bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Insights Regarding the Operational Characteristics and Teleological Purpose of the Renal Na+-K+-Cl2 Cotransporter (NKCC2s) Splice Variants
Authors: Brunet Geneviève M. , Gagnon Edith , Simard Charles F. , Daigle Nikolas D. , Caron Luc , Noël Micheline , Lefoll Marie-Hélène , Bergeron Marc J. , Isenring Paul
Published: September 2005

The absorptive Na+-K+-Cl− cotransporter (NKCC2) is a polytopic protein that forms homooligomeric complexes in the apical membrane of the thick ascending loop of Henle (TAL). It occurs in at least four splice variants (called B, A, F, and AF) that are identical to one another except for a short region in the membrane-associated domain. Although each of these variants exhibits unique functional properties and distributions along the TAL, their teleological purpose and structural organization remain poorly defined. In the current work, we provide additional insight in these regards by showing in mouse that the administration of either furosemide or an H2O-rich diet, which are predicted to alter NKCC2 expression in the TAL, exerts differential effects on mRNA levels for the variants, increasing those of A (furosemide) but decreasing those of F and AF (furosemide or H2O). Based on a yeast two-hybrid mapping analysis, we also show that the formation of homooligomeric complexes is mediated by two self-interacting domains in the COOH terminus (residues 671 to 816 and 910 to 1098), and that these complexes could probably include more than one type of variant. Taken together, the data reported here suggest that A, F, and AF each play unique roles that are adapted to specific physiological needs, and that the accomplishment of such roles is coordinated through the splicing machinery as well as complex NKCC2–NKCC2 interactions.
Three‐Dimensional Computed Tomography Angiography of the Internal Carotid Artery for Preoperative Evaluation of Sinonasal Lesions and Intraoperative Surgical Navigation
Authors: Leong Jern‐Lin , Batra Pete S. , Citardi Martin J.
Published: September 2005

Introduction: Three‐dimensional computed tomographic angiography (3DCTA) demonstrates the spatial relationships of the internal carotid artery (ICA) and adjacent skull base. This imaging modality may be incorporated into intraoperative surgical navigation during endoscopic skull base surgery.

Methods: The charts of patients who had undergone 3DCTA imaging between July 2002 and February 2005 were reviewed. For 3DCTA, 1 mm axial computed tomography (CT) scan images were obtained with simultaneous intravenous contrast bolus on a multidetector CT scanner (Somatom Sensation 16, Siemens, Munich, Germany). The CBYON Suite version 2.6 to 2.8 (Med‐Surgical Services, Mountain View, CA) was also used for creating CTA images through its volume‐rendering protocols.

Results: A total of 22 3DCTA studies were performed for diagnostic evaluation or preoperative planning. In 18 instances, the 3DCTA images were used during intraoperative surgical navigation. The specific indications for obtaining the 3DCTA study included neoplasm (11 cases), cerebrospinal fluid leak (3 cases), fibro‐osseous lesion (2 cases), mucocele (2 cases), and other (4 cases). Images generated by 3DCTA facilitated the definition of the anatomic relationships between the ICA and skull base lesion. During intraoperative surgical navigation, the 3DCTA provided critical information about the ICA location and adjacent skull base anatomy in the operative field.

Conclusions: 3DCTA is a useful means for assessing the ICA and its relationship to skull base lesions. Incorporation of 3DCTA into intraoperative surgical navigation facilitates the comprehension of operative field anatomy in the ICA region. As a result, this imaging technique, especially when combined with intraoperative surgical navigation, may extend the applications of minimally invasive endoscopic approaches to the skull base.
Radial Forearm Osteocutaneous Free Flap in Maxillofacial and Oromandibular Reconstructions
Authors: Kim J H. , Rosenthal E L. , Ellis T , Wax M K.
Published: September 2005

Objectives/Hypothesis: The radial forearm osteocutaneous free flap is an excellent reconstructive modality for oromandibular and maxillofacial reconstruction in certain well‐defined circumstances. The initial concern over donor site morbidity and the ability of the bone to reconstruct mandibular defects have led to only a few published series.

Study Design: Retrospective study of the experience of two tertiary medical centers with radial forearm osteocutaneous free flap.

Methods: Retrospectively, 52 patients were studied who underwent radial forearm osteocutaneous free flap reconstruction for cancer (49 cases) and trauma (3 cases). Bone length and skin paddle harvested, general morbidity (hematoma, wound infection, and dehiscence), recipient site morbidity (nonunion of neomandible, flap failure, and bone or plate exposure), and donor site morbidity (radius bone fracture, plate exposure, and skin graft failure) were reviewed.

Results: The average skin paddle size was 55.1 cm2 (range, 15–112 cm2). The average radius bone harvest length was 6.3 cm (range, 2.5–11 cm). Donor site complications included tendon exposure (3 cases), radius bone fracture (1 case), and exposure of the plate (0). Recipient site complications included nonunion of the mandible (4), exposed mandible (1), exposed mandibular plates (2), exposed maxillary plates or bone (0), venous compromise (1), and flap failure (1). Two patients had perioperative deaths.

Conclusion: Radial forearm osteocutaneous free flap is a valuable and viable option for oromandibular and maxillofacial reconstruction.
Level IIb Lymph Nodes Metastasis in Elective Supraomohyoid Neck Dissection for Oral Cavity Squamous Cell Carcinoma: A Molecular‐Based Study
Authors: Elsheikh Mohamed Nasser , Mahfouz Magdy Elsayed , Elsheikh Ehab
Published: September 2005

Objectives: To identify the incidence of level IIb lymph nodes metastasis in elective supraomohyoid neck dissection (SOHND) as a treatment for patients with squamous cell carcinoma (SCC) of the oral cavity.

Study Design: Prospective analysis of a case series.

Methods: Forty‐eight patients with SCC of the oral cavity and with no palpable lymph nodes at the neck who underwent an elective SOHND were prospectively studied. The incidence of micrometastasis to level IIb lymph nodes after performing elective SOHND was evaluated by pathologic examination and molecular analysis.

Results: Of the 48 patients, 15 (31%) by pathologic analysis and 22 (46%) by molecular analysis had lymph nodes positive for metastatic SCC. By molecular analysis, 5 (10%) of the 48 patients had involvement of level IIb lymph nodes. All patients with metastasis to level IIb lymph nodes have their primary lesions in the tongue and constituted 22% of patients with tongue lesions. There was no instance of isolated metastasis to level IIb lymph nodes without involvement of other nodes in the SOHND specimens.

Conclusions: In this study, level IIb lymph node metastasis was only found in association with tongue carcinoma. Although this region may be preserved in elective SOHND in patients with SCC of the oral cavity, it should be included whenever the tongue is the primary site.
Species richness vs evenness: independent relationship and different responses to edaphic factors
Authors: Ma Maohua
Published: September 2005

The main components of species diversity are richness and evenness. Species richness has been the subject of biodiversity studies more often than species evenness. Some simulated models suggest that the relationship between these two components is equal and predictable in a stable environment. This study was conducted in a human‐shaped and unstable environment in arable field boundaries. A total of 30 sampling plots were laid out at random in the boundaries. Vegetation at the sites was of meadow type. The relationship between species richness and evenness, and their responses to edaphic nutrient factors in the field boundaries, were examined by correlation analysis and an ordination method. Correlation analyses demonstrated that no consistent pattern was present in the relationship between species evenness and species richness in this human‐shaped ecosystem. Species richness and evenness also had different responses to edaphic factors, with species richness being negatively correlated with phosphorus, and species evenness negatively correlated with the ratio of organic carbon to total nitrogen in soil. The results indicate that different and independent ecological processes determine species richness and evenness. The relationship between these two components may be site‐specific, reflecting variation in resource utilization by plant species. The pattern thus reflects the spatial heterogeneity of disturbances or ‘patchiness’ of resources between sites in a semi‐natural ecosystem.
Scale dependent effects of predatory fish on stream benthos
Authors: Englund Göran
Published: September 2005

In open predation experiments the effects of predators on prey densities can be influenced by predator consumption and by prey movements in to and out of experimental arenas. A published model predicts that the predator effects observed in such experiments are scale dependent over the scale range where there is a transition from movement control (of prey densities) to consumption control. The scale dependence follows from the assumption that per capita rate of emigration out of an experimental arena decreases with increasing arena size.

To test this model the effects of a small benthic fish (Cottus gobio) on densities of stream invertebrates was investigated in instream channels of different length (0.5, 2 and 8 m). The effect of fish predation was scale dependent for four prey taxa. For three of these taxa predator effects increased with experimental scale, which is in agreement with model predictions. However, this proved to be a case of “making the right prediction for the wrong reason” as the basic assumption of scale dependent emigration rate was not upheld. By analyzing the behaviour of the model, parameterized with emigration and consumption rates observed in the experimental channels, it was found that observed scale effects occurred because prey emigration in response to the predator treatment was modified by the experimental scale. Further analysis of the parameterized model suggested that the densities of most prey taxa were controlled by prey movements and not by consumption by the sculpins.
Inbreeding depression and family variation in a social insect,Bombus terrestris(Hymenoptera: Apidae)
Authors: U. Gerloff Christine , Schmid‐Hempel Paul
Published: September 2005

In bumble bees, only a few colonies dominate reproduction at the end of a season such that successful families are likely to experience episodic inbreeding. We compared the fitness of 33Bombus terrestriscolonies resulting from 188 brother–sister matings (“inbred”) to that of 43 outbred colonies (from 183 outbred matings). Large among‐family variation appears to be a key feature that characterizes all main steps in the life cycle ofB. terrestris. Hibernation survival and colony foundation of queens varied strongly among maternal lines (P>0.001). Mating with a brother significantly reduced a queen's hibernation survival and colony foundation success by varying degrees (6% vs 23%). The effect on hibernation survival was not found when including the date of mating into the analysis: queens that mated and hibernated early in the season survived hibernation more frequently than those that mated later (P=0.001) irrespective of mating treatment. Furthermore, maternal family lines differed significantly in their response to inbreeding with respect to colony size, caloric investment into sexuals, and the Shaw‐Mohler fitness index. Some family lines even showed outbreeding depression. On average, inbred colonies were significantly smaller (coefficient of depression δ=0.20) but there was no inbreeding depression for reproductive output or cumulative fitness. Thus, as a population,B. terrestrisis likely to cope with inbreeding episodes. However, among‐family variation in the effects of inbreeding intensifies the differential representation of families in the population.
Effects of local density on insect visitation and fertilization success in the narrow‐endemicCentaurea corymbosa(Asteraceae)
Authors: Kirchner Florian , Luijten Sheila H. , Imbert Eric , Riba Miquel , Mayol Maria , González‐Martínez Santiago C. , Mignot Agnès , Colas Bruno
Published: September 2005

Plant population size and density can influence the interactions between plants and pollinators, and affect plant reproductive success. We investigated the effects of local conspecific density on insect visitation and fertilization success in the rare, cliff‐dwelling, self‐incompatibleCentaurea corymbosa, in which fecundity plays a key role in variation in population growth rate among years and among the six extant populations. Plant size, capitulum size, the abundance of co‐flowering species, and the weather conditions were added as covariates in the analyses. Over three populations and two years (1995 and 2002), fertilization rate was positively related to the number of flowering conspecifics within 10 m. Fertilization rate varied among populations, but this variation over all six populations in 2002 could not be attributed to differences in population size. Observations in one population in 2003 showed that there was no difference in insect visitation per capitulum between plants in sparse vs dense patches whilst plants from sparse patches suffered reduced fertilization rate. Visitation and fertilization rates were not affected by plant size and the abundance of co‐flowering species, but weather conditions at the time of flowering had a strong effect on both variables. Capitulum size had a positive effect on visitation rate, but an effect on fertilization rate only in 1995 and 2002 and not in 2003. Our results suggest that pollen limitation affects fertilization rate inC. corymbosadue to limited compatible mate availability rather than pollinator limitation. They agree with previous genetic results derived from paternity analysis. Whether or not the benefits of local spatial agregation to reproductive success result in increased individual fitness will depend on the relative reduction in survival of vegetative stages due to intra‐specific competition.
Experimental testing and numerical modeling of human skin
Authors: Ren X. J. , Smith C. W. , Evans K. E. , Dooling P. J. , Burgess A. N. , Zahlan N. , Rawlings A. V. , Wiechers J. W.
Published: September 2005

In this feasibility study, changes in the elastic properties of skin following various skin treatments (e.g. water, skincare ingredients, etc.) were investigated using a new in vivo tensile testing method. The properties of human skin in vivo, and in particular the properties of the stratum corneum, were studied using a combined experimental‐numerical approach. Tests on a small number of volunteers indicated that the in vivo testing method was sensitive to the effect of skin treatments and could distinguish their relative performances. The ingredients tested effectively made skin less stiff. Preliminary results showed that stratum corneum is highly anisotropic and that the data was in general agreement with some but not all published in vitro results, although this is the first time ever that Poisson ratios for human stratum corneum are presented.
Assessment of different methods of inhalation from salbutamol metered dose inhalers by urinary drug excretion and methacholine challenge
Authors: Tomlinson Heather S. , Corlett Sarah A. , Allen Martin B. , Chrystyn Henry
Published: September 2005

Aims

Methods to determine the lung delivery of inhaled bronchodilators from metered dose inhalers include urinary drug excretion 30 min post inhalation and methacholine challenge (PD20). We have compared these two methods to differentiate lung delivery of salbutamol from metered dose inhalers using different inhalation methods.

Methods

In phase 1 of the study, on randomized study days, 12 mild asthmatics inhaled placebo, one and two 100 µg salbutamol doses from a breath actuated metered dose inhaler, in randomized fashion on different days. In phase 2, they inhaled one 100 µg salbutamol dose from a metered dose inhaler using a SLOW (20 l min−1) and a FAST (60 l min−1) inhalation technique and a slow inhalation delayed until after they had inhaled for 5 s (LATE). Urinary excretion of salbutamol (0–30 min) and PD20 were measured after each dose.

Results

Following placebo, one and two 100 µg salbutamol doses, the geometric mean for PD20 was 0.10, 0.41 and 0.86 mg respectively and the mean (SD) urinary drug excretion after one and two doses was 2.25 (0.65) and 5.37 (1.36) µg, respectively. After SLOW, FAST and LATE inhalations the geometric mean for PD20 was 0.50, 0.40 and 0.42 mg, respectively, and mean (SD) salbutamol excretion was 2.67 (0.84), 1.90 (0.70) and 2.72 (0.67) µg, respectively. Only the amount of drug excreted during the FAST compared with the SLOW and LATE inhalations showed a statistical difference (95% confidence interval on the difference 0.12, 1.54 and 0.06,1.59 µg, respectively).

Conclusions

Urinary salbutamol excretion but not PD20 showed differences between the inhalation methods used. When using a metered dose inhaler slow inhalation is better and co‐ordination is not essential if the patient is inhaling when they actuate a dose of the drug.
Children with stable asthma have reduced airway matrix metalloproteinase‐9 and matrix metalloproteinase‐9/tissue inhibitor of metalloproteinase‐1 ratio
Authors: Doherty G. M. , Kamath S. V. , De Courcey F. , Christie S. N. , Chisakuta A. , Lyons J. D. , Heaney L. G. , Ennis M. , Shields M. D.
Published: September 2005

Background
Childhood asthma is characterized by inflammation of the airways. Structural changes of the airway wall may also be seen in some children early in the course of the disease. Matrix metalloproteinases (MMPs) are key mediators in the metabolism of the extracellular matrix (ECM).

Objective
To investigate the balance of MMP‐8, MMP‐9 and tissue inhibitor of metalloproteinases (TIMP)‐1 in the airways of children with asthma.

Methods
One hundred and twenty‐four children undergoing elective surgical procedures also underwent non‐bronchoscopic bronchoalveolar lavage (BAL). MMP‐8, MMP‐9 and TIMP‐1 were measured by ELISA.

Results
There was a significant reduction in MMP‐9 in atopic asthmatic children (n=31) compared with normal children (n=30) [median difference: 0.57 ng/mL (95% confidence interval: 0.18–1.1 ng/mL)]. The ratio of MMP‐9 to TIMP‐1 was also reduced in asthmatic children. Levels of all three proteins were significantly correlated to each other and to the relative proportions of particular inflammatory cells in BAL fluid (BALF). Both MMP‐8 and MMP‐9 were moderately strongly correlated to the percentage neutrophil count (r=0.40 and 0.47, respectively, P<0.001).

Conclusions
An imbalance of MMPs and their inhibitors occurs in children with well‐controlled asthma, which may indicate early derangement of the metabolism of the ECM.
The predictive value of the skin prick test weal size for the outcome of oral food challenges
Authors: Verstege A. , Mehl A. , Rolinck‐Werninghaus C. , Staden U. , Nocon M. , Beyer K. , Niggemann B.
Published: September 2005

Background
The skin prick test (SPT) is regarded as an important diagnostic measure in the diagnostic work‐up of food allergy.

Objective
To evaluate the diagnostic capacity of the SPT in predicting the outcome of oral food challenges, and to determine decision points for the weal size and the skin index (SI) that could render double‐blind, placebo‐controlled food challenges unnecessary.

Methods
In 385 children (median age 22 months), 735 controlled oral challenges were performed with cow's milk (CM), hen's egg (HE), wheat and soy. Three hundred and thirty‐six of 385 (87%) children suffered from atopic dermatitis. SPT was performed in all children. Diagnostic capacity, receiver–operator characteristics (ROC) curves and predictive decision points were calculated for the mean weal size and the calculated SI.

Results
Three hundred and twelve of 735 (43%) oral food challenges were assessed to be positive. Calculation of 95% and 99% predicted probabilities using logistic regression revealed predictive decision points of 13.0 and 17.8 mm for HE, and 12.5 and 17.3 mm for CM, respectively. However, using the SI, the corresponding cut‐off levels were 2.6 and 3.7, respectively, for HE, and 2.7 and 3.7 for CM. For wheat, 95% and 99% decision points of 2.2 and 3.0 were found in children below 1 year of age.

Conclusion
Predictive decision points for a positive outcome of food challenges can be calculated for HE and CM using weal size and SI. They may help to avoid oral food challenges.
Anti-endomysial antibody of IgG1 isotype detection strongly increases the prevalence of coeliac disease in patients affected by type I diabetes mellitus
Authors: Picarelli A , Sabbatella L , Di Tola M , Vetrano S , Casale C , Anania M C , Porowska B , Vergari M , Schiaffini R , Gargiulo P
Published: September 2005

A strong association between type 1 insulin-dependent diabetes mellitus (IDDM1) and coeliac disease (CD) is well documented, but it is known that prevalence values are underestimated. Serum anti-endomysial antibodies (EMA), considered diagnostic for CD because of their high sensitivity and specificity, belong to the IgA class, but the existence of EMA of IgG1 isotype in the presence or absence of IgA deficiency was reported. In order to re-evaluate the occurrence of CD in IDDM1 patients we performed a screening in IDDM1 patients using EMA of both isotypes. Ninety-four adults affected by IDDM1 (unaffected by CD before enrolling) were enrolled and 83 blood donors as controls. All subjects were on a gluten-containing diet. Histology and biopsy culture were performed. EMA IgA and IgG1 in sera and culture supernatants were detected. Serum EMA were positive in 13 of 94 IDDM1 patients (13·8%). Six of 13 presented IgA-EMA, seven of 13 presented IgG1-EMA. No EMA were found in the control population. Total intestinal atrophy was found in all six patients with serum IgA-EMA and in five of seven with serum IgG1-EMA. Diagnosis of CD was confirmed by histology and organ culture in all 13 patients with serum EMA. The prevalence of CD in the patients affected by IDDM1 was 6·4% for IgA-EMA-positive and 7·4% for IgG1-EMA-positive patients. We confirmed the prevalence of CD in the IDDM1 population obtained with IgA-EMA screening only (6·4%). This prevalence value increases dramatically to 13·8% when IgG1-EMA are also used in the screening. We conclude that IgG1-EMA should also be sought whenever an IDDM1 patient undergoes screening for CD.
Defining the role of T cell-derived leptin in the modulation of hepatic or intestinal inflammation in mice
Authors: Fantuzzi G , Sennello J A , Batra A , Fedke I , Lehr H A , Zeitz M , Siegmund B
Published: September 2005

The role of leptin in the immune system has been well established. While adipocytes represent the major source, leptin production by lymphocytes, infiltrating at the site of inflammation, was recently demonstrated. However, the significance of this locally released leptin remains unresolved. In the present study, two models in which absence of leptin-signalling is associated with protection were employed: the model of ConA-induced hepatitis and the CD4+CD45Rbhigh transfer model of colitis. For the ConA model, scid mice were reconstituted with either WT or leptin-deficient (ob/ob) CD4+ T cells. Eight weeks post transfer, ConA was injected and serum ALT, TNFα, leptin as well as liver mononuclear cell activation and histological signs of inflammation were evaluated. No difference between recipients of WT or ob/ob cells was observed for any of the parameters evaluated. In the second model, either WT or ob/ob CD4+CD45Rbhigh cells were transferred into scid mice. No histological differences were detected, although recipients of ob/ob cells showed higher weight loss compared to recipients of WT cells. Spontaneous production of IL-6 from colon cultures obtained from recipients of ob/ob cells was reduced compared to recipients of WT cells, whereas stimulation of lamina propria lymphocytes with leptin resulted in a higher IFNγ release in recipients of ob/ob cells compared to recipients of WT cells. In conclusion, the present study provides evidence that T cell-derived leptin does not play a major role in the regulation of the inflammatory process, indicating that the adipose tissue is the critical player in the immune-modulating effects of leptin.
Increase of CCL20 expression by human gingival fibroblasts upon stimulation with cytokines and bacterial endotoxin
Authors: Hosokawa Y , Hosokawa I , Ozaki K , Nakae H , Matsuo T
Published: September 2005

We have demonstrated recently that CCL20 was expressed in periodontal diseased tissues and abundant CCR6 positive T cells infiltrated in periodontally diseased tissue. However, it is uncertain which cells can elicit CCL20 production. In the present study, we examined the properties of CCL20 production by human gingival fibroblasts (HGF) culture. Here, we report that interleukin-1 beta (IL-1β), tumour necrosis factor-alpha (TNF-α) and Escherichia coli lipopolysaccharide (LPS) can significantly induce the production of CCL20 by HGF. We found that TNF-α and E. coli LPS enhanced the production of CCL20 by HGF treated with IL-1β. In contrast, interferon-gamma (IFN-γ) dramatically diminished CCL20 production induced by IL-1β. Moreover, we demonstrated that nuclear factor-kappaB (NF-κB), p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases (ERK) play an important role in mediating the production of CCL20 induced by IL-1β and TNF-α. On the other hand, we found that not only NF-κB, p38 MAPK and ERK but also c-Jun NH2-terminal kinase (JNK) are involved in CCL20 production induced by E. coli LPS. Finally, we found that HGF express CCR6, CCL20 receptor, and CCL20 induced vascular endothelial growth factor (VEGF) by HGF. Taken together, these findings that HGF will be a source of CCL20 in periodontal tissue, and the CCL20 production will be controlled by proinflammatory cytokine and bacterial LPS in periodontally diseased tissue. Thus, CCL20 by HGF might be involved in inflammatory cells infiltration, and promote the progression of periodontal disease.
Inhibition of human immunodeficiency virus-1 (HIV-1) by β-chemokine analogues in mononuclear cells from HIV-1-infected patients with active tuberculosis
Authors: Toossi Z , Mayanja-Kizza H , Baseke J , Peters P , Wu M , Abraha A , Aung H , Okwera A , Hirsch C , Arts E
Published: September 2005

Tuberculosis (TB) enhances human immunodeficiency virus-1 (HIV-1) activity in patients with dual HIV-1/TB infection. Therapies that control augmentations of HIV-1 activity at sites of Mycobacterium tuberculosis (MTB) infection may be useful in inhibition of viral expansion. Regulated upon activation, normal T-cell expressed and secreted (RANTES) analogues (AOP and NNY) are potent in inhibiting the entry of primary HIV-1 isolates into host mononuclear cells. These analogues were used to inhibit MTB-induced HIV-1 entry in blood monunuclear cells (PBMC) from patients with pulmonary TB, and pleural fluid mononuclear cells (PFMC) from patients with pleural TB. PBMC or PFMC were cultured with and without MTB in presence and absence of RANTES analogues. HIV-1 strong stop DNA was assessed by real-time polymerase chain reaction (PCR) as a measure of infection. CCR5 mRNA was assessed by real-time reverse transcription (RT)-PCR and by immunostaining and FACS analysis. HIV-1 infection was induced by MTB in vitro in PBMC from the majority (14 of 20) of HIV-1/TB subjects, and new infection was inhibited by AOP- or NNY-RANTES. HIV-1 infection was also inhibited by these reagents in MTB-induced PFMC from three of three patients with pleural TB. Expression of CCR5 mRNA was significantly induced by MTB in PBMC from patients with pulmonary TB. Further, expression of CCR5 was higher in PFMC compared to PBMC from patients with pleural TB. Also, CCR5 was fourfold higher on CD14+ pleural mononuclear cells than on CD4+ lymphocytes. Blocking new HIV-1 infection of mononuclear cells may be useful in control of HIV-1 during dual HIV-1/TB infection.
Optimumin vitroexpansion of human antigen-specific CD8+ T cells for adoptive transfer therapy
Authors: Montes M , Rufer N , Appay V , Reynard S , Pittet M J , Speiser D E , Guillaume P , Cerottini J-C , Romero P , Leyvraz S
Published: September 2005

Increasing evidence suggests that adoptive transfer of antigen-specific CD8+ T cells could represent an effective strategy in the fight against chronic viral infections and malignancies such as melanoma. None the less, a major limitation in the implementation of such therapy resides in the difficulties associated with achieving rapid and efficient expansion of functional T cells in culture necessary to obtain the large numbers required for intravenous infusion. Recently, the critical role of the cytokines interleukin (IL)-2, IL-7 and IL-15 in driving T cell proliferation has been emphasized, thus suggesting their use in the optimization of expansion protocols. We have used major histocompatibility complex (MHC) class I/peptide multimers to monitor the expansion of antigen-specific CD8 T lymphocytes from whole blood, exploring the effect of antigenic peptide dose, IL-2, IL-7 and IL-15 concentrations on the magnitude and functional characteristics of the antigen-specific CD8+ T cells generated. We show here that significant expansions of antigen-specific T cells, up to 50% of the CD8+ T cell population, can be obtained after a single round of antigen/cytokine (IL-2 or IL-15) stimulation, and that these cells display good cytolytic and interferon (IFN)-γ secretion capabilities. Our results provide an important basis for the rapid in vitro expansion of autologous T cells from the circulating lymphocyte pool using a simple procedure, which is necessary for the development of adoptive transfer therapies.
The effects of atorvastatin on gluten-induced intestinal T cell responses in coeliac disease
Authors: Ráki M , Molberg Ø , Tollefsen S , Lundin K E A , Sollid L M
Published: September 2005

Various experimental models suggest that the cholesterol-lowering drugs statins may also modulate immune responses. Cellular level studies on human disorders are needed, however, to provide a rational basis for clinical testing of statins as immune therapy. Coeliac disease, a chronic small intestinal inflammation driven by HLA-DQ2 restricted mucosal T cells that are specific for ingested wheat gluten peptides, is in many ways ideal for this purpose. In addition, there is a need for alternative treatment to the gluten-free diet in this disorder. Here we have assessed the effects of atorvastatin on gluten-reactive T cells, dendritic cells and the coeliac mucosa by in vitro culture of biopsies. Atorvastatin inhibited gluten-induced proliferation and specific cytokine production of human intestinal gluten-reactive T cell clones and lines. Dendritic cells exposed to atorvastatin displayed a reduced expression of the costimulatory molecule CD83 upon maturation with lipopolysaccharide. Incubation of intestinal biopsy specimens with atorvastatin in vitro, however, did not influence gluten-induced cytokine release. In conclusion, atorvastatin has specific effects on isolated gluten-reactive T cells and dendritic cells, but does not shut down the gluten-induced production of proinflammatory cytokines in intestinal biopsies.
Altered phenotype and function of dendritic cells in children with type 1 diabetes
Authors: Angelini F , Del Duca E , Piccinini S , Pacciani V , Rossi P , Manca Bitti M L
Published: September 2005

The importance of dendritic cells (DC) in the activation of T cells and in the maintenance of self-tolerance is well known. We investigated whether alterations in phenotype and function of DC may contribute to the pathogenesis of Type 1 diabetes (T1DM). Mature DC (mDC) from 18 children with T1DM and 10 age-matched healthy children were tested. mDC, derived from peripheral blood monocytes cultured for 6 days in presence of interleukin (IL)-4 and granulocyte-macrophage colony stimulating factor (GM-CSF) and stimulated with lipopolysaccharide (LPS) for the last 24 h, were phenotyped for the expression of the co-stimulatory molecules B7·1 and B7·2. In six patients and six controls allogenic mixed leucocyte reaction (AMLR) was performed using mDC and cord blood-derived naive T cells at a DC/T naive ratio of 1 : 200. Proliferation was assessed on day 7 by [3H]-thymidine incorporation assay. Mature DC derived from patients showed, compared with controls, a reduced expression of B7·1 [mean of fluorescence intensity (MFI): 36·2 ± 14·3 versus 72·9 ± 34·5; P = 0·004] and B7·2 (MFI: 122·7 ± 67·5 versus 259·6 ± 154·1; P = 0·02). We did not find differences in the HLA-DR expression (P = 0·07). Moreover, proliferative response of allogenic naive T cells cultured with mDC was impaired in the patients (13471 ± 9917·2 versus 40976 ± 24527·2 cpm, P = 0·04). We also measured IL-10 and IL-12 concentration in the supernatant of DC cultures. Interestingly, we observed in the patients a sevenfold higher level of IL-10 (P = 0·07) and a ninefold lower level of IL-12 (P = 0·01). Our data show a defect in the expression of the co-stimulatory molecules and an impairment of DC priming function, events that might contribute to T1DM pathogenesis.
Temporal cytokine gene expression patterns in subjects with trachoma identify distinct conjunctival responses associated with infection
Authors: Faal N , Bailey R L , Sarr I , Joof H , Mabey D C W , Holland M J
Published: September 2005

Ocular chlamydial disease is clinically diagnosed by the appearance of characteristic inflammatory changes and development of lymphoid follicles in the conjunctiva. Nucleic acid amplification tests and relatively non-invasive methods of sampling the conjunctival surface can be used to quantify the expression of chlamydial and host genes. Using quantitative real-time polymerase chain reaction to detect the presence of Chlamydia trachomatis (CT) 16S rRNA and human interleukin (IL)-1β, IL-10, IL-12p40, interferon (IFN)-γ and tumour necrosis factor (TNF)-α transcripts we examined the immune response at the conjunctival surface in a cohort of children living in a trachoma-endemic village in The Gambia. Elevated cytokine transcript levels were associated with the presence of CT 16S rRNA. Subclinical infection (CT infection without clinical signs of disease) elicited an immune response that is proinflammatory in nature, with elevations in the transcription of IL-1β, IFN-γ and IL-12p40. Clinically apparent infections were associated with the elevation of mRNA for the multi-functional cytokine TNF-α (fibrotic, type 1 inflammatory and regulatory) and the counter regulatory cytokine, IL-10, in addition to the other proinflammatory cytokines. A positive correlation between IFN-γ transcript levels and the amount of CT 16S rRNA expressed in conjunctiva was found.
Molecular manipulation with the arthritogenic epitopes of the G1 domain of human cartilage proteoglycan aggrecan
Authors: Murad Y M , Szabó Z , Ludányi K , Glant T T
Published: September 2005

Systemic immunization of BALB/c mice with human cartilage proteoglycan (PG) aggrecan induces progressive polyarthritis. The G1 domain of the PG aggrecan molecule contains most of the T cell epitopes, including three immunodominant (‘arthritogenic’) and at least six subdominant T cell epitopes. The three dominant T cell epitopes (P49, P70 and P155) were deleted individually or in combination by site directed mutagenesis, and the recombinant human G1 (rhG1) domain (wild type and mutated) proteins were used for immunization. Close to 100% of BALB/c mice immunized with the wild-type (nonmutated) rhG1 domain developed severe arthritis, which was 75% in the absence of P70 (5/4E8) epitope, and very low (< 10% incidence) when all three dominant T cell epitopes were deleted. The onset was delayed and the severity of arthritis reduced in animals when dominant T cell epitopes were missing from the immunizing rhG1 domain. The lack of T cell response to the deleted epitope(s) was specific, but the overall immune response against the wild-type rhG1 domain of human PG was not significantly affected. This study helped us to understand the dynamics and immune-regulatory mechanisms of arthritis, and supported the hypothesis that the development of autoimmune arthritis requires a concerted T cell response to multiple epitopes, rather than the immune response to a single arthritogenic structure.
Anti-phospholipid antibodies following vaccination with recombinant hepatitis B vaccine
Authors: Martinuč Porobič J , Avčin T , Božič B , Kuhar M , Čučnik S , Zupančič M , Prosenc K , Kveder T , Rozman B
Published: September 2005

This study was undertaken to evaluate the possible role of hepatitis B recombinant vaccine inducing the synthesis of IgG and IgM anti-cardiolipin antibodies (aCL), antibodies against β2GPI (anti-β2GPI), lupus anti-coagulant (LA), anti-nuclear antibodies and antibodies against extractable nuclear antigens (anti-ENA). The study population consisted of 85 healthy students (63 female, 22 male; mean age 20·8 years), vaccinated with three doses of recombinant DNA hepatitis B vaccine. One month after vaccination with the first dose of hepatitis B vaccine a minority of vaccinated individuals showed changes in IgG or IgM aCL or anti-β2GPI or LA activity (P < 0·001). Among subjects in whom changes of IgG anti-β2GPI were observed, a significantly higher number of increased (8/85) than decreased (2/85) values were found (P < 0·01). Analyses of paired data showed that differences in aCL or anti-β2GPI levels before vaccination or 1 month later did not reach statistical significance. In two people aCL transitorily reached medium positivity after the first dose of hepatitis B vaccine with a drop 5 months later. Similar evident anti-β2GPI fluctuation was also observed in one person. Another participant was initially low positive for IgG anti-β2GPI and the levels were increasing after vaccination. Two participants became positive for anti-nuclear antibodies during 6 months’ follow-up. There were no sex-dependent differences in tested antibodies observed and no associations between levels of aPL and levels of anti-HBV antibodies. We conclude that HBV can induce aPL, although rarely. In genetically susceptible individuals or together with some other triggers such combination might confer the risk of developing a continuous autoimmune response in an individual.
Cellular immunity in healthy volunteers treated with an octavalent conjugatePseudomonas aeruginosavaccine
Authors: Zuercher A W , Imboden M A , Jampen S , Bosse D , Ulrich M , Chtioui H , Lauterburg B H , Lang A B
Published: September 2005

Humoral immunity in response to an octavalent O-polysaccharide-toxin A conjugate Pseudomonas aeruginosa vaccine is well studied, and a Phase III clinical study in cystic fibrosis (CF) patients is currently ongoing. In contrast, little is known about cellular immunity induced by this vaccine. Fifteen healthy volunteers were immunized on days 1 and 60. Parameters of cellular immunity were studied before vaccination on day 1, and on day 74. Analyses included flow cytometry of whole blood, and antigen-induced proliferation of and cytokine production by lymphocyte cultures. The effects of immunization on the composition of peripheral blood lymphocytes as determined by flow cytometry were minor. In contrast, after immunization a highly significant increase of proliferation in response to stimulation with detoxified toxin A was noted: the stimulation index rose from 1·4 on day 1 to 42·2 on day 74 (restimulation with 0·4 µg/ml; P = 0·003). Immunization led to significant production of interferon (IFN)-γ and tumour necrosis factor (TNF)-α by antigen-stimulated lymphocytes. In contrast, no significant induction of interleukin (IL)-4 or IL-10 was observed. In conclusion, immunization of healthy volunteers led to activation of cellular immunity including strong antigen-specific proliferation and cytokine production. In CF patients priming of the cellular immune system towards a Th1-like pattern would be of potential advantage. Therefore, confirmatory analyses in immunized CF patients with and without chronic infection with P. aeruginosa are foreseen.
Dancing with the enemy: the interplay of herpes simplex virus with dendritic cells
Authors: Novak N , Peng W M
Published: September 2005

Herpes simplex virus (HSV) represents a smart pathogen, which displays both lytic and latent modes of interaction with its natural human host. In order to be optimally equipped for immune evasion and to reply to any attacks of the host during reactivation, HSV has developed a multitude of cleverly devised defence strategies. Dendritic cells (DC) as antigen-presenting cells located at the border zones of the body and the environment have been shown to play a crucial role as one of the first cells interacting with HSV beside epithelial cells, on one hand, and as important controllers of the viral spreading on the other hand. Here, we provide a research update about the interaction of HSV with DC and summarize the latest proceedings in this field.
Stable, soluble, high-affinity, engineered T cell receptors: novel antibody-like proteins for specific targeting of peptide antigens
Authors: Boulter J M , Jakobsen B K
Published: September 2005

The recent development of T cell receptor phage display opens up the possibility of engineering human T cell receptors with antibody-like binding properties for cell-surface peptide antigens. In this review we briefly discuss recent developments in molecular targeting of peptide antigens. We then discuss potential clinical applications of engineered high-affinity T cell receptors in autoimmunity and cancer.
Defective signalling in salivary glands precedes the autoimmune response in the non-obese diabetic mouse model of sialadenitis
Authors: Rosignoli F , Roca V , Meiss R , Leceta J , Gomariz R P , Pérez Leirós C
Published: September 2005

The spontaneous non-obese diabetic (NOD) mouse model of Sjögren's syndrome provides a valuable tool to study the onset and progression of both the autoimmune response and secretory dysfunction. Our purpose was to analyse the temporal decline of salivary secretion in NOD mice in relation to the autoimmune response and alterations in various signalling pathways involved in saliva secretion within each salivary gland. A progressive loss of nitric oxide synthase activity in submandibular and parotid glands started at 12 weeks of age and paralleled the decline in salivary secretion. This defect was associated with a lower response to vasoactive intestinal peptide in salivary flow rate, cAMP and nitric oxide/cGMP production. No signs of mononuclear infiltrates or local cytokine production were detectable in salivary glands in the time period studied (10–16 weeks of age). Our data support a disease model for sialadenitis in NOD mice in which the early stages are characterized by defective neurotransmitter-mediated signalling in major salivary glands that precedes the autoimmune response.
Correlations among measles virus-specific antibody, lymphoproliferation and Th1/Th2 cytokine responses following measles–mumps–rubella-II (MMR-II) vaccination
Authors: Dhiman N , Ovsyannikova I G , Ryan J E , Jacobson R M , Vierkant R A , Pankratz V S , Jacobsen S J , Poland G A
Published: September 2005

Immunity to measles is conferred by the interplay of humoral and cellular immune responses, the latter being critical in maintaining long-term recall response. Therefore, it is important to evaluate measles-specific humoral and cellular immunity in populations several years after vaccination and understand the correlations among these measures of immunity. We examined measles-specific antibodies, lymphoproliferation and the Th1/Th2 signature cytokines, interferon (IFN)-γ and interleukin (IL)-4, in a population-based cohort of healthy children from Olmsted County, Minnesota after two doses of measles–mumps–rubella-II (MMR-II) vaccine. We detected positive measures of measles-specific cellular and humoral immunity in the majority of our study population. However, a small proportion of subjects demonstrated an immune response skewed towards the Th2 type, characterized by the presence of either IL-4 and/or measles-specific antibodies and a lack of IFN-γ production. Further, we observed a significant positive correlation between lymphoproliferation and secretion of IFN-γ (r = 0·20, P = 0·0002) and IL-4 (r = 0·15, P = 0·005). Measles antibody levels were correlated with lymphoproliferation (r = 0·12, P = 0·03), but lacked correlation to either cytokine type. In conclusion, we demonstrated the presence of both long-term cellular and humoral responses after MMR-II vaccination in a significant proportion of study subjects. Further, a positive correlation between lymphoproliferation and IL-4 and IFN-γ suggests that immunity to measles may be maintained by both Th1 and Th2 cells. We speculate that the Th2 biased response observed in a subset of our subjects may be insufficient to provide long-term immunity against measles. Further examination of the determinants of Th1 versus Th2 skewing of the immune response and long-term follow-up is needed.
Selective expansion of CD16highCCR2– subpopulation of circulating monocytes with preferential production of haem oxygenase (HO)-1 in response to acute inflammation
Authors: Mizuno K , Toma T , Tsukiji H , Okamoto H , Yamazaki H , Ohta K , Ohta K , Kasahara Y , Koizumi S , Yachie A
Published: September 2005

Monocytes are composed of two distinct subpopulations in the peripheral blood as determined by their surface antigen expressions, profiles of cytokine production and functional roles played in vivo. We attempted to delineate the unique functional roles played by a minor CD16highCCR2– subpopulation of circulating monocytes. They produced significant levels of interleukin (IL)-6 and tumour necrosis factor (TNF)-α, but very low levels of IL-10 upon in vitro stimulation. Characteristic profiles of cytokine production were confirmed by stimulating purified subpopulations of monocytes after cell sorting. It was noteworthy that freshly isolated CD16highCCR2– monocyte subpopulations produced significant levels of haem oxygenase (HO)-1, whereas the major CD16lowCCR2+ subpopulation produced little. These results were contrary to the generally accepted notion that the CD16highCCR2– monocyte subpopulation plays a predominantly proinflammatory role in vivo. The CD16highCCR2– subpopulation increased in Kawasaki disease and influenza virus infection. In accord with this, HO-1 mRNA expression by mononuclear cells was significantly increased in these illnesses. These results indicate that CD16highCCR2– subpopulations are of a distinct lineage from CD16lowCCR2+ monocytes. More importantly, they may represent a monocyte subpopulation with a unique functional role to regulate inflammation by producing HO-1 in steady state in vivo.