bluuebunny/crossref_metadata_2025 huggingface dataset:
Paclitaxel (Taxol) Inhibits the Arylamine N‐Acetyltransferase Activity and Gene Expression (mRNA NAT1) and 2‐Aminofluorene‐DNA Adduct Formation in Human Bladder Carcinoma Cells (T24 and TSGH 8301)
Authors: Yang Ching‐Chiang , Chen Guang‐Wei , Lu Hsueh‐Fu , Wang Der‐Yuan , Chen Yi‐Shuan , Chung Jing‐Gung
Published: May 2003

Abstract: Acetylator polymorphism in man results from differential expression of human liver N‐acetyltransferase. N‐Acetyltransferase enzyme activity has been demonstrated to be involved in some types of chemical carcinogenesis. Paclitaxel (taxol) had been shown to affect N‐acetyltransferase activity of human lung cancer cells. In this study, paclitaxel was chosen to investigate the effects of arylamine N‐acetyltransferase activity (N‐acetylation of substrate), gene expresssion and 2‐aminofluorene‐DNA adduct formation in human bladder carcinoma cell lines (T24 and TSGH 8301). The N‐acetyltrasnsferase activity (N‐acetylation of substrates) was determined by high performance liquid chromatography assaying for the amounts of acetylated 2‐aminofluorene and p‐aminobenzoic acid and nonacetylated 2‐aminofluorene and p‐aminobenzoic acid. Intact human bladder carcinoma T24 and TSGH 8301 cells were used for examining N‐acetyltransferase activity, gene expression and 2‐aminofluorene‐DNA adduct formation. The results demonstrated that the N‐acetyltransferase activity, gene expression (NAT1 mRNA) and 2‐aminofluorene‐DNA adduct formation in intact human bladder carcinoma cells were inhibited and decreased by paclitaxel in a dose‐dependent manner. The effects of paclitaxel on the apparent values of Km and Vmax of N‐acetyltransferase enzyme from intact human bladder carcinoma cells were also determined in these cell lines. A marked influence of paclitaxel was observed on the decreasing apparent values of Km and Vmax from intact human bladder carcinoma cells (T24 and TSGH 8301). Thus, paclitaxel is an uncompetitive inhibitor to the NAT enzyme.
Gene Expression of Cytochrome P450 1B1 and 2D6 in Leukocytes in Human Pregnancy
Authors: Lind Anna‐Britta , Wadelius Mia , Darj Elisabeth , Finnström Niklas , Lundgren Stefan , Rane Anders
Published: May 2003

Abstract: We investigated the influence of human pregnancy on gene expression of two cytochrome P450 enzymes in white blood cells. Cytochrome P450 1B1 (CYP1B1) catalyses oestradiol 4‐hydroxylation, and may participate in the endocrine regulation of oestrogens. Cytochrome P450 2D6 (CYP2D6) metabolises many commonly used drugs, and previous studies have suggested that it is induced during pregnancy. CYP1B1 and CYP2D6 were therefore considered to be of interest in human pregnancy. As it is not ethically possible to take liver biopsies from healthy mothers during pregnancy, easily accessible cells that express the genes were used as a surrogate tissue. White blood cells were collected from eighteen pregnant women, and were used to measure CYP1B1 and CYP2D6 ribonucleic acid (RNA). The analysis was repeated after pregnancy, the women, thus, serving as their own controls. Real‐time reverse transcriptase – polymerase chain reaction methods were used with 18S ribosomal RNA as an internal control. A slight, but not significant, increase in gene activity of CYP1B1 was detected during pregnancy. Expression of CYP2D6 in blood was extremely low, and induction of CYP2D6 during pregnancy could not be confirmed. In conclusion, gene expression of CYP1B1 and CYP2D6 in leukocytes was not significantly up‐regulated in the third trimester of pregnancy, but a trend indicating an altered metabolism during pregnancy was detected.
Identification of candidate genes for schizophrenia based on natural resistance to infectious diseases
Authors: Brown James S
Published: May 2003

Background:

Identification of candidate genes for schizophrenia may be more successful than genome screens as the latter have not found consistent linkages.

Objective:

To assist in the gene search, a model of schizophrenia based on resistance to infectious diseases, rather than susceptibility, is proposed. The theory blends the geography of schizophrenia with the assumption that genes that cause schizophrenia likely evolved and persist from selection pressure. The theory includes the notion that schizophrenia enhances biological survival at the cost of psychological and social functioning.

Method:

To demonstrate the utility of using this model, the Medline literature was searched for resistance genes, mostly identified in mice.

Results:

Based on homologous locations in the human genome, these resistance genes are shown to be located in human chromosome regions linked significantly, in at least one genome screen, with schizophrenia or some physiologically related function or condition associated with schizophrenia.

Conclusions:

The infectious disease resistance theory of schizophrenia is offered as a viable model for understanding the origins of schizophrenia. The theory also allows for the inclusion of persistent infections, seasonal variability and translational pathophysiology to contribute to the etiology of schizophrenia.
Magnetic resonance imaging studies on autism and childhood-onset schizophrenia in children and adolescents – a review
Authors: Lahuis Bertine , Kemner Chantal , Van Engeland Herman
Published: May 2003

Objective:

To find out whether the neurodevelopmental disorders autism and childhood-onset schizophrenia have a common developmental pathway and whether the abnormalities detected are ‘disorder-specific’, by reviewing magnetic resonance imaging (MRI) studies.

Methods:

As a result of a Medline search, we were able to access 28 studies on autism and 12 studies on childhood-onset schizophrenia, which focused on children and adolescents.

Results:

Larger lateral ventricles were found to be a common abnormality in both disorders. ‘Disorder-specific’ abnormalities in patients with autism were larger brains, a larger thalamic area, and a smaller right cingulate gyrus. Subjects with childhood-onset schizophrenia were found to have smaller brains, a smaller amygdalum and thalamus, and a larger nucleus caudatus. In subjects with childhood-onset schizophrenia, abnormalities appeared to progress over a limited period of time.

Conclusions:

Because the study designs varied so much, the results should be interpreted cautiously. Before abnormalities found in the disorders can be designated as equal or ‘disorder-specific’, it will be essential to perform large longitudinal and cross-sectional follow-up studies.
Diversity of rodents and shrews along an elevational gradient in Bwindi Impenetrable National Park, south‐western Uganda
Authors: Kasangaki Aventino , Kityo Robert , Kerbis Julian
Published: June 2003

Small mammal species diversity in the major vegetation zones of Bwindi Impenetrable National Park is discussed in relation to altitude. Species richness of the small mammals was found to decrease with an increase in altitude. The main factors accounting for the observed diversity are the wide altitudinal variation and a complex array of vegetation types. Sixty‐seven species of rodents and shrews were found to exist in the Park; 47 of which were rodents and 20 shrews. Of these, 26 species are new to the Bwindi Park list. Three species have probably not been described before. The study found 10 species of small mammals to be Albertine Rift endemics. Three genera are recorded in Uganda for the first time: Rwenzorisorex, Suncus and Paracrocidura. Five species are new records for East Africa. These are Crocidura stenocephala, Lophuromys rahmi, L. medicaudatus, Paracrocidura maxima and Hylomyscus aeta. Because of the high endemism of plants, butterflies, birds and now of small mammal species, Bwindi forest is a unique biodiversity hotspot and is among the highest conservation priorities in the Albertine Rift.
Relationship between grass species richness and ecosystem stability in Kruger National Park, South Africa
Authors: Kennedy A. D. , Biggs H. , Zambatis N.
Published: June 2003

The relationship between grass species richness and ecosystem stability was investigated in the Kruger National Park. A total of 135 489 individual grasses were identified from 189 sites spread across 19 485 km2 of savanna biome, making this one of the largest studies of its kind. At each site, grass percentage abundance and standing crop were measured at one year intervals to provide an index of ecosystem function. A severe drought that affected the region between 1991 and 1993 provided a convenient perturbation. At the height of the drought, mean species richness declined by 12.7% while standing crop declined by 38.1%, from 3199 to 1979 kg ha−1. Percentage grass abundance declined to 87.5% of its pre‐drought value. After the drought had passed species richness, standing crop and percentage abundance recovered to 92.1%, 113.8% and 92.8% of their pre‐perturbation values, respectively. Statistical analysis of these responses revealed that grass assemblages of low species richness were more resistant to drought than assemblages of high species richness. Species‐poor sites also showed better recovery from perturbation after the drought had passed. These findings suggest that ecosystem stability may be negatively related to grass species richness in South African savanna grasslands.
Feeding ecology and dietary differences between blue monkey (Cercopithecus mitis stuhlmanni Matschie) groups in logged and unlogged forest, Budongo Forest Reserve, Uganda
Authors: Fairgrieve Chris , Muhumuza Geresomu
Published: June 2003

Several studies have investigated the effects of logging on primate population density, and attempted to relate density differences to changes in vegetation composition. As population density in some forest primates may be considerably influenced by dietary quality, it is important to understand how the effects of vegetation changes commonly associated with logging influence primate feeding ecology.

Results are presented from a study carried out in the Budongo Forest Reserve in Uganda to investigate differences in blue monkey (Cercopithecus mitis stuhlmanni) feeding ecology in forest blocks with different logging history.

Dietary composition differed between logged and unlogged forest, with a higher proportion of unripe fruit consumed in logged forest. By contrast, the proportion of young leaves, invertebrates and seeds in the diet was higher in unlogged forest. Variations in the intake of fruit was also higher in unlogged forest, but feeding patterns on other dietary items were more selective. These differences in diet reflect differences in availability of different categories of food items, as determined by tree species composition, abundance and plant phenological patterns. The observed differences between feeding ecology in logged and unlogged forest are discussed in relation to primate feeding ecology and habitat composition in Budongo.
Primate census and habitat evaluation in the Tana delta region, Kenya
Authors: Muoria P. K. , Karere G. M. , Moinde N. N. , Suleman M. A.
Published: June 2003

Nineteen indigenous forest patches in the Tana River delta region, Kenya were surveyed between October and November 2000 for primates and habitat disturbance. Special emphasis was placed on the endangered Tana River red colobus (Procolobus rufomitratus Peters) and crested mangabeys (Cercocebus galeritus galeritus Peters), both of which are endemic to the region. Habitat disturbances evident in the forests included cutting of trees, harvesting of thatching material, firewood collection, dyke construction, cultivation, palm wine tapping and charcoal burning. A total of 85 groups of five primate species were counted. These comprised eighteen, ten, 22, 31 and four groups of red colobus, crested mangabey, baboons (Papio cynocephalus L.), sykes monkeys (Cercopithecus mitis Wolf) and vervet monkeys (Cercopithecus aethiops L.), respectively. A wider distribution of red colobus and crested mangabeys than was documented previously was noted, implying that they are probably more abundant than hitherto reported. It is hypothesized that extensive studies on some fauna considered endangered world‐wide would probably redefine their conservation status. Future studies in the lower Tana River region should cover the previously unsurveyed forests and focus on ways of curbing forest destruction.
Assessment of the performance of exotic and indigenous tree and shrub species for rehabilitating saline soils of Northern Kenya
Authors: Olukoye Godfrey A. , Wamicha Wellington N. , Kinyamario Jenesio I.
Published: June 2003

In the semi‐desert zone of North Horr, northern Kenya, soil salinity reaches toxic levels, with extensive salt crusts at the surface. Environmental groups are involved in tree and shrub planting trials with the objective of stabilizing sand dunes and rehabilitating land. However, the performance of the various indigenous and exotic tree and shrub species in rehabilitating these saline soils has not been examined. Therefore this study aimed at evaluating the performance of various exotic and indigenous tree and shrub species in rehabilitating saline soils in North Horr. Mortality and relative growth rates for both exotic and indigenous tree and shrub species were assessed. Compared to indigenous species, exotic tree species (not naturally growing in the Marsabit ecosystem) failed to sustain an improved growth performance in saline soils of North Horr. The results of this study demonstrate that survival rates for indigenous plants in North Horr are higher than those of exotic ones. Thus, to rehabilitate saline soils and stabilize sand dunes, the use of indigenous tree and shrub species adapted to such saline conditions should be encouraged.
The morphology, distribution and conservation implications of introduced rats, Rattus spp. in the granitic Seychelles
Authors: Hill M. J. , Vel T. , Shah N. J.
Published: June 2003

There are no native land mammals in the Seychelles archipelago other than bats. Introduced rats have reduced the conservation value of most islands. This paper compares the results of rat‐trapping carried out on eight islands in the granitic Seychelles, between July 1999 and April 2000. Trapping was carried out in both the dry and wet seasons. Three introduced rodent species were caught, including two species of rat (ship rat Rattus rattus Linnaeus and Norway rat R. norvegicus Berkenhout), but only one Rattus species occurred on each island. Both rat species were smaller than European or Asian conspecifics, and there were variations in the size and appearance of rats on different islands. Inter‐island differences in size and pelage colour are discussed.
Short‐ and long‐term therapy for reflux oesophagitis in the elderly: a multi‐centre, placebo‐controlled study with pantoprazole
Authors: Pilotto A. , Leandro G. , Franceschi M. ,   (ON BEHALF OF THE AGEING and ACID‐RELATED DISEASE STUDY GROUP)
Published: June 2003

Background: No placebo‐controlled clinical trials have yet been published on the efficacy of therapy in older subjects with oesophagitis.

Aim: To evaluate the efficacy of pantoprazole in preventing the recurrence of oesophagitis in elderly subjects.

Methods: One hundred and sixty‐four patients aged 65 years and over with acute oesophagitis were treated openly with pantoprazole, 40 mg daily, for 8 weeks. Patients with documented healing of erosive oesophagitis were then treated with pantoprazole, 20 mg daily, for 6 months. Thereafter, cured patients were randomized to receive pantoprazole, 20 mg daily, or placebo for the following 6 months. Clinical evaluations were performed every 2 months, and endoscopy was repeated after 8 weeks and after 6 and 12 months and/or whenever symptoms suggested a relapse of oesophagitis.

Results: After 8 weeks, the healing rates of oesophagitis were 81.1% (75.1–87.1%) and 93.7% (89.7–97.7%) by intention‐to‐treat and per protocol analyses, respectively. After 6 months, the corresponding values were 82% (75.4–88.5%) and 92.4% (87.6–97.2%), respectively. After 12 months, the per protocol and intention‐to‐treat healing rates of oesophagitis were 95.1% (88.5–100%) and 79.6% (68.3–90.9%), respectively, in the treatment group vs. 32.7% (19.9–45.4%) and 30.4% (18.3–42.4%), respectively, in the placebo group (P = 0.0001). Heartburn, acid regurgitation and chest pain were significantly associated with the relapse of oesophagitis (P = 0.0001), whereas hiatus hernia, Helicobacter pylori infection, concomitant diseases and treatments were not.

Conclusion: In the elderly, pantoprazole was highly effective in healing and reducing the relapse of oesophagitis; discontinuing active treatment after 6 months was associated with a significant increase in the relapse rate.
An open‐label trial of the selective cyclo‐oxygenase‐2 inhibitor, rofecoxib, in inflammatory bowel disease‐associated peripheral arthritis and arthralgia
Authors: Reinisch W. , Miehsler W. , Dejaco C. , Harrer M. , Waldhoer T. , Lichtenberger C. , Vogelsang H.
Published: June 2003

Background: Conventional non‐steroidal anti‐inflammatory drugs have been associated with an increased risk of exacerbation of inflammatory bowel disease.

Aim: To evaluate, in a prospective, open‐label study, the safety and efficacy of a 20‐day regimen of the selective cyclo‐oxygenase‐2 inhibitor, rofecoxib, 12.5–25 mg/day, in inflammatory bowel disease patients with associated peripheral arthropathy and/or arthritis.

Methods: Patients with clinically inactive to mild inflammatory bowel disease and a joint pain score of at least two points on a scale ranging from zero (none) to four (very poor) were eligible. Response was defined by a decrease of at least two points in the arthralgia score.

Results: Of the 32 patients included, 26 (81%) were treated with rofecoxib, 25 mg/day, and six (19%) with rofecoxib, 12.5 mg/day. In three patients (9%), rofecoxib had to be withdrawn after a few days due to gastrointestinal complaints which ceased immediately after drug discontinuation. No flare of inflammatory bowel disease occurred. Thirteen of the 32 patients (41%) were responders and, overall, the arthralgia score decreased from two to one (P = 0.0001).

Conclusions: This is the first prospective study on the use of a selective cyclo‐oxygenase‐2 inhibitor in inflammatory bowel disease patients with peripheral arthropathy and/or arthralgia. The promising safety and efficacy profile warrants further evaluation in controlled trials.
Activation of human intraepithelial lymphocytes reduces CD3 expression
Authors: EBERT E C
Published: May 2003

The aim of this study was to examine in detail the low functional capacity of human intraepithelial lymphocytes (IELs) in response to phytohaemagglutinin (PHA) and CD3 ligation. Human IELs were extracted from jejunal mucosa obtained from patients undergoing gastric bypass operations for morbid obesity and compared to peripheral blood (PB) lymphocytes composed predominantly of CD8+ T cells. Calcium influx ([Ca2+]i) was analysed using Fura-2-loaded cells; IL-2 receptor expression was measured by immunofluorescence and flow cytometry; IL-2 binding was determined using radiolabelled IL-2; IL-2 production was quantified by ELISA; and apoptosis was detected with Apo 2·7 staining. Compared to naive PB CD8+ T lymphocytes, calcium influx by IELs was only transient with CD3 ligation and low in amplitude with PHA. IL-2 receptor expression was reduced after CD3 ligation, yet normal in numbers and affinity after PHA stimulation. Both cell types secreted similar amounts of IL-2. CD3 expression on IELs, but not PB CD8+ T cells, declined upon activation, due partly to incomplete reexpression after modulation. Little apoptosis was found. The partial activation of IELs in response to PHA and CD3 ligation, as manifested by diminished [Ca2+]i, resulted in a decline in CD3 expression.
Staphylococcal superantigens and T cell expansions in Wegener's granulomatosis
Authors: POPA E R , STEGEMAN C A , BOS N A , KALLENBERG C G M , TERVAERT J W COHEN
Published: May 2003

In Wegener's granulomatosis (WG), a form of autoimmune systemic vasculitis, chronic carriage of Staphylococcus aureus constitutes a risk factor for the development of exacerbations. Circulating T cells in this disease are persistently activated, suggesting the presence of a chronic stimulus. A causal link between chronic carriage of S. aureus and chronic T cell activation in WG is conceivable, because S. aureus produces superantigens (SAg), which are potent T cell stimulators. Superantigenic stimulation of T cells results in expansion of T cell subsets expressing SAg-binding T cell receptor V-beta (Vβ) chains. In the present study we hypothesized that in WG the presence of staphylococcal SAg is accompanied by expansion of SAg-reacting T cell subsets. We tested our hypothesis in a cross-sectional and a longitudinal study in which the association between seven staphylococcal SAg genes [typed by poplymerase chain reaction (PCR)], eight SAg-binding Vβ chains and four SAg-non-binding Vβ chains (assessed by flow-cytometry) was assessed. Both studies showed that T cell expansions were present at a significantly higher rate in WG patients than in healthy individuals, but were not associated with the presence of either S. aureus or its SAg. Moreover, T cell expansions were generally of small extent, and did not appear simultaneously in both CD4 and CD8 subsets. We conclude that in WG S. aureus effects its supposed pathogenic function by a mechanism other than superantigenic T cell activation.
The role of p53 in the immunobiology of cutaneous squamous cell carcinoma
Authors: BLACK A P B , OGG G S
Published: May 2003

Cutaneous squamous cell carcinoma is typically characterized by the over-expression of the tumour suppressor protein p53. Considerable evidence suggests that immune competence is important in the control of cutaneous SCC. We discuss the immunobiology of p53 and its relevance to cutaneous SCC, including the potential interaction with human papillomavirus.
Circulating concentrations of soluble granzyme A and B increase during natural and experimental Plasmodium falciparum infections
Authors: HERMSEN C C , KONIJNENBERG Y , MULDER L , LOÉ C , VAN DEUREN M , VAN DER MEER J W M , VAN MIERLO G J , ELING W M C , HACK C E , SAUERWEIN R W
Published: May 2003

Release of soluble Granzymes (sGranzymes) is considered to reflect activation of cytotoxic T lymphocytes and NK cells. sGranzymes and a number of pro-inflammatory cytokines were measured in plasma of malaria patients with natural or experimentally induced Plasmodium falciparum infections. Concentrations of sGranzyme A and B, IL-10, IL-12p70 and CRP were significantly increased in African children presenting with clinical malaria; IL-10 and CRP concentrations were significantly correlated with disease severity. In nonimmune Dutch volunteers which were experimentally infected by P. falciparum-infected mosquitoes, sGranzyme A increment started 1–2 days prior to clinical symptoms and microscopically detectable parasitaemia. This coincided with increases in IFNγ, IL-12p40 and IL-8, while sGranzyme B and IL-10 levels increased 24–48 h later. The elevation of sGranzyme A and IFNγ in nonimmune volunteers suggests that NK cells are activated upon release of parasites by infected liver cells and subsequently during blood stage infection; thus, NK cells are likely involved innate immune human host resistance in the early phase of a malaria infection.
Evidence for intramolecular B-cell epitope spreading during experimental immunization with an immunogenic thyroglobulin peptide
Authors: THRASYVOULIDES A , LYMBERI P
Published: May 2003

Thyroglobulin (Tg) is a target autoantigen in autoimmune thyroid diseases, such as Graves’ disease (GD) and Hashimoto's thyroiditis. In a previous study we identified three 20mer Tg peptides bearing epitopes of autoantibodies associated with GD (TgP15, TgP26 and TgP41: sequences 2339–2358, 2471–2490 and 2651–2670 of human Tg, respectively). In the present study, we investigated the antigenicity of the above peptides in experimental immunization with Tg, the immunogenicity of antigenic peptides and the possibility of intramolecular B-cell epitope spreading during peptide immunization. For this purpose, two rabbits were injected with human Tg in CFA six times, every three weeks. Two control animals were injected only with CFA. Testing of antisera and of affinity-purified antibodies, by ELISA against the three peptides, revealed reactivity only to TgP41. This synthetic peptide was subsequently administered to two rabbits, in its free form (100µg in CFA six times, every two weeks). A strong serological response was developed not only against TgP41, but also to intact human and rabbit Tg. Immunization with TgP41 induced intramolecular B-cell epitope spreading, i.e. production of antibodies to sites on Tg other than that corresponding to TgP41, as revealed by immunoadsorption and competitive ELISA. Histopathological studies did not reveal any infiltration in thyroid glands. We conclude that peptide TgP41 encompasses not only an epitope of disease-associated autoantibodies, but also a dominant immunogenic epitope of experimentally induced Tg-specific antibodies, able to drive B-cell epitope spreading.
Mucosal and systemic immune responses to plasmid protein pgp3 in patients with genital and ocular Chlamydia trachomatis infection
Authors: GHAEM-MAGHAMI S , RATTI G , GHAEM-MAGHAMI M , COMANDUCCI M , HAY P E , BAILEY R L , MABEY D C W , WHITTLE H C , WARD M E , LEWIS D J M
Published: May 2003

The circulating and cervical B cell responses to Chlamydia trachomatis plasmid protein pgp3 were characterized in children and adults with ocular or genital chlamydial infection using the enzyme-linked immunospot assay (ELISPOT) and ELISA. No pgp3-specific ASCs were detected in healthy controls, but predominantly IgA ASCs were detected in UK adults with uncomplicated cervicitis or urethritis (P = 0·03, 0·019). In patients with extragenital complications or pelvic inflammatory disease a mixed response with more IgG and IgM ASCs was evident, suggesting a breach of mucosal immune compartmentalization with more extensive infection. In women with chlamydial cervicitis, ASCs secreting predominantly IgA, but also IgG, to pgp3 were present in cervix at presentation, with a frequency 30–50 times higher than blood. Cervical ASC numbers, especially IgG, fell markedly six weeks after antibiotic treatment. We detected principally IgA pgp3-specific antibody secreting cells (ASCs) in children resident in a Gambian endemic area, with a trend towards suppression of IgA responses during intense trachomatous inflammation (P = 0·06), as previously reported for other chlamydial antigens, and in keeping with the findings in genital disease. These data provide a rationale for further studies of immune responses to pgp3 in humans and animal models of chlamydia-induced disease, and its potential use in diagnostic assays and protective immunization strategies.
Th2 cytokines, IgE and mast cells play a crucial role in the induction of para-phenylenediamine-induced contact hypersensitivity in mice
Authors: YOKOZEKI H , WU M-H , SUMI K , IGAWA K , MIYAZAKI Y , KATAYAMA I , TAKEDA K , AKIRA S , NISHIOKA K
Published: May 2003

We previously reported the establishment of a mouse model system of contact hypersensitivity (CHS) to paraphenylemediamine (PPD). In order to analyse the functional contribution of Th2 cytokines, IL-4 and IL-5, in PPD induced CHS, STAT6 deficient (STAT6–/–) and wild-type control (WT) mice (C57BL/6) were immunized by the topical application of a PPD solution, and then the subsequent skin reactions were examined. Ear swelling was significantly reduced with a delayed peak response in STAT6–/– mice as compared with that of WT mice. A histological analysis showed the infiltration of both eosinophils and neutrophils in the skin of STAT6–/– mice challenged 24 h previously to significantly decrease in comparison with that in the WT mice. The expression of Th2 cytokines (IL-4, IL-5) by ELISA in the PPD-challenged skin tissue specimens as well as the IgE and IgG1 response after challenge were also profoundly reduced in the STAT6–/– mice. The adoptive transfer of the serum obtained from sensitized WT mice for the putative IgE transfer induced a peak response at 3 h and 24 h after challenge. To further investigate the role of mast cells in the induction of PPD-CHS, mast cell deficient W/Wv mice were sensitized with PPD and then were challenged. Maximal ear swelling was detected from 12 to 24 h and another small peak response was observed at 1 h in+/+mice, whereas only a small peak response at 24 h was detected in W/Wv mice. These data indicate that not only Th2 cytokines and IgE but also mast cells play an essential role in the induction of PPD-CHS.
Anti-Saccharomyces cerevisiae antibodies (ASCA) and autoimmune liver diseases
Authors: MURATORI P , MURATORI L , GUIDI M , MACCARIELLO S , PAPPAS G , FERRARI R , GIONCHETTI P , CAMPIERI M , BIANCHI F B
Published: May 2003

Antibodies to the baker's yeast Saccharomyces cerevisiae (ASCA), recently proposed as a serological marker of Crohn's disease, have also been detected in other autoimmune disorders. The aim of this study was to determine prevalence and clinical significance of ASCA in autoimmune liver disease. The presence of IgG and IgA ASCA was evaluated using a commercially available immunoassay in 215 patients with autoimmune liver disease (primary biliary cirrhosis, PBC, 123 cases; autoimmune hepatitis, AIH, 67 cases; primary sclerosing cholangitis, PSC, 25 cases), 48 with inflammatory bowel disease and 19 healthy blood donors. Anti neutrophil cytoplasmic antibodies with the perinuclear pattern (p-ANCA) were assessed by indirect immunofluorescence in PSC patients. The main clinical and biochemical parameters between ASCA-positive and negative patients were analysed and compared. ASCA are predominant in Crohn's disease (70%); among liver patients, PSC and AMA-negative PBC show the highest ASCA prevalence (53% and 44%). In PBC ASCA correlate with higher levels of circulating IgA (P < 0·05). In PSC the detection of either ASCA or p-ANCA is neither associated with any clinical or biochemical feature, nor with an underlying inflammatory bowel disease. ASCA can not be considered an additional serological marker of autoimmune liver disease, but the possibility of detecting such a reactivity in autoimmune liver disorders should be considered; their correlation with elevated IgA in PBC suggests that ASCA may be an indirect sign of enhanced mucosal immunity; in PSC patients neither ASCA nor p-ANCA predict the occurrence of a concomitant inflammatory bowel disease.
Oral administration of type II collagen suppresses pro-inflammatory mediator production by synoviocytes in rats with adjuvant arthritis
Authors: DING C H , LI Q , XIONG Z Y , ZHOU A W , JONES G , XU S Y
Published: May 2003

The objective of this study was to investigate the effect of the oral administration of type II collagen (CII) on pro-inflammatory mediator production by synoviocytes in rats with adjuvant arthritis (AA). Sprague-Dawley rats were fed with bovine CII either before immunization with Complete Freund's adjuvant (CFA) or after initiation of arthritis. Hind paw secondary swelling was measured and synoviocytes were harvested. Sera from portal vein of oral tolerized rats were collected and in vitro synoviocytes culture or synoviocytes-Peyer's Patches (PP) cells coculture system were developed. Interleukin (IL)-1 activity was measured by a mouse thymocyte activation assayed by MTT dye reduction and tumour necrosis factor (TNF) activity was measured by an L929 cytotoxicity bioassay. Nitric oxide (NO) and malondialdehyde (MDA) levels were measured by biochemical methods. We found that feeding with CII (5, 50 and 500 µg/kg) for 7 days before immunization significantly suppressed hind paw secondary swelling measured at day 16, 20, 24 and 28 (all P < 0·01) and pro-inflammatory mediator (IL-1, TNF, NO and MDA) production by synoviocytes (all P < 0·01) in rats with AA. Feeding with CII (5, 50 and 500 µg/kg) for 7 days after initiation of arthritis had a similar effect. CII (1, 10, 100 µg/ml) had no effect on IL-1 and TNF production by synoviocytes in vitro, but CII 10 µg/ml suppressed IL-1 and TNF production by synoviocytes-PP cells coculture system (P < 0·01), which was antagonized by anti-TGF-β antibody (10 µg/ml) (P < 0·01). Portal serum (1 : 10) from oral tolerized rats suppressed IL-1 and TNF production by synoviocytes (P < 0·01), which was also antagonized by anti-TGF-β antibody (10 µg/ml) (P < 0·01). We conclude that oral administration of CII had prophylactic and therapeutic effects on AA and over-production of IL-1, TNF, NO and MDA by synoviocytes was suppressed. Bystander active suppression may be the main mechanism of oral CII in the suppression of synoviocyte function.
Measurement of soluble Fcγ receptor type IIIa derived from macrophages in plasma: increase in patients with rheumatoid arthritis
Authors: MASUDA M , MORIMOTO T , KOBATAKE S , NISHIMURA N , NAKAMOTO K , DONG X H , KOMIYAMA Y , OGAWA R , TAKAHASHI H
Published: May 2003

FcγRIII (CD16) is found in two alternative forms, a transmembrane FcγRIIIa expressed on NK cells and macrophages, and a glycosylphosphatidylinositol-linked FcγRIIIb present on neutrophils. Previously, we measured soluble FcγRIIIa (sFcγRIIIa) in plasma of NA(1 +, 2-) phenotyped donors with the anti-FcγRIII monoclonal antibody (MoAb) GRM1, which recognizes NA2-FcγRIIIb and FcγRIIIa. The level of sFcγRIIIa, as well as the total sFcγRIII (sFcγRIIIa plus sFcγRIIIb) in patients with rheumatoid arthritis (RA) was significantly higher than that in healthy controls. In this study, we measured sFcγRIIIaMφ in plasma with a newly developed anti-FcγRIII MoAb, MKGR14 (mIgM), which recognizes FcγRIIIaMφ specifically. From the recovery of purified sFcγRIIIaMφ, the amount of sFcγRIIIaMφ present was about half that of sFcγRIIIaNK, and that of sFcγRIIIa was about 50 times lower than that of sFcγRIIIb in pooled plasma from healthy NA(1 +, 2-) phenotyped donors. The level of sFcγRIIIaMφ in RA patients was about four times higher than that in healthy controls. In RA patients, both the sFcγRIIIaMφ and sFcγRIIIa levels were increased as proportionally as the Lansbury Index. The sFcγRIIIa, but not sFcγRIIIaMφ levels, were increased directly proportional to C-reactive protein. sFcγRIIIaMφ may be a novel marker of disease activity in RA.
Interleukin-12 alone can not enhance the expression of the cutaneous lymphocyte associated antigen (CLA) by superantigen-stimulated T lymphocytes
Authors: SIGMUNDSDÓTTIR H , GUDJÓNSSON J E , VALDIMARSSON H
Published: May 2003

It has been reported that bacterial superantigens induce interleukin (IL)-12 dependent expression of the cutaneous lymphocyte associated antigen (CLA) and that this may be relevant to the association between certain skin diseases and infections including psoriasis and streptococcal tonsillitis. We have confirmed that the streptococcal pyrogenic superantigen C (SpeC) increases CLA expression by both CD4+ and CD8+ T cells when PBMCs are incubated in medium enriched with fetal calf serum (FCS). However, such an increase could not be induced in medium enriched with human serum (HS) even when recombinant IL-12 was added to the PBMCs cultures. Strikingly, CD4+ T cells incubated with SpeC in HS showed a marked reduction in CLA expression, which was not due to apoptosis. In contrast, SpeC did induce T cell proliferation and expression of CD25, CD54 and CD103 in the presence of HS indicating that the absence of SpeC induced CLA expression in HS was not due to SpeC inhibitors. Although addition of low amounts of lipopolysaccharide endotoxin (LPS) caused a highly significant increase in CLA expression in the absence of SpeC in cultures enriched with HS, a combination of LPS and SpeC did not increase CLA expression beyond that induced by LPS alone. The superantigen-induced CLA expression in FCS was partially inhibited by anti-IL-12 but not by anti-IL-18 or antibodies to transforming growth factor (TGF)-β. It is concluded that IL-12 alone can not increase CLA expression but requires the help of other factor(s) present in FCS but not in HS. Although LPS can induce CLA expression it does not seem to be the factor that interacts with IL-12 to induce superantigen-mediated CLA expression in cultures enriched with FCS.
Insight into antibody responses induced by plasmid or adenoviral vectors encoding thyroid peroxidase, a major thyroid autoantigen
Authors: GUO J , PICHURIN P , NAGAYAMA Y , RAPOPORT B , MCLACHLAN S M
Published: May 2003

Plasmid and adenoviral vectors have been used to generate antibodies in mice that resemble human autoantibodies to the thyrotrophin receptor. No such studies, however, have been performed for thyroid peroxidase (TPO), the major autoantigen in human thyroiditis. We constructed plasmid and adenovirus vectors for in vivo expression of TPO. BALB/c mice were immunized directly by intramuscular injection of TPO-plasmid or TPO-adenovirus, as well as by subcutaneous injection of dendritic cells (DC) infected previously with TPO-adenovirus. Intramuscular TPO-adenovirus induced the highest, and TPO-plasmid the lowest, TPO antibody titres. Mice injected with TPO-transfected DC developed intermediate levels. Antibodies generated by all three approaches had similar affinities (Kd ∼10−9m) and recognized TPO expressed on the cell-surface. Their epitopes were analysed in competition assays using monoclonal human autoantibodies that define the TPO immunodominant region (IDR) recognized by patients with thyroid autoimmune disease. Surprisingly, high titre antibodies generated using adenovirus interacted with diverse TPO epitopes largely outside the IDR, whereas low titre antibodies induced by DNA-plasmid recognized restricted epitopes in the IDR. This inverse relationship between antibody titre and restriction to the IDR is likely to be due to epitope spreading following strong antigenic stimulation provided by the adenovirus vector. However, TPO antibody epitope spreading does not occur in Hashimoto's thyroiditis, despite high autoantibody levels. Consequently, these data support the concept that in human thyroid autoimmunity, factors besides titre must play a role in shaping an autoantibody epitopic profile.
The production of tumour necrosis factor-alpha is decreased in peripheral blood mononuclear cells from multidrug-resistant tuberculosis patients following stimulation with the 30-kDa antigen of Mycobacterium tuberculosis
Authors: LEE J-S , SONG C-H , LIM J-H , KIM H-J , PARK J-K , PAIK T-H , KIM C-H , KONG S-J , SHON M-H , JUNG S-S , JO E-K
Published: May 2003

The clearance of intracellular bacteria requires the appropriate induction of proinflammatory cytokines and chemokines to recruit macrophages and T cells to the site of infection. In this study, we investigated the production of tumour necrosis factor (TNF)-α, interleukin (IL)-8 and interferon (IFN)-γ by the peripheral blood mononuclear cells (PBMC) of patients with multidrug-resistant tuberculosis (MDR-TB) in response to in vitro stimulation with the 30-kDa antigen of Mycobacterium tuberculosis. The results were compared with those from cases of newly diagnosed TB (N-TB) and TB with treatment failure (TF-TB), and healthy tuberculin reactors (HTR). The most significantly depressed TNF-α levels were found in MDR-TB patients. IFN-γ production was depressed significantly in all groups of TB patients compared with the HTR group. TNF-α secretion in response to the 30-kDa antigen was unchanged by coculturing with recombinant human interferon (rhIFN)-γ, and was increased dramatically following IL-10 neutralization with an anti-human IL-10 antibody. The IL-8 levels were depressed significantly in MDR-TB patients compared with N-TB patients, but were similar to the IL-8 levels in TF-TB patients. Furthermore, rhTNF-α directly increased IL-8 secretion, and neutralizing antibody to TNF-α inhibited IL-8 production by the PBMC of MDR-TB patients that were stimulated with the 30-kDa antigen. Taken together, these data suggest that the PBMC of MDR-TB patients typically show TNF-α depression in response to the 30-kDa antigen, and this effect is modulated by IL-10. In addition, we highlight the role of TNF-α in IL-8 secretion in MDR-TB patients.
Release of endogenous anti-inflammatory complement regulators FHL-1 and factor H protects synovial fibroblasts during rheumatoid arthritis
Authors: FRIESE M A , MANUELIAN T , JUNNIKKALA S , HELLWAGE J , MERI S , PETER H H , GORDON D L , EIBEL H , ZIPFEL P F
Published: May 2003

Rheumatoid arthritis is a chronic inflammatory disease of unknown aetiology predominantly affecting cells and tissues of synovial joints. Here we show that the two important complement regulators FHL-1 and factor H play a protective anti-inflammatory role in rheumatoid arthritis. Expression analyses at the mRNA- and protein level show in vitro expression and secretion of both regulators by synovial fibroblasts derived from patients with rheumatoid arthritis. Similarly the two regulators are synthesized in vivo in diseased synovial tissue, and in particular synovial lining cells express high levels of FHL-1. The anti-inflammatory role of these regulators in rheumatoid arthritis is highlighted by their induction with IFN-γ and dexamethasone, whilst the pro-inflammatory cytokine TNF-α had no effect. Transient transfection experiments with various FHL-1/factor H promoter-luciferase reporter constructs into cells of distinct origin show independent cell and tissue specific promoter regulated transcription of these two regulators. The inducible expression, specifically of FHL-1 has physiological consequences. By binding directly to surfaces the released proteins protect cells from inflammatory damage and complement-mediated cell lysis. This study shows a novel protective and anti-inflammatory role of the two important complement regulators FHL-1 and factor H in rheumatoid arthritis and suggests a disease controlling role of the two proteins.
Difference in antigenic determinant profiles between human and rat myeloperoxidase
Authors: PATRY Y C , NACHMAN P H , AUDRAIN M A P , FALK R J , MEFLAH K , ESNAULT V L M
Published: May 2003

We tested whether rat and human MPO have similar antigenic determinants using 36 human MPO-ANCA positive sera, one mouse anti-rat MPO and four mouse anti-human MPO monoclonal reagents. Purified rat and human MPO were used in ELISA, with or without crossinhibition by preincubation with human MPO or irrelevant antigen in the liquid phase. Only one human MPO ANCA positive serum exhibited significant binding in rat MPO ELISA. This binding was poorly inhibited by preincubation with human MPO in the liquid phase, but was conserved after adsorption of non specific anti-rat activity in a chromatography column. Three mouse anti-human MPO IgG monoclonal antibodies did not recognize rat MPO. Only one mouse anti-human MPO IgA monoclonal antibody bound to rat MPO. This binding was poorly inhibited by preincubation with human MPO (35% at 2 µg/ml). Conversely, the mouse anti-rat MPO monoclonal did not bind human MPO. We have concluded that: (1) Most human MPO-ANCA recognize antigenic determinants on human MPO which are absent on rat MPO. Therefore, human auto-antibodies bind to epitopes which recently appeared after species evolution; (2) Inversely, the mouse anti-rat MPO monoclonal do not bind human MPO. Therefore, rat MPO epitopes have been altered during species evolution; (3) Mice injected with human MPO preferentially develop antibodies against xeno-epitopes which are not present in rodents. Therefore, human MPO may not be the best antigen to raise ANCA in animal models and (4) A comparison of the amino acid sequences of rat and human MPO may help elucidate the major antigenic epitopes.
Specific lytic activity against mycobacterial antigens is inversely correlated with the severity of tuberculosis
Authors: DE LA BARRERA S S , FINIASZ M , FRIAS A , ALEMÁN M , BARRIONUEVO P , FINK S , FRANCO M C , ABBATE E , SASIAIN M DEL C
Published: May 2003

The ability of peripheral blood mononuclear cells (PBMC) from patients with active tuberculosis to display cytotoxic responses against autologous Mycobacterium tuberculosis (Mtb)-pulsed macrophages was evaluated. Non-MHC restricted cell-dependent lytic activity was observed in ex vivo effector cells from tuberculosis patients and was mediated mainly by CD3+γδ TCR+ T (γδ T) cells bearing CD56 and/or CD16 molecules. MHC-restricted and non-MHC restricted cytotoxic T cells (CTL) were differentially expanded upon stimulation with Mtb in tuberculosis patients and normal controls (N). Class-I restricted CD8+ CTL and class-II restricted CD4+ CTL were generated in PPD+N and to a lesser extent in PPD–N. Mtb-stimulated effector cells from tuberculosis patients became progressively non-MHC restricted CD4–CD8–γδ T cells, while lytic activity of CD4+ and CD8+CTL decreased gradually as the disease became more severe. On the other hand, target cells were lysed by ex vivo cells from tuberculosis patients through the Fas-FasL and perforin pathways. Mtb-induced CD4+ CTL from tuberculosis patients and N controls preferentially employed the Fas-FasL mechanism. Mtb-induced CD8+ CTL effector cells from patients used the perforin-based mechanism while cells from N controls also used the Fas-FasL pathway. While Mtb-induced γδ CTL from patients and PPD–N employed the latter mechanism cells from PPD+N individuals also used the perforin pathway. It can be concluded that shifts in the CTL response and the cytolytic mechanisms take place as the pulmonary involvement becomes more severe.
Absence of CD4+CD25+ regulatory T cells is associated with a loss of regulation leading to increased pathology inHelicobacter pylori-infected mice
Authors: RAGHAVAN S , FREDRIKSSON M , SVENNERHOLM A-M , HOLMGREN J , SURI-PAYER E
Published: May 2003

Helicobacter pylori induces symptomatic chronic gastritis in a subpopulation of infected individuals. The mechanism(s) determining the development and severity of pathology leading to symptoms are not fully understood. In a mouse model of H. pylori infection we analysed the influence of immunoregulatory CD4+CD25+ T cells on H. pylori colonization and gastritis. Athymic C57BL/6 nu/nu mice were reconstituted with (a) lymph node (LN) cells (b) LN cells depleted of CD25+ T cells (CD25– LN) or (c) not reconstituted at all. Mice were then infected orally with 3 × 108H. pylori SS1 bacteria. At 2 and 6 weeks after the inoculation there was a significant (P < 0·001) reduction in H. pylori colonization in athymic mice transferred with CD25– LN cells compared to mice transferred with LN cells. Colonization was still reduced at 12 weeks after inoculation. Mice transferred with CD25– LN cells showed an earlier onset and increased severity of gastritis as compared to mice receiving LN cells. Splenic cells isolated from mice receiving CD25– LN cells produced the highest level of IFN-γ on stimulation with H. pylori antigens in vitro, had a higher H. pylori-specific DTH response and increased infiltration of CD4+ T cells and macrophages in the gastric mucosa. Athymic mice not transferred with T cells had persistent high H. pylori colonization and displayed a normal gastric epithelium without inflammatory cells. In conclusion, CD4+CD25+ cells reduce immunopathology in H. pylori infection, possibly by reducing the activation of IFN-γ producing CD4+ T cells, even at the expense of a higher H. pylori load in the gastric mucosa.