OBJECTIVETo compare the ability of the polysaccharide from various Porphyromonas gingivalis (Pg) type and clinical strains in inducing THP-1 cells to produce cytokines interleukin(IL)-1β, IL-8, and tumor necrosis factor(TNF)-α, in order to analyze the immunogenicity of Pg polysaccharide components and the virulence-associated factors of this periodontal pathogen.

METHODSThe bacterial polysaccharide was extracted from high virulent Pg strains, W83, SJD2, SJD12 and low virulent Pg, ATCC33277, SJD4, SJD5, and SJD11 by phenol-water extraction. The extracted polysaccharide was used to stimulate the THP-1 cells with different simulation periods and doses. The level of the cytokines, including IL-1β,IL-8 and TNF-α in the cell culture suspension was measured by enzyme-linked immunosorbent assay(ELISA).

RESULTSThe polysaccharide extraction of Pg strains was composed of lipopolysaccharide(LPS) and capsular polysaccharide. The secretion of IL-1β, IL-8 and TNF-α, produced by the THP-1 cells showed in a time- and dose-dependent manner in the medium containing 10% fetal bovine serum. The level of these cytokines of the high virulent strains was higher than that of the low virulent strains in medium containing 1% fetal bovine serum.Four hours after stimulation with polysaccharide extracted from high virulent strains, the levels of IL-1β,IL-8, and TNF-α in the cell suspension were (1 639 ± 497), (1 648 ± 513) and (140 ± 48) µg/L, respectively, whereas for low virulent strains, the levels of IL-1β, IL-8, and TNF-α were (773 ± 382), (892 ± 400) and (67 ± 33) µg/L, respectively.

CONCLUSIONSPolysaccharide extracted from Pg could induced the THP-1 cells to secrete the cytokines of IL-1β, IL-8 and TNF-α. The level of the cytokines produced by the THP-1 cells associates with the bacterial virulent properties.

Cytokines ; metabolism ; Interleukin-1beta ; Interleukin-8 ; Lipopolysaccharides ; physiology ; Porphyromonas gingivalis ; metabolism ; Tumor Necrosis Factor-alpha

BACKGROUNDKeratinocytes play a crucial role in the biological function of skin barrier. The relationship between sodium lauryl sulfate (SLS) and keratinocytes has been studied. However, the cytotoxicity and effects of sodium dodecyl benzene sulfonate (SDBS), a common detergent similar to SLS, on keratinocytes are still not known. This study aimed to investigate the effects of SDBS on cytotoxicity and expression of proinflammatory cytokines in cultured human keratinocytes.

METHODSThis study was carried out using the keratinocytes cell line, HaCaT cells. The cytotoxicity of SDBS on HaCaT cells was evaluated with cell counting kit-8 (CCK-8) and phase-contrast microscopy. After exposure to different concentrations of SDBS, the total RNA of the HaCaT cells was extracted for evaluating the relative mRNA expression of IL-1α, IL-6, IL-8, and TNF-α by qPCR. The supernatants of cells were collected for measuring the levels of IL-6 and IL-8 by enzyme-linked immunosorbent assay (ELISA).

RESULTSSDBS at concentrations of 20 µg/ml and over showed direct cytotoxicity and induced morphological changes of the HaCaT cells. The mRNA expressions of IL-1α, IL-6, IL-8, and TNF-a in different concentrations of SDBS at different time were comparable with that of controls. SDBS at concentrations of 5, 10, and 15 µg/ml had no significant effects on IL-6 and IL-8 excretion from HaCaT cells after 24-hour exposure. Moreover, no significant effects on the IL-6 and IL-8 excretion were found after 10 and 15 µg/ml SDBS stimulations for 6, 12, and 24 hours, respectively.

CONCLUSIONSDBS at higher concentrations had cytotoxicity on HaCaT cells but had no effects on the mRNA expression of IL-1α, IL-6, IL-8, and TNF-a, that was different from SLS.

Benzenesulfonates ; pharmacology ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-1alpha ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Keratinocytes ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the role of CCL20/CCR6/Th17 axis in vascular invasion and metastasis of primary hepatocellular carcinoma (HCC).

METHODSExpression levels of CCL20 mRNA in the normal human liver cell line L-02, and human hepatocellular carcinoma cell lines Hep3B, Huh7 and HepG2 were quantified by using SYBR green real time PCR. CCL20 secretions from these cell lines were quantified by using ELISA. The chemotactic effect of HCC cell line Hep3B on human peripheral blood mononuclear cells was determined by using transwell chemotaxis assay. Pre-therapy serum levels of IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-17, IL-23, IFN-γ, TNF-α and CCL20 in 93 patients with HCC were measured by using 9-plex array and ELISA. All the patients were chronic hepatitis B virus associated HCC, and 51 cases were those with vascular invasion and metastasis (metastasis group) and 42 cases were not (non-metastasis group). CCL20 and CCR6 mRNA expressions in the HCC and tumor-adjacent tissues were determined by using SYBR Green real time PCR in 41 patients, among them, 20 cases were from the group of patients with metastasis and 21 cases were from the group of patients without metastasis. The CCL20 expression was further determined by immunohistochemistry.

RESULTSThe HCC cell lines expressed and secreted higher amount of CCL20, which effectively recruited CCR6(+) T cells. Pre-therapy serum levels of CCL20 in 93 HCC patients were (38.2 ± 28.4)pg/ml, significantly increased than those with benign hepatic hemangiomas [(7.8 ± 17.8)pg/ml, P < 0.01]. In addition, the serum levels of CCL20 were positively correlated with the tumor diameters in HCC patients (r = 0.32, P = 0.0018). CCL20 was dominantly expressed in the cytoplasm in HCC cells, and it was also expressed by some infiltrating immune cells. The mRNA expression levels of CCL20 of the tumor tissues were significantly higher than that in the tumor-adjacent tissues (P < 0.05). Multivariate logistic regression analysis showed that serum levels of IL-17 and CCL20 were independent risk factors of metastasis in HCC patients (P < 0.05 for both). CCL20 mRNA showed no statistically significant differences between patients with metastasis and without metastasis in both tumor tissues and tumor-adjacent tissues (P > 0.05 for both). But the patients with metastasis showed significantly higher expressions of CCR6 both in their tumor [5.75 (1.79, 19.13)]and tumor-adjacent tissues [7.99 (4.49, 19.54)] than those with non-metastasis [1.69 (0.76, 2.87) and 3.58 (1.84, 4.32), P < 0.05 for both].

CONCLUSIONCCL20/CCR6/Th17 axis may promote vascular invasion and metastasis hepatocellular carcinoma.

Bile Duct Neoplasms ; Carcinoma, Hepatocellular ; metabolism ; Chemokine CCL20 ; metabolism ; Humans ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Interleukin-23 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocytes, Mononuclear ; Liver Neoplasms ; metabolism ; RNA, Messenger ; Th17 Cells ; Tumor Necrosis Factor-alpha ; metabolism

Castleman's disease (CD) is a rare lymphoproliferative disorder. The etiology of CD may involve viral infection, abnormal modulation of cytokines, and angiogenesis. Human herpes virus (HHV) -8 infection and interleukin-6 (IL-6) overexpression may play key roles in the development of CD. Treatment options include surgical excision, radiation therapy, chemotherapy, antiviral therapy, and targeted therapy. No standardized treatment has been established for multicentric CD and the treatment efficacy usually is poor. Among newly available agents, the effectiveness of antiviral therapy against HHV-8 is unclear; anti-CD20 and anti-IL-6 receptor monoclonal antibodies have shown promising efficacy; thalidomide and bortezomib have shown their initial efficacy.
Castleman Disease ; etiology ; metabolism ; therapy ; Herpesvirus 8, Human ; Humans ; Interleukin-6 ; metabolism

OBJECTIVE: Schizophrenia is a disabling disorder of unknown aetiology, lacking definite diagnostic method and cure. A reliable biological marker of schizophrenia is highly demanded, for which traceable immune mediators in blood could be promising candidates. We aimed to gather the best findings of neuroinflammatory markers for first-episode psychosis (FEP). METHODS: We performed an extensive narrative review of online literature on inflammation-related markers found in human FEP patients only. RESULTS: Changes to cytokine levels have been increasingly reported in schizophrenia. The peripheral levels of IL-1 (or its receptor antagonist), soluble IL-2 receptor, IL-4, IL-6, IL-8, and TNF-α have been frequently reported as increased in FEP, in a suggestive continuum from high-risk stages for psychosis. Microglia and astrocytes establish the link between this immune signalling and the synthesis of noxious tryptophan catabolism products, that cause structural damage and directly hamper normal neurotransmission. Amongst these, only 3-hydroxykynurenine has been consistently described in the blood of FEP patients. CONCLUSION: Peripheral molecules stemming from brain inflammation might provide insightful biomarkers of schizophrenia, as early as FEP or even prodromal phases, although more time- and clinically-adjusted studies are essential for their validation.
Astrocytes ; Biomarkers ; Encephalitis ; Humans ; Interleukin-1 ; Interleukin-4 ; Interleukin-6 ; Interleukin-8 ; Metabolism ; Methods ; Microglia ; Polytetrafluoroethylene ; Psychotic Disorders ; Receptors, Interleukin-2 ; Schizophrenia ; Synaptic Transmission ; Tryptophan

OBJECTIVETo investigate the effect of different patterns of intermittent hypoxia on the levels of interleukin-6 (IL-6) and interleukin-8 (IL-8) of vascular endothelial cell.

METHODSThe human umbilical vein endothelial cells of the line ECV304 were used to set up the cells model. The experiment cell strains contained one control group and eight experimental groups, 1 constance normoxia group (control group), 3 different intermittent hypoxia (IH) degree groups (10% O₂ group, 1.5% O₂ group, 0.5% O₂ group with 12 times/h), 4 different IH frequency groups (40 times/h, 20 times/h, 12 times/h, 9 times/h and 6 times/h with 0.5%O2). The specimen were put into a program-controlled gas delivery system with different level and frequency of IH respectively. Then enzyme linked immunosorbent assay (ELISA) was used to examine the levels of IL-6 and IL-8.

RESULTSHigher level of IL-6 and IL-8 were found in 3 different IH degree groups than in control group (F were 1961.100 and 103.855 respectively, P all < 0.001). The level of IL-6 and IL-8 was gradually increased with the decreasing of IH degree. The difference of IL-6 and IL-8 levels between each two different groups in IH degree was significant (P all < 0.05). The level of IL-6 and IL-8 groups was significantly different in severe IH exposure (F were 544.396 and 149.328 respectively, P all < 0.001 in both groups). There was a trend as the IH frequency decreased, the level of IL-6 and IL-8 was tenderly increased in 20/hour group (P < 0.05), highest in 12/hour group (P < 0.05), decreased in 9/hour group (P < 0.05), and decreased further in 6/hour group (P < 0.05) in severe IH expose. There was statistical significance between each two different frequency IH groups (P < 0.05).

CONCLUSIONSIntermittent hypoxia caused significant inflammatory reaction in vascular endothelial cells. The secreted level of IL-6 and IL-8 depended on the degree of hypoxia. There was a trend as the level of IH frequency decreased, the level of IL-6 and IL-8 gradually increased at first, and then decreased in the same way.

Cell Hypoxia ; Cell Line ; Endothelium, Vascular ; metabolism ; Humans ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Umbilical Veins ; cytology ; metabolism

OBJECTIVETo investigate the effect of one of the acute-phase proteins, fibrinogen, on the release of IL-1beta and -8 by human peripheral polymorphonuclear leukocytes (PMN) and the possible role of fibrinogen during the destruction of periodontium.

METHODSPeripheral PMN were isolated by discontinuous density gradient centrifuging technique. The freshly isolated PMN were suspended in Hank's balanced saline solution (1 x 10(9)/L) supplemented with 0.5% BSA and 0.1% glucose. The levels of IL-1beta and -8 in the supernatants produced by cultured cells upon the addition of human fibrinogen at different concentrations were measured by ELISA technique.

RESULTSIncubated with human fibrinogen at 2 g/L or 10 g/L for different time periods, human peripheral PMN released significantly greater amount of IL-1beta [(10.41 +/- 0.37) - (35.86 +/- 0.30) ng/L or (22.81 +/- 0.45) - (57.77 +/- 2.08) ng/L] and IL-8 [(93.90 +/- 13.95) - (2045.66 +/- 53.03) ng/L or (115.02 +/- 10.61) - (3858.69 +/- 25.65) ng/L] than PMN without the stimulation of fibrinogen (IL-1beta, P < 0.001, and IL-8, P < or = 0.016). The higher concentration of fibrinogen or the longer treatment time, the higher levels of IL-1beta and -8 were released by PMN (P < 0.001).

CONCLUSIONSFibrinogen induced the secretion of pro-inflammatory cytokines IL-1beta and -8 by PMN and may be involved in magnification of the inflammatory response of periodontium and bone resorption.

Cells, Cultured ; Fibrinogen ; pharmacology ; Humans ; Interleukin-1beta ; metabolism ; Interleukin-8 ; metabolism ; Middle Aged ; Neutrophils ; drug effects ; secretion

OBJECTIVETo evaluate the effect of nitric oxide (NO), TNF alpha, IL-6, IL-8 on the pathogenesis of infertile men.

METHODSNO, TNF alpha, IL-6 and IL-8 levels were assayed in 80 infertile men and 26 normal men.

RESULTSThe levels of NO, TNF alpha, IL-6 and IL-8 in infertile men were all significantly higher than those in controls (P < 0.01 or P < 0.05). NO was positively correlated with TNF alpha, IL-6 and IL-8 (P < 0.01).

CONCLUSIONNO, TNF alpha, IL-6 and IL-8 might play an important role in the pathogenic process of infertile.

Adult ; Cytokines ; metabolism ; Humans ; Infertility, Male ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Male ; Middle Aged ; Nitric Oxide ; metabolism ; Semen ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVEIt is demonstrated that excessive activation of NF-κB is central to the pathogenesis of inflammatory bowel disease (IBD). Zinc finger protein A20 (A20) is a key player in the negative feedback regulation of NF-κB signaling in response to multiple stimuli and has been described as central gatekeeper in inflammation and immunity. Mice genetically deficient in A20 develop severe intestinal inflammation and have increased susceptibility to dextran sodium sulfate (DSS)-induced colitis. Few studies have been done to explore the role of A20 in the pathogenesis of IBD. To clarify the relationship between intestinal inflammation and the expression level of A20 in IBD patients, the expression level of A20 and a series of inflammatory cytokines, such as NF-κB, IL-6, and IL-8, in children with IBD and controls were examined.

METHODTerminal ileal mucosal samples were obtained via endoscopy. Fifty-seven mucosal samples were divided into 4 groups: normal control group (n = 16), IBD remission group (n = 12), IBD active group (n = 13) and non-IBD enteritis group (n = 16). According to disease activity index scores, the IBD patients were divided into IBD remission group and IBD active group. Normal control group was consisted of patients with functional bowel disorders or intestinal polyps. Non-IBD enteritis was defined as changes in which endoscopy and histological examination showed inflammatory changes but could not be diagnosed as IBD. Real-time PCR was adopted for detecting the mRNA levels of A20, IL-6 and IL-8. Meanwhile immunohistochemistry was performed to measure the expression of A20 and NF-κB.

RESULT(1) The expression of A20 and NF-κB were very low in normal control group, but significantly up-regulated in IBD active group and non-IBD enteritis group (P < 0.01 for both); (2) Compared with normal control group, expression of NF-κB [(9.35 ± 4.84)% vs. (0.57 ± 0.44)%, P < 0.01], IL-6 (t' = 1.34, P > 0.05), IL-8 (t = 1.38, P > 0.05) increased in IBD remission group, while the expression of A20 in both mRNA (t = 1.03, P > 0.05) and protein levels [(0.36 ± 0.18)% vs. (0.87 ± 0.29)%, P < 0.01] decreased; (3) Compared with non-IBD enteritis group, although the expression of NF-κB [(24.17 ± 11.27)% vs. (55.29 ± 21.84)%, P < 0.01], IL-6 (t = 2.22, P < 0.05), IL-8 (t = 2.97, P < 0.01) were highly increased in IBD active group, the expression of A20 in both mRNA(t = 2.26, P < 0.05) and protein levels [(29.23 ± 11.70)% vs. (16.81 ± 5.90)%, P < 0.01]significantly decreased; (4) The expression of IL-6, IL-8 were similar in IBD remission group and non-IBD enteritis group (both P > 0.05), but the expression of A20 was much lower in both mRNA (t = 4.42, P < 0.01) and protein levels [(29.23 ± 11.70)% vs. (0.47 ± 0.25)%, P < 0.01] in IBD remission group.

CONCLUSIONThe results demonstrate that there is an excessive inflammatory response but insufficient up-regulation of A20 expression in IBD patients. Low levels expression of A20 may play an important role in the pathogenesis of IBD.

Case-Control Studies ; Child ; Endopeptidases ; metabolism ; Female ; Humans ; Inflammatory Bowel Diseases ; metabolism ; pathology ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Intestinal Mucosa ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism

BACKGROUNDVulvovaginal candidiasis is caused by Candida albicans. The vaginal epithelium, as the first site of the initial stage of infection by pathogens, plays an important role in resisting genital tract infections. Moreover, lactobacilli are predominant members of the vaginal microbiota that help to maintain a normal vaginal microenvironment. Therefore, Lactobacillus crispatus was explored for its capacity to intervene in the immune response of vaginal epithelial cells VK2/E6E7 to C. albicans.

METHODSWe examined the interleukin-2 (IL-2), 4, 6, 8, and 17 produced by VK2/E6E7 cells infected with C. albicans and treated with L. crispatus in vitro. The capacity of L. crispatus to adhere to VK2/E6E7 and inhibit C. albicans growth was also tested by scanning electron microscopy (SEM) and adhesion experiments.

RESULTSCompared with group VK2/E6E7 with C. albicans, when treated with L. crispatus, the adhesion of C. albicans to VK2/E6E7 cells decreased significantly by 52.87 ± 1.22%, 47.03 ± 1.35%, and 42.20 ± 1.55% under competition, exclusion, and displacement conditions, respectively. SEM revealed that the invasion of C. albicans into VK2/E6E7 cells was caused by induced endocytosis and active penetration. L. crispatus could effectively protect the cells from the virulence of hyphae and spores of C. albicans and enhance the local immune function of the VK2/E6E7 cells. The concentrations of IL-2, 6, and 17 were upregulated significantly (P < 0.01) and that of IL-8 were downregulated significantly (P < 0.01) in infected VK2/E6E7 cells treated with L. crispatus. The concentration of IL-4 was similar to that of the group VK2/E6E7 with C. albicans (24.10 ± 0.97 vs. 23.12 ± 0.76 pg/ml, P = 0.221).

CONCLUSIONSL. crispatus can attenuate the virulence of C. albicans, modulate the secretion of cytokines and chemokines, and enhance the immune response of VK2/E6E7 cells in vitro. The vaginal mucosa has a potential function in the local immune responses against pathogens that can be promoted by L. crispatus.

Candida albicans ; pathogenicity ; Cell Line, Tumor ; Epithelial Cells ; immunology ; metabolism ; microbiology ; ultrastructure ; Female ; Humans ; Interleukin-17 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Lactobacillus crispatus ; physiology ; Microscopy, Electron, Scanning ; Vagina ; cytology