bluuebunny/crossref_metadata_2025 huggingface dataset:
Oral, colonic‐release low‐molecular‐weight heparin: an initial open study of Parnaparin‐MMX for the treatment of mild‐to‐moderate left‐sided ulcerative colitis
Authors: PASTORELLI L. , SAIBENI S. , SPINA L. , SIGNORELLI C. , CELASCO G. , DE FRANCHIS R. , VECCHI M.
Published: October 2008

Background  Efficacy of heparin and low‐molecular‐weight heparins (LMWHs) in inflammatory bowel disease (IBD) treatment has been suggested. The multimatrix oral formulation MMX releases active drugs in the colon, avoiding systemic absorption. Parnaparin sodium is the LMWH chosen to be carried in the MMX formulation.

Aim  To assess the safety of three different oral dosages (70, 140 and 210 mg once daily) of Parnaparin‐MMX (CB‐01‐05) in left‐sided ulcerative colitis (UC).

Methods  Left‐sided UC patients, with a mild‐to‐moderate relapse were enrolled. All patients received Parnaparin‐MMX for 8 weeks. Clinical Activity Index (CAI), Disease Activity Index (DAI), Endoscopic Activity Index and IBD‐QoL were assessed throughout the study. A strict clinical and laboratory follow‐up, including assessment of anti‐factor Xa activity, was performed. Clinical remission was defined as CAI <4.

Results  Ten UC patients were enrolled. One patient retired for clinical deterioration. No relevant side effects, including either interference with haemostasis parameters or increased bleeding, were observed. At the end of the treatment, seven patients (70%) were in clinical remission, only one achieving endoscopic healing. Mean final CAI, DAI and IBD‐QoL scores were significantly improved from baseline.

Conclusions  Parnaparin‐MMX appears to be a safe treatment option in mild‐to‐moderate UC. Controlled studies are warranted.
Meta‐analysis: tumour invasion‐related genetic polymorphisms and gastric cancer susceptibility
Authors: GAO L. , NIETERS A. , BRENNER H.
Published: October 2008

Background  Host genetic susceptibility has been suggested as one of the most important possible explanations for interindividual difference in gastric cancer (GC) risk.

Aim  To evaluate the impact of tumour invasion‐related gene polymorphisms, which may be involved in a variety of processes during GC development, such as cell adhesion and angiogenesis, on the risk of GC.

Methods  We reviewed published studies on tumour invasion‐related gene polymorphisms and GC susceptibility until 31 March 2008, and then quantitatively summarized associations of the most widely‐studied polymorphism, CDH1−160C>A, with GC using meta‐analysis.

Results  Twenty‐seven eligible studies were included in this review. Fourteen polymorphisms significantly related to GC in at least one study were identified. For several polymorphisms, heterogeneous results were observed and associations in opposite directions were seen among Asian and Caucasian populations. In meta‐analysis, CDH1−160C>A showed an inverse association with GC among Asians (OR, 0.76; 95% CI, 0.55–1.05) and a positive association among Caucasians (OR, 1.40; 95% CI, 0.95–2.04).

Conclusions  This review suggests that genetic polymorphisms in tumour invasion could be candidate biomarkers of GC risk. However, differences between populations and stages of cancer need to be taken into account and may explain some of the inconsistencies found in previous studies.
Clinical trial: peg‐interferon alfa‐2b and ribavirin for the treatment of genotype‐1 hepatitis C recurrence after liver transplantation
Authors: LODATO F. , BERARDI S. , GRAMENZI A. , MAZZELLA G. , LENZI M. , MORELLI M. C. , TAME M. R. , PISCAGLIA F. , ANDREONE P. ,   BOLOGNA LIVER TRANSPLANTATION GROUP (BLTG), BALLARDINI G. , BERNARDI M. , BIANCHI F. B. , BISELLI M. , BOLONDI L. , CESCON M. , COLECCHIA A. , D’ERRICO A. , DEL GAUDIO M. , ERCOLANI G. , GRAZI G. L. , GRIGIONI W. , LORENZINI S. , PINNA A. D. , RAVAIOLI M. , RODA E. , SAMA C. , VIVARELLI M.
Published: October 2008

Background  Treatment of hepatitis C virus (HCV) recurrence after liver transplantation (LT) is difficult with low response rates.

Aim  To assess the safety and efficacy of pegylated‐interferon (PEG‐IFN) alfa‐2b + ribavirin (RBV) in patients with post‐LT recurrent genotype‐1 HCV and to establish stopping rules according to response.

Methods  Fifty‐three patients with post‐LT HCV recurrence were enrolled. Patients received PEG‐IFN alfa‐2b 1.0 μ/kg/week plus RBV 8–10 mg/kg/day for 24 weeks. Those with ‘early virological response at week 24’ (EVR24) continued treatment for 24 weeks (group A). Patients without EVR24 were randomized to continue (group B) or to discontinue (group C).

Results  Overall sustained virological response (SVR) was 26% (14/53). Alanine aminotransferase, rapid virological response, EVR12, EVR24, undetectable serum HCV‐RNA at weeks 12 (cEVR12) and 24 (cEVR24) were related to SVR. cEVR12 and cEVR24 (OR: 14.7; 95% CI: 2.02–106.4) were independent predictors of SVR. All patients with SVR, had cEVR12. No patient in groups B and C achieved end‐of‐treatment response. One patient in group B had SVR.

Conclusions  Pegylated‐interferon alfa‐2b was effective in one of four of patients with HCV genotype 1 after LT. Treatment should be discontinued in patients with no virological response at week 12. Further studies are needed to evaluate whether a longer treatment period may be beneficial in patients with ≥2 log10 drop in HCV‐RNA at week 24.
Clinical trial: effectiveness of chewing‐gum in accelerating capsule endoscopy transit time – a prospective randomized, controlled pilot study
Authors: APOSTOLOPOULOS P. , KALANTZIS C. , GRALNEK I. M. , LIATSOS C. , TSIRONIS C. , KALANTZIS N.
Published: October 2008

Background  Capsule endoscopy (CE) fails to reach the caecum in approximately 20% of patients. Data suggest that chewing‐gum, simulating sham feeding, provokes the cephalic phase of gastrointestinal (GI) motor response and may increase GI motility.

Aim  To determine whether chewing‐gum increases the ability of CE reaching the caecum.

Methods  Prospective, randomized, single‐blinded controlled trial. Ninety‐three consecutive patients were randomized either to use chewing‐gum (n = 47) or not (n = 46). All patients received the identical bowel preparation. Patients chewed one piece of gum for approximately 30 min every 2 h. Two blinded gastroenterologists examined all studies. The number of CE that reached the caecum within 8‐h, gastric transit time (GTT) and small bowel transit time (SBTT) were evaluated in all patients.

Results  The CE percentage passed into the caecum was higher in the chewing‐gum group compared with those in the other (83.0% vs. 71.7% respectively, P = 0.19). Both GTT and SBTT were significantly shorter in the chewing‐gum vs. control group [40.8 min (interquartile range: 21–61 min) vs. 56.1 min (interquartile range: 22–78 min) (P = 0.045) and 229.1 min (interquartile range: 158–282 min) vs. 266.2 min (interquartile range: 204–307 min) (P = 0.032) respectively]. Chewing‐gum did not adversely affect CE image quality.

Conclusions  Chewing‐gum significantly reduces GTT and SBTT during CE. Its use may improve the likelihood of the capsule reaching the caecum without affecting CE image quality.
Meta‐analysis: the outcome of anti‐viral therapy in HCV genotype 2 and genotype 3 infected patients with chronic hepatitis
Authors: ANDRIULLI A. , MANGIA A. , IACOBELLIS A. , IPPOLITO A. , LEANDRO G. , ZEUZEM S.
Published: October 2008

Background  Anti‐viral therapy seems more successful in HCV genotype 2 than genotype 3‐infected patients.

Aim  To report sustained virological response (SVR) rates for HCV‐2 and HCV‐3 infection.

Methods  Meta‐analyses were carried out on SVR data on 2275 patients treated for 24 weeks in eight individual trials and on 968 patients with rapid virological response (RVR) treated for 12–16 weeks or 24 weeks in four studies.

Results  After 24 weeks of therapy, SVR rates were 74% and 68%, respectively, for HCV‐2 and HCV‐3 genotype patients. Among high viraemics, SVR rate in HCV‐2 infection (75%) differed from the 58% value in HCV‐3 infection. Among low viraemic patients, respective rates were 79% and 75%. In RVR patients treated for 12–16 or 24 weeks, SVR rates in HCV‐2 infection were 83% and 84%, respectively, and in HCV‐3 infection 84% and 86%. In patients without RVR treated for 24 weeks, SVR was higher in HCV‐2, with a 17.8% weighted difference.

Conclusions  Twenty‐four weeks of therapy should remain standard duration for HCV‐2 and low viraemic HCV‐3 patients. In RVR patients, HCV‐3 patients respond to short‐treatment as well as HCV‐2 patients, irrespective of basal viraemia. Patients without RVR may need longer treatment than the recommended 24 weeks.
Long‐term effect of tacrolimus therapy in patients with refractory ulcerative colitis
Authors: YAMAMOTO S. , NAKASE H. , MIKAMI S. , INOUE S. , YOSHINO T. , TAKEDA Y. , KASAHARA K. , UENO S. , UZA N. , KITAMURA H. , TAMAKI H. , MATSUURA M. , INUI K. , CHIBA T.
Published: October 2008

Background  Little is known about long‐term outcome of tacrolimus therapy for ulcerative colitis.

Aim  To evaluate long‐term efficacy and safety of tacrolimus in Japanese patients with refractory ulcerative colitis.

Methods  Twenty‐seven patients with UC refractory to conventional therapy were administered tacrolimus with trough whole‐blood levels of 10–15 ng/mL to induce remission and 5–10 ng/mL to maintain remission. Median treatment duration was 11 months (1–39 months) and median follow‐up duration was 17 months (2–65 months). Evaluation of the clinical response was based on a modified Truelove–Witts severity index (MTWSI).

Results  Tacrolimus produced a clinical response in 21 patients (77.8%), and remission was achieved in 19 of these 21 (70.4%) within 30 days. Overall cumulative colectomy‐free survival was estimated as 62.3% at 65 months. In 18 of 19 patients treated with corticosteroids at the initiation of tacrolimus therapy, corticosteroids were discontinued or tapered. Adverse events were tremor (25.9%), renal function impairment (18.5%), infectious disease (14.8%), hot flashes (11.1%), hyperkalaemia (7.4%), headache (7.4%), epigastralgia (7.4%) and nausea (3.7%). No mortality occurred.

Conclusion  Long‐term administration of tacrolimus appears to be an effective and well‐tolerated treatment for Japanese patients with refractory ulcerative colitis.
Clinical trial: comparison of ibuprofen‐phosphatidylcholine and ibuprofen on the gastrointestinal safety and analgesic efficacy in osteoarthritic patients
Authors: LANZA F. L. , MARATHI U. K. , ANAND B. S. , LICHTENBERGER L. M.
Published: October 2008

Background  Chronic use of NSAIDs is associated with gastrointestinal (GI) toxicity that increases with age.

Aim  To evaluate the GI safety and therapeutic efficacy of ibuprofen chemically associated with phosphatidylcholine (PC) in osteoarthritic (OA) patients.

Methods  A randomized, double‐blind trial of 125 patients was performed. A dose of 2400 mg/day of ibuprofen or an equivalent dose of ibuprofen‐PC was administered for 6 weeks. GI safety was assessed by endoscopy. Efficacy was assessed by scores of analgesia and anti‐inflammatory activity. Bioavailability of ibuprofen was pharmacokinetically assessed.

Results  Ibuprofen‐PC and ibuprofen provided similar bioavailability/therapeutic efficacy. In the evaluable subjects, a trend for improved GI safety in the ibuprofen‐PC group compared with ibuprofen that did not reach statistical significance was observed. However, in patients aged >55 years, a statistically significant advantage for ibuprofen‐PC treatment vs. ibuprofen in the prevention of NSAID‐induced gut injury was observed with increases in both mean Lanza scores and the risk of developing >2 erosions or an ulcer. Ibuprofen‐PC was well tolerated with no major adverse events observed.

Conclusion  Ibuprofen‐PC is an effective osteoarthritic agent with an improved GI safety profile compared with ibuprofen in older OA patients, who are most susceptible to NSAID‐induced gastroduodenal injury.
Clinical trial: a primary‐care‐based model for the delivery of anti‐viral treatment to injecting drug users infected with hepatitis C
Authors: JACK K. , WILLOTT S. , MANNERS J. , VARNAM M. A. , THOMSON B. J.
Published: October 2008

Background  Injecting drug use is the main risk factor for hepatitis C virus (HCV) infection. Secondary‐care‐based strategies for the management of HCV do not effectively target this vulnerable population.

Aims  To evaluate the feasibility, safety and efficacy of a primary‐care‐based model for the delivery of HCV services including anti‐viral therapy to injecting drug users.

Methods  A partnership between a clinical nurse specialist employed by, and working under the supervision of, a secondary‐care‐based hepatitis service and drug workers and general practitioners. Three hundred and fifty‐three clients attending opiate substitution clinics in primary care were evaluated. Outcomes were: number of new diagnoses of HCV infection, number of clients assessed as suitable for anti‐viral treatment, and number of patients treated.

Results  174 HCV antibody positive clients were identified. Of these, 124 were chronically infected with HCV of whom only six had been previously identified. Of 118 new chronically‐infected individuals, 86 entered the care pathway, 43 were assessed as suitable for anti‐viral treatment and 30 have so far been treated. Outcomes of anti‐viral treatment are comparable with those obtained in secondary care settings.

Conclusion  A primary‐care‐based model offers a new paradigm for the treatment of HCV in injecting drug users.
Cyclosporin A increases the pulmonary eosinophilia induced by inhaledAspergillusantigen in mice
Authors: WANG J M , DENIS M , FOURNIER M , LAVIOLETTE M
Published: October 2008

We evaluated the effects of anti-inflammatory drugs in a murine model of allergic bronchopulmonary aspergillosis(ABPA). Mice instilled with 100μg of Aspergillus fumigatus antigen (intranasally, 3 days a week for 3 weeks) developed pulmonary lesions, characterized by a perivascular and peribronchial eosinophil infiltration, a bronchoalveolar lavage (BAL) eosinophilia, and elevated levels of total IgE, total IgGI and A. fumigatus–specific IgGl. Under the same conditions, groups of mice receiving a daily dose of 2 mg/kg dexamethasone showed decreased numbers of eosinophils and total cells in BAL, had less numerous eosinophils in their pulmonary infiltrates, and had lower levels of serum and BAL fluid total IgE, total IgGl and A. fumigatus–specific IgGl. Conversely, groups of mice pretreated with an immunosuppressive agent, cyclosporin A (CsA) at a dose of 50 mg/kg, three times per week, developed pulmonary lesions with enhanced lung eosinophilic influx and increased total IgE levels, both in serum and in BAL fluid. These findings show that dexamethasone potently prevents the murine immunopathologic response to A. fumigatus. The effect of CsA on this inflammatory response was paradoxical, insofar as it suggests an activation of the T helper 2 subset, which up–regulates eosinophil recruitment and IgE production.
A longitudinal study of the IgG antibody response to HIV-1 pl7gagprotein in HIV-1+ patients with haemophilia: titre and avidity
Authors: CHARGELEGUE D , O'TOOLE C M , COLVIN B T
Published: October 2008

The IgG response to HIV-1 p17 gag protein was studied for up to 6 years in 12 HIV-1-infected patients with haemophilia, who had seroconverted between 1982 and 1985. To assess any prognostic value, pl7 IgG titres were compared with p24 IgG titres. CD4 cell counts and p24 antigenacmia. p17 IgG avidity index was also examined. A strong similarity was found between the IgG titre to HIV-1 pl7 and that to p24. In patients who developed AIDS the decline in pl7 IgG titres could precede by several years the drop in CD4 cells to under 200 cells/μl; whereas some long-term asymptomatic patients (CDCII) had increasing p17 IgG titres and stable CD4 cell counts. Declining p17 and p24 IgG titres were not always associated with an increase in p24 antigenaemia. IgG titres were found to be better predictors of disease progression than CD4 cell counts or p24 antigenaemia. Patients who developed AIDS during the study were also characterized by a lower p17 IgG avidity than patients who remained asymptomatic This result suggests that IgG avidity could have prognostic relevance and be of importance for host resistance to AIDS onset.
Parainfluenza type 1 virus-infected cells are killed by both CD8+ and CD4+ cytotoxic T cell precursors
Authors: SLOBOD K S , ALLAN J E
Published: October 2008

The memory cytotoxic T lymphocyte (CTL) response to human parainfluenza type 1 virus (hPIV-1), a prominent cause ol'respiratory infection in young children, has been analysed for a panel of healthy adults. The CTL response to the parainfluenza viruses has not been investigated previously. Precursor CTL (CTLp) with activity against hPI V-1 -infected Epstein Barr virus (EBV)-transformed B lymphoblastoid target cells were found at a relatively high precursor frequency (approximately 1/ 2500–1/4700 CD8+ and CD4+ subsets respectively) in peripheral blood. Both CD4+ and CD8+ CTLp were delected by the analysis of individual microcultures set up under limiting dilution conditions from freshly isolated blood, the phenotype of the responder cell from individual wells being determined by flow cytometry. Further characterization of the CTL response demonstrated MHC restriction by the HLA-A2 glycoprotein in 3/4 HLA-A2+ donors. The presence of effective, hPIV-1-directed T cell memory may explain, in part, the protection observed in the adult population.
T cell memory specific for self and non-self antigens in rats persistently infected with Borna disease virus
Authors: ROTT O , HERZOG S , CASH E
Published: October 2008

We have studied CD4+ Thl T cell responses in Borna disease (BD). a virus-mediated immune disease of the central nervous system (CNS). and demonstrate the priming of virus-specific as well as autoreacti ve T cells specific for myelin antigens in the course of viral infection. The fate of these in vivo generated T cells was subsequently assessed by in vitro proliferation assays with lymphocytes from different lymphoid organs of diseased animals over a long period of time. Virus-specific T cell responses continuously decreased during the establishment of persistent infection and could no longer be detected after 5–6 months post infectionem, when inflammatory reactions in the brain had ceased. By contrast, autoantigen-specific T cells kept their ability to mount characteristic secondary responses—although at an overall rather low level—over long periods of time; these autoreactive T cells homed to a specific lymphoid organ, the perithymic lymph node. Our study thus describes for the first time a complete decline of virus-specific T cell memory in a persistent viral infection, and raises the question how long-lasting T cell aulorcaclivily is controlled.
Immunogenicity of the meningococcal stress protein MSP63 during natural infection
Authors: PANNEKOEK Y , SCHUURMAN I G A , DANKERT J , van PUTTEN J P M
Published: October 2008

Acute- and convalesccnt-phasc sera from 40 patients with meningococcal disease were evaluated for immunorcactivity with the meningococcal member of the hsp60 stress protein family. The IgG response was measured by ELISA, using bacterial cell lysate of the corresponding patients' strain, and purified hsp60 proteins from Neisseria meningitldis (MSP63), Escherichia coli (GroEL) and Mycobacterium bemis BCG (65K) as antigens. Analysis of the antibody responses revealed that 24/35 patients (69%) with elevated anti-meningococcal titres. generated anti-MSP63 antibodies during the time course of infection. Twelve of these patients generated antibodies specific for MSP63. in six patients anti-MSP63 levels exceeded anli-GroEL/65K antibodies. In the remaining six patients, equal levels of anti-MSP63 and anti-GroEL/65K were measured. We conclude that MSP63 is expressed and immunogenic during natural meningococcal infection, and that individual subjects have a restricted response to the antigen, resulting in the recognition of Neisseria-sp hsp60 epitopes and/or cross-reactive hsp60 determinants.
Successful primate immunization with peptides conjugated to purified protein derivative or mycobacterial heat shock proteins in the absence of adjuvants
Authors: PERRAUT R , LUSSOW A R , GAVOILLE S , GARRAUD O , MATILE H , TOLGNE C , VAN EMBDEN J , VAN DER ZEE R , LAMBERT P-H , GYSIN J , DEL GIUDICE G
Published: October 2008

We have previously shown in mice that antibodies can be induced to synthetic malaria peptides conjugated to mycobacterial antigens, such as purilied protein derivative (PPD) or heal shock proteins (hsp), and given in the absence of adjuvants after a previous priming with bacille Calmette-Guerin (BCG). In the present study we investigated this model of immunization in the non-human primates. Saimiri sciureus monkeys. Monkeys primed with BCG subcutaneously and then immunized subcutaneously with the Plasmodium falciparum sporozoite (NANP)40, synthetic peptide conjugated to PPD or mycobacterial hsp of 65 or 70 kD. in the absence of adjuvants, produced anti-peptide and anti-sporozoite IgG anybodies. Interestingly, the carrier effect of the hsp of 70 kD for the induction of anti-(NANP)40 antibodies was also observed in the absence of a previous priming with BCG. These data suggest that such a vaccination strategy may be applied to humans.
Contrasting levels ofin vitrocytokine production by rheumatoid synovial tissues demonstrating different patterns of mononuclear cell infiltration
Authors: YANNI G , WHELAN A , FEIGHERY C , QUINLAN W , SYMONS J , DUFF G , BRESNIHAN B
Published: October 2008

Synovial membrane samples obtained at knee arthroplasty from 22 patients with rheumatoid arthritis (RA) were characterized histologically. Two groups were identified. Tissue samples from 15 patients demonstrated multiple focal lymphoid aggregates of mononuclear cells (group A). Samples from the remaining seven patients demonstrated diffuse mononuclear cell infiltration (group B). Samples of each synovial membrane (0·25 g) were cultured for cytokine production. The highest levels of IL-1β and IL-6 were produced by group A tissues: 19·1 ± 19·6 ng/ml IL-1β (mean ± s.d.) and 264·4 ± 301·9 ng/ml IL-6, versus 3·8±6·6 ng/ml and 54·7±42·6 ng/ml respectively. Small quantities of IL-2 and IL-4 were measured in both groups: the levels of IL-2 in group A cultures were highest (P=0·04). Moreover, using MoAbs, the most intense cytokine staining in the tissues was detected in group A. Similar total numbers of each cell subpopulation and similar quantities of immunoglobulin and rheumatoid factor synthesis were measured in both groups. It is suggested that the presence of multiple focal lymphoid aggregates associated with higher levels of cytokine production observed in group A represent a greater degree of immunological activation, and may represent a subgroup of patients with a greater potential for articular destruction.
Antibody penetration of viable human cells. II. Anti-RNP antibodies binding to RNP antigen expressed on cell surface, which may mediate the antibody internalization
Authors: MA J , KING N , CHEN S-L , PENNY R , BREIT S N
Published: October 2008

As U1 small nuclear ribonucleoprotein (U1 snRNP2) has a crucial role in pre-mRNP splicing, the interaction of anti-RNP antibody with snRNP within viable lymphocytes may profoundly influence cell functions. We have shown that antibody can penetrate viable human lymphocytes, and anti-RNP antibodies enter more cells than other anti-nuclear antibodies or control IgG. In order to study the in vitro interaction of anti-RNP antibodies with viable cells. T lymphocytes were metabolically labelled with 35S-methionine, then incubated with the antibodies and washed. A set of 35S-labelled cell-associated snRNP polypeptides A, B′/B, C and D were found to bind to both monospecific human polyclonal anti-RNP IgG (human anti-RNP IgG) and a mouse monoclonal anti-RNP antibody (2·73), indicating that anti-RNP antibodies interacted with RNP antigen inside or/and on the surface of viable cells. To investigate antibody binding to RNP antigen on the cell surface, the cell surface proteins were either iodinated with 125I or the cells processed for immunoelectron microscopic studices after incubation with MoAb. At least seven 125I-labelled polypeptides on the cell surface were found to be immunoprecipitated by the anti-RNP MoAb which have similar molecular weights to U snRNP polypeptidcs 70K, A, B, D, E, F, and G. The immunoelectron microscopic studies showed that the gold particles formed clustered patches on the cell membrane. Further studies suggested that RNP antigen bound to the cell surface, and the RNP binding structure was probably a heterodimer receptor. This study provides evidence to suggest that anti-RNP antibody entry into viable cells may be mediated by interaction with RNP antigen expressed on the cell surface.
Relevance of classic anti-neutrophil cytoplasmic autoantibody (C-ANCA)-mediated inhibition of proteinase 3-α1-antitrypsin complexation to disease activity in Wegener's granulomatosis
Authors: DOLMAN K M , STEGEMAN C A , van de WIEL B A , HACK C E , Kr von dem BORNE A E G , KALLENBERG C G M , GOLDSCHMEDING R
Published: October 2008

In the sera of patients with Wegener's granulomatosis (WG), C-ANCA can be detected that are directed against proteinase 3 (PR3). We have previously observed that C-ANCA interfere with PR3 proteolytic activity and with complexation of PR3 with its major physiologic inhibitor, α1antitrypsin (α1AT). In the present study we investigated whether this inhibitory effect of C-ANCA on PR3-α1 AT complexation correlates with clinical activity of WG. Serial serum samples of eight consecutive patients with histologically proven relapses of WG were tested. At the moment of relapse all sera revealed inhibitory activity towards PR3-α-1 AT complexation (median 22%, range 10–59%). Disease activity score (r=0·87, P<0·02) and C-rcactive protein (CRP) levels (r= 0·66. P<0·1) correlated with C-ANCA inhibition of PR3-α1 AT complexation, while they did not correlate with the C-ANCA titre detected by indirect immunofluorcsccnce (IIF) nor with IgG anti-PR3 antibody level measured by ELISA. The inhibitory effect of C-ANCA on PR3-α1 AT complexation had risen significantly at the moment of relapse compared with values 3 months (P<0·05) and 6 months (P<0·01) before relapse. Eight patients with established WG and positive for C-ANCA but without clinical evidence of relapse served as controls. In this group no inhibitory effect of C-ANCA on PR3-α1AT complexation was observed in 7/8 patients sera. Sera of one control patient contained moderate C-ANCA inhibitory activity towards PR3-α1AT complexalion, which remained at a constant level during the 6 months period of observation. Thus, disease activity in WG appears to be more closely related to C-ANCA inhibitory activity towards PR3-α1AT complexation.
C3 and C4 gene expression and interferon-gamma-mediated regulation in human glomerular mesangial cells
Authors: SACKS S , ZHOU W , CAMPBELL R D , MARTIN J
Published: October 2008

The glomerular mesangial cell (GMC) plays a key role in the maintenance of glomerular structure and function and in the mediation of glomerular injury. To explore the potential of this cell to produce complement and react to local inflammatory signals, we studied the synthesis and regulation of the third and fourth components of complement in cultured human GMC. Using metabolic labelling and immunoprecipitation, we found that C3 and C4 polypeptide chains were synthesized and secreted by GMC. Interferon-gamma (IFN-γ) led to an increase in C4 protein synthesis, but not C3 synthesis. There was a corresponding increase in C4 mRNA in IFN-γ-activated cells, but no increase in C3 mRNA, as determined by semi-quantitative polymerase chain reaction (PCR) estimation. These results demonstrate that human GMC can synthesize C3 and C4 proteins, and that regulation of expression of the C4 gene is mediated by IFN-γ. We hypothesize that GMC production of complement could influence the clearance of immune aggregates by the kidney and the mediation of glomerular injury.
Serum glomerular binding activity is highly correlated with renal disease in MRL/lprmice
Authors: BERNSTEIN K A , BOLSHOUN D , LEFKOWITH J B
Published: October 2008

The pathogenesis of lupus nephritis is felt to be mediated by anti-DNA antibodies. However, the anti-DNA response and renal disease do not entirely correspond. We recently developed a new assay which detects immune elements based on their ability to bind glomeruli as an alternative approach to understanding the pathogenesis of this disorder. The glomerular binding activity (GBA) defined by this assay consists of immune elements containing IgG which interact specifically with renal (issue, the binding of which is DNase-inhibitable, but which do not bind to DNA directly. In the current study we assessed the relationship between GBA and renal disease in MRL/lpr mice (both untreated and cyclophosphamide-treated) and compared it with the anti-DNA assay. Both assays were highly correlated with renal disease in untreated mice in terms of proteinuria. In cyclophosphamide-treated mice, however, only a weak correlation between the anti-DNA assay and proteinuria was apparent. GBA. in contrast, was more strongly correlated with proteinuria in treated mice. This correlation improved substantially when the DNase-sensitive component of the GBA was used. GBA appeared related to, but not covariant with, the anti-DNA response. These results demonstrate that GBA is a better correlate of murine lupus nephritis than the anti-DNA assay, and suggest that the immune elements detected by this assay, the DNase-sensitive component in particular, may be pathogenically important.
The paraproteins in systemic capillary leak syndrome
Authors: ZHANG W , EWAN P W , LACHMANN P J
Published: October 2008

Systemic capillary leak syndrome (SCLS) is a rare disease characterized by episodes of collapse due to rapid transfer of considerable volumes of plasma from the intravascular to the extravascular compartment. The pathogenesis of this disease is unknown. The diagnosis is made largely on clinical grounds, and investigations arc unhelpful. The only consistent abnormality is that an IgG paraprotein is found in most patients, raising the possibility that the paraprotein may be involved in the pathogencsis of the disease. Reduction of the paraprotein level in our patient was associated with remission. Blood samples from three SCLS patients and one probable SCLS have been studied. All patients had monoclonal IgG paraproteins. The purified paraproteins were all of IgG1 subclass and had k light chains. However, they differed in size and charge. Antibodies against each of the paraproteins were raised in rabbits. Affinity-purified anti-idiotypic antibodies were tested for cross-reactivity against the other paraproteins using immunoblotting and Ouchtcrlony assay. These assays showed that the anti-idiotypic antibodies reacted only with the immunizing paraprotein and not with any of the other paraproteins. i.e. that the paraproteins do not share a common idiotype. Paraproteins did not bind to cultured endothelial cells, either unaclivated or following activation with interferon-gamma (IFN-γ). IL-2 or IL-6. In addition, we were unable to demonstrate any cytotoxicity towards cultured human endothelial cells by paraprolein alone, or in the presence of neutrophils (pronounced neutrophilia being a feature of attacks). The relationship between the paraproteins and the disease remains unclear. It is likely that additional, as yet unidcntilied. factors arc required for the paraprotein to lead to capillary leak.
The presence of immunosuppressive ‘p15E-like’ factors in the serum and urine of patients suffering from malign and benign breast tumours
Authors: STÖGER H , WILDERS-TRUSCHNIG M , SAMONIGG H , SCHMID M , BAUERNHOFER T , TIRAN A , TAS M , DREXHAGE H A
Published: October 2008

Certain types of tumours are capable of producing factors inhibiting mononuclear phagocyte chemotaxis which may contribute to defects in imtnunosurveillance. In head and neck cancer these factors are said to be related to the retroviral protein p15E. This study examines the presence of p15E-like factors in serum and urine of patients with malign and benign breast tumours. Thirty patients with breast cancer, 29 patients with benign breast masses, and 28 healthy controls were tested blindly with the monocyte polarization assay, using N-formyl-methionyl-leueylphenylalanine as chemoattractant. The low molecular weight fractions prepared of sera of the malign tumour patients inhibited the monocytc polarization significantly (mean inhibition 34%, s.d. = 12) compared with those of benign tumour patients (15%, s.d. = 7) and of controls (14%, s.d. = 6). The observed inhibitory effects on the monocyte polarization could be compensated by MoAbs reactive to p15E-related antigens. The mean difference between the polarization inhibition with and without anti-p15E adsorption (the ‘p15E-likc factor-induced inhibition’) was 25% (s.d. = 13) in the breast cancer group, compared with 7% (s.d. = 5) in the benign tumour patients and 5% (s.d. = 4) in the healthy control group. Surgical removal of the tumours resulted in a restoration of the monocytc polarization in 20/23 (87%) patients of the breast cancer group. Results testing preoperative urine samples correlated well with those of corresponding sera. These data give additional support to the concept that tumour-derived p15E-like factors are responsible for the inhibitory effect on monocytc chemolaxis in breast cancer patients, and that these factors can be found in serum as well as in urine.
IgG inhibits the increase of platelet-associated C3 stimulated by anti-platelet antibodies
Authors: NOMURA S , MIYAZAKI Y , MIYAKE T , YAMAGUCHI K , KIDO H , KAWAKATSU T , FUKUROI T , KAGAWA H , SUZUKI M , YANABU M , KOKAWA T
Published: October 2008

We investigated the increase of platelet-associated IgG and complement component 3 (C3) caused by the in vitro action of anti-platelet MoAbs, and the effect of mouse and human IgG on these events. Anti-glycoprotein IIb/IIIa and anti-glycoprotein Ib MoAbs caused a slight increase of C3. but not of platelet-associated IgG. In contrast, anti-CD9and anti-Fcγ II receptor MoAbs caused an increase of both platelet-associated C3 and IgG. In particular, three MoAbs which activated the complement system caused a marked increase of C3. When platelet-rich plasma was treated with aspirin and prostaglandin E1 before incubation with antibodies, the increase of platelet-associated IgG was inhibited in all cases. In contrast, the increase of platelet-associated C3 was scarcely influenced. These results suggest that the binding to platelets of platelet-activating antibodies caused the increased expression of IgG molecules on the platelet surface and a possible increase of platelet-associated IgG. However, the increase of platelet-associated C3 appeared to depend on specific characteristics of the antibodies tested, such as a complement-activating effect. In addition, intact mouse or human IgG inhibited the increase of platelet-associated C3 caused by complement-activating antibodies, while F(ab′)2 mouse or human IgG had no such effect. This suggested that the Fc portion of IgG may block the increase of C3 mediated by anti-platelet antibodies.
A novel flow cytometric assay for the quantification of adhesion of subsets within a heterogeneous cell population; analysis of lymphocyte function-associated antigen-1 (LFA-l)-mediated binding of bone marrow-derived primary tumour cells of patients with multiple myeloma
Authors: AHSMANN E J M , BENSCHOP R J , de GRUYL T D , FABER J A J , LOKHORST H M , BLOEM A C
Published: October 2008

In a previous study the expression of the adhesion molecule LFA-1 on tumour cells in patients suffering from multiple myeloma (MM) was correlated with growth of the malignant plasma cells in vivo. Here we describe a novel in vitro flow cytometric adhesion assay (FCAA) which, based on scatter and fluorescence properties, was used to analyse the contribution of the LFA-1/intercellular adhesion molecule-1 (ICAM-1) adhesion pathway in the binding of bone marrow (BM)-dcrived LFA-1-positive primary tumour cells of patients with MM to interferon-gamma (IFN-γ)-activated, ICAM-1-positive, human venous umbilical endothclial cells (huVEC) in vitro. To validate the FCAA. cells from different myeloma cell lines were labelled with the fluorescent dye CFDA or stained for CD38 expression, and LFA-1-mediated adhesion to IFN-γ-activated endothclial cells was quantified. Results obtained with the FCAA were compared with a conventional adhesion assay employing 51Cr-labelled cells. Statistical analysis revealed that both assays gave similar results. This allowed analysis of the contribution of LFA-1 to the adhesive potential of malignant plasma cells in bone marrow mononuclcar cells (BMMC) from MM patients to IFN-γ-activated endothclial cells. The results prove that LFA-1 expressed on bone marrow-derived plasma cells from MM patients can be used for cellular adhesion to ICAM-1 expressed on adherent growing cells, and are suggestive fora role of the LFA-1/ICAM-1 adhesion pathway in the pathophysiology of MM. The FCAA described in this study is a generally applicable assay, allowing analysis of the interaction of distinct subpopulations with in vitro grown adherent cells of different origin.
IL-2 receptor gene expression and IL-2 production by human preterm newborns' cells
Authors: BESSLER H , SIROTA L , NOTTI I , MILO T , DJALDETTI M
Published: October 2008

IL-2 receptor (IL-2R) gene expression in human umbilical cord blood mononuclear cells (CBMC) of preterm and term newborns was examined following stimulation for 18 h with phytohaemagglutinin (PHA) and compared with that of adult peripheral blood mononuclear cells (PBMC: mothers and control group). mRNA for IL-2R could not be detected in CBMC of preterm infants, whereas the mRNA levels for IL-2R found in full term neonales were similar to those observed in PBMC of adults. IL-2 activity in conditioned medium (CM) of mononuclear cells stimulated with either optimal or suboptimal PHA concentrations for 24 h and 48 h was also determined. At 24 h of stimulation, IL-2 activity found in CM obtained from CBMC of preterm and term newborns was significantly higher than that found in CM of adults' PBMC. A further enhancement of IL-2 activity (six to eight times) was observed in CM of preterm and term cells stimulated for 48 h. whereas no significant difference was found in IL-2 activity in CM from adult cells tested at the two incubation periods. The present findings may provide an additional explanation for the impaired function of the immune system, and the high susceptibility to infections observed in preterm newborns.
Increased frequency of CD4+ CD25+ regulatory T cells in the cerebrospinal fluid but not in the blood of multiple sclerosis patients
Authors: Feger U , Luther C , Poeschel S , Melms A , Tolosa E , Wiendl H
Published: October 2008

Naturally occurring CD4+ CD25+ regulatory T cells (nTreg) play a major role in controlling autoimmunity by suppressing self-reactive T cells. Multiple sclerosis (MS) is an inflammatory demyelinating disorder of the central nervous system (CNS), where T cells play a key role in orchestrating tissue damage. While CD4+ CD25+ nTreg have been investigated in peripheral blood from MS patients, little is known about their presence and possible function within the target organ, the CNS. In order to study whether these cells are present in the cerebrospinal fluid (CSF) under pathological conditions, we have analysed the frequency of CD4+ CD25+ nTreg in peripheral blood and CSF from MS patients (n = 14), patients with other neurological disorders (OND; n = 9) and compared peripheral levels with healthy controls (n = 40). We found that the frequency of CD4+ CD25+ forkhead transcription factor 3 (FOXP3)+ nTreg was significantly elevated in the CSF from MS patients (mean CSF = 4·05 ± 1·54% versus mean peripheral blood = 2·93 ± 0·94%) but not from patients with other neurological disorders (mean CSF = 3·78 ± 1·26% versus mean peripheral blood = 3·74 ± 1·4%). The frequency of nTreg in the periphery did not differ between MS patients and healthy donors; however, nTreg from MS patients showed reduced suppressive capacity.
Cytokine responses in acute and persistent human parvovirus B19 infection
Authors: Isa A , Lundqvist A , Lindblom A , Tolfvenstam T , Broliden K
Published: October 2008

The aim of this study was to characterize the proinflammatory and T helper (Th)1/Th2 cytokine responses during acute parvovirus B19 (B19) infection and determine whether an imbalance of the Th1/Th2 cytokine pattern is related to persistent B19 infection. Cytokines were quantified by multiplex beads immunoassay in serum from B19-infected patients and controls. The cytokine responses were correlated with B19 serology, quantitative B19 DNA levels and clinical symptoms. In addition to a proinflammatory response, elevated levels of the Th1 type of cytokines interleukin (IL)-2, IL-12 and IL-15 were evident at time of the initial peak of B19 viral load in a few patients during acute infection. This pattern was seen in the absence of an interferon (IFN)-γ response. During follow-up (20–130 weeks post-acute infection) some of these patients had a sustained Th1 cytokine response. The Th1 cytokine response correlated with the previously identified sustained CD8+ T cell response and viraemia. A cross-sectional study on patients with persistent B19 infection showed no apparent imbalance of their cytokine pattern, except for an elevated level of IFN-γ response. No general immunodeficiency was diagnosed as an explanation for the viral persistence in this later group. Neither the acutely infected nor the persistently infected patients demonstrated a Th2 cytokine response. In conclusion, the acutely infected patients demonstrated a sustained Th1 cytokine response whereas the persistently infected patients did not exhibit an apparent imbalance of their cytokine pattern except for an elevated IFN-γ response.
Activated signal transducer and activator of transcription-3 (STAT3) is a poor regulator of tumour necrosis factor-α production by human monocytes
Authors: Prêle C M , Keith-Magee A L , Murcha M , Hart P H
Published: October 2008

Signal transducer and activator of transcription-3 (STAT3) activation has been associated with suppressed inflammatory processes in experimental animals, murine myeloid cells and macrophage cell lines. Manipulation of STAT3 activity may therefore be a focus for pharmacological intervention of inflammatory diseases in humans. However, the ability of STAT3 to reduce the production of inflammatory mediators by activated human monocytes and macrophages has been characterized inadequately. To establish this, we used a recently optimized adenoviral approach to study the effect of overexpressed STAT3 or a transcriptionally inactive mutant STAT3 in lipopolysaccharide (LPS)-stimulated human monocytes. STAT3 activated by LPS did not directly regulate inhibitor of kappa B α (IκBα) activation or tumour necrosis factor (TNF)-α production, a process dependent on the transcriptional activity of nuclear factor kappa B (NFκB), although the transcriptional activity of STAT3 contributed to the mechanism by which interleukin (IL)-10 suppressed LPS-induced TNF-α levels. This contrasted with the efficient block in IL-10 induction of suppressor of cytokine signalling-3 (SOCS3) in monocytes infected with an adenovirus expressing mutant STAT3. These results indicate that STAT3 activation cannot directly regulate LPS-signalling in human monocytes and represents only part of the mechanism by which IL-10 suppresses TNF-α production by activated human monocytes. This study concludes that pharmacological manipulation of STAT3 transcriptional activity alone would be insufficient to control NFκB-associated inflammation in humans.
Factors involved in the generation of memory CD8+ T cells in patients with X-linked lymphoproliferative disease (XLP)
Authors: Belmonte L , Parodi C , Baré P , Malbrán A , Ruibal-Ares B , de Bracco María M de E
Published: October 2008

We have analysed the phenotype of T lymphocytes in two X-linked lymphoproliferative disease (XLP) patients with the same SH2D1A mutation differing in initial exposure to Epstein–Barr virus (EBV) and treatment. While memory T lymphocytes (with low CCR7 and CD62L expression) prevailed in both XLP patients, in patient 9, who developed acute infectious mononucleosis (AIM) and received B cell ablative treatment, the predominant phenotype was that of late effector CD8 T cells (CD27–, CD28–, CCR7–, CD62L–, CD45 RA+, perforin+), while in patient 4 (who did not suffer AIM) the prevalent phenotype of CD8 T lymphocytes was similar to that of normal controls (N) or to that of adult individuals who recovered from AIM: CD27+ , CD28+, CCR7–, CD62L–, CD45 RO+ and perforin–. CD57 expression (related to senescence) was also higher in CD8 T cells from patient 9 than in patient 4, AIM or N. Persistently high EBV viral load was observed in patient 9. The results obtained from this limited number of XLP patients suggest that events related to the initial EBV encounter (antigen load, treatment, cytokine environment) may have more weight than lack of SH2D1A in determining the long-term differentiation pattern of CD8 memory T cells.
Depletion of CD4+CD25+ regulatory T cells exacerbates sodium iodide-induced experimental autoimmune thyroiditis in human leucocyte antigen DR3 (DRB1*0301) transgenic class II-knock-out non-obese diabetic mice
Authors: Flynn J C , Meroueh C , Snower D P , David C S , Kong Y M
Published: October 2008

Both genetic and environmental factors contribute to autoimmune disease development. Previously, we evaluated genetic factors in a humanized mouse model of Hashimoto's thyroiditis (HT) by immunizing human leucocyte antigen DR3 (HLA-DR3) and HLA-DQ8 transgenic class II-knock-out non-obese diabetic (NOD) mice. DR3+ mice were susceptible to experimental autoimmune thyroiditis (EAT) induction by both mouse thyroglobulin (mTg) and human (h) Tg, while DQ8+ mice were weakly susceptible only to hTg. As one environmental factor associated with HT and tested in non-transgenic models is increased sodium iodide (NaI) intake, we examined the susceptibility of DR3+ and/or DQ8+ mice to NaI-induced disease. Mice were treated for 8 weeks with NaI in the drinking water. At 0·05% NaI, 23% of DR3+, 0% of DQ8+ and 20% of DR3+DQ8+ mice had thyroid destruction. No spleen cell proliferation to mTg was observed. Most mice had undetectable anti-mTg antibodies, but those with low antibody levels usually had thyroiditis. At 0·3% NaI, a higher percentage of DR3+ and DR3+DQ8+ mice developed destructive thyroiditis, but it was not statistically significant. However, when DR3+ mice had been depleted of CD4+CD25+ regulatory T cells prior to NaI treatment, destructive thyroiditis (68%) and serum anti-mTg antibodies were exacerbated further. The presence of DQ8 molecules does not alter the susceptibility of DR3+DQ8+ mice to NaI-induced thyroiditis, similar to earlier findings with mTg-induced EAT. Susceptibility of DR3+ mice to NaI-induced EAT, in both the presence and absence of regulatory T cells, demonstrates the usefulness of HLA class II transgenic mice in evaluating the roles of environmental factors and immune dysregulation in autoimmune thyroid disease.
Translational Mini-Review Series on Vaccines: Development and evaluation of improved vaccines against tuberculosis
Authors: Sander C , McShane H
Published: October 2008

Tuberculosis (TB) continues to be a major global health disaster, despite the widespread use of BCG and effective drug therapies. The development of an efficacious new TB vaccine would be an important component of disease control in the future. Many approaches are being utilised to enhance understanding of the requirements of a successful vaccine. Numerous vaccines are being designed and assessed in a series of animal models, with a few progressing to clinical trials. Here, the steps involved in the development and evaluation of TB vaccines will be discussed, including description of the most frequently used animal models and the processes involved in advancing vaccines to phase III trials.