Serum proteins TGFBI, PCSK9, and CCL14 are potential biomarkers for different traditional Chinese medicine syndromes of multidrug-resistant tuberculosis.

Patients with multidrug-resistant tuberculosis (MDR-TB) tend to have a long course of anti-TB treatment and severe side effects. Traditional Chinese Medicine (TCM) has a synergistic effect in attenuation of MDR-TB. However, the lack of objective biological standards to classify and diagnose MDR-TB TCM syndromes could result in less effective TCM treatment. Therefore, in this study, we identified differentially expressed proteins (DEPs) in serum of individuals with MDR-TB TCM syndromes by applying isobaric tags for relative and absolute quantification coupled with two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2DLC-MS/MS) method and bioinformatics analysis. The functional analysis of DEPs was also performed. Additionally, DEPs among three different TCM syndromes of MDR-TB were validated by enzyme-linked immunosorbent assay (ELISA). Finally, a receiver operating characteristic (ROC) curve was performed to estimate the diagnostic ability of DEPs. A total of 71 DEPs were identified in the three different MDR-TB TCM syndrome groups such as the pulmonary Yin deficiency (PYD) syndrome group, the Hyperactivity of Fire due to Yin deficiency (HFYD) syndrome group, and the deficiency of Qi and Yin (DQY) syndrome group. The results showed that the expression level of transforming growth factor-beta-induced protein ig-h3 (TGFBI) was lower in the PYD syndrome group (p = .002), the proprotein convertase subtilisin/kexin type 9 (PCSK9) was overexpressed in the HFYD syndrome group (p < .0001), and the C-C motif chemokine ligand 14 (CCL14) expression level was reduced in the DQY syndrome group (p = .004). Our study demonstrated that serum TGFBI, PCSK9, and CCL14 may serve as potential novel biomarkers for PYD syndrome, HFYD syndrome and DQY syndrome of MDR-TB, respectively. The study provides a biological basis for MDR-TB TCM syndromes classification and can be of great significance for the treatment of different TCM syndromes.

The effect of wasabi rhizome extract on atopic dermatitis-like symptoms in HR-1 hairless mice.

We investigated the effect of wasabi rhizome extract on atopic dermatitis (AD) model mice. The wasabi extract was fed to the HR-1 hairless mice, which develop AD-like symptoms with a special diet (HR-AD diet). The extract was expected to reduce the symptoms induced. Wasabi rhizome-containing HR-AD diet (5% and 10%) reduced the scratching behavior, and the 10% wasabi rhizome HR-AD diet significantly reduced scratching behavior on days 28, 35 and 42. Plasma components (histamine, eotaxin, IgE and thymus and activation-regulated chemokine (TARC)) were decreased in the 10% wasabi rhizome HR-AD diet. In histopathological examinations (toluidine blue (T.B.), major basic protein (MBP), CD4, IL-4, IL-5, eotaxin, TARC and IgE), the wasabi rhizome-containing HR-AD diet (5% and 10%) significantly reduced the number of positive stained cells. These results suggested that the wasabi rhizome extract improved the AD-like symptoms of HR-1 hairless mice.

Preclinical safety and pharmacology of an anti-human interleukin-13 monoclonal antibody in normal macaques and in macaques with allergic asthma.

Interleukin-13 (IL-13) plays a central role in chronic airway diseases, including asthma. These studies were conducted to evaluate the safety of administration of a human anti-IL-13 monoclonal antibody (mAb) to normal macaques and in macaques with allergic asthma. In addition, serum and bronchioalveolar lavage fluid were collected from allergic cynomolgus macaques in order to identify potential surrogate markers of IL-13 pharmacology that could be useful for subsequent clinical trials. In vitro studies demonstrated that the anti-IL-13 mAb inhibited the pharmacological actions of both human and cynomolgus macaque IL-13. Allergic macaques were treated systemically with 10 mg/kg anti-IL-13 mAb 1 day prior to inhaled Ascaris suum antigen challenge. Normal macaques were dosed intravenously with anti-IL-13 once per week for 3 weeks at doses of 10 or 50 mg/kg. Treatment of macaques with the anti-IL-13 mAb was not associated with any toxicologically significant findings. A slight treatment-related but nonadverse decrease in platelet counts was observed in both the normal and allergic macaques. In allergic macaques, the anti-IL-13 mAb treatment did not affect lung function, lung eosinophilia, or serum or BAL immunoglobulin E (IgE) concentrations but did produce a reduction in BAL and serum eotaxin concentrations (p < .05) at 6 h post antigen challenge. This study shows that administration of an anti-IL-13 mAb was well tolerated in both normal and allergic asthmatic macaques and that serum eotaxin concentrations may be a useful early in vivo marker for evaluating IL-13 inhibition in patients with asthma.

Changes in plasma TARC levels during Japanese cedar pollen season and relationships with symptom development.

BACKGROUND: Japanese cedar pollinosis (JCPsis) is an immunoglobulin E-mediated type I allergy caused by exposure to Japanese cedar pollen (JCP). Blood thymus and activation-regulated chemokine (TARC) levels are well known as an objective parameter for disease severity for several allergic disorders. The present study aims to evaluate the relationship between TARC levels and disease symptoms during the pollen season. METHODS: Analysis was performed of results of symptom scores and blood parameters obtained from 42 JCPsis patients who participated in a probiotic (Bifidobacterium longum BB536) intake trial in the JCP season of 2005 (January to April), using a randomized, double-blind, placebo-controlled design. RESULTS: Significant increases in plasma TARC levels were observed in subjects receiving placebo (p < 0.05 in February and p < 0.01 in March), but not in subjects receiving BB536. Increased plasma TARC levels were markedly greater in subjects who experienced severe symptoms and were thus excluded early from the intervention (placebo group: n = 8; BB536 group: n = 2). Significant differences were found in changes from baseline TARC levels in February and March between the subjects where treatment was terminated early and the remaining ones. Among the remaining subjects, significant positive correlations were found as regards changed values of TARC compared to baseline in March and April with symptom scores recorded in the pollen season. CONCLUSION: Changed values of blood TARC in the pollen season may offer promising parameters for assessing disease severity and monitoring treatment.

The effect of fexofenadine on pruritus in a mouse model (HR-ADf) of atopic dermatitis.

Fexofenadine, a histamine H1-receptor antagonist, is approved for the treatment of pruritus associated with atopic dermatitis. The effects of fexofenadine on scratching behaviour, and plasma levels of histamine and eotaxin were assessed in a new model of atopic dermatitis. Mice fed a diet low in Mg2+ and Zn2+ (special diet S) were compared with mice on a normal diet (N) or diet S plus fexofenadine HCl for weeks 0-10 (S + F(0-10)), 0-5 (S + F(0-5)) or 6 - 10 (S + F(6-10)) (seven mice per group). Compared with group N, group S mice showed significantly greater scratching frequency, and plasma histamine and eotaxin concentrations; these three variables were significantly lower in group S + F(0-10) than in group S. Scratching frequency increased when fexofenadine was discontinued. Fexofenadine significantly reduced mast cell and eosinophil numbers. Histamine may be important in the pathological changes seen in this model of atopic dermatitis, suggesting that it might aid future development of antihistamines for the treatment of atopic dermatitis.

Long-term erythropoietin therapy decreases CC-chemokine levels and intima-media thickness in hemodialyzed patients.

BACKGROUND: CC-chemokines are now widely accepted in the recruitment of leukocytes from the blood compartment into tissues, and their role in the progression of atherosclerosis has been documented. Recombinant human erythropoietin (EPO) has become widely used to treat anemic HD patients. However, little is known about the effect of EPO on the plasma CC-chemokine levels and intima-media thickness (IMT) in HD patients. METHODS: Assessment of CC-chemokines: monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory proteins (MIP-1alpha, MIP-1beta), regulated upon activation, normal T cell expressed and secreted (RANTES) and IMT were performed in 26 stable HD patients and 15 healthy controls. The patients were divided into 3 groups: group I (n = 8, without EPO), group II (n = 9, EPO at a mean dose of 76 +/- 48 U/kg/week for more than 4 months), and group III (n = 9, EPO at a mean dose of 110.5 +/- 21 U/kg/week for more than 12 months), none of them on iron therapy. RESULTS: MCP-1, MIP-1alpha, MIP-1beta and IMT values were significantly higher, whereas RANTES were significantly lower in HD patients without EPO therapy than those in healthy controls. CC-chemokine levels were found to be significantly lower in patients administered EPO when compared to subjects without EPO. In the patients treated with EPO for more than 12 months IMT values were significantly decreased compared to patients not receiving this hormone. CONCLUSION: These results suggest that long-term EPO therapy decreased CC-chemokine and IMT values in patients undergoing regular HD in the absence of concomitant iron supplementation.

Short-term effect of miglustat in every day clinical use in treatment-naïve or previously treated patients with type 1 Gaucher's disease.

In a prospective, open-label study, 25 patients with mild-to-moderate type 1 Gaucher's disease (GD1) were treated with miglustat (Zavesca), an oral glucosylceramide synthase inhibitor, over 12 months. Of the 25 patients, 10 were therapy-naïve and 15 had previously received enzyme replacement therapy (ERT). Clinical status, blood parameters, biomarkers, and organomegaly were assessed at baseline at 6 months and at 12 months. At 6 months the previously untreated patients showed a mean increase in hemoglobin of 0.77 g/dL, platelet counts improved or remaining stable, chitotriosidase and CCL18 decreased. These results were similar to those observed in 40 Spanish GD1 patients on ERT. Bone marrow infiltration cleared at 12 months. In the previously treated group, clinical and hematologic parameters and biomarkers were maintained/ improved at 12 months. Miglustat was well tolerated. The efficacy of miglustat treatment after 6 months was comparable to that of ERT.

A new synbiotic, Lactobacillus casei subsp. casei together with dextran, reduces murine and human allergic reaction.

We studied the development of atopic dermatitis-like skin lesions in NC/Nga mice and the allergic symptoms and blood patterns of healthy volunteers during the cedar (Cryptomeria japonica) pollen season in Japan following oral administration of a new synbiotic, Lactobacillus casei subsp. casei together with dextran. The combination of L. casei subsp. casei and dextran significantly decreased clinical skin severity scores and total immunoglobulin E levels in sera of NC/Nga mice that had developed picryl chloride-induced and Dermatophagoides pteronyssinus crude extract-swabbed atopic dermatitis-like skin lesions. During the most common Japanese cedar pollen season, synbiotic L. casei subsp. casei and dextran in humans led to no significant changes in total nasal and ocular symptom scores, in the levels of cedar pollen-specific immunoglobulin E, interferon-gamma and thymus and activation regulated chemokine or in the number of eosinophils in sera, whereas the placebo group showed a tendency for increased levels of cedar pollen-specific immunoglobulin E, thymus and activation regulated chemokine and number of eosinophils, and a decrease in interferon-gamma levels. Thus, the oral administration of synbiotic L. casei subsp. casei together with dextran appears to be an effective supplement for the prevention and treatment of allergic reactions.

Elevated plasma chemokine CCL18/PARC in beta-thalassemia.

Plasma CCL18/PARC, a member of the CC chemokine family, has been found to be several ten-fold increased in symptomatic Gaucher type I patients. Elevated plasma chitotriosidase levels are a well-known abnormality in Gaucher patients, however, its diagnostic use is limited by the frequent genetic deficiency in the protein. Like the situation in Gaucher disease, lipids accumulate in macrophages of patients suffering from beta-thalassemia, and, in both conditions, increased chitotriosidase levels occur. We here report that plasma CCL18/PARC is also significantly increased in patients with beta-thalassemia major (range 76.8-4977.8, median=650.8 ng/ml, n=36 and control range 10-72, median=33 ng/ml n=36 respectively, P<0.001). The CCL18/PARC levels are lower than in Gaucher patients (range 174.8-10798.7, median 2538.2 ng/ml, n=28, P<0.001). In our cohort of beta-thalassemic patients, CCL18/PARC showed a significant negative correlation to iron chelation therapy and a significant positive correlation to ferritin and chitotriosidase levels, the latter only in the patients with the wild type genotype for the enzyme. Our study demonstrates that beta-thalassemic patients have increased CCL18/PARC levels that could be of value in monitoring iron overload and compliance to therapy.

Long-term treatment with nifedipine modulates procoagulant marker and C-C chemokine in hypertensive patients with type 2 diabetes mellitus.

In type 2 diabetes mellitus, there is increased risk of nephropathy and cardiovascular complications and the incidence of renal failure increases in advanced stages of the disease. Nifedipine, a dihydropyridine-type calcium antagonist, improves endothelial function in hypercholesterolemia by enhancing nitric oxide function, and increases endothelial nitric oxide bioavailability by antioxidative mechanisms. We administered nifedipine, 50 mg/day, to the hypertensive patients for 12 months. There were no other changes in any of the patient's pharmacologic regimen during nifedipine treatment. Clinical and biochemical data obtained before and after nifedipine administration were compared. All markers were measured by ELISA. The levels of platelet activation markers (CD62P, CD63, PAC-1, and Annexin V), microparticles (PDMP and MDMP), RANTES and soluble adhesion markers (sP-selectin and sVCAM-1) differed in the control group and the hypertension group. The levels of these markers were also different in hypertensive patients with and without type 2 diabetes but were unchanged in patients without diabetes in comparison to the control group. However, the concentrations of MDMPs, chemokines, and soluble adhesion markers in hypertensive patients without type 2 diabetes decreased significantly following nifedipine treatment, although the level of RANTES was unchanged. Systolic blood pressure correlated with CD62P, CD63, annexin V, and RANTES levels, and diastolic blood pressure with CD62P and annexin V levels. The effect of nifedipine on platelet activation markers and C-C chemokines in the present study indicates potential effectiveness of calcium antagonist therapy for hypertensive patients with type 2 diabetes.

Influence of age and gender on serum eotaxin concentration in healthy and allergic people.

BACKGROUND: Eotaxin is one of the important chemokines that modulate allergic inflammation. In many studies a correlation between an elevated serum concentration of eotaxin, allergen exposure and allergic symptoms has been confirmed. Influence of other factors on eotaxin concentration is feebly recognized. We made an attempt to assess the influence of age and gender on the serum eotaxin level in healthy people and in patients with intermittent IgE-mediated rhinoconjunctivitis (AR). METHODS: The serum eotaxin level was measured in 245 healthy people and 241 patients with AR before the pollen season with the ELISA technique (KITS, R&D USA, pg/ml). The parametric tests and linear regression analysis were used in statistical calculations. RESULTS: There were no differences between the allergic group and the healthy one in the mean age (accordingly: 31.3 +/- 11.6 yrs. vs. 31.6 +/- 12.5 yrs.; p=0.1) and the mean serum eotaxin content (118.1 +/- 44.9 pg/ml vs. 116.3 +/- 34.8 pg/ml; p=0.3). A significant relationship between the serum eotaxin level, gender and age was revealed in both groups and regression models were derived. A linear correlation between age (semi-partial correlation beta = 0.47, p = 0.0000001) and gender (semi-partial correlation beta = 0.3, p = 0.0000001), on the one side, and the serum eotaxin level, on the other, was found for the allergic people. In the control group a similar relationship between the serum eotaxin level and age (semi-partial correlation coefficient beta = 0.63, p = 0.0000001) and gender (semi-partial correlation factor beta = 0.23, p = 0.000006) was observed. CONCLUSIONS: Age and sex significantly influence the serum eotaxin content in healthy people and patients with IgE-mediated rhinoconjunctivitis.

Release of inflammatory mediators in irradiated cell salvage blood and their biological consequences in human beings following transfusion.

BACKGROUND AND OBJECTIVE: Irradiation of intraoperative cell salvage blood has recently been used to inactivate tumour cells before retransfusion, during cancer surgery. No information is available about a potential inflammatory response of the recipient to the retransfusion of irradiated intraoperative cell salvage blood. This pilot study was conducted to investigate the possible release of the pro-inflammatory mediators, tumour necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), eotaxin and monocyte chemo-attractant protein-1 (MCP-1), in the serum of recipients by intraoperative retransfusion of irradiated intraoperative cell salvage blood. METHODS: Nine patients undergoing gynaecological cancer surgery were included in this study. Intraoperative cell salvage blood was irradiated with 50 Gy and retransfused to the patient. Serum and intraoperative cell salvage blood concentrations of TNF-alpha, IL-1beta, eotaxin and MCP-1 were repeatedly analysed before and after retransfusion, respectively before and after irradiation. RESULTS: Traces of mediators were detected in intraoperative cell salvage blood but no increase due to irradiation was observed. Following transfusion of intraoperative cell salvage blood, minute quantities (all < 30 pg mL(-1) of mediators were detected in the serum of patients. However, there was no significant upregulation compared to serum values before retransfusion. CONCLUSIONS: These results provide evidence that retransfusion of irradiated intraoperative cell salvage blood might represent a blood-saving strategy in cancer surgery without an immunological inflammatory response as shown by a lack of upregulation of inflammatory mediators.

Cc-chemokine eotaxin as a marker of efficacy of specific immunotherapy in patients with intermittent IgE-mediated allergic rhinoconjunctivitis.

BACKGROUND: Allergen-specific immunotherapy (SIT) is believed to be a valuable remedy in several allergic diseases; however, an accurate immunological marker of the efficacy of this treatment method has not been found yet. Cc-chemokine eotaxin, owing to its selective action on eosinophils, seems to play an important role in the pathophysiology of allergic response. The purpose of this study was to assess the usefulness of eotaxin in monitoring of SIT efficacy in patients with IgE-mediated allergic rhinoconjunctivitis. METHODS: One hundred and twenty-two patients with seasonal IgE-mediated allergic rhinoconjunctivitis due to tree- (birch, n = 42; hazel/alder, n = 14) or grass/cereal- (n = 66) pollen received allergen-specific immunotherapy. Serum eotaxin levels were determined four times in every patient, shortly before immunotherapy (Evaluation 0), immediately after the treatment (Evaluation 1), in the height of pollen season (Evaluation 2) and at least 2 weeks after the pollen season (Evaluation 3). Serum eotaxin levels were simultaneously measured in 59 healthy people from the control group. Changes in serum eotaxin levels were assessed in the healthy and allergic groups. Clinical symptoms of IgE-mediated rhinoconjunctivitis were evaluated and compared with serum eotaxin concentration changes. RESULTS: Mean values of eotaxin concentrations in serum during Evaluations 0-3 did not significantly differ in the healthy subjects and the patients with IgE-mediated rhinoconjunctivitis (P > 0.05). Moreover, no statistically significant differences in the serum eotaxin levels between the visits were observed in the patients who received immunotherapy (P > 0.05); however, immediately after immunotherapy (Evaluation 1) the mean serum level of eotaxin was lowest and closest to the serum eotaxin concentration in the control group at the same visit. No significant correlation between the mean value of the serum eotaxin level in the height of pollen season (r = -0.12, P > 0.05) or mean changes of the eotaxin levels between Evaluations 2 and 1 (r = -0.03, P > 0.05), on the one hand, and the cumulative score of symptoms and drug, on the other, were found. CONCLUSIONS: The results allow to conclude that although eotaxin plays a significant role in the mechanism of antigen-specific immunotherapy, its serum expression remains a poor marker of SIT efficacy.

Gene cloning of a new plasma CC chemokine, activating and attracting myeloid cells in synergy with other chemoattractants.

Chemokines are important mediators of cell migration during inflammation and normal leukocyte trafficking. Inflammatory chemokines are induced in multiple cell types at sites of infection. Here, we describe a novel bovine CC chemokine, designated regakine-1, that is constitutively present at high concentrations in plasma. Cloning of its gene revealed an expected two intron/three exon organization, with a rather long first intron. In addition to a 21-residue signal peptide, the coding sequence corresponded to a 71-residue secreted protein. However, the natural regakine-1 protein missed the COOH-terminal lysine residue. Regakine-1 has only weak sequence similarity (<50% identical residues) with other animal or human chemokines. Northern blot analysis demonstrated regakine-1 RNA expression in spleen and lung. At physiological concentrations (30-100 ng/mL), natural 7.5 kDa regakine-1 stimulated gelatinase B release from neutrophils and chemoattracted immature myeloid HL-60 cells, as well as mature granulocytes. Regakine-1 was more potent on human myeloid cells than the human plasma CC chemokine hemofiltrate CC chemokine-1 (HCC-1). Moreover, regakine-1 synergized with the bacterial peptide N-formylmethionylleucylphenylalanine (fMLP), yielding a 10-fold increase in neutrophil chemotactic response above their additive effect. Regakine-1 did not compete with interleukin-8 (IL-8) for binding to neutrophils, nor did it affect fMLP-induced calcium signaling, suggesting that regakine-1 recognizes a different receptor. In view of its high constitutive plasma concentration, regakine-1 is believed to recruit myeloid cells into the circulation, whereas its synergy with other neutrophil chemoattractants suggests that it also enhances the inflammatory response to infection.

[The effect of specific immunotherapy on serum eotaxin level in patients with pollinosis: preliminary studies]. / Wplyw swoistej immunoterapii na zachowanie sie stezen eotaksyny w surowicy u chorych na pylkowice--doniesienie wstepne.

Eotaxin belongs to CC chemokines with selective activity for eosinophils and basophils. Specific immunotherapy (SIT) represents the only curative approach for allergic inflammation. The aim of this study was an evaluation of the effect of SIT on serum eotaxin level in patients with seasonal allergy. 40 patients (mean age 26 +/- 8.15 with seasonal allergy to birch, 25 with seasonal allergy to grasses) and 35 persons (mean age 27 +/- 8) of the control group took part in this study. Preseasonal immunotherapy (Allergovit, Germany) was performed in conventional schedule in patients with pollinosis. Clinical assessment of symptoms in season (score) was done by all of the patients studied. ELISA test was used to measure serum eotaxin level (pg/ml, kits from R&D) before, after immunotherapy, in pollen season and after season in patients with disease. There was observed increased eotaxin concentration in patients with allergy to birch (140.6 +/- 53.7) as compared to the eotaxin level in the control group (102.1 +/- 53.7) (p < 0.05). There was observed decreased eotaxin level after immunotherapy (115.9 +/- 49.9) (p < 0.05) in patients with allergy to birch, but no with allergy to grasses. When patients with clinical improvement were taken into account, the significant difference (p < 0.05) in eotaxin concentration was showed before (126.7 +/- 52.4) and after immunotherapy (102.5 +/- 44.8). The eotaxin level in pollen season of patients with clinical improvement was significantly (p < 0.05) decreased (116.5 +/- 54.3) as compared to the eotaxin level in patients without clinical improvement (139.8 +/- 46.3). The results suggest an involvement of eotaxin in pathomechanism of SIT. The effect depends on kind of allergen evaluated. Further longitudinal controlled investigations should establish the role measuring serum eotaxin level in the clinical evaluation of SIT.