bluuebunny/crossref_metadata_2025 huggingface dataset:
Specific patterns of responsiveness to microbial antigens staphylococcal enterotoxin B and purified protein derivative by cord blood mononuclear cells are predictive of risk for development of atopic dermatitis
Authors: Sharp M. J. , Rowe J. , Kusel M. , Sly P. D. , Holt P. G.
Published: December 2004

Background Mononuclear cells from children with active atopic dermatitis (AD) have been reported to be hyper‐responsive to certain microbial stimuli, in particular staphylococcal enterotoxin B (SEB). However, it is not known whether this responsiveness is acquired during disease development, or is inherent. We investigated this question in a cohort of children at high risk of atopy followed prospectively from birth to age 3 years. We asked whether their cord blood mononuclear cell (CBMC) cytokine responses to SEB, to an unrelated microbial stimulus purified protein derivative (PPD), or to common allergens, were predictive of risk for subsequent AD development during infancy.

Methods Children at high risk of developing atopy were randomly selected from an ongoing prospective cohort. Cord blood was collected at birth. The children were seen at 6 months, 1, 2 and 3 years and examined for the development of AD. IFN‐γ, IL‐5, IL‐10 and IL‐13 production by CBMC cultured in the presence of SEB, PPD, PHA, house dust mite (HDM) allergen, ovalbumin (OVA) and cat allergen was determined.

Results SEB‐induced IL‐5 production by CBMC was elevated in children who developed AD at 6 months (P = 0.01) and 2 years (P = 0.009). PPD‐induced IL‐5 responses were also elevated in CBMC from children who developed AD at 6 months, 2 years and 3 years (P = 0.05, P = 0.06 and P = 0.06, respectively), as were PPD‐induced IL‐10 responses (P = 0.05 at 1 years, P = 0.007 at 2 years, P = 0.003 at 3 years) and corresponding IFN‐γ responses (P = 0.05 at 6 months, P = 0.003 at 2 years, P = 0.0004 at 3 years). Increased IL‐10 responses to HDM allergen were also observed throughout the observation period in CBMC from children who developed AD.

Conclusion Children who develop infantile AD appear to have a predisposition to respond to SEB in a Th2‐dominant manner involving selective stimulation of IL‐5 production. The increased IL‐10 and IFN‐γ induced in response to PPD by children with AD may point to additional intrinsic differences in responses to microbial stimuli between those at high vs. those at low risk for AD, which merit more detailed investigations.
Onset of allergy and asthma symptoms in extra‐European immigrants to Milan, Italy: possible role of environmental factors
Authors: Tedeschi A. , Barcella M. , Bo G. A. Dal , Miadonna A.
Published: December 2004

Background Allergy and asthma are typical disorders of the affluent societies. Migrants from developing to industrialized countries seem to be at increased risk of allergy and asthma development.

Objective To evaluate time of onset, spectrum of sensitization and clinical features in a population of extra‐European immigrants to Milan, Italy, complaining of allergy and asthma symptoms.

Methods Data regarding 243 extra‐European immigrants checked at an allergy clinic from 1994 to 2000 were collected retrospectively. The demographic data were compared with those of the extra‐European immigrants living in Milan at the end of 1999.

Results The patients were complaining of asthma (63.7%), rhinoconjunctivitis (56.7%), rhinitis alone (21%) or urticaria (3%). One hundred and eighty‐seven out of 222 patients (84.3%) declared they were healthy before migrating and allergy/asthma symptoms started to appear after their arrival in Italy, namely after an average period of 4 years and 7 months. The proportion of male patients was lower than the proportion of men in the extra‐European immigrant population (48% vs. 55%), suggesting that in adult immigrants allergy and asthma are more common in women than in men. Furthermore, there was an over‐representation of Central‐South Americans attending the clinic, which seemed to be due to a genetic predisposition to allergy/asthma development. When data were analysed for single countries, a trend towards an increased risk of allergy and asthma was found in immigrants from all Central‐South American countries. A skin test positivity for at least one inhalant allergen was found in 196 out of 232 patients (81%), and the spectrum of allergic sensitization was similar to that of the Italian population living in the North of Italy.

Conclusion Most extra‐European immigrants declared that they were healthy at home and that allergy and asthma symptoms had appeared after immigration to Milan; lifestyle and environmental factors in a western industrialized city seem indeed to facilitate allergy/asthma onset in immigrants from developing countries. Allergy/asthma risk seems to be different in different ethnic groups.
CD14 and development of atopic disease at 2 years of age in children with atopic or non‐atopic mothers
Authors: Holmlund U. , Höglind A. , Larsson A.‐K. , Nilsson C. , Sverremark Ekström E.
Published: December 2004

Background CD14, a myeloid cell marker and LPS receptor has been acclaimed to play a role in development and manifestation of atopic allergy, as the gene encoding CD14 is located in a chromosomal region linked to total IgE levels and atopic disease.

Objective To investigate the levels of soluble (s) and membrane bound (m) CD14 in cord blood and at 2 years of age from children with atopic or non‐atopic mothers and relate these parameters to atopy development at 2 years of age.

Methods Blood samples were collected at delivery (cord blood) and at 2 years of age among infants with atopic (n = 41) and non‐atopic (n = 32) mothers. Blood samples were also obtained from mothers at the same occasions. Levels of sCD14 and total IgE were measured in plasma, and percentages of CD14+ cells were measured in cord and peripheral blood mononuclear cells.

Results We observed significant differences in sCD14 levels in cord blood, where children with atopic mothers had the highest levels. The same pattern could be observed in the mothers at delivery. At 2 years of age no significant differences in sCD14 levels were observed between children with atopic mothers and children with non‐atopic mothers and no association between sCD14 and atopic disease was found. Further, we observed large differences in sCD14 and mCD14 with respect to age, where newborns displayed a higher frequency of CD14+ cells compared with the 2‐year‐olds and the mothers. The reverse was observed for sCD14, with significantly lower values in cord blood than those seen in the 2‐year‐olds and mothers.

Conclusion Based on our findings, we suggest that CD14 could be involved in the regulation of IgE production, but that it might also be important for the maturation and development of the neonatal immune system.
A single nasal allergen challenge increases induced sputum inflammatory markers in non‐asthmatic subjects with seasonal allergic rhinitis: correlation with plasma interleukin‐5
Authors: Beeh K. M. , Beier J. , Kornmann O. , Meier C. , Taeumer T. , Buhl R.
Published: December 2004

Background Seasonal allergic rhinitis (SAR) is a risk factor for asthma in affected individuals. Nasal allergic inflammation enhances bone‐marrow eosinophil production, mainly via IL‐5, and rhinitis patients have increased airway inflammation during the pollen season.

Objective To assess the impact of nasal allergy on sputum inflammatory markers.

Methods In an open‐labelled, randomized, placebo‐controlled cross‐over study with 16 non‐asthmatic SAR patients (median age 25 years, 56% males), the effect of a single nasal allergen challenge performed out of season on induced sputum inflammatory parameters was evaluated. SAR patients were identified by history, skin‐prick test and specific IgE. All patients had normal lung function/bronchial hyper‐responsiveness out of season and a negative asthma/wheezing history. Sputum cells and supernatant levels of ECP, sICAM, IL‐5 and IL‐10, and plasma levels of IL‐5 and ECP, were measured before and 24 h after nasal allergen challenge. After a washout period of at least 4 weeks, the procedure was repeated with placebo challenge (diluent).

Results Nasal allergen challenge led to an increase in sputum ECP (pre = 60 ± 12, post = 212 ± 63 µg/L, P = 0.02 vs. placebo), and sICAM (4.8 ± 2.7 to 6.5 ± 2.9 ng/mL, P = 0.02 vs. placebo), whereas IL‐10 decreased after provocation (44 ± 11 to 29 ± 6 pg/mL, P = 0.06 vs. placebo). Sputum IL‐5 was undetectable in all patients. The absolute number of blood and sputum eosinophils did not change significantly after allergen or placebo challenge (P > 0.07, both comparisons). Plasma levels of IL‐5 increased after allergen challenge (8.7 ± 2.9 to 14.5 ± 3.9 pg/mL, P = 0.001), and the increase in plasma IL‐5 was positively correlated with the rise in sputum ECP in a subgroup of ‘responders’ (n = 12, r = 0.71, P = 0.01).

Conclusions A single nasal allergen challenge in SAR patients increased markers of allergic inflammation in the lower respiratory tract, possibly via pronounced activation of inflammatory cells through circulating immediate‐type reaction cytokines like IL‐5. These findings may provide additional explanatory data for the high susceptibility of SAR patients to incident asthma.
Dendritic cells exposedin vitroto TGF-β1 ameliorate experimental autoimmune myasthenia gravis
Authors: YARILIN D , DUAN R , HUANG Y-M , XIAO B-G
Published: December 2004

Experimental autoimmune myasthenia gravis (EAMG) is an animal model for human myasthenia gravis (MG), characterized by an autoaggressive T-cell-dependent antibody-mediated immune response directed against the acetylcholine receptor (AChR) of the neuromuscular junction. Dendritic cells (DC) are unique antigen-presenting cells which control T- and B-cell functions and induce immunity or tolerance. Here, we demonstrate that DC exposed to TGF-β1 in vitro mediate protection against EAMG. Freshly prepared DC from spleen of healthy rats were exposed to TGF-β1 in vitro for 48 h, and administered subcutaneously to Lewis rats (2 × 106DC/rat) on day 5 post immunization with AChR in Freund’s complete adjuvant. Control EAMG rats were injected in parallel with untreated DC (naive DC) or PBS. Lewis rats receiving TGF-β1-exposed DC developed very mild symptoms of EAMG without loss of body weight compared with control EAMG rats receiving naive DC or PBS. This effect of TGF-β1-exposed DC was associated with augmented spontaneous and AChR-induced proliferation, IFN-γ and NO production, and decreased levels of anti-AChR antibody-secreting cells. Autologous DC exposed in vitro to TGF-β1 could represent a new opportunity for DC-based immunotherapy of antibody-mediated autoimmune diseases.
Immunoglobulins in children with epilepsy: the Dutch Study of Epilepsy in Childhood
Authors: CALLENBACH P M C , JOL-VAN DER ZIJDE C M , GEERTS A T , ARTS W F M , VAN DONSELAAR C A , PETERS A C B , STROINK H , BROUWER O F , VAN TOL M J D
Published: December 2004

In an unselected cohort of 282 children, serum immunoglobulin (Ig) concentrations were determined shortly after the first presentation with one or more unprovoked epileptic seizures and before the start of treatment with anti-epileptic drugs (AEDs), and after 9–18 months of AEDs use. At intake, IgA, IgG1, IgG2 and IgG4 concentrations were significantly higher than published reference values in healthy age-matched controls. In a subset of 127 children, Ig levels at intake were compared with those after AEDs use for 9–18 months. IgA and IgG4 levels had decreased significantly to normal concentrations, but IgG1 and IgG3 levels increased significantly. To determine the influence of AEDs, Ig levels in children who used carbamazepine or valproic acid monotherapy were analysed separately. The use of carbamazepine was associated with a significant decrease of IgA and IgG4 levels, and the use of valproic acid with a significant decrease of IgA and increase of IgG1 levels. In conclusion, humoral immunity is already altered in children shortly after the first presentation with epileptic seizures. Whether this is the consequence of an exogenous event, and to what extent this is related to an interaction of the central nervous system and the immune system, remains to be evaluated. Treatment with AEDs, such as carbamazepine and valproic acid, is associated with significant changes of Ig (sub)class concentrations.
Differences in MHC-class I presented minor histocompatibility antigens extracted from normal and graft-versus-host disease (GVHD) mice
Authors: ROSSET M BRULEY , TIENG V , CHARRON D , TOUBERT A
Published: December 2004

Graft-versus-host disease (GVHD) may develop after allogeneic bone marrow transplantation (BMT) between donors and recipients incompatible for minor histocompatibility antigens (mHAg). Here, we examined the possible relationship between tissue-specific distribution of dominant mHAg peptides and specific organ destruction caused by GVHD. In the B6 anti-Balb/b (H-2b) strain combination, a GVHD developed against Balb/b mHAgs. Despite the high number of incompatible mHAgs between these two strains, both cytotoxic T lymphocyte (CTL) response and GVHD could be attributed to a limited number of dominant mHAgs. We studied CTL-defined expression of dominant mHAgs in normal tissues and their GVHD-related modifications. mHAg peptides were prepared by acid elution and reversed-phase high pressure liquid chromatography fractionation from the spleen, liver, gut and skin as GVHD target tissues and from the heart and kidney as control tissues. Peptidic fractions extracted from normal and GVHD tissues were incubated with RMA-S targets and analysed using bulk B6 anti-Balb/b CTL. In each tissue several fractions were recognized with a given pattern of mHAg expression. GVHD induced qualitative and quantitative changes in antigenic peptide expression. Modifications in mHAg presentation during GVHD concerned preferentially GVHD target organs as opposed to non-GVHD target organs. In addition, when immunizing tissues were derived from GVHD mice instead of normal mice, the profile of CTL recognition was different. In conclusion, these data indicate that broad differences could exist in peptide presentation between various normal and GVHD-target organs.
Elevated angiogenin levels in synovial fluid from patients with inflammatory arthritis and secretion of angiogenin by cultured synovial fibroblasts
Authors: LIOTÉ F , CHAMPY R , MOENNER M , BOVAL-BOIZARD B , BADET J
Published: December 2004

Angiogenesis is a key process in the pathogenesis of inflammatory arthritis. Angiogenin is one of the most potent inducers of neovascularization in experimental models in vivo. To look for evidence that angiogenin is involved in inflammatory joint disease, we examined plasma and synovial fluid (SF) samples from rheumatology patients and synovial fibroblast cell culture supernatants. Angiogenin levels were determined by radioimmunoassay and ELISA. Plasma angiogenin concentrations ranged from 96 to 478 ng/ml, with no significant difference between patients and normal controls. In SF, angiogenin concentrations were significantly higher in patients with acute or chronic synovitis (rheumatoid arthritis (RA): median, 104 ng/ml; range 13–748, n= 14; crystal-induced arthritis (CIA): median, 149 ng/ml; range, 37–616, n= 14, and other chronic inflammatory arthritis: median, 42 ng/ml; range, 15–205; n= 9) than in the 18 patients with osteoarthritis (OA) (median, 20 ng/ml; range 8–116) (P < 0·0001, anova). Angiogenin levels in SF from RA patients in remission with secondary OA were similar to those achieved in primary OA, and decreased in parallel with the resolution of acute gout. Angiogenin protein was released by cultured synovial fibroblasts from OA and RA patients, and reached 1·18 ng/106 cells/day. These data suggest that angiogenin may mediate local inflammation in arthritis via effects on angiogenesis and leucocyte regulation.
Evaluation of CD4+ T cells proliferating to grass pollen in seasonal allergic subjects by flow cytometry
Authors: RIMANIOL A C , GARCIA G , TILL S J , CAPEL F , GRAS G , BALABANIAN K , EMILIE D , HUMBERT M
Published: December 2004

Our objective was to characterize T-cell responses to Phleum pratense in grass pollen allergic individuals and healthy controls using the fluorescent dye PKH26. Peripheral blood mononuclear cells were stimulated with P. pratense, or with recall antigens, and CD3+/CD4+ and CD3+/CD8+ T-cells that had proliferated were analysed by flow cytometry. In the presence of P. pratense CD4+/CD3+ T-cells proliferated more in grass pollen sensitive atopic patients than in nonallergic controls or in nongrass pollen sensitive atopic subjects. PPD and TT recall antigens elicited uniformly high proliferation in all T-cell subsets. Only half of pollen sensitive patients also had an increased proliferation of CD3+/CD8+ T-cells in response to P. pratense. We determined precursor frequency of CD4+ T cells in the original population that responded to P. pratense and found values ranging from 1 × 10−3 to 0·6 × 10−1, in the same range as those measured for PPD and TT. In conclusion, grass pollen sensitive atopic patients show enhanced CD4+ T-cell reactivity to P. pratense, and this could be related to the presence of elevated numbers of circulating allergen-specific CD4+ T cells. This flow cytometric method should allow the identification of other phenotypic markers such as intracellular cytokines in allergen specific responding CD4+ T cells.
Clearance of red cells by monoclonal IgG3 anti-Din vivois affected by the VF polymorphism of FcγRIIIa (CD16)
Authors: KUMPEL B M , DE HAAS M , KOENE H R , VAN DE WINKEL J G J , GOODRICK M J
Published: December 2004

Human red cells (RBC) coated with IgG anti-D are cleared from the circulation to the spleen by macrophages which express IgG receptors (Fcγ R). Polymorphisms of Fcγ RIIa and Fcγ RIIIa affect IgG binding in vitro, and may alter the efficiency of clearance of immune complexes in vivo. In a RBC clearance study, 22 Rh D-negative subjects were given 100–400 µg human monoclonal or polyclonal IgG anti-D i.m. followed 48 h later by 51Cr-labelled D+ RBC. The half lives of the infused D+ RBC were determined, together with the coating levels of anti-D on the D+ RBC. Fcγ RIIA and FcγRIIIA genotyping was performed. Large ranges of phagocytosis and extracellular lysis of RBC in vitro, and of half lives of RBC in vivo, were observed. Clearance of RBC coated with monoclonal IgG3 anti-D (BRAD-3) was more rapid in five subjects homozygous for Fcγ RIIIa-F/F158 than in three subjects expressing the Fcγ RIIIa-V158 allele (P = 0·024). This effect was not observed, however, for those individuals given polyclonal anti-D. There was also no significant difference in the efficiency of RBC destruction in vitro or of RBC clearance in vivo between the subjects analysed for individual genotypes or alleles or combinations of alleles. In conclusion, the presence of the Fcγ RIIIa-V158 allele compromised the efficiency of removal of RBC coated with IgG3 anti-D.
Relevance of differential immunogenicity of human and mouse recombinant desmoglein-3 for the induction ofAcantholyticautoantibodies in mice
Authors: KAITHAMANA S , FAN J-L , MEMAR O , LI K , UITTO J , SEETHARAMAIAH G S , PRABHAKAR B S
Published: December 2004

Pemphigus vulgaris (PV) is an antibody-mediated autoimmune disease of the skin and mucous membranes. Desmoglein-3 (dsg-3) expressed in the suprabasal layer of the skin serves as an autoantigen in PV. Passive transfer of sera, either from patients with PV or from experimental animals immunized with a recombinant human dsg3 (hdsg3) into neonatal BALB/c mice results in blister formation, suggesting strongly that there is significant cross-reactivity between the mouse dsg3 (mdsg3) and the hdsg3. However, efforts to induce disease in adult mice through active immunization using hdsg-3 have not been successful, suggesting that the epitopes required for the induction of pathogenic antibodies in adult mice might not be present in hdsg3. Therefore, in this study, we expressed a full-length mdsg3 in insect cells and compared its serological reactivity with that of the hdsg3 using species specific polyclonal sera and a panel of seven monoclonal antibodies (MoAbs) with unique binding specificities to hdsg3. Studies using sera demonstrated a considerable cross-reactivity, while studies using MoAbs exhibited specific epitope differences between the two proteins. Because of these differences, we reasoned that immunization with mdsg3 might induce disease in adult mice. Immunization of four strains of mice (i.e. BALB/c, DBA/1, HRS/J and SJL/J) with mdsg3 resulted in considerable antibody response, but failed to induce lesions. However, sera from immunized BALB/c mice induced acantholysis of neonatal mouse skin in vitro. These studies indicated that our inability to induce lesions in adult mice through active immunization is not due to differences in the ability of mouse and human dsg3 to induce acantholytic antibodies, but due probably to structural differences between adult and neonatal mouse skin. Alternatively, immunization with a combination of dsg3 protein along with other proteins might be necessary to induce pemphigus disease in adult mice. Nevertheless, our current studies show that molecular mechanisms leading to the production of acantholytic antibodies in mice can now be studied using homologous mdsg3.
Predicting death from tumour necrosis factor-alpha and interleukin-6 in 80-year-old people
Authors: BRUUNSGAARD H , LADELUND S , PEDERSEN A N , SCHROLL M , JØRGENSEN T , PEDERSEN B K
Published: December 2004

Ageing is associated with low-grade inflammation and markers such as IL-6 possess prognostic value. Tumour necrosis-alpha (TNF-α) initiates the inflammatory cascade and has been linked to several age-associated disorders. It remains, however, unknown if TNF-α is associated with mortality in old populations. The aim of the present study was to investigate if serum levels of TNF-α were associated with all-cause mortality independently of interleukin (IL)-6 in a prospective study of 333 relatively healthy 80-year-old people. A Cox regression model was used to explore effects of TNF-α and IL-6 on survival in the following 6 years. A total of 133 participants died during this follow-up period. TNF-α was associated with mortality in men, but not in women, whereas low-grade elevations in IL-6 were associated strongly with mortality in both sexes. TNF-α explained only 7% of the variability in IL-6 and effects of the two cytokines were independent of each other as well as of other traditional risk factors for death [smoking, blood pressure, physical exercise, total cholesterol, co-morbidity, body mass index (BMI) and intake of anti-inflammatory drugs]. These findings indicate that at least in old populations chronic elevated levels of TNF-α and IL-6 have different biological functions that trigger age-associated pathology and cause mortality.
Calcium signalling is altered in myeloid cells with a deficiency in NADPH oxidase activity
Authors: RADA B K , GEISZT M , VAN BRUGGEN R , NÉMET K , ROOS D , LIGETI E
Published: December 2004

The relation of O2.−-production and Ca2+ homeostasis was investigated in PLB-985 cell lines and neutrophilic granulocytes from peripheral blood. In differentiated wild-type PLB-985 cells, a high level of O2.−-production was associated with a significant decrease in the membrane potential and the inhibition of capacitative Ca2+ entry. These correlations were not observed in gp91phox –/– cells or in cells transfected with a non-functional mutant of gp91phox (Thr341Lys). Membrane depolarization and inhibition of Ca2+ entry reappeared in cells transfected with wild-type gp91phox. These experiments demonstrate that inhibition of Ca2+ entry depends on the presence of a functional NADPH oxidase. The Ca2+ signal induced by stimulation of chemotactic receptors also showed remarkable differences: [Ca2+]ic in the sustained phase was higher in gp91phox–/– than in wild-type cells. Alteration of the Ca2+ signal was reproduced by treating peripheral blood neutrophils with the NADPH oxidase inhibitor diphenylene-iodonium. It is concluded that the deficiency in O2.−-production is accompanied by significant alterations of Ca2+ homeostasis in myeloid cells.
Gonadotropin treatment restoresin vitrointerleukin-1β and tumour necrosis factor-α production by stimulated peripheral blood mononuclear cells from patients with idiopathic hypogonadotropic hypogonadism
Authors: MUSABAK U , BOLU E , OZATA M , OKTENLI C , SENGUL A , INAL A , YESILOVA Z , KILCILER G , OZDEMIR I C , KOCAR I H
Published: December 2004

In the present study, we aimed to investigate the effects of testosterone deficiency and gonadotropin therapy on the in vitro production of tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) by peripheral blood mononuclear cells (PBMCs) from patients with idiopathic hypogonadotropic hypogonadism (IHH) in order to elucidate the modulatory role of androgen in cytokine production. Fifteen male patients with untreated IHH and 15 age-matched healthy male subjects were enrolled in the study. Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), free testosterone (FT), sex hormone binding globulin (SHBG), prolactin, and IL-2 and IL-4 levels were also measured. In unstimulated cultures, IL-1β and TNF-α secretion were not significantly different between patient and control groups. However, after stimulation with lipopolysaccharide (LPS), secretion of IL-1β and TNF-α was significantly higher in cultures from untreated patients with IHH than in control subjects. Mean FSH, LH and FT levels were significantly lower, whereas SHBG, IL-2 and IL-4 levels were significantly higher in patients with IHH compared than in controls. In patients with IHH, FT negatively affected the serum levels of IL-4 and in vitro secretion of IL-1β and TNF-α. In addition, IL-2 and IL-4 affected the in vitro secretion of IL-1β in a positive manner. Gonadotropin therapy decreased both TNF-α and IL-1β in PBMCs from patients with IHH. The levels of serum IL-2 and IL-4 were also decreased by therapy. In conclusion, in the present study, gonadotropin treatment restored the in vitro production of IL-1β and TNF-α by PBMCs from patients with IHH, suggesting that androgen modulates proinflammatory cytokine production, at least directly through its effects on PBMCs. It seems probable that this effect plays an important role in the immunosuppressive action of androgens.
Discordant effects of interleukin-2 on viral and immune parameters in human immunodeficiency virus-1-infected monocyte-derived mature dendritic cells
Authors: BAHR G M , DARCISSAC E C A , MOUTON Y
Published: December 2004

Use of interleukin-2 (IL-2) in the immunotherapy of human immunodeficiency virus (HIV) has frequently resulted in the restoration of CD4 lymphocyte counts but not of virus-specific responses. We reasoned that the absence of reconstituted functional immune parameters could be related to the inability of IL-2 to correct HIV-induced dysfunctions in antigen-presenting cells. In this study, we used in vitro-differentiated monocyte-derived macrophages (MDMs) and mature dendritic cells (MDDCs), acutely infected with primary HIV-1 isolates, to analyse the effects of IL-2 on virus replication, co-receptor expression, and cytokine or chemokine release. Stimulation of MDMs with IL-2 had no measurable effect on HIV-1 replication, on cytokine secretion, or on CD4 and CXCR4 gene expression. Moreover, although a significant down-regulation of CCR5 mRNA expression could be repeatedly detected in MDMs, this IL-2-mediated effect was not of substantial magnitude to affect virus replication. On the other hand, IL-2 stimulation of MDDCs dramatically increased HIV-1 replication and this effect was highly evident on low-replicating, CXCR4-dependent isolates. Nevertheless, the HIV-enhancing activity of IL-2 in MDDCs was not accompanied by any measurable change in cytokine or chemokine release, in virus receptor and co-receptor mRNA accumulation, or in the surface expression of a battery of receptors implicated in virus entry, cell activation or costimulatory function. Taken together, these findings point to a role for IL-2 in inducing virus purging from dendritic cell reservoirs but indicate no relevant potential of the cytokine in restoring defective elements of innate immunity in HIV infection.
Normal production of inflammatory cytokines in chronic fatigue and fibromyalgia syndromes determined by intracellular cytokine staining in short-term cultured blood mononuclear cells
Authors: AMEL KASHIPAZ M R , SWINDEN D , TODD I , POWELL R J
Published: December 2004

It has been proposed that cytokines play a role in the pathogenesis of chronic fatigue syndrome (CFS) and fibromyalgia syndrome (FMS). However, different studies have reported conflicting results using enzyme-linked immunosorbent assay or polymerase chain reaction to detect cytokines in these conditions. In the present study, for the first time, the production of inflammatory [interleukin (IL)-1α, IL-6, and TNF-α] and anti-inflammatory (IL-10) cytokines by CD14+ and CD14– peripheral blood mononuclear cells (PBMC) from chronic fatigue syndrome (CFS) and fibromyalgia syndrome (FMS) patients and sex- and age-matched normal subjects was investigated at the level of individual cells using the technique of intracellular cytokine staining and flow cytometry. Cultures were carried out in the presence of polymyxin B to inhibit the effect of endotoxins on cytokine production by monocytes. The mean intensity of fluorescence (MIF) and percentage of CD14+ (monocytes) and CD14– (lymphocytes) cytokine-producing mononuclear cells were comparable in patients and controls in either unstimulated or IFN-γ-stimulated conditions. Our study indicates that dysregulation of cytokine production by circulating monocytes or non-monocytic cells (lymphocytes) is not a dominant factor in the pathogenesis of CFS/FMS.
Functional and molecular aspects of transient T cell unresponsiveness: role of selective interleukin-2 deficiency
Authors: KÖLLER M D , KIENER H P , ARINGER M , GRANINGER W B , MEUER S , SAMSTAG Y , SMOLEN J S
Published: December 2004

Defects of T cell (Tc) proliferation have been demonstrated in several autoimmune diseases. Detailed mechanisms governing activation and proliferation of Tc are still not completely known. Here we show that under certain conditions human peripheral blood lymphocytes, once activated by anti-CD3, fail to respond to a subsequent restimulation via the Tc-receptor. Peripheral blood mononuclear cells (PBMC) were preactivated by anti-CD3 for 96 h following restimulation by anti-CD3, interleukin (IL)-2 and other mitogens. In control experiments unstimulated PBMC were incubated in medium alone. Immunophenotypes were analysed by flow cytometry. Cytokine production was determined by reverse transcription-polymerase chain reaction and intracellular signalling protein contents of Tc were compared by Western blotting. Furthermore, apoptosis was detected by terminal deoxyribose transferase-mediated deoxyuridine triphosphate nick end labelling assay. Unstimulated PBMC proliferate well after subsequent stimulation with anti-CD3, whereas IL-2 induces only limited proliferation. In contrast, preactivated cells respond only minimally to restimulation with anti-CD3, but IL-2 induces a marked proliferation. Both preactivated and unstimulated Tc respond well to restimulation by phytohaemagglutinin (PHA). In contrast, preactivated Tc show only a weak response to concanavalin A. Interestingly, when cells have been allowed to rest for 168 h, the responsiveness of preactivated Tc is restored. Immunoblots reveal that preactivated cells have a higher intracellular content of ζ-chain and p56lck. No differences are found concerning apoptosis after restimulation with anti-CD3 or the expression of ERK 1/2. The unresponsiveness to restimulation is due to an impairment of the transcription of the IL-2 gene and this defect is temporary. Despite the lack of proliferation, preactivated Tc phenotypically maintain an intermediate stage of activation. These data show how the same cell population can change its functional phenotype into a non-responder state.
HIV-gp160 modulates differentially the productionin vitroof IgG, IgA and cytokines by blood and tonsil B lymphocytes from HIV-negative individuals
Authors: COGNASSE F , BÉNIGUEL L , El HABIB R , SABIDO O , CHAVARIN P , GENIN C , GARRAUD O
Published: December 2004

HIV1-gp160 holds promises in anti-HIV vaccinal strategies. However, this molecule has been described to exhibit superantigenic activities. The present study aimed at examining the effect(s) of HIV1-gp160 on human B cells and in particular on B cells originating from HIV– donors. We purified human B cells of various origins, i.e. from blood and from tonsils (representing a mucosal-type origin), and we tested these cells (stimulated with a polyclonal B cell activator, interleukin (IL)-2 and IL-10 as cytokines, and recombinant HIV1-gp160) for the production of IgG and IgA in an in vitro model. Gp160 induced significantly less total IgG by blood − but not tonsil-originating − B cells and did not affect total IgA production. Further, HIV1-gp160 up-regulated IL-2-, IL-4- and IL-10-mRNA levels in stimulated blood B cells (these cytokines are known to be active on B cell activation and differentiation). Interestingly, HIV1-gp160 also up-regulated IL-1β-, transforming growth factor (TGF)-β-, interferon (IFN)-γ- and IL-12-mRNA levels in stimulated mucosal-type, tonsil-originating, B cells. As these latter cytokines are involved in proinflammatory activities, HIV-gp160 delivery at the mucosal sites would be compatible with an adjuvant activity.
CCR4 is an up-regulated chemokine receptor of peripheral blood memory CD4+ T cells in Crohn's disease
Authors: JO Y , MATSUMOTO T , YADA S , FUJISAWA K , ESAKI M , ONAI N , MATSUSHIMA K , IIDA M
Published: December 2004

Several chemokine receptors are expressed selectively on the surface of T cells depending on their polarization. The aim of this study was to characterize chemokine receptor expression in peripheral blood memory T cells in Crohn's disease (CD) and ulcerative colitis (UC), and to correlate the expression with disease activity. Peripheral blood mononuclear cells (PBMCs) were obtained from 24 patients with CD, 30 patients with UC, 24 normal controls and 10 disease controls. PBMCs were stained by anti-CCR3, CCR4, CCR5, CXCR3, CD4, CD8, CD45RO and β 7 integrin, and the expression of the chemokine receptors were determined by flow cytometry. CCR4 expression on memory T cells was significantly lower in UC than in CD or normal controls, and that of memory CD4+ T and β 7high memory CD4+ T cells was significantly higher in CD than in UC or normal controls. CCR4 expression on memory CD4+ T cells exhibited significant positive correlation with disease activity in CD, and this decreased significantly after treatment. Such a decrease was not found in the disease controls. CCR5 and CXCR3 expression on memory CD8+ T cells was significantly lower in CD than in normal controls. CXCR3 expression on β 7high memory CD4+ T and CXCR3 expression on memory CD8+ T cells were lower in UC than in normal controls. These findings suggest that in peripheral blood memory T cells, chemokine receptor expression is different between CD and UC. Enhancement of CCR4 and suppression of CCR5 and CXCR3 seem to be the characteristic chemokine receptor profile in peripheral blood memory T cells of CD.
Dendritic cells, T cell tolerance and therapy of adverse immune reactions
Authors: MOREL P A , FEILI-HARIRI M , COATES P T , THOMSON A W
Published: December 2004

Dendritic cells (DC) are uniquely able to either induce immune responses or to maintain the state of self tolerance. Recent evidence has shown that the ability of DC to induce tolerance in the steady state is critical to the prevention of the autoimmune response. Likewise, DC have been shown to induce several type of regulatory T cells including Th2, Tr1, Ts and NKT cells, depending on the maturation state of the DC and the local microenvironment. DC have been shown to have therapeutic value in models of allograft rejection and autoimmunity, although no success has been reported in allergy. Several strategies, including the use of specific DC subsets, genetic modification of DC and the use of DC at various maturation stages for the treatment of allograft rejection and autoimmune disease are discussed. The challenge for the future use of DC therapy in human disease is to identify the appropriate DC for the proposed therapy; a task made more daunting by the extreme plasticity of DC that has recently been demonstrated. However, the progress achieved to date suggests that these are not insurmountable obstacles and that DC may become a useful therapeutic tool in transplantation and autoimmune disease.
Evaluation of amastigote reactive cells in human cutaneous leishmaniasis caused byLeishmania aethiopica
Authors: MAASHO K , MCMAHON-PRATT D , RAITA J , RAUD M , BRITTON S , SOONG L , AKUFFO H
Published: December 2004

Lymphoproliferative responses to three affinity chromatography purified amastigote antigens of Leishmania pifanoi, P-2, P-4 and P-8, were evaluated in peripheral blood mononuclear cells (PBMC) from patients with Ethiopian cutaneous leishmaniasis. Antigen-stimulated cells were analysed for the percentage of CD4+, CD8+ and CD16/56+ cells and the expressed levels of gamma interferon (IFNγ) and interleukin (IL)-10 were determined in culture supernatants. The amastigote antigens induced cellular responses in leishmaniasis patients with heterologous Leishmania parasite infection. These responses were compared to those of freeze-thawed L. aethiopica promastigote antigen stimulation. The membrane protein (P-8), and to a lesser extent the megasomal/cytoplasmic cysteine proteinase(P-2), induced proliferation with high levels of IFNγ and IL-10 production in cells from patients with active L. aethiopica lesions. CD16/56+ NK cells were the main cell types induced to proliferate in response to P-8 and P-2 stimulation, followed by CD8+ cell populations. P-4 had no such effect. This contrasts from previous studies of New World human leishmaniasis where P-4 and P-8 were stimulatory. The success of a particular molecule in the induction of a response with a protective phenotype may be dependent on the infecting Leishmania spp. To our knowledge, there are no studies that directly compare the New versus Old World cutaneous leishmaniasis in respect of NK cell and IL-10 responses. Our studies indicate that some leishmanial molecules are recognized across the species, while others are apparently more species specific.
Up-regulation of ICAM-1, CD11a/CD18 and CD11c/CD18 on human THP-1 monocytes stimulated byStreptococcus suisserotype 2
Authors: Al-NUMANI D , SEGURA M , DORÉ M , GOTTSCHALK M
Published: December 2004

Streptococcus suis serotype 2 is known to be a major pathogen of swine, causing mainly meningitis. It is also a zoonotic agent leading predominantly to meningitis in humans working in close contact with pigs. In this study, we investigated the ability of S. suis to up-regulate the expression of adhesion molecules involved in inflammation, using an enzyme-linked immunosorbent assay. S. suis serotype 2 stimulated the up-regulation of the expression of intercellular adhesion molecule-1 (ICAM-1, CD54), CD11a/CD18 and CD11c/CD18 on human THP-1 monocytes, but did not change that of ICAM-1, vascular cell adhesion molecule-1 (VCAM-1, CD106) and E-selectin (CD62E) on human endothelial cells. The up-regulation of adhesion molecules was time- and bacterial concentration-dependent, and cell wall components were largely responsible for such stimulation. To a lesser extent, purified haemolysin of S. suis also stimulated adhesion molecule expression. Stimulation of monocytes with strains of different origin showed that there was no clear tendency for human strains to induce a higher expression of adhesion molecules than strains from diseased pigs. Finally, monocytes stimulated with S. suis also showed an increase in adherence to endothelial cells. Hence, S. suis is capable of up-regulating important adhesion molecules involved in inflammation, which may result in an increased leucocyte recruitment into sites of infection, thus providing a possible mechanism for some of the inflammatory features of meningitis caused by this pathogen.
Validation of the interleukin-10 knockout mouse model of colitis: antitumour necrosis factor-antibodies suppress the progression of colitis
Authors: SCHEININ T , BUTLER D M , SALWAY F , SCALLON B , FELDMANN M
Published: December 2004

Advances in understanding pathogenesis and developing new therapies are hastened by the use of effective animal models of disease. In inflammatory bowel disease, such as Crohn's disease, a variety of models have been used, including the IL-10 knockout mouse. However, in order to be truly valuable, the models need to respond to existing therapy in a way which resembles the human disease. In the light of recent developments, in which refractory Crohn's disease responds well to anti-TNF antibody therapy, we set out to validate the IL-10 knockout model of Crohn's disease by examining its response to anti-TNF therapy. We developed a new scoring system for IL-10 knockout mice, similar to the Crohn's Disease Activity Index in humans, analysed stool samples for cytokines and compared the findings with histology. We found that anti-TNF antibody therapy starting at 4 weeks markedly ameliorated the disease, as judged by the clinical score or by histological analysis of the gut. Furthermore, analysis of stool samples for cytokines revealed a marked diminution of inflammatory cytokines, adding a further accurate measure of the improvement. This model may thus be useful for evaluating other therapeutic modalities of relevance to Crohn's disease.
Autologous apoptotic T cells interact with dendritic cells, but do not affect their surface phenotype or their ability to induce recall immune responses
Authors: NEWTON P J , WELLER I V D , KATZ D R , CHAIN B M
Published: December 2004

Dendritic cells (DCs) play an important role in determining immunogenicity and the subsequent immune response. They may also have a role in maintaining peripheral tolerance to self-antigens by initiating an immune response only in the context of danger signals released from cells during stress, damage or death. These signals may originate from surrounding T cells as well as from other cells. Therefore, in this study the effect of autologous T cell injury on DC morphology and function has been investigated. Co-incubation of apoptotic or necrotic T cells with immature DCs altered their morphology towards a more mature appearance, with more cells showing activation as judged by spreading and formation of arborizing long processes. The apoptotic autologous T cells were rarely phagocytosed by immature DCs, compared to macrophages. The DC surface phenotype was not affected by the co-incubation with autologous injured T cells. The ability of DCs to elicit a secondary immune response was not altered by exposure to autologous injured T cells. These findings suggest that DC can continue to function in T cell activation, rather than in tolerogenic mode, even in the presence of large numbers of dying autologous T cells, such as may be present in the aftermath of an acute antiviral response.
Characterization of bronchoalveolar lavage T cell subsets in sarcoidosis on the basis of CD57, CD4 and CD8
Authors: KURUMAGAWA T , SEKI S , KOBAYASHI H , KOIKE Y , KANOH S , HIRAIDE H , MOTOYOSHI K
Published: December 2004

T cells expressing CD57 (a natural killer cell marker) with interferon-γ (IFN-γ) producing capacity increase under various conditions. CD57+ T cells are also present in the bronchoalveolar lavage fluid (BALF) of sarcoidosis, and several phenotypical and functional analyses of these cells have been reported. In the present study, BALF T cells obtained from 52 patients with sarcoidosis were classified further into CD4+CD57+ T cells, CD4+CD57– T cells, CD8+CD57+ T cells and CD8+CD57– T cells and their phenotypes and functional characteristics were assessed. Substantial proportions of these T cell subsets expressed natural killer cell markers CD161 and CD122. The biased expansion of Vβ2 T cells was observed in both CD4+CD57+ T cells and CD4+CD57– T cells in BALF from most patients, while the expansion of other Vβ T cells was also observed in some patients. Unexpectedly, the biased expansion of certain Vβ T cells was also seen in either CD8+CD57+ T cells or CD8+CD57– T cells, while the expanded Vβ T cells in CD8+ T cells differed substantially among individuals. BALF T cells showed a remarkably lower T cell receptor (TCR) intensity than that of peripheral blood T cells. Both CD8+ T cell subsets in BALF of sarcoidosis expressed the intracellular perforin/granzyme B, while all four subsets expressed intracellular IFN-γ after in vitro activation, and CD4+ T cells, especially CD4+CD57+ T cells, expressed tumour necrosis factor-α. These findings indicate that CD57+ T cells as well as CD57– T cells in the BALF are phenotypically and functionally different from peripheral blood T cells and may play an important role in the Th1 dominant state and inflammation in pulmonary sarcoidosis.
In vitroco-stimulation with anti-CD28 synergizes with IL-12 in the generation of T cell immune responses to leukaemic cells; a strategy forex-vivogeneration of CTL for immunotherapy
Authors: ORLEANS-LINDSAY J K , DERU A , CRAIG J I O , PRENTICE H G , LOWDELL M W
Published: December 2004

The existence of an immune based graft-versus-leukaemia (GvL) effect highlighted the prospect of managing relapsed leukaemias with T cell-based adoptive immunotherapy. Thus, various strategies have been explored for the in vitro expansion of acute myeloid leukaemia (AML)-specific T cells. In a popular approach, AML blasts have been genetically modified to express co-stimulatory molecules essential for effective T cell priming. One such tactic has been the modification of AML cells to express the B7/CD80 co-stimulatory molecule that binds to CD28 on T cells initiating events that culminate in enhanced cytokine production, proliferation and development of effector functions by T cells. The success of these strategies has been limited by difficulties in attaining sufficient transduction efficiencies and associated high levels of CD80 expression. We demonstrate that these problems can be circumvented by using anti-CD28 monoclonal antibody. Furthermore, we show that the synergistic relationship between CD80/CD28 pathway and interleukin 12 cytokine (IL-12), documented in the generation of cytotoxic T lymphocytes (CTL) for solid tumours, also applies to AML. CD28/IL-12 synergy facilitated the proliferation of allogeneic T cells in response to stimulation with primary AML blasts. The synergy also favoured generation of a Th1-type immune response, evidenced by gamma interferon (IFN-γ) secretion and facilitated naive and memory T cell proliferation. Unlike some methods of in vitro T cell expansion, use of CD28/IL-12 synergy left T cells in the physiologically appropriate CD45RA–/CCR7– subsets known to be associated with immediate cytotoxic functions.