OVA-sensitization increased number of IL-18 receptor (R)(+) mast cells. IL-18 and tryptase induced dramatic increase in IL-18R(+) mast cells and mean fluorescence intensity (MFI) of IL-18R on mast cells. Moreover, while IL-18 induced an increase in PAR-2(+) mast cells in nonsensitized mice, IL-18 and tryptase provoked increases in IL-4 and thymic stromal lymphopoietin (TSLP) in the peritoneum of OVA-sensitized mice. In summary, the correlation between IL-18 and tryptase in plasma of patients with asthma indicates close interactions between them, which should be considered for development of anti-IL-18 and antitryptase therapies. Interactions between IL-18 and tryptase may contribute to mast cell recruitment in asthma.

[Up-regulated expression of NK1R in eosinophil-enriched blood cells from patients with chronic spontaneous urticaria].

Objective To investigate the expressions of substance P (SP) and neurokinin-1 receptor (NK1R) in eosinophil-enriched blood cells from patients with chronic spontaneous urticaria (CSU). Methods Peripheral venous blood samples were collected from patients with CSU and healthy controls (HCs), and then stimulated with crude extracts of Artemisia pollen, dust mite, and Platanus pollen (all at concentrations of 0.1 and 1.0 µg/mL). The expressions of SP and NK1R in eosinophil-enriched blood cells were detected by flow cytometry. Results Compared with HCs, eosinophil proportion in peripheral blood of CSU patients increased 1.2-fold. Percentage of NK1R+ eosinophils in the patients with CSU was elevated up to 66% compared with HCs when cultured in the medium only. However, the level of SP decreased by 40% in the CSU patients. In eosinophil-enriched blood cells from the CSU patients, the crude extract of dust mite at 0.1 µg/mL induced approximately 1.11-fold increase of NK1R expression. Conclusion Expression of NK1R increases in the eosinophils of CSU patients. Blockers of NK1R might be used for CSU treatment.

[Increased expressions of substance P and neurokinin/tachykinin receptor 1 in eosinophils of patients with psoriasis].

Objective To investigate the expressions of substance P (SP) and its receptor neurokinin/tachykinin receptor 1 (NK1R) in peripheral blood eosinophils of patients with psoriasis. Methods The levels of SP and NK1R in the peripheral blood of both patients with psoriasis and healthy people were detected by flow cytometry. This method was again used to detect the levels of SP and NK1R in the peripheral blood eosinophils of patients with psoriasis after stimulated with the crude extracts of Artemisia pollen, dust mite and Platanus pollen (all at concentrations of 0.1 and 1.0 µg/mL). Results Compared with the healthy controls, the percentages of SP+ and NK1R+ eosinophils in psoriasis patients increased up to 2.7 and 0.5 folds, respectively. Moreover, the mean fluorescence intensity (MFI) of SP+ and NK1R+ eosinophils of psoriasis patients were elevated by 1.5 and 0.2 folds, respectively. The percentage of SP+ eosinophils in psoriasis were down-regulated by 60% after the stimulation with Platanus pollen extract (1 µg/mL), while 0.1 µg/mL Platanus pollen extract induced a 0.6-fold increase in the percentage of NK1R+ eosinophis. Conclusion The expressions of SP and NK1R are up-regulated in peripheral blood eosinophils of patients with psoriasis.

Analysis of murine and human Treg subsets in inflammatory bowel disease.

Previous studies have indicated that regulatory T cells serve essential roles in maintaining intestinal homeostasis, however, the role of different Treg subsets in modulating inflammatory bowel disease has still not been addressed clearly. In the present study, the authors measured the percentage of Foxp3+ IL­10+ TGF­ß+ natural Tregs, Foxp3­ IL­10+ TGF­ß­ induced Tregs, CD127­ induced Tregs and CD8+ Tregs at different time points in DSS­induced experimental colitis model in murine lamina propria lymphocytes, mesenteric lymph node and peripheral blood. In addition, the authors compared the frequency of four Treg subsets in patients diagnosed of ulcerative colitis at different stages with enrolled healthy controls. The percentage of Foxp3+ IL­10+ TGF­ß+ natural Tregs decreased in acute stage of both human and mice was observed, but proliferated significantly during remittent stage. Foxp3­ IL­10+ TGF­ß­ inducible (i) Treg and CD127­ iTreg was observed as being significantly decreased percentage in LPL at 4 and 7 days, the frequency of Foxp3­ IL­10+ TGF­ß­ iTreg cells became decreased and CD127­ iTreg only slightly increased at the chronic stage following DSS induction. However, the proportion of both Foxp3­ IL­10+ TGF­ß­ iTreg and CD127­ iTreg was nearly unchanged in human IBD. Although intestinal inflammation decreased the percentage of CD8+ Tregs, it remained lower in the remittent stage of human IBD. Only enhanced proliferation of lamina propria lymphocytes­derived CD8+ Treg was reported at 7 days in dextran sodium sulfate­induced murine colitis. The results demonstrated that Foxp3+ IL­10+ TGF­ß+ natural Tregs may serve an essential role in exhibiting suppressive and protecting from immune­related mucosal injury during chronic stage in inflammatory bowel disease.

[Expression of LRG-1 in clinical specimens and Tca8113 cell line of tongue carcinoma].

OBJECTIVE: To investigate the expression of LRG-1 in clinical specimens and Tca8113 cell line of tongue carcinoma and analyze the relationship between LRG-1 expression and the clinicopathological parameters. METHODS: LRG-1 expression was detected in 40 tongue squamous cell carcinoma (TSCC) tissues and paired normal adjacent tissues, 20 atypical hyperplasia tissues of the tongue, and 20 tissues of tongue cancer in situ using immunohistochemical method. The expression of LRG-1 in Tca8113 cell line was detected using flow cytometry. The expression of LRG-1 was also detected in human TSCC tissues and Tca8113 cells with Western blotting. The effect of LRG-1 on the proliferation of HUVECs was determined using MTT assay, and its effect on angiogenesis was evaluated with Matrigel tube formation assays. RESULTS: Human TSCC tissues had a significantly higher rate of positive expression for LRG-1 (85%, 34/40) than the adjacent tissues (10%, 4/40), invasive tongue cancer (30%, 6/20), and tongue cancer in situ (50%, 10/20) (P<0.05). LRG-1 expression was correlated with the degree of tumor differentiation, clinical stage and lymph node metastasis of the tumor (P<0.05) but not with the patients' age or gender. In the in vitro experiment, LRG-1 promoted HUVEC proliferation and angiogenesis. CONCLUSION: Abnormal LRG-1 expression is present in the human TSCC tissue and Tca8113 cells. LRG-1 can promote HUVEC proliferation and angiogenesis in vitro, suggesting its possible role in promoting tumor angiogenesis.

Pre-treatment mean platelet volume associates with worse clinicopathologic features and prognosis of patients with invasive breast cancer.

BACKGROUND: Mean platelet volume (MPV) is one of the four platelet parameters (platelet count, MPV, platelet distribution width and plateletcrit), which indicates the activation of platelet. We aim to investigate the associations between pre-treatment MPV levels and clinical hematology parameters, pathology parameters and prognosis of patients with invasive breast cancer (IBC). METHODS: Medical records of 340 breast tumor patients (170 IBC vs. 170 breast benign tumor) were retrospectively reviewed. Patients in two groups were matched for age, body mass index, smoking status and complications. To analyze: differences in pre-treatment MPV levels between IBC group and breast benign tumor group; differences between pre- and postoperative MPV levels in IBC patients; correlations between pre-treatment MPV and clinical hematology parameters, clinicopathologic parameters and prognosis in IBC patients. RESULTS: As we analyzed, pre-treatment MPV levels of IBC patients were significantly higher than the controls (8.65 ± 0.98 vs 8.34 ± 0.78, P = 0.002), and preoperative MPV levels were significantly higher than the postoperative in IBC patients (8.65 ± 0.98 vs 8.44 ± 0.91, P = 0.042). In IBC group, pre-treatment MPV level associated, significantly, with clinical hematology parameters (platelet, fibrinogen, albumin, fasting blood glucose, P = 0.003, 0.042, 0.032, 0.046, respectively) and with clinicopathological parameters (distant metastasis, primary tumor size, tumor node metastasis stages, P = 0.039, 0.002, 0.001, respectively). Furthermore, univariate and multivariate survival analysis demonstrated that MPV was significant prognostic factor (P = 0.035, HR 1.86, 95 % confidence interval 1.06-3.25). CONCLUSION: High pre-treatment MPV level in IBC patients was a potential predictive factor and significant independent prognostic factor.

Bioconcentration of polycyclic aromatic hydrocarbons in roots of three mangrove species in Jiulong River Estuary.

The polycyclic aromatic hydrocarbons(PAHs) concentrations were determined in the root of three mangrove species (Kandelia candel, Avicennia marina and Bruguiera gymnorrhiza) and their growing environment (sediment) in mangrove wetlands of Jiulong River Estuary, Fujian, China. The total PAHs (16 parent PAHs) in mangrove sediments ranged from 193.44 to 270.53 ng/g dw, with a mean value of 231.76 +/- 31.78 ng/g dw. Compared with other mangrove and coastal marine sediments, the PAHs concentrations of all the sampling areas in this study were at relatively lower level. The total PAHs (13 parent PAHs) values varied from 30.83 to 62.73 ng/g dw in mangrove roots. Benzo[a]pyrene(five-ring), fluoranthene(four-ring) and pyrene(four-ring) dominated in mangrove sediments. Based on ratios of phenathrene/anthracene, fluoranthene/pyrene and fluoranthene/pyrene + fluoranthene, the main possible sources of surface sediment PAHs were identified as grass, wood or coal combustion for mangrove wetlands of Jiulong River Estuary. Naphthalene(two-ring) and phenathrene(three-ring) were the most abundant compounds in mangrove roots. Sediment-to-vegetation bioconcentration factors (BCF(SV) S) were calculated and their relationships with PAHs' physico-chemical properties were investigated. The average BCF(SV) S of PAHs for three mangrove species roots were almost all under the level of 1 except for naphthalene. Good linear relationship between BCF(SV) values for mangrove roots and PAHs water solubility, octanol-water partitioning coefficients was derived in present study. The solubility and the octanol-water partition coefficient were proved to be good predictors for the accumulation of PAHs in mangrove roots, respectively.

[Absorption and distribution of K, Na and Mg in Avicennia marina seedlings under cadmium stress].

In this paper, mangrove seedlings Avicennia marina were treated with various contents of cadmium (0, 0.5, 5, 25, 50, 100, 150 mg · L(-1)). These seedlings were cultivated by man-made seawater with a salinity of 15 in sand for 90 days in a greenhouse. The absorption and distribution of elements contents (K, Na and Mg) under cadmium stress were investigated at 45th and 90th day, respectively. The results showed that the enrichment of cadmium in the different components of seedlings increased with the increasing cadmium stress level and exposure time. The cadmium contents in roots and cotyledons were relatively higher than in the other components, accounting for 66.9% and 16.3% of cadmium in the seedlings under the 150 mg · L(-1) cadmium stress, respectively. The fall of cotyledons could reduce the damage of cadmium stress to the whole seedlings. The Na contents increased in roots and stems and decreased in leaves and cotyledons after cadmium stress for 90 days. The K content decreased in roots and cotyledons, while had no significant change in stems and leaves. The Mg content in roots, stems, leaves and cotyledons of seedlings treated with cadmium for 90 days were lower than those of the control, and were negatively related to the cadmium content.