bluuebunny/crossref_metadata_2025_split huggingface dataset:
Limited role of the Toll‐like receptor‐2 in resistance to Mycobacterium avium
Authors: Gomes M. Salomé , Flórido Manuela , Cordeiro João V. , Teixeira C. Miguel , Takeuchi Osamu , Akira Shizuo , Appelberg Rui 
Published: January 2004

SummaryThe role of the Toll‐like receptor (TLR)‐2 in the generation of protective immunity to Mycobacterium avium was evaluated using gene‐disrupted mice. TLR‐2−/− mice were more susceptible than wild‐type C57Bl/6 mice to M. avium strains that were able to proliferate in vivo before the development of protective immunity and mycobacteriostasis. In contrast, the elimination of non‐virulent strains was not affected by the mutation. The generation of interferon‐γ (IFN‐γ)‐producing T cells and the expression of the interleukin‐12 p40 gene were reduced in TLR‐2‐deficient mice as compared to C57Bl/6 mice early during infection with M. avium strain 2447. The generation of protective CD4+ T cells was also compromised in the mutated mice as compared with the controls. Our data show that TLR‐2 is required for optimal immunity against certain virulent M. avium strains.
Fluvastatin treatment inhibits leucocyte adhesion and extravasation in models of complement-mediated acute inflammation
Authors: FISCHETTI F , CARRETTA R , BOROTTO G , DURIGUTTO P , BULLA R , MERONI P L , TEDESCO F 
Published: January 2004

SUMMARYComplement activation plays a relevant role in the development of tissue damage under inflammatory conditions, and clinical and experimental observations emphasize its contribution to inflammatory vasculitides. Statins have recently been shown to reduce cardiovascular morbidity independently of plasma cholesterol lowering and in vitro studies support a direct anti-inflammatory action of these drugs. The aim of this study was to verify the in vivo effect of fluvastatin on complement-mediated acute peritoneal inflammation. The effect of oral treatment with fluvastatin was investigated in normo-cholesterolaemic rats that received intraperitoneal injection of either yeast-activated rat serum (Y-act RS) or lipopolysaccharide to induce peritoneal inflammation monitored by the number of PMN recruited in peritoneal fluid washes. In addition, vascular adherence and extravasation of leucocytes were evaluated by direct videomicroscopy examination on mesentery postcapillary venules topically exposed to Y-act RS. The number of PMN in the peritoneal washes of rats treated with fluvastatin was 38% lower than that of untreated animals (P &lt; 0·05) 12  h after LPS injection, and was even lower (56%) in rats treated with Y-act RS already 8 h after injection (P &lt; 0·02). Firm adhesion to endothelium and extravasation of leucocytes evaluated under direct videomicroscopy observation were significantly inhibited in fluvastatin treated rats (77% and 72%, respectively; P &lt; 0·01), 120 min after treatment with Y-act RS. Our results demonstrate that fluvastatin inhibits in vivo complement-dependent acute peritoneal inflammation and suggest a role for statins in preventing the inflammatory flares usually associated with complement activation in chronic diseases, such as SLE or rheumatoid arthritis.
Qualitative and quantitative abnormalities in splenic dendritic cell populations in NOD mice
Authors: VASQUEZ A C , FEILI-HARIRI M , TAN R J , MOREL P A 
Published: January 2004

SUMMARYThe phenotype and function of splenic DC populations from diabetes-prone NOD mice were chara-cterized and compared to DC from diabetes-resistant strains in the presence or absence of Flt3 ligand (FL) treatment. NOD mice were found to have significantly fewer CD8α+ DC than both B10.BR and C57BL/6 mice, and this defect was reversed by FL treatment. Freshly isolated CD8α+ and CD8α– DC from all three strains were found to express similar levels of costimulatory molecules and this was similar in both FL-treated and untreated animals. IL-12 p40 production was significantly lower in purified CD11c+ DC from NOD mice compared to DC from C57BL/6 or B10.BR mice. CD8α+ DC isolated from NOD mice produced lower levels of IL-12p40 than CD8α+ DC from C57CBL/6 and this was dependent on the nature of the stimulus given. In contrast both CD8α+ and CD8α– DC from FL-treated mice produced high levels of IL-12p40 following activation, but only the CD8α– DC produced IL-12p70. Functionally, freshly isolated CD8α– DC were more stimulatory than CD8α+ DC in a primary allogeneic mixed lymphocyte reaction. However, DC maturation resulted in increased T cell stimulatory capacity for both DC subsets, and this pattern was seen in all strains. These results demonstrate significant differences in phenotype and function of splenic NOD CD8α+ DC, and further suggest that FL treatment may reverse some of these abnormalities.
Stimulation of innate immunity by susceptible and multidrug-resistant Pseudomonas aeruginosa: an in vitro and in vivo study
Authors: GIAMARELLOS-BOURBOULIS E J , PLACHOURAS D , TZIVRA A , KOUSOULAS V , BOLANOS N , RAFTOGIANNIS M , GALANI I , DONTAS I , DIONYSSIOU-ASTERIOU A , GIAMARELLOU H 
Published: January 2004

SUMMARY
In attempt to investigate the stimulatory effect of Pseudomonas aeruginosa on innate immunity and to correlate it to its level of resistance to antimicrobials, 20 isolates were applied; 8 isolates were susceptible and 12 multidrug-resistant. Genetic diversity was defined by PFGE. Human monocytes of two healthy volunteers were in vitro stimulated by the isolates for the production of pro-inflammatory (TNF-α, IL-1β, IL-6, IL-8 and IL-12) and anti-inflammatory cytokines (IL-10), of malondialdehyde and of procalcitonin. Cytokines were estimated by EIA, malondialdehyde by the thiobarbiturate assay and procalcitonin by an immunochemiluminometric assay. Survival of 48 Wistar rats was recorded after induction of sepsis by the intraperitoneal injection of three susceptible and three multidrug-resistant isolates. To test whether comparative effect of the latter isolates on survival correlates with any difference of monocyte-mediated release of pro-inflammatory mediators, monocytes of two rats were in vitro stimulated for the production of TNF-α and of malondialdehyde. In vitro stimulation of human monocytes by the susceptible isolates elicited elevated production of malondiadeheyde, of IL-1β and of IL-6 compared to stimulation by multidrug-resistant isolates. Similar differences were found for TNF-α and IL-8, but they were not statistically significant. Production of IL-10 and IL-12 was not detected after stimulation with any isolate. Levels of procalcitonin were similar after induction with either susceptible or multidrug-resistant isolates. Mean survival of animals was 7·56, 21·80 and 55·20 h, respectively, after challenge by the susceptible isolates and 28·89, 61·8 and more than 120 h, respectively, after challenge by the multidrug-resistant isolates. Differences of survival were accompanied by greater rodent monocyte-release of TNF-α and malondialdehyde after stimulation by the susceptible isolates compared to multidrug-resistant ones. It is concluded that considerable differences are encountered on the stimulation of human monocytes by susceptible and resistant isolates of Pseudomonas aeruginosa. These results correlate with in vivo evidence and might influence decision on therapeutics.
Detection of HLA class II-dependent T helper antigen using antigen phage display
Authors: SOMASUNDARAM R , SATYAMOORTHY K , CAPUTO L , YSSEL H , HERLYN D 
Published: January 2004

SUMMARYMajor histocompatibility complex (MHC) class II-dependent antigens not only activate CD4+ T helper (Th) cells, but also cytolytic T lymphocytes and effector cells of the innate immune system. These antigens therefore are candidate vaccines against cancer and infectious agents. We have developed a novel approach using a model antigen, tetanus toxoid (TT), which provides the basis for the establishment of a novel strategy of cloning Th antigens. In the TT model system, a cDNA library encoding part of the TT light chain which contained a TT-associated Th epitope recognized by TT-specific Th clones was displayed on a phage vector (TT-phage) and presented to TT-specific Th cells by autologous Epstein-Barr virus-transformed B cells (APC). These TT-phages were able to specifically activate two different TT-specific CD4+ Th cell lines as demonstrated both in [3H]thymidine incorporation and cytokine release assays. Th cell stimulation by TT-phages was significant at a ratio of one TT-phage in 50 irrelevant phages. The described approach provides the basis for the development of a novel strategy of cloning MHC class II-dependent Th antigens, using available Th cells. This strategy has several potential advantages over existing antigen cloning methods or biochemical peptide isolation.
Expression of CTLA-4 in nonhuman primate lymphocytes and its use as a potential target for specific immunotoxin-mediated apoptosis: results ofin vitrostudies
Authors: PALMISANO G L , TAZZARI P L , COZZI E , BOLOGNESI A , POLITO L , SEVESO M , ANCONA E , RICCI F , CONTE R , STIRPE F , FERRARA G B , PISTILLO M P 
Published: January 2004

SUMMARYT-cell-mediated immunoregulation is one of the main mechanisms implicated in induction and maintenance of transplantation tolerance. In this regard, deletion or modulation of xeno/alloantigen-specific T cells, as well as blocking of their interactions with other cell populations, are currently being pursued for tolerance induction in humans as well as nonhuman primates. In order to investigate whether cytotoxic T-lymphocyte antigen-4 (CTLA-4) may represent a suitable target for a T cell depletion approach in nonhuman primate models, we analysed CTLA-4 expression in peripheral blood mononuclear cells (PBMCs) from nonhuman primates and the potential role of two anti-CTLA-4 saporin-conjugated immunotoxins. The analysis was performed in PBMCs from 8 cynomolgus monkeys from Philippines and from Mauritius both at protein level by flow cytometry and at transcriptional level by RT-PCR. In addition, the apoptotic role of the immunotoxins was investigated. The results showed that CTLA-4 was expressed at variable levels depending on the origin of the cynomolgus monkeys and the resting or activated cell condition. CTLA-4 was not expressed on resting Mauritius PBMCs and showed a lower up-regulation upon PMA/PHA activation compared to the Philippines PBMCs that expressed CTLA-4 also before activation. Two CTLA-4 RNA transcripts (672 and 550 bp) were detected with levels variations after cell stimulation. Two anti-CTLA-4 immunotoxins induced in vitro apoptosis of activated PBMCs from both sources of cynomolgus monkeys. This is the first report that documents CTLA-4 expression both at protein and transcriptional level by nonhuman primate PBMCs and provides novel perspectives of xeno/allograft rejection immunotherapy based on CTLA-4 targeting.
IL-12 enhances the generation of tumour antigen-specific Th1 CD4 T cells duringex vivoexpansion
Authors: KNUTSON K L , DISIS M L 
Published: January 2004

SUMMARYCD4+ T cells are essential for the immune response against cancer. Vaccination against cancer will likely only be effective at preventing growth of micrometastatic disease while adoptive T cell therapy will be better suited for eradication of bulky pre-existing disease (Knutson et al. Expert Opin Biol Ther 2002; 2:55–66). Problems with the use of adoptive T cell therapy include lack of CD4+ T cell help, low frequency of antigen-specific T cells, and lack of effective ex vivo expansion techniques. In this study, we focused on improving ex vivo expansion of CD4+ T helper cells. The effects of IL-12, along with IL-2, on the ex vivo generation of HER-2/neu antigen-specific T cells were examined. Patients were immunized with a peptide-based vaccine that contained a helper epitope, p776–790, derived from the intracellular domain of HER-2/neu. While T cell immunity to p776–790, assessed by proliferation assays, could be readily measured in short-term cultures, cell line generation by multiple in vitro stimulation with peptide and IL-2 as the only added cytokine resulted in loss of antigen-specific proliferation. The inclusion of IL-12, along with IL-2, restored antigen-specific proliferation in a dose-dependent fashion. The resulting p776–790-specific T cells responded readily to antigen by proliferating and producing type I cytokines (IFN-γ and TNF-α). The increased proliferative response of the cultures was due in part to an increase in the number of HER-2/neu-specific T cells. These results suggest that IL-12 is an important cytokine for ex vivo recovery and maintenance of antigen-specific CD4+ T lymphocytes that would otherwise be lost by using IL-2 alone in combination with antigen. Furthermore, these results have important implications for ex vivo expansion of CD4+ T cell for use in anti-tumour adoptive immunotherapy.
Increased Toll-like receptor 4 expression in infants with respiratory syncytial virus bronchiolitis
Authors: GAGRO A , TOMINAC M , KRŠULOVIĆ-HREŠIĆ V , BAĆE A , MATIĆ M , DRAŽENOVIĆ V , MLINARIĆ-GALINOVIĆ G , KOSOR E , GOTOVAC K , BOLANČA I , BATINICA S , RABATIĆ S 
Published: January 2004

SUMMARYThe fusion protein of the respiratory syncytial virus (RSV) binds to the pattern recognition receptors, TLR4 and CD14, and initiates innate immunity response to the virus. The aim of the study was to investigate the expression of TLR4 on peripheral blood lymphocytes and monocytes in peripheral blood of infants in both acute and convalescent phase of RSV bronchiolitis (n = 26). In addition, TNF-α expression in lipopolysaccharide-stimulated monocytes was also assessed. The results showed TLR4 to be expressed predominantly by monocytes in both sick infants and controls. During the acute phase of infection monocytes up-regulated TLR4 in eight infants, which returned to the levels recorded in controls 4–6 weeks from infection. There was no difference in the percentage of TNF-α secreting monocytes. Of the clinical parameters tested, minimal oxygen saturation was found to correlate negatively with this expression in the group of infants with increased TLR4. Additional studies are under way to correlate this finding with the outcome of the immune response to RSV.
Low plasma level of adiponectin is associated with stavudine treatment and lipodystrophy in HIV-infected patients
Authors: LINDEGAARD B , KELLER P , BRUUNSGAARD H , GERSTOFT J , PEDERSEN B K 
Published: January 2004

SUMMARY
This study tested the hypothesis that in patients with HIV-associated lipodystrophy, adiponectin levels were related to insulin resistance, TNF-α and IL-6 and treatment with nucleoside analaogues. HIV seropositive men undergoing highly active antiretroviral treatment were enrolled into three predetermined clinical groups: lipodystrophy with central fat accumulation (n = 12); lipodystrophy without central fat accumulation (n = 15); no lipodystrophy (n = 15). HIV-negative healthy men served as controls (n = 12). Both lipodystrophic groups had a low percentage of limb fat compared to the two control groups. Patients with lipodystrophy with fat accumulation had increased truncal fat compared with controls. Levels of adiponectin did not correlate with either TNF-α or IL-6. Low levels of adiponectin were found in both lipodystrophic groups and were associated with current or previous treatment with stavudine. Furthermore, the adiponectin level correlated with the percentage of limb fat. Patients with lipodystrophy with fat accumulation were more insulin resistant, measured by HOMA-IR, compared with controls. However, HOMA-IR did no correlate to adiponectin or other cytokines. In conclusion, the finding of no difference between the two lipodystrophic groups with regard to adiponectin, indicates that low levels of adiponectin reflects fat atrophy, whereas the insulin resistance was best explained by increased truncal fat mass.
Cellular immunity induced by the recombinant Plasmodium falciparum malaria vaccine, RTS,S/AS02, in semi-immune adults in The Gambia
Authors: PINDER M , REECE W H H , PLEBANSKI M , AKINWUNMI P , FLANAGAN K L , LEE E A M , DOHERTY T , MILLIGAN P , JAYE A , TORNIEPORTH N , BALLOU R , MCADAM K P M J , COHEN J , HILL A V S 
Published: January 2004

SUMMARY
Vaccination of malaria-naive humans with recombinant RTS,S/AS02, which includes the C-terminus of the circumsporozoite protein (CS), has been shown to induce strong T cell responses to both the whole protein antigen and to peptides from CS. Here we show that strong T cell responses were also observed in a semi-immune population in The Gambia, West Africa. In a Phase I study, 20 adult male volunteers, lifelong residents in a malaria-endemic region, were given three doses of RTS,S/AS02 at 0, 1 and 6 months. Responses to RTS,S, hepatitis B surface antigen and peptides from CS were tested using lymphocyte proliferation, interferon (IFN)-γ production in microcultures, and IFN-γ ex vivo and cultured ELISPOT, before and after vaccination. Cytotoxic responses were tested only after vaccination and none were detected. Before vaccination, the majority of the volunteers (15/20) had detectable responses in at least one of the tests. After vaccination, responses increased in all assays except cytotoxicity. The increase was most marked for proliferation; all donors responded to RTS,S after the third dose and all except one donor responded to at least one peptide after the second or third dose. There was a lack of close association of peptide responses detected by the different assays, although in microcultures IFN-γ responses were found only when proliferative responses were high, and responses by cultured ELISPOT and proliferation were found together more frequently after vaccination. We have therefore identified several peptide-specific T cell responses induced by RTS,S/AS02 which provides a mechanism to investigate potentially protective immune responses in the field.
Inter‐observer reliability of radiological signs of necrotising enterocolitis in a population of high‐risk newborns
Authors: Di Napoli Anteo , Di Lallo Domenico , Perucci Carlo A. , Schifano Patrizia , Orzalesi Marcello , Franco Francesco , De Carolis Maria Pia 
Published: January 2004

SummaryRadiological examination of the abdomen is critical to the diagnosis of necrotising enterocolitis (NEC). Previous studies on the reproducibility of radiological findings have been limited in size and based only on infants with the disease. We conducted a study among a sample of high‐risk infants with and without the diagnosis of NEC: (1) to measure the degree of inter‐observer agreement of NEC radiological diagnosis and signs and (2) identify a profile of radiological signs which led the observers to make a diagnosis of NEC.We collected 297 X‐rays from a sample of 57 newborns admitted in 1999 to neonatal intensive care and neonatal surgery units in Rome, Italy. Three specialists in paediatric radiology examined the films independently and without any clinical information about patients. The analyses were conducted on a total of 891 forms filled in by the observers. Kappa values were calculated to measure the inter‐observer reliability. To identify the profiles of radiological signs, a multidimensional analysis, binary segmentation, was carried out. The reproducibility of radiographic signs was 0.55 (P < 0.01) for diffuse gaseous intestinal distention, 0.22 (P < 0.01) for bowel wall thickening, 0.10 (P < 0.01) for presence of portal venous gas and 0.29 (P < 0.01) for pneumatosis intestinalis. The agreement for radiographic diagnosis suspected/confirmed of NEC was 0.31 (P < 0.01). Among the 23 possible combinations of radiographic signs, the three radiologists indicated four profiles that produced a diagnosis of NEC containing, respectively, two, three, four and five signs.Our study found, in a large sample of radiographs selected from a population of infants with and without NEC, a poor reliability for NEC diagnosis and individual radiological signs among three expert radiologists. Clinical information and the presence of more than one radiological sign can reduce the margin of observer's error that inevitably exists when dealing with a diagnosis as difficult as NEC.
Repeated validation of parental self‐reported smoking during pregnancy and infancy: a prospective cohort study of infants at high risk for allergy development
Authors: Christensen Anne E. , Tobiassen Mette , Jensen Tina K. , Wielandt Hanne , Bakketeig Leiv , Høst Arne 
Published: January 2004

SummaryExposure to environmental tobacco smoke (ETS) during fetal life and infancy is closely related to the smoking habits of the parents. Estimates of exposure to ETS require valid and detailed information on changes in cigarette smoking over time. The objective was to test the validity of self‐reported smoking among parents during pregnancy and early childhood in a cohort of children at high risk for allergy development by measurement of exhaled carbon monoxide (CO). The cohort comprised 117 families enrolled from the general population of pregnant women at admission to antenatal care.Data on parental tobacco smoking were obtained by interview and exhaled CO was measured (Micro‐Smokerlyzer®) in parents twice during pregnancy and when the child was 6 and 18 months old. The median (range) exhaled CO levels were 3 (0–10) parts per million (ppm) for non‐smokers and 15 (1–39) ppm for smokers (P < 0.0005). A receiver operating characteristic (ROC) analysis was performed at each examination. The areas under the ROC curve were high for both mothers (between 0.88 and 0.99) and fathers (between 0.87 and 0.89), indicating exhaled CO as a good diagnostic tool for determining smoking status. Comparing the ROC areas obtained for mothers from late pregnancy and during infancy with the area from early pregnancy showed no statistical differences (P = 0.21, 0.43 and 0.44 respectively) and the same was true for fathers during infancy (P = 0.81). The level of 8 ppm was used as the cut‐off between smokers and non‐smokers, based on data from a pilot study. Using CO as a diagnostic tool for smoker status showed very high specificity (between 97 and 100%), indicating that very few persons claiming to be non‐smokers had CO levels higher than 8 ppm.In conclusion, the validity of interview‐obtained self‐reported smoking among parents during pregnancy and early childhood was high. Repeated interviews and CO measurements in a prospective study design did not change the validity, indicating a low risk of information bias. A structured interview combined with measurement of exhaled CO is a valid and reliable method for estimating ETS exposure to the fetus and young infant.