Leukocyte Migration Inhibition Research Articles

Prognostic significance of tissue oxygenation changes and early signaling responses in fasciocutaneous advancement flap healing

Regional axial flaps have become mainstays of reconstructive and plastic surgery aimed at restoring function, form and integrity of various body parts after traumatic injuries, burns, tumor resection or the repair of birth defects. Such flaps remain attached to the donor site via single intact vascular pedicle, which serves as a conduit for supplying nutrients and removing waste during wound healing process. Despite advances in surgical techniques, complications associated with partial or full flap loss remains a significant clinical problem, particularly in the angiopathic population. In addition to inducing debilitating patient stress, reconstructive failures inordinately contribute to health care costs due to prolonged hospitalization, readmission and revision surgery. The knowledge about the diagnostic biomarkers that may predict the fate of a flap can greatly improve the rate and quality of ischemic flap healing, as it permits early therapeutic interventions to reduce the risk of flap loss. Twenty three 10–12 months old Sprague‐Dawley rats underwent bilateral epigastric flap reconstruction surgery. Tissue oxygen saturation was measured in vivo by optical photoacoustic imaging between 24 and 48 hours post‐surgery and 0.75×2.5cm skin biopsies as well as wound fluid (if present) were taken from the distal part of each flap. Tissue oxygenation changes, intracellular and/or wound exudate levels of matrix metalloproteinases (MMPs), various growth factors/cytokines, as well as the activation of major cell signaling pathways (JAK/STATs, c‐Src/FAK, PI3‐kinase/Akt/eNOS, PLC‐γ/PKC/H3, HO‐1/COX‐2, Ras/ERK, SAPK/JNK, MKK/p38 MAPK, Rac/PAK) including the induction of key transcription factors (Fra‐1, c‐Myc, c‐Fos, CREB1, EGR1, Sp1) and wound healing‐associated heat shock proteins (HSP27, HSP90, HSP47), have been correlated to the degree of survival of 45 superficial inferior epigastric artery (SIEA)‐based fasciocutaneous advancement flaps as assessed by planimetry analysis, Western blotting and H&E/Masson's Trichrome staining at 9th day after the surgery. Although the level of heme oxygenase‐1 (HO‐1), a cytoprotective enzyme, which inhibits leukocyte migration to inflammatory sites, and can be induced by hypoxia or various pro‐inflammatory stimuli, was uniformly increased on average by 2.77 fold in distal segments of all surviving and partially failed flaps, there was a significant correlation between the positive or negative changes in oxygen saturation percentage of middle‐to‐distal flap sections and functional flap outcomes. Impaired flap healing was significantly associated only with a loss of HSPs. Poor outcome, to a lesser extent, correlated with increased phosphorylation of p54 JNK isoform and expression of cyclooxygenase‐2 (COX‐2) as well as inducible, but not endothelial, nitric oxide synthase (NOS).Support or Funding InformationThis Research was supported by Department of Otolaryngology ‐ Head & Neck Surgery at Thomas Jefferson University

Toxicological analysis and antihyperalgesic, antidepressant, and anti-inflammatory effects of Campomanesia adamantium fruit barks

Objective: This study evaluates the anti-inflammatory, antihyperalgesic, and antidepressive potential of the hydroalcoholic extract of Campomanesia adamantium fruit barks (CAE) on rodents and determines the safety of this plant.Methods: The acute toxicity of CAE was evaluated by oral administration to female rats as single doses of 0, 500, 1000, or 2000 mg/kg body weight. General behavior and toxic symptoms were observed for 14 days. In the subacute toxicity test, male and female rats received 125 or 250 mg/kg body weight of CAE for 28 days. The oral anti-inflammatory activity of CAE was evaluated in carrageenan-induced pleurisy in male mice. The effect of treatment with CAE (100 mg/kg) for 15 days was evaluated in mechanical hyperalgesia (electronic von Frey), depressive behavior (forced swimming test), and cold hypersensitivity in spared nerve injury (SNI) model in rats.Results: No clinical signs of toxicity were observed in animals from the experimental groups during acute and subacute exposure to CAE. At pleurisy test, the oral administration of CAE significantly inhibited leukocyte migration and protein leakage at all doses tested when compared to control. Oral administration of CAE for 3–15 days significantly inhibited SNI-induced mechanical hyperalgesia and increased immobility in the forced swim test. Finally, on the 15th day, oral treatment with CAE prevented the increase in sensitivity to a cold stimulus induced by SNI.Discussion: The present study shows that C. adamantium extract has anti-inflammatory, antihyperalgesic, and antidepressive properties in rodents without causing toxicity.

Anti-Inflammatory Activity of Thymol and Estragole

The inflammatory process is usually a physiological response to injury or infection. However, this may appear exacerbated in some diseases requiring control. Promising alternative molecules are isolated from natural sources, such as Thymol, and Estragole. In this sense, the objective of this research was to assess the safety of these compounds and determine their anti-inflammatory activity. One group of mice received the compounds orally and were monitored during 30 days. Then, two group of animals received Thymol or Estragole and underwent the paw edema or air pouch assay. It was observed that the compounds do not exhibit toxicity at the doses tested and both showed significant anti-inflammatory activity, which revealed estragole in novel ways the ability to inhibit leukocyte migration. These results encourage further research of Thymol and Estragole as important molecules in situations where inflammation can cause harm to patients.

Features of influence adenosine, AMP and hyperadrenalinemiya on the immune status, metabolic enzymes of purine nucleotides and the antioxidant defense system

Administration of a large dose of adrenaline (4 mg/kg 60 min before analysis) increased blood levels of total leukocytes, lymphocytes, decreased T-cell suppressors, leukocyte migration inhibition reaction (LMIR) and NBT test, but increased the level of conjugated dienes (CD). Administration of AMPand adenosine increased levels of total leukocytes, lymphocytes, T- lymphocytes, T-helpers, decreased the level of malondialdehyde (MDA), LMIR, and T-cell suppressors. Sympathetic hyperactivation induced by administration of a large dose of adrenaline (4 mg/kg 60 min before analysis) was accompanied by an increase in heart and liver activities of glutathione peroxidase (GPx), catalase, AMP deaminase (AMPD), and adenosine deaminase (AD). Administration of AMP or adenosine caused a decrease in activities of glutathione reductase (GR), GPx, catalase, a decrease in the MDA level and an increase in activities of AMPD and AD in the heart. In the liver AMP and adenosine also caused a decrease in activities of glutathione reductase (GR), GPx, a decrease in the MDA level and an increase in activities of AMPD and AD. The data obtained suggest that administration of adrenaline, AMP, and adenosine influences activity of enzymes involved in purine nucleotide metabolism. However, in contrast to adrenaline, administration of AMP or adenosine does not provoke stress reaction.

Study the Ability of Enterotoxigenic E.coli Antigens To Induce Cellular Immune Response by Erythrocyte-Rosette Formation and Leukocyte Migration Inhibition Factor in Rabbits

Objectives: This study was aimed to investigate the ability of Enterotoxigenic E.coli and their enterotoxins to induce cellular immune response by means of erythrocyte-rosette formation and leukocyte migration inhibition factor tests. Methodology: Total of 30 stool sample was collected of patient with severe diarrhea whom attended to Al-Sadder teaching hospital in Al-Najaf city in period extended of April to June 2013. All bacterial isolates was identified according to morphological and biochemical testes and by API 20 E system, which further conformational tested for heat stable enterotoxin production by Suckling Mice Assay (SMA). three antigens (heat killed bacterial cell, crude enterotoxin and purified heat stable enterotoxin ETECST) was prepared and injected in experimental animals(rabbit) and tested in the level of systemic and mucosa (appendix and duodenum). Statistical analysis was done by least significant difference (LSD). Results: The results showed that 20% (6 isolates) was Enterotoxigenic E.coli and the immune experiments showed significant increase level of E-rosette formation in rabbit injected with heat killed antigen (50.1%, 49.7% and 53.0%) compared with control (23.1%, 31.8% and 29.6%) in blood appendix and duodenum consecutively, while high level appeared in crude enterotoxin and ETECST in appendix (41.3%) and (47.4%) consecutively. The assessment of ETECST LIF in migration percent to sensitizer was showing high significant LIF (54.5%, 47.6% and 43.3%) compared with control (82.0%, 79.5% and 79.5%) in blood appendix and duodenum consecutively. Conclusions: crud and partially purified enterotoxins have ability to induce cellular immune response in blood and mucosa. Recommendation: prepare a carrier molecule model for partially purified enterotoxins to make a vaccine against Enterotoxigenic E.coli .

Evaluation of cell mediated immune response in rabbits fed with Cucurbita maxima seeds

Cucurbita maxima seed was tested for its immunomodulatory effect by comparing with immunostimulant, Levamisole HCL using dexamethasone induced immuno suppression model in rabbits in terms of assessing cell mediated immune response. Thirty six rabbits were randomly divided into six groups of six animals in each. Group I was the untreated control. Group II was dexamethasone sodium treated. Group III was levamisole HCL treated. Group IV was treated with Cucurbita maxima seeds. Group V was levamisole and dexamethasone treated and Group VI was treated with dexamethasone and Cucurbita maxima. Levamisole HCL was given at the rate of 2.5 mg/kg subcutaneously thrice a week, dexamethasone sodium was given at the rate of 2 mg/kg intramuscularly for seven days and Cucurbita maxima was given at the rate of 1000 mg/kg orally for 10 days. To assess the cell mediated immune response, leukocyte migration immigration test, delayed type hypersensitivity test and haematological indices were carried out. The body weight and spleen weight were also recorded. The results indicated that there were significant increase in % leukocyte migration inhibition in Cucurbita maxima and dexamethasone + Cucurbita maxima treated groups as compared to control. In tuberculin sensitivity, Cucurbita maxima, dexamethasone + Cucurbita maxima, levamisole, dexamethasone + levamisole showed significant increase in skin thickness as compared to control. This study suggested that the Cucurbita maxima seed has the potential to reverse dexamethasone induced cell mediated immunosuppression in rabbits.

Toxicological analysis and anti-inflammatory effects of essential oil from Piper vicosanum leaves

This study assessed the anti-inflammatory effects of the essential oil from Piper vicosanum leaves (OPV) and evaluated the toxicological potential of this oil through acute toxicity, genotoxicity and mutagenicity tests. The acute toxicity of OPV was evaluated following oral administration to female rats at a single dose of 2 g/kg b.w. To evaluate the genotoxic and mutagenic potential, male mice were divided into five groups: I: negative control; II: positive control; III: 500 mg/kg of OPV; IV: 1000 mg/kg of OPV; V: 2000 mg/kg of OPV. The anti-inflammatory activity of OPV was evaluated in carrageenan-induced pleurisy and paw edema models in rats. No signs of acute toxicity were observed, indicating that the LD50 of this oil is greater than 2000 mg/kg. In the comet assay, OPV did not increase the frequency or rate of DNA damage in groups treated with any of the doses assessed compared to that in the negative control group. In the micronucleus test, the animals treated did not exhibit any cytotoxic or genotoxic changes in peripheral blood erythrocytes. OPV (100 and 300 mg/kg) significantly reduced edema formation and inhibited leukocyte migration analyzed in the carrageenan-induced edema and pleurisy models. These results show that OPV has anti-inflammatory potential without causing acute toxicity or genotoxicity.

Ferulic acid ethyl ester diminished Complete Freund's Adjuvant-induced incapacitation through antioxidant and anti-inflammatory activity.

Ferulic acid ethyl ester (FAEE) is a derivate from ferulic acid which reportedly has antioxidant effect; however, its role on inflammation was unknown. In this study, we investigated the orally administered FAEE anti-inflammatory activity on experimental inflammation models and Complete Freund's Adjuvant (CFA)-induced arthritis in rats. CFA-induced arthritis has been evaluated by incapacitation model and radiographic knee joint records at different observation time. FAEE (po) reduced carrageenan-induced paw edema (p < 0.001) within the 1st to 5th hours at 50 and 100 mg/kg doses. FAEE 50 and 100 mg/kg, po inhibited leukocyte migration into air pouch model (p < 0.001), and myeloperoxidase, superoxide dismutase, and catalase activities (p < 0.001) increased total thiol concentration and decreased the TNF-α and IL-1β concentrations, NO, and thiobarbituric acid reactive species. In the CFA-induced arthritis, FAEE 50 and 100 mg/kg significantly reduced the edema and the elevation paw time, a joint disability parameter, since second hour after arthritis induction (p < 0.001). FAEE presented rat joint protective activity in radiographic records (p < 0.001). The data suggest that the FAEE exerts anti-inflammatory activity by inhibiting leukocyte migration, oxidative stress reduction, and pro-inflammatory cytokines.

Anti-inflammatory and antinociceptive effects of racemic goniothalamin, a styryl lactone.

The present study aimed to further investigate the anti-inflammatory activity of goniothalamin (GTN), a styryl lactone, as well as its antinociceptive effects. The anti-inflammatory activity was evaluated in models of paw edema induced by different mediators in mice and carrageenan-induced peritonitis. Evaluation of the antinociceptive effect was performed through acetic acid-induced writhing test and formalin test. Activity of GTN on gene expression levels of interleukin-1beta (IL-1β), induced nitric oxidase synthase (iNOS) and cyclooxygenase-2 (COX-2) were evaluated in vitro in lipopolysaccharide (LPS)-stimulated macrophage (RAW 264.7), as well as gene expression and protein levels of tumor necrosis factor-alpha (TNF-α). Pretreatment with GTN (300 mg/kg) significantly reduced paw edema induced by compound 48/80, prostaglandin E2, phospholipase A2 and bradykinin. GTN (10, 30 and 100mg/kg) inhibited leukocyte migration in the peritonitis model and gene expression levels of IL-1β, iNOS and TNF-α, as well as TNF-α protein levels, in LPS-stimulated macrophages, without affecting COX-2 gene expression levels. GTN inhibited nociception induced by acetic acid in the writhing model and in the formalin test, when both neurogenic and inflammatory phases were inhibited. For the first time the acute anti-inflammatory profile of GTN is characterized and its antinociceptive activity reported. The current study shows that GTN inhibits both vascular and cellular phases of inflammation, with bradykinin and PLA2 induced inflammation being the most affected by GTN. Its anti-inflammatory effects also involved the in vitro inhibition of gene expression of alarm cytokines and mediators as IL-1β, iNOS and TNF-α.

Paracrine in vivo inhibitory effects of adipose tissue–derived mesenchymal stromal cells in the early stages of the acute inflammatory response

Background aimsExcessive or unresolved inflammation leads to tissue lesions. Adipose tissue–derived mesenchymal stromal cells (AMSCs) have shown protective effects that may be dependent on the modulation of inflammation by secreted factors. MethodsWe used the zymosan-induced mouse air pouch model at two time points (4 h and 18 h) to evaluate the in vivo effects of AMSCs and their conditioned medium (CM) on key steps of the early inflammatory response. We assessed the effects of AMSCs and CM on leukocyte migration and myeloperoxidase activity. The levels of chemokines, cytokines and eicosanoids in exudates were measured by use of enzyme-linked immunoassay or radio-immunoassay. In addition, the expression of cyclooxygenase-2 and microsomal prostaglandin E synthase-1 (mPGES-1) was studied by use of Western blotting and the phosphorylation of p65 nuclear factor-κB (NF-κB) by immunofluorescence. ResultsAll inflammatory parameters were significantly reduced by CM and AMSCs to a similar extent at 4 h after zymosan injection with lower effects at 18 h. The observed inhibition of leukocyte migration was associated with reduced levels of chemokines and leukotriene B4. Interleukin-1β, interleukin-6, tumor necrosis factor-α and tumor necrosis factor–stimulated gene 6 levels were significantly decreased. The downregulation of mPGES-1 was associated with inhibition of prostaglandin E2 production. Our results suggest that these anti-inflammatory effects are related, in part, to the inhibition of NF-κB activation. ConclusionsAMSCs dampen the early process of inflammation in the zymosan-induced mouse air pouch model through paracrine mechanisms. These results support the potential utility of these cells as a source of novel treatment approaches for inflammatory pathologies.

Randomized clinical trials for mice

Stroke Treatment To ensure valid conclusions for formal drug approval, the design and analysis of clinical trials are very stringent. Llovera et al. applied the criteria of the gold-standard randomized controlled clinical trial to a preclinical investigation in mice. They tested an antibody to CD49d, which inhibits leukocyte migration into the brain, in two mouse models of stroke. Their six-center randomized controlled study showed that the antibody reduced both leukocyte invasion and infarct volume after a small cortical stroke, but had no effect on larger injuries. Sci. Transl. Med. 7 , 299ra121 (2015).

Antinociceptive and anti-inflammatory activities of the ethanolic extract of Annona vepretorum Mart. (Annonaceae) in rodents.

BackgroundAnnona vepretorum Mart. (Annonaceae) is a native tree from Caatinga (Brazilian Northeastern savanna biome), popularly known as “araticum” and “pinha da Caatinga”. In this study, we investigated the effects of the crude ethanolic extract (Av-EtOH) in models of pain and inflammation in rodents.MethodsThe evaluation of antinociceptive activity was carried out by the acetic acid-induced writhing, formalin, hot plate and tail flick tests, while paw edema induced by carrageenan or histamine, and leukocyte migration to the peritoneal cavity were used for anti-inflammatory profile. Histological analyses also were carried out.ResultsAv-EtOH (25, 50 and 100 mg/kg, p.o) significantly reduced the number of writhing (P < 0.01) and decreased (P < 0.01) the paw licking time in both phases of the formalin test. In the hot plate and tail flick tests, this extract increased the reaction time, consequently reduced painful behavior. The effects in the formalin and hot plate tests were antagonized by naloxone. Av-EtOH inhibited significantly (P < 0.01) the increase in the edema volume after administration of carrageenan and histamine. In the peritonitis test, acute pre-treatment with Av-EtOH inhibited leukocyte migration. Histological analysis showed less inflammation in the groups treated with the extract when the inflammation was induced by carrageenan or histamine.ConclusionThus, Av-EtOH has significant antinociceptive and anti-inflammatory properties, which are related probably with the activation of opioid receptors and inhibition of release of mediators of the inflammatory process. This specie is a potential target for drug discovery.

Effect of Kalpaamruthaa on immunosuppression of both humoral and cell-mediated immunity in DMBA-induced mammary tumours of rats

The present study was carried out to elucidate the protective effect of Kalpaamruthaa on improving 7,12-dimethylbenz(a)anthracene (DMBA)-induced immunosuppression of both humoral and cell-mediated immunity in mammary carcinoma-induced rats. Breast cancer was induced in rats by administering DMBA orally (25 mg/rat) as a single dose. After 90 days of induction, SA (200 mg/kg body weight) and KA (300 mg/kg body weight) were administered for 14 days, by gastric intubation. Several immunotoxicological assays such as T cell rosette delayed type hypersensitivity (DTH) response, migration inhibition factor (MIF) assay, lymphocyte proliferation assay, plaque forming cell (PFC) assay and haemagglutination assay, plaque forming cell (PFC) assay, serum soluble immune complex and cytokine production, T and B cell mitogenesis induced by Con A and nonspecific cell-mediated immunity were evaluated using phagocytosis activity and NBT reduction. In cancer-induced animals (group II), the leukocyte migration inhibition declined markedly (p < 0.001), the levels of cytokines IFN-γ and IL-2 were significantly decreased (p < 0.001) and also the antibody titre level (p < 0.001) was significantly reduced when compared with control rats. A marked decline in PFC (p < 0.001) and serum soluble immune complex (PEG) formation (p < 0.001) was also observed. Hence, the present study clearly demonstrates the immunoprotective effect of KA.

Analysis of the anti-inflammatory and chemopreventive potential and description of the antimutagenic mode of action of the Annona crassiflora methanolic extract

Context: Annona crassiflora Mart. (Annonaceae) is a medicinal plant that is widely used in folk medicine, which leads to its investigation as a potential source of new pharmacological principles.Objective: This study describes the anti-inflammatory, antiallodynic, and antimutagenic/chemopreventive activities of the leaves A. crassiflora methanolic extract. Its antimutagenic mode of action was analyzed in a plant or animal experimental model.Materials and methods: Total flavonoids were quantified by spectrophotometry at 415 nm and its composition was analyzed by 1H NMR spectra. Animals received orally, 30, 100, and 300 mg/kg of extract in both tests, carrageenan-induced paw edema and myeloperoxidase activity. Animals were treated with 100 and 300 mg/kg, in all the analyzed tests, pleural cell migration and protein exudation, carrageenan-induced cell migration into the pouch, induction of joint inflammation and carrageenan-induced allodynia response in the mouse paw. To evaluate the antimutagenic/chemopreventive activity through the Allium cepa test, we used 5, 10, and 15 mg/L of extract, and for the micronucleus test in the peripheral blood, we used the dose of 15 mg/kg.Results: The fractionation of the ethyl acetate (EA) fraction, resulting from the partition of the methanol extract of the A. crassiflora, afforded through chromatographic methods resulted in the isolation of kaempferol 3-O-β-glucoside and kaempferol 3-O-β-diglucoside. Oral treatment with 100 and 300 mg/kg of extract significantly inhibited the carrageenan-induced edema formation, with inhibitions of 53 ± 7% and 47 ± 10%; in MPO activity, the observed inhibitions were 60 ± 7% for 100 mg/kg treatment and 63 ± 7% for 300 mg/kg. The ACME reduced significantly the total leukocytes (an inhibition of 78 ± 9% with 100 mg/kg and 90 ± 7% with 300 mg/kg) and protein levels (approximately 100% inhibition with both doses) in the pleurisy model. In carrageenan-induced leukocyte migration into the pouch, the extract inhibited leukocyte migration only when administered 300 mg/kg per dose (the reduction was 43 ± 5%). Pretreatment with extract failed to reduce the zymosan-induced edema formation and did not inhibit the carrageenan-induced mechanical allodynia. Damage reduction in Allium cepa tested with different concentrations (5, 10, and 15 mg/L) was 66.17, 75.75, and 69.19% for the pre-treatment; 72.72, 33.33, and 22.22% for the simple simultaneous treatment; 100.50, 93.93, and 102.52% for the simultaneous treatment with pre-incubation; 89.39, 79.79, and 84.34%; for the post-treatment, and 86.36, 81.31, and 93.43% for the continuous treatment. The antimutagenic evaluation in the micronucleous test showed a damage reduction of 75.00 and 64.58% for the pre-treatment and simultaneous protocols, respectively. The post-treatment protocol increased the cyclophosphamide effects in 45.83%.Conclusion: These results suggest that this medicinal plant has chemopreventive and anti-inflammatory therapeutic potential.

Effect of Camphor on the Behavior of Leukocytes &lt;i&gt;In vitro&lt;/i&gt; and &lt;i&gt;In vivo&lt;/i&gt; in Acute Inflammatory Response

Purpose: To evaluate the effect of camphor on acute inflammatory response by leukocytes on chemotaxis, antiedematogenic and phagocytic activities. Methods: The effect of camphor in acute inflammatory response evaluated by neutrophil chemotaxis, ear edema induced by croton oil, activity of the enzyme myeloperoxidase (MPO) and phagocytic activity of macrophages in Swiss mice. Results: Camphor treatment did not show any cytotoxicity. Camphor at 3, 10, and 30 µg/ml doses exhibited significant (p < 0.01) reduction on leukocyte migration toward N-formyl methionyl leucyl phenylalanine fMLP. Topical treatment with camphor did not reduce significant ear edema or MPO activity at any of the doses tested. However, in contrast, oral treatment with 100 and 200 mg/kg camphor significantly (p < 0.01) reduced ear edema and myeloperoxidase (MPO) activity. Additionally, the phagocytic activity of macrophages was not affected by camphor. Conclusions: These results indicate that the anti-inflammatory activity of camphor may be related to the inhibition of leukocyte migration and antiedematogenic activity.

P-101 Anti-MAdCAM Antibody PF-00547659 in Crohnʼs Disease

The fully human anti-MAdCAM monoclonal antibody PF-00547659 is being developed to treat inflammatory bowel disease. It interrupts the a4b7 integrin-MAdCAM-1 interaction, thereby selectively inhibiting leukocyte migration from the bloodstream into the intestinal wall but not the central nervous system Patients with moderate to severe Crohn's disease (HBI >8 and hsCRP ≥5 mg/L or intestinal stoma and active disease on imaging or endoscopy) who failed prior treatment with anti-TNF and immunosuppressant (IS) agents underwent a lumbar puncture (LP) followed by 3 doses of 225 mg PF-00547659 at 4 week intervals. A 2nd LP was performed after the last dose and CSF analyzed by flow cytometry. Clinical Crohn's disease activity was measured throughout using the Harvey Bradshaw Index (HBI). The efficacy outcomes of interest were the rate of response (decrease in HBI ≥3 points) and the rate of remission (HBI <5) at week 12 in evaluable patients. All the responders in the study were eligible for enrollment into an open label extension study with up to 19 months of treatment Forty-nine patients were enrolled (28 women, 21 men) in 8 European centers; mean (SD) age = 38.1 (11.8) yr, CD duration = 12.7 (6.2) yr, hsCRP = 16.3 (19.9) mg/L (Geometric mean hsCRP = 7.0). Fourteen patients had stomas and hence were not evaluable for HBI; 29 (59%) were seropositive for antibody to the JC virus. All had failed previous treatment with thiopurines (TP) and either infliximab and/or adalimumab. Twenty-six had also failed methotrexate (MTX). Ten patients (7 evaluable) were taking IS at study entry: 7 TP and 4 MTX (1 took both TP and MTX). As observed efficacy results are shown in the Table. At week 12, the geometric mean of hsCRP declined from baseline 7.0 mg/L to 5.7 mg/L (-16%). A full induction course of the highest clinical dose of PF-00547659 did not affect CSF lymphocytes while it resulted in increased circulating b7+ lymphocytes indicating that it is pharmacologically active. Overall good safety and tolerability was observed with low rates of adverse events. All subjects have completed study treatment and 4 subjects were in follow up at the time of the targeted data extraction. Selective inhibition of white blood cell translocation to the gut with PF-00547659 in patients with moderate to severe Crohn's disease and prior treatment with anti-TNF and IS drugs is associated with high rates of disease response and remission by week 12 in the open label TOSCA study. Administration was safe, and side effects were generally those seen in this population. The results of the TOSCA study support the gut selectivity and the CNS-sparing mechanism proposed for PF-00547659.

Characterization of hyaluronan, hyaluronidase PH20, and HA synthase HAS2 in inflammation and cancer

Hyaluronan (HA, an extracellular matrix glycosaminoglycan) has been reported to have a variety of biological activities, including regulation of inflammation. While high molecular weight HA (HMW HA) is recognized to be anti-inflammatory, the activity of low molecular weight HA (LMW HA, the degradation catabolite of hyaluronidase) in inflammation is less clear. Because of reagent contamination of both hyaluronidase and HA used in many studies, many reported aspects of the associated biology need reinvestigation. There are many reports which have shown that LMW HA is pro-inflammatory; however, some recent publications raised serious doubts that LMW HA and hyaluronidase PH20 are not pro-inflammatory. Endotoxin and other contaminants in the reagents used in previous reports (i.e., bovine testicular hyaluronidase [BTH] Hyal type I-S and IV-S from Sigma) may be responsible for the observed inflammation. Further investigation has shown that the most purified BTH (type VI-S from Sigma) contains no endotoxin, but has substantial level of peptidoglycan, which is also pro-inflammatory. Caution should be taken when conducting studies of inflammation using HA and hyaluronidase that may contain endotoxin and peptidoglycan, as well as other pro-inflammatory contaminants. We have characterized HA affinity to its receptor CD44. While HMW HA binds to CD44 strongly, LMW HA degraded by PH20 hyaluronidase does not bind to CD44 on the cell surface. This may explain the observation that recombinant human hyaluronidase PH20 (rHuPH20) inhibits leukocyte migration upon inflammatory stimulation by interrupting CD44 interaction with HA, mediated by HMW HA. PEGPH20, a pegylated rHuPH20, has the same inhibitory activity on leukocyte transmigration as rHuPH20. Further investigation of leukocyte-inhibiting activity of rHuPH20 may reveal further clinical applications, such as wound healing and arthritis. Another report which is difficult to understand is that the ultra-high molecular weight HA produced by naked mole rat hyaluronan synthase-2 (nmrHAS2) contributes to cancer resistance, probably due to its specific anti-inflammatory activity. In our study, nmrHAS2 is cancer-promoting in human cancer cells, to a similar extent as human HAS2. The proposed cancer resistant activity of nmrHAS2 may apply selectively to its host animal species, the naked mole rat.

Airway remodeling is reversed by aerobic training in a murine model of chronic asthma.

The aim of this study was to investigate if the aerobic training (AT) reverses airway remodeling (AR) in an asthma model. BALB/c were divided into four groups: control (unsensitized and untrained); ovalbumin (OVA: sensitized and untrained); AT (unsensitized and trained) and OVA + AT. Allergic inflammation was induced with intraperitoneal and OVA inhalation. AT (low intensity; 5×/week; 60 min/session) was performed at 7, 15, and 30 days. Leukocyte counting in the bronchoalveolar lavage fluid; the expression of IL-5, eotaxin, RANTES, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1); AR features (airway smooth muscle, epithelium thickness, collagen and elastic fibers, mucus production); and AR inducers (transforming growing factor-beta, osteopontin, vascular endothelial growth factor). OVA induced an increase in leukocyte airway migration and increased AR features (P < 0.05). After 7 days, AT reversed the OVA-induced eosinophil and macrophage airway migration, the expression of IL-5, eotaxin, RANTES, ICAM-1, VCAM-1, and all AR inducers. However, total reversion of the AR features and inducers and airway inflammation occurred only after 15 days of AT compared with the OVA groups (P < 0.05) and the effects were maintained until the 30th day. AT reverses AR after 15 days and this effect is preceded by the inhibition of leukocyte migration and occurs simultaneously with the reduction in the expression of inflammatory mediators and AR inducers.

Is there a role for vedolizumab in the treatment of ulcerative colitis and Crohn's disease?

Inflammatory bowel disease (IBD) is an important cause of morbidity and mortality for millions of patients worldwide. Current treatment options include corticosteroids, 5-aminosalicylates, immunosuppressants, and TNFα antagonists. However, these are frequently ineffective in achieving sustained response and remission over time. At present, gastroenterologists lack safe and effective treatments if patients fail anti-TNF therapy. Vedolizumab is a promising new agent for IBD patients refractory to anti-TNF therapy. Vedolizumab is an integrin antagonist which is thought to act by reducing inflammation by selectively inhibiting leukocyte migration in the gut. Emerging evidence from clinical trials suggests a potential role for vedolizumab in both ulcerative colitis (UC) and Crohn’s disease (CD), particularly in patients who have previously failed biological therapy. The safety profile of vedolizumab appears reasonable, possibly because it has a “gut-selective” mode of action, with no reported cases of progressive multifocal leukoencephalopathy, a condition which has been linked to another integrin antagonist, natalizumab. This review discusses the available evidence for integrin antagonists and their potential role in the management of IBD.

Anethole and eugenol reduce in vitro and in vivo leukocyte migration induced by fMLP, LTB4, and carrageenan

The aim of this study was to investigate the effect of anethole (AN) and eugenol (EUG) on leukocyte migration using in vitro chemotaxis and in situ microcirculation assays. BALB/c mice were used for the in vitro chemotaxis assay, and Wistar rats for the in situ microcirculation assay. We evaluated (a) the in vitro leukocyte migration in response to chemotactic factors (formyl-methionyl-leucyl-phenylalanine [fMLP] and leukotriene B4 [LTB4]) and (b) the rolling, adhesion, and migration of leukocytes induced by an injection of carrageenan (100µg/cavity) into the scrotum of the animal. In the in vitro chemotaxis assay, AN and EUG at doses of 1, 3, 9, and 27µg/ml significantly inhibited leukocyte migration when stimulated by the chemotactic agents fMLP and LTB4. In the in situ microcirculation assay, AN at doses of 125 and 250mg/kg and EUG at a dose of 250mg/kg significantly decreased the number of leukocytes that rolled, adhered, and migrated to perivascular tissue. The results indicate that AN and EUG exert inhibitory effects on leukocyte migration, highlighting their possible use to diminish excessive leukocyte migration in the inflammatory process.