To investigate the effect of cetirizine hydrochloride on the expression of neurokinin 1 receptor (NK-1R) and cytokines production induced by substance P (SP) in HaCaT cells (a human epidermal keratinocyte cell line) and dermal fibroblasts. The effect of cetirizine on the expression of NK-1R protein was detected by flow cytometry and Western blotting analysis. The modulation of cetirizine on the production of interferon (IFN)-gamma, interleukin (IL)-1beta, IL-6 and IL-8 in HaCaT cells and fibroblasts was measured by ELISA. The results showed that cetirizine significantly inhibited the expression of NK-1R in HaCaT cells and fibroblasts. SP induced the production of IFN-gamma, IL-1beta and IL-8 in both cell types. Cetirizine 1-100 micromol x L(-1) inhibited SP-induced IL-1beta and IL-8 production in HaCaT cells and fibroblasts, while had no effect on the production of IFN-gamma in both cells. Both SP and cetirizine had no effect on the secretion of IL-6 in HaCaT cells and fibroblasts. These findings suggest that cetirizine may be involved in the treatment of SP-induced skin inflammation by inhibiting the expression of substance P receptor and regulation the production of IL-1beta and IL-8 in epidermal keratinocyte and dermal fibroblasts.
Anti-Allergic Agents ; pharmacology ; Cell Line ; Cetirizine ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Histamine H1 Antagonists, Non-Sedating ; pharmacology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-8 ; metabolism ; Keratinocytes ; cytology ; metabolism ; Receptors, Neurokinin-1 ; metabolism ; Substance P ; pharmacology

BACKGROUNDT cell immune abnormalities in patients with dilated cardiomyopathy (DCM) has been intensively studied over the past 10 years. Our previous study has suggested that immunization of mice with the peptides derived from human adenine nucleotide translocator (ANT) result in the production of autoantibodies against the ANT and histopathological changes similar to those in human DCM. The ANT peptides can induce autoimmune cardiomyopathy like DCM in Balb/c mice. In this study we aimed to focus on the molecular mechanism of T cells in the autoimmune cardiomyopathy mouse model by detecting the expression of the two T cell signaling molecules.

METHODSThe ANT peptides were used to cause autoimmune cardiomyopathy in Balb/c mice. Anti-L3T4 or rat anti-mouse IgG was administered to the mice (n = 6 in each group) simultaneously immunized with ANT. ELISA analysis was used to detect autoantibodies against the ANT peptides and the percentages of interferon-gamma and interleukin-4 producing cells among splenic CD4(+) lymphocytes was determined by using flow cytometry analysis. The expression of CD45 in spleen T cells was determined by immunohistochemistry and the mRNAs of T cell signaling molecules were detected by real-time PCR.

RESULTSTreatment of ANT immunized Balb/c mice with anti-CD4 mAb caused a reduction in the gene expression of P56lck and Zap-70 and a lower level of CD45 expression by spleen T cells. Also, a reverse of the Th1/Th2 ratio that results in the reduced production of antibodies against ANT was found in the anti-CD4 monoclonal antibodies (mAb) group. Whereas irrelevant antibody (rat anti-mouse IgG) did not suppress T cell signaling molecules nor inhibit CD45 expression, and control-antibody mice did not show any significant differences compared with the DCM group.

CONCLUSIONThe results show that anti-CD4 mAb is a powerful inhibitor of the early initiating events of T cell receptor (TCR) signal transduction in mouse autoimmune dilated cardiomyopathy.

Adenine Nucleotide Translocator 1 ; immunology ; Animals ; Antibodies, Monoclonal ; therapeutic use ; Autoantibodies ; blood ; Autoimmune Diseases ; therapy ; CD4 Antigens ; immunology ; Cardiomyopathy, Dilated ; immunology ; therapy ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; biosynthesis ; Leukocyte Common Antigens ; analysis ; Mice ; Mice, Inbred BALB C ; Receptors, Antigen, T-Cell ; antagonists & inhibitors ; physiology ; Signal Transduction

CD137, which is expressed on activated T cells, plays a critical role in inflammatory responses. However, the exact role that CD137 plays in monocytes is not fully known. Here we studied the expression and function of CD137 in human monocytic THP-1 cells, which we found constitutively expresses CD137 at the mRNA and protein level. Cross-linking of CD137 increased the secretion of IL-8 and TNF-alpha, promoted the expression of CD54 and CD11b, and increased adhesion to extracellular matrix (ECM) proteins. In particular CD137-induced adhesion of THP-1 cells was inhibited by an inhibitor of mitogen-activated protein kinase kinase (MEK), but not by a p38 kinase inhibitor. Taken together, these results show that the adhesion and cytokine production of THP-1 cells induced by CD137 occur via activation of MEK, which results in the activation of ERK-1/2 signaling pathways. Therefore, this study suggests that CD137 induces an activating and migrating signal during inflammatory processes.
Antigens, CD/biosynthesis/*immunology ; Antigens, CD11/biosynthesis ; *Cell Adhesion ; Cell Adhesion Molecules/biosynthesis ; Cell Line ; Cytokines/*biosynthesis ; Enzyme Activation ; Extracellular Matrix Proteins/metabolism ; Flow Cytometry ; Humans ; Immunity, Natural ; Intercellular Adhesion Molecule-1/biosynthesis ; Interleukin-8/biosynthesis ; Mitogen-Activated Protein Kinase 1/metabolism ; Mitogen-Activated Protein Kinase 3/metabolism ; Mitogen-Activated Protein Kinase Kinases/antagonists & inhibitors/metabolism ; Monocytes/metabolism/*physiology ; Phosphorylation ; Protein Binding ; Receptors, Nerve Growth Factor/biosynthesis/*immunology ; Receptors, Tumor Necrosis Factor/biosynthesis/*immunology ; Research Support, Non-U.S. Gov't ; Signal Transduction ; Tumor Necrosis Factor-alpha/biosynthesis ; p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors

OBJECTIVETo identify an interaction between the interleukin-1 receptor antagonist gene polymorphism and risk of Alzheimer's disease.

METHODSThe study included 117 healthy controls, 85 patients with Alzheimer's disease in a Northeastern Chinese population of Han nationality. Genotypes were determined by a polymerase chain reaction amplification of the intron 2 fragment, harbouring a variable number of short tandem nucleotide sequences. Amplification products were separated on a 2% agarose gel.

RESULTSThe allele 2 frequency was 27% in healthy controls, and 21% in patients with Alzheimer's disease. Thus for allele 2 as well as for all other alleles, genotypes, or carriage rates, no significant differences compared with controls.

CONCLUSIONSNo association of interleukin-1 receptor antagonist gene polymorphism with Alzheimer's disease was identified in this population. It is also possible that the increased risk and disease modifying effects are caused by linkage disequilibrium with other genomic variants in other nearby genes.

Aged ; Alleles ; Alzheimer Disease ; genetics ; Asian Continental Ancestry Group ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Introns ; Male ; Middle Aged ; Polymorphism, Genetic ; Receptors, Interleukin-1 ; antagonists & inhibitors

OBJECTIVETo investigate the anti-HIV effects of ampelopsin and its interaction with HIV-1 coreceptor CXCR4.

METHODSThrough anti-virus experiments in vitro, the inhibitory effect of ampelopsin on HIV-1 infection was verified. Chemotaxis assay was performed to show the ability to induce PBMCs migration by ampelopsin, RANTES and SDF-1alpha. Fluorescence labelling monoclonal antibody was utilized to observe the interaction of ampelopsin and CXCR4. Mice immunosuppressant model was also established to detail the role ampelopsin played in regulating cellular immunological functions.

RESULTSAmpelopsin could protect sensitive cells against HIV-1 infection and dramatically reduce HIV-1 antigen P24 expression. HIV-1SF33 attaching to MT-4 cells was interfered by ampelopsin, and the EC50 was 0.175 mg/mL for cellular protection and 0.024 mg/mL for P24 inhibition. At co-cultivating phase, EC50 was 0.229 mg/mL and 0.197 mg/mL respectively. Furthermore, the EC50 was 0.179 mg/mL and 0.348 mg/mL in acute infection. Human PBMCs migration was induced after being challenged with ampelopsin or chemokines, and synergistic action was observed during co-treatment. Ampelopsin alone resulted in maximal chemotaxis at 1 mg/mL. HIV-1 co-receptor CXCR4 on the surface of PBMCs was decreased by internalization, which indicated the effect of ampelopsin on CXCR4. About 70% CXCR4 was reduced by ampelopsin at 1 mg/mL. Ampelopsin also augmented cellular immunological functions in immunosuppressive mice.

CONCLUSIONAmpelopsin displays a strong inhibitive role during HIV-1 absorption, incubation and acute infection. These results are coincident with its immune enhancement.

Ampelopsis ; chemistry ; Animals ; Anti-HIV Agents ; pharmacology ; Cell Line ; Chemokine CCL5 ; pharmacology ; Chemokine CXCL12 ; Chemokines, CXC ; pharmacology ; Chemotaxis, Leukocyte ; Down-Regulation ; Drugs, Chinese Herbal ; Flavonoids ; economics ; isolation & purification ; pharmacology ; HIV Infections ; virology ; HIV-1 ; drug effects ; metabolism ; pathogenicity ; Humans ; Interleukin-2 ; biosynthesis ; Leukocytes, Mononuclear ; drug effects ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Plant Roots ; chemistry ; Receptors, CXCR4 ; antagonists & inhibitors ; drug effects ; Spleen ; immunology ; T-Lymphocytes ; immunology

AIMTo study the effects of human recombinant interleukin-1 receptor antagonist (IL-1ra) on isolated trachea smooth muscle (TSM) of the guinea pig.

METHODSChanges of the tension of isolated trachea were measured by force-displacement transducer and MedLab recording system in vitro.

RESULTSIL-1ra showed direct relaxed effect on TSM in normal and ovalbumin sensitized guinea pig. The EC50 values were 8.06 x 10(-8) and 5.88 x 10(-7) mol.L-1 respectively. IL-1ra (1 x 10(-7)-1 x 10(-5) mol.L-1) concentration-dependently inhibited the contraction of TSM induced by 1 x 10(-3) mol.L-1 histamine (His), 1 x 10(-3) mol.L-1 acetylcholine (ACh) and 1 x 10(-6) mol.L-1 5-hydroxytryptamine (5-HT) (P < 0.05 or 0.01). When IL-1ra was given in advance, the contractile actions of His, ACh and 5-HT were antagonized by IL-1ra (1 x 10(-7)-1 x 10(-5) mol.L-1), the pD'2 value for His was 6.6 +/- 0.3. However, the contractile action of ACh was enhanced by IL-1ra at low concentration of 1 x 10(-9)-1 x 10(-8) mol.L-1. IL-1ra significantly prevented and inhibited the contraction of sensitized TSM induced by antigen ovalbumin, the IC50 value was 4.48 x 10(-7) mol.L-1.

CONCLUSIONThe results indicate that IL-1ra, within certain concentration, can relax the normal, contracted and sensitized ISM of the guinea pig in vitro.

Animals ; Female ; Guinea Pigs ; In Vitro Techniques ; Interleukin 1 Receptor Antagonist Protein ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; Receptors, Interleukin-1 ; antagonists & inhibitors ; Recombinant Proteins ; pharmacology ; Sialoglycoproteins ; pharmacology ; Trachea ; cytology

AIMTo study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.

METHODSThe model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.

RESULTSThe model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.

CONCLUSIONThe TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.

Animals ; Ankle Joint ; pathology ; Arthritis, Experimental ; immunology ; pathology ; Interleukin-1 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; metabolism ; Male ; Peptides ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

AIMTo evaluate the effect of interleukin-1 receptor antagonist(IL-1ra) on apoptosis and associated mechanism of eosinophil in guinea pig with asthma.

METHODSA model of guinea pig with asthma was established. After inhalation of different concentrations of IL-1ra, asthma was induced in the guinea pig for 8 days, the concentration of eosinophil cationic protein (ECP) in serum and bronchoalveolar lavage fluid (BALF), IL-5 in serum, the eosinophil counts and apoptosis were assayed by radioimmunology, enzyme-linked immunosorbent assay(ELISA), fluoromicroscope and light microscope.

RESULTSIL-1ra indirectly decreased the level of IL-5 in serum, improved the apoptosis of eosinophil(EOS) in lung, then decreased the level of ECP in serum and BALF.

CONCLUSIONInhalation of nebulized IL-1ra showed protective effect against asthma through change of the activity and infiltration of EOS in lung.

Animals ; Apoptosis ; Asthma ; blood ; chemically induced ; pathology ; Blood Proteins ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Eosinophil Granule Proteins ; Eosinophils ; drug effects ; pathology ; Female ; Guinea Pigs ; Interleukin 1 Receptor Antagonist Protein ; Interleukin-5 ; blood ; Leukocyte Count ; Male ; Ovalbumin ; Random Allocation ; Receptors, Interleukin-1 ; antagonists & inhibitors ; Ribonucleases ; blood ; metabolism ; Sialoglycoproteins ; pharmacology

OBJECTIVEThe purpose of this study was to investigate methods and expression of the recombinant human interleukin-1 receptor antagonist gene transfected into chondrocytes of temporomandibular joints (TMJ).

METHODSChondrocytes derived from the TMJ of 5-7 months old human fetus were transfected by recombinant human interleukin-1 receptor antagonist gene via cationic liposome (Lipefectine) as a medium. The expressing level of hIL-1ra protein was detected using immunocytochemistry and enzyme-linked immunosorbent assay (ELISA).

RESULTSThe positive results of the immunocytochemistry were found. The positive expression was detected in the cell plasma and the cell culture supernatant, and there was significant difference between the cells with and without gene transfection (P < 0.01).

CONCLUSIONThis feasible method provides experimented evidence for the future gene therapy of temporomandibular joint osteoarthrosis.

Chondrocytes ; cytology ; Enzyme-Linked Immunosorbent Assay ; Fetus ; Phosphatidylethanolamines ; Receptors, Interleukin-1 ; antagonists & inhibitors ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Temporomandibular Joint ; cytology ; Transfection

BACKGROUND: The human immune response to Mycobacterium tuberculosis is mediated by macrophages and T-lymphocytes. The alveolar macrophage phagocyting mycobacterium produces interleukin (IL) -1 as an inflammatory mediator, and IL-8 as a cytokine for leukocyte recruitment and granuloma formation. Interleukin-1 receptor antagonist (IL-1ra) is an internal antagonist of IL-1. METHODS: Plasma levels of IL-1ra and IL-8 and other serologic markers were measured in 18 patients with active tuberculosis before treatment and after 2 months and 6 months of treatment. RESULTS: During treatment with antituberculous medication, patients showed significant changes in hemoglobin, hematocrit, white blood cells (WBC), platelet, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), ferritin and plasma IL-1ra. After 2 months of treatment, ESR and CRP diminished significantly; after 6 months, hemoglobin increased while WBC, platelet, ESR, CRP and ferritin decreased significantly compared to their pre-treatment levels. There were two groups: patients with delayed therapeutic responses, and patients with early responses. At each point of observation, the former group of patients showed lower body weight and lower levels of hemoglobin and hematocrit, and higher levels of WBC, platelet, ESR, IL-8 and IL-1ra than the latter group. During the course of the treatment, we observed considerable differences in body weight, body mass index, hemoglobin, hematocrit, WBC and platelet counts, ESR, CRP and ferritin in both the early-response and delayed-response groups. CONCLUSION: We believe that the plasma concentrations of IL-1ra and IL-8, which showed different peaks during the course of treatment, reflected their different functions and patterns of secretion. Moreover the concentrations did not seem as sensitive as other inflammatory markers to evaluate disease activity during antituberculosis treatment. However, IL-1ra can be considered a marker for disease activity and response to treatment.
Adult ; Aged ; Aged, 80 and over ; Antitubercular Agents/therapeutic use ; Biological Markers/*blood ; C-Reactive Protein/analysis ; Comparative Study ; Female ; Human ; Interleukin-8/*blood ; Male ; Middle Aged ; Receptors, Interleukin-1/*antagonists & inhibitors ; Sialoglycoproteins/*blood ; Tuberculosis, Pulmonary/*blood/*drug therapy