bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytokine mRNA expression in patients with mild allergic asthma following low dose or cumulative dose allergen provocation
Authors: Prieto J. , Van Der Ploeg I. , Roquet A. , Gigliotti D. , Dahlén B. , Eklund A. , Grunewald J.
Published: March 2003

Background Allergen provocation is a very useful way to study the inflammatory response in asthmatic patients. Although cumulative dose regimens are most often applied, another provocation model with repeated inhalations of low doses of allergens has recently come into use.

Objective We were interested to compare these two allergen provocation models. To evaluate the inflammation induced by either model, we examined the mRNA expression of several cytokines that are implicated in the orchestration of the inflammatory response observed in asthma.

Methods Interleukin (IL)‐4, IL‐5, IL‐13 and interferon (IFN)‐γ mRNA expression was analysed in bronchoalveolar lavage (BAL) cells and peripheral blood (PB) CD4+ and PB CD8+ T cells following any of the two provocation regimens. IL‐4 and IFN‐γ mRNA expression was analysed by a competitive reverse transcriptase‐polymerase chain reaction (RT‐PCR) method, while IL‐5 and IL‐13 were analysed semiquantitatively, before and after allergen provocation with either model.

Results After low dose provocations none of the cell populations studied showed a clear change in the pattern of IL‐4 or IFN‐γ gene expression. In contrast, after cumulative dose provocation there was a clear tendency towards an increased IL‐4 mRNA expression in BAL cells, correlating with a significant increase in IL‐4 mRNA in PB CD4+ as well as in CD8+ T cells (P = 0.005 and P = 0.04, respectively). Regardless of the allergen provocation method used, in PB IL‐4 mRNA was preferentially expressed by CD4+ cells while IFN‐γ was expressed more by CD8+ cells. IL‐5 transcripts increased after low dose provocations in PB CD4+ T cells in six of eight patients, while after cumulative dose provocation IL‐5 mRNA increased in BAL cells in six out of nine patients but decreased especially in PB CD8+ T cells in six out of eleven patients, suggesting an accumulation of IL‐5 expressing cells to the lungs.

Conclusion Thus, the cumulative dose provocation regimen can induce a more pronounced Th2‐like immune response in asthmatic patients than the low dose provocation model.
The interaction of the polymorphisms in transporter of antigen peptides (TAP) and lymphotoxin α (LT‐α) genes and atopic diseases in the Czech population
Authors: Izakovičová Hollá L. , Vašků A. , Izakovič V. , Znojil V.
Published: March 2003

Background Transporter antigen peptide gene (TAP) products are involved in antigen processing. These genes, inducible by interferon γ, as well as lymphotoxin α (LT‐α), are located in the HLA region. Their involvement in immune response regulation makes them candidate atopy susceptibility genes.

Objective This study investigates a possible association between previously identified polymorphisms within the TAP‐1 and LT‐α genes and clinically manifested atopic diseases in the Czech population.

Methods Caucasian subjects of Czech nationality (n = 427) were included in our study. We examined 184 healthy controls and 243 patients with histories of atopic asthma, allergic rhinitis and atopic dermatitis or their combinations. We used the amplification refractory mutation system polymerase chain reaction to determine TAP‐1 gene polymorphisms. LT‐α genotypes were determined by PCR and restriction analysis by NcoI.

Results No significant differences were found in allele or genotype frequencies of the LT‐α gene, as well as in polymorphisms for Val→Ile at codon 333 and Gly→Asp at codon 637 in the TAP‐1 gene between controls and patients. However, analysis of the concurrence of the double genotypes of the TAP‐1 polymorphism at codon 333 and the LT‐α genes showed differences between controls and atopic patients (P < 0.02).

Conclusion Several reports have indicated that different HLA products and genes may be risk factors for or protective factors against the development of atopy. We report no association between polymorphisms in the LT‐α and TAP‐1 genes alone and atopic diseases in the central Europe Caucasian (Czech) population, but there was an interesting interaction between the TAP333 and LT‐α polymorphisms.
T cell responses to recombinant isoforms, synthetic peptides and a mutant variant of Lep d 2, a major allergen from the dust mite Lepidoglyphus destructor
Authors: Eriksson T. L. J. , Gafvelin G. , Elfman L. H. M. , Johansson C. , Van Hage‐Hamsten M. , Olsson S.
Published: March 2003

Background The dust mite Lepidoglyphus destructor is an important cause of allergic reactions to dust, especially in farming environments. Two isoforms, recombinant (r)Lep d 2.01 and rLep d 2.02, of the major allergen Lep d 2, have previously been expressed as recombinant proteins. These isoforms differ 10.4% at the amino acid level. Furthermore, a mutant form of Lep d 2.01 (rLep d 2.6Cys) with a highly reduced IgE reactivity, has also been produced.

Objective To investigate the T cell responses to the recombinant isoforms of Lep d 2, the Lep d 2.6Cys mutant and peptides of Lep d 2, in allergic and non‐allergic individuals.

Methods Peripheral blood mononuclear cells from 18 allergic and 16 non‐allergic individuals were stimulated with the different antigens and the proliferative responses were measured. The cytokine production (interleukin (IL)‐4, IL‐5 and interferon (IFN)‐γ) were measured by ELISA.

Results Higher T cell proliferation was measured to isoform 01 than to 02 in 28/34 subjects. The responses to rLep d 2.6Cys were lower than to isoform 01 in most subjects, but higher than to Lep d 2.02. Two immuno‐dominant peptides, corresponding to amino acid residue 11–25 and 61–75 were identified. The atopic subjects produced significantly lower IFN‐γ in response to Lep d 2.01 as compared to the non‐atopics.

Conclusions There was a significant difference in T cell response between the two isoforms of rLep d 2. The hypoallergenic mutant rLep d 2.6Cys was able to evoke a T cell response with a magnitude which is between the two isoforms. Amino acid residue 11–25 and 61–75 are the most frequently recognized parts of Lep d 2 and are likely to contain the immuno‐dominant T cell epitopes.
Inhalation of a harmless antigen (ovalbumin) elicits immune activation but divergent immunoglobulin and cytokine activities in mice
Authors: Swirski F. K. , Gajewska B. U. , Alvarez D. , Ritz S. A. , Cundall M. J. , Cates E. C. , Coyle A. J. , Gutierrez‐Ramos J.‐C. , Inman M. D. , Jordana M. , Stämpfli M. R.
Published: March 2003

Background Exposure to aerosolized harmless antigen such as ovalbumin (OVA) has previously been shown to induce inhalation tolerance, a state characterized by inhibition of IgE synthesis and airway inflammation, upon secondary immunogenic antigen encounter. Immune events associated with this phenomenon are still poorly understood.

Objective The aim of this study was to investigate cellular and molecular mechanisms underlying this state of ‘unresponsiveness’.

Methods; After initial repeated OVA exposure, mice were subjected to a protocol of antigen‐induced airway inflammation, encompassing two intraperitoneal injections of OVA adsorbed to aluminium hydroxide followed by airway challenge. We assessed immune events in the draining lymph nodes after sensitization, and in the lungs after challenge.

Results In animals initially exposed to OVA, we observed, at the time of sensitization, considerable expansion of T cells, many of which expressed the activation markers CD69 and CD25, as well as increased numbers of antigen‐presenting cells, particularly B cells. While these animals produced low levels of IgE, the observed elevated levels of IgG1 signified isotype switching. Splenocytes and lymph node cells from OVA‐exposed mice produced low levels of IL‐4, IL‐5, IL‐13 and IFN‐γ, indicating aborted effector function of both T helper (Th)2‐ and Th1‐associated cytokines. Real time quantitative polymerase chain reaction (PCR) (TaqMan) analysis of costimulatory molecules in the lungs after in vivo challenge showed that B7.1, B7.2, CD28 and CTLA‐4 mRNA expression was low in animals initially exposed to OVA. Ultimately, these events were associated with abrogated airway inflammation and attenuated airway hyper‐responsiveness. The decreased inflammation was antigen‐specific and independent of IL‐10 or IFN‐γ.

Conclusion Initial exposure to OVA establishes a programme that prevents the generation of intact, fully functional inflammatory responses upon secondary antigen encounter. The absence of inflammation, however, is not associated with categorical immune unresponsiveness.
Type I allergy to natural rubber latex and type IV allergy to rubber chemicals in health care workers with glove‐related skin symptoms
Authors: Nettis E. , Assennato G. , Ferrannini A. , Tursi A.
Published: March 2003

Background It has been established that there are type I and type IV allergens in latex gloves.

Objective The purpose of the study was to establish the prevalence of rubber glove‐induced skin symptoms among health care workers in one Italian hospital.

Methods Health care workers (n = 1584) were evaluated using a written questionnaire and 295 respondents with glove‐induced skin symptoms were tested. We performed: skin prick test with latex glove extract and commercial latex, and environmental and food allergens; glove use test; patch tests with a rubber additive series; and RASTs.

Results Hospital employees who used or had used latex gloves at work were 1294. Three hundred and sixteen (24.4%) reported glove‐induced symptoms, namely, cutaneous symptoms in all the cases and non‐cutaneous symptoms in 105 subjects (8.1%). Twenty‐seven of the 295 symptomatic employees tested (9.1%) were latex sensitive. Thirty‐one patients (10.5%) exhibited positive patch test to rubber‐related allergens. The most positive readings were obtained from the Thiuram mix and the Carba mix, with 12 and 9 positivities, respectively. The risk factors for latex skin sensitization were: a previous history of atopy and asthma; history of surgery; pre‐existing hand dermatitis; work‐related symptoms; and positive skin tests to common inhalant and certain foods (P < 0.05). Subjects who exhibited positive patch test were significantly more likely to have a prior hand dermatitis (P < 0.001). Of the 295 cases, 275 (93.2%) were contact dermatitis (CD), 13 (4.4%) contact urticaria (CU) (including protein CD) and 7 (2.4%) CD associated with CU.

Conclusions Our results show a high prevalence of rubber glove‐induced dermatoses among the employees in one Italian hospital. The majority of skin complaints of latex gloves are related to skin irritation rather than to allergy. The immediate allergy to latex and the delayed allergy to rubber chemicals suggest that all the health care workers with glove‐related dermatitis should undergo both skin prick test and glove use test to detect type I hypersensitivity to latex, and patch test to detect type IV hypersensitivity to rubber chemicals.
Repeated ozone exposures enhance bronchial allergen responses in subjects with rhinitis or asthma
Authors: Holz O. , Mücke M. , Paasch K. , Böhme S. , Timm P. , Richter K. , Magnussen H. , Jörres R. A.
Published: March 2003

Background Single exposures to > 200 p.p.b. of ozone are capable of enhancing the early‐phase lung function response to allergen.

Objective The aim of the present study was to compare the effect of single vs. repeated exposures to ozone on early and late‐phase allergen responses.

Methods Eleven subjects with allergic asthma and 22 subjects with allergic rhinitis underwent single exposures to filtered air, 125 p.p.b. and 250 p.p.b. ozone, as well as repeated exposures to 125 p.p.b. ozone on four consecutive days. Twenty hours after the (final) exposure, subjects inhaled a single dose of allergen and a sputum induction was performed 6–7 h later.

Results In the subjects with rhinitis, the mean early‐phase response of FEV1 and the number of ≥ 20% reductions were significantly greater after exposure to 250 or 4 × 125 p.p.b. ozone compared with filtered air. In addition, most of the ≥ 15% late‐phase responses in FEV1 occurred after exposure to 4 × 125 p.p.b., as well as the strongest effects on sputum parameters. The rise in the number of eosinophils was statistically significant in both groups. Regarding the number of lymphocytes and the concentrations of mast cell tryptase, histamine or LDH, significance was, however, only reached in the asthma group.

Conclusion Our data suggest that repeated exposure to ozone, at a peak ambient air level, can enhance both functional and inflammatory responses to inhaled allergen in subjects with pre‐existing allergic airway diseases, and that these effects might reach a clinically relevant magnitude.
Sputum processing: a new method to improve cytospin quality
Authors: Ronchi M. C. , Galli G. , Zonefrati R. , Tanini A. , Scano G. , Duranti R.
Published: March 2003

Background  Sputum examination is being increasingly used as a non‐invasive method for studying airway inflammation. However, the application of sputum still presents some methodological problems and the results of sputum analysis may be substantially flawed by salivary contamination, cell and mucus debris. In addition, much work is needed to deepen the possibility of extensive application of cell and molecular biology techniques to sputum analysis.

Objective  In an attempt to improve the technique of sputum processing, we investigated the effect of: (i) 20 and 11 µm filtration in addition to 40 µm on salivary contamination; (ii) Percoll density gradient centrifugation on sputum slides quality; (iii) a culture medium (Minimum Essential Medium containing HEPES 22 mm, pH 7.4: MEM) as washing and suspension solution compared to PBS on cell viability.

Methods  Induced sputum samples were obtained in 37 asthmatics. 21 samples were processed as selected sputum and 16 samples as entire expectorates. After dithiotreitol (DTT) homogenization, each specimen was aliquoted in two parts of equal volume. One portion was processed with the usual method, the other using a modified method: cell pellet was suspended in sterile MEM, filtered through 40, 20 and 11 µm net filters and separated from the residual debris by Percoll gradient centrifugation.

Results  As compared to the current sputum processing this method resulted in: (i) no selective bronchial cellular loss; (ii) a significant decrease of salivary contamination, particularly in entire expectorates in which squamous cells were reduced from 47 (36) to 15.5% (20) as median values and interquartile range; (iii) a higher proportion of good quality cytospins; (iv) maintenance of cell viability over the time (88% vs. 81% in MEM and PBS, respectively) 1 h after sample collection.

Conclusion  In the present study we demonstrated that the proposed method is feasible and makes it possible to overcome most of the technical limits met with the commonly used method, pointing to a potential extension of induced sputum application for more sophisticated techniques.
Are cysteinyl leukotrienes involved in allergic responses in human skin?
Authors: Church M. K. , Griffiths T. J. , Jeffery S. , Ravell L C. , Cowburn A. S. , Sampson A. P. , Clough G. F.
Published: March 2003

Background Cysteinyl leukotrienes have been suggested to be involved in producing the symptoms of both the early and late phases of the allergic response in the lung and other tissues.

Objective To use scanning laser Doppler imaging, microdialysis and immunocytochemistry to explore the mediator and cellular mechanisms of the dermal allergic response.

Methods Thirteen atopic volunteers received intradermal injections into the forearm of grass pollen or D. pteronyssinus extract. Changes in dermal blood flow up to 8 h were monitored by scanning laser Doppler imaging. The release of histamine, PGD2 and LTC4/D4/E4 was assessed by dermal microdialysis. Skin biopsies were taken at 6 h to determine numbers of mast cells, eosinophils, basophils, Langerhans' cells, and monocytes/macrophages, and the expression of COX‐1, COX‐2, 5‐LO and FLAP.

Results Allergen provocation produced an immediate weal and flare response followed by an erythematous induration peaking at 6 h. During the first hour, c. 84 pmoles of histamine and c. 0.3 pmoles of PGD2 were recovered by microdialysis (both P < 0.001) but LTC4/D4/E4 was undetectable. No histamine, PGD2 or LTC4/D4/E4 was detectable at later times. Immunocytochemical examination of biopsies taken at 8 h showed increased numbers of eosinophils and basophils and in COX‐2, 5‐LO and FLAP, but not COX‐1. Expression of 5‐LO and FLAP was associated primarily with eosinophils.

Conclusions These findings suggest that inflammatory cells recruited to the site of allergen injection are not activated to release detectable amounts of cysteinyl leukotrienes. Hence, it is unlikely that the late‐phase erythematous induration is mediated by this autocoid.
German cockroach extract increases bronchial epithelial cell interleukin‐8 expression
Authors: Bhat R. K. , Page K. , Tan A. , Hershenson M. B.
Published: March 2003

Background Cockroach exposure has been recognized as a common trigger for asthma. While dust mite and Aspergillus fumigatus aeroallergens have been noted to have direct effects on airway epithelium, direct effects of cockroach proteins have not been determined.

Objective The purpose of this study was to investigate whether cockroach extract has a direct pro‐inflammatory effect on airway epithelium.

Methods We examined the effect of crude German cockroach (Blattella germanica) extract on IL‐8 expression in a human bronchial epithelial cell line (16HBE14o‐cells) and primary human bronchial epithelial cells. Transcription from the IL‐8 promoter and protein abundance were determined by reporter assay and enzyme‐linked immunosorbent assay (ELISA), respectively. Endotoxin levels in the crude cockroach extracts were determined using the Limulus Amebocyte Lysate assay. Protease activity was assessed using Azocoll as a substrate.

Results We found that crude cockroach extract induced a synergistic increase in TNF‐α‐induced transcription from the IL‐8 promoter. The synergistic effect was observed with as little as 0.3 µg/mL of crude cockroach extract, while larger concentrations (30 µg/mL) approximately doubled TNF‐α‐induced IL‐8 promoter activity. Similar effects of cockroach extract on IL‐8 protein abundance were observed in both 16HBE14o‐ and primary human bronchial epithelial cells. Addition of endotoxin at concentrations found in the cockroach extract had no effect on TNF‐α‐mediated IL‐8 expression. The serine protease inhibitors aprotinin and phenylmethylsulphonyl fluoride abolished cockroach‐induced synergy, while the cysteine protease inhibitors E64 and leupeptin had little effect. Measurement of protease activity using Azocoll as a substrate confirmed the presence of protease activity in cockroach extracts. Addition of recombinant Bla g 2, Bla g 3 and Bla g 5 had no effect on TNF‐α‐induced IL‐8 promoter activation. Finally, cockroach extract also increased TNF‐α‐induced transcription from the IL‐6 promoter.

Conclusions German cockroach extract contains novel serine protease activity which has a direct pro‐inflammatory effect on airway epithelial cells
Is the increase in allergic respiratory disease caused by a cohort effect?
Authors: Linneberg A. , Nielsen N. H. , Madsen F. , Frølund L. , Dirksen A. , Jørgensen T.
Published: March 2003

Background Changes in lifestyle or environmental factors are responsible for the increasing prevalence of allergic respiratory disease. Establishing the time at which the increase began may provide a clue as to what factors possibly could have contributed to the increase. Many cross‐sectional studies have shown that the prevalence of allergic sensitization decreases with increasing age. This could reflect the natural course of allergic sensitization. Alternatively, this could reflect that the increase in sensitization is caused by a cohort effect, i.e. an increase among subjects born during recent decades.

Objective The aim was to investigate age‐specific changes in the prevalence of allergic sensitization in a cohort of adults.

Methods A total of 599 subjects aged 15 to 69 years participated in a cross‐sectional general population study in 1990. In 1998 they were invited to a follow‐up, and 64.4% (386/599) were re‐examined. Serum samples obtained from the participants in 1990 and 1998 were analysed for specific IgE to six common inhalant allergens with the same assay.

Results The prevalence of allergic sensitization (specific IgE to at least one allergen) increased among subjects who were less than c. 30 years at baseline (1990), i.e. subjects born during the 1960s or later, while the prevalence was unchanged among subjects who were more than c. 30 years at baseline.

Conclusions The results support the notion that the increasing prevalence of allergic respiratory disease is caused by a cohort effect. Thus, changes in lifestyle or environmental factors that occurred around or after 1960 may have contributed to this increase.
Constitutive secretion of the granule chymase mouse mast cell protease‐1 and the chemokine, CCL2, by mucosal mast cell homologues
Authors: Brown J. K. , Knight P. A. , Wright S. H. , Thornton E. M. , Miller H. R. P.
Published: March 2003

Background The mucosal mast cell (MMC) granule‐specific β‐chymase, mouse mast cell protease‐1 (mMCP‐1), is released systemically into the bloodstream early in nematode infection before parasite‐specific IgE responses develop and TGF‐β1 induces constitutive release of mMCP‐1 by homologues of MMC in vitro. Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein‐1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated.

Objective To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP‐1, and the chemokine, CCL2.

Methods MMC homologues were generated by culturing bone marrow cells in the presence of TGF‐β1, IL‐3, IL‐9 and stem cell factor (SCF). The intracellular distribution of mMCP‐1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP‐1 and CCL2 was investigated using the Golgi‐disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA.

Results mMCP‐1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP‐1 was significantly inhibited (≈ 40%) by cycloheximide and by the Golgi‐disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP‐1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2.

Conclusion The culture conditions that promote mMCP‐1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators.
Immunoadjuvant properties of glove cornstarch powder in latex‐induced hypersensitivity
Authors: Barbara J. , Santais M.‐C. , Levy D. A. , Ruff F. , Leynadier F.
Published: March 2003

Background Cornstarch powder present in medical gloves plays an important role in latex‐induced hypersensitivity as allergen carrier either, by the inhalation route, by skin contact or by direct contact with mucous membranes.

Objective Our objective was to test the hypothesis that cornstarch could act as an immunoadjuvant in immediate type‐I latex‐induced hypersensitivity.

Methods Guinea‐pigs were sensitized by intraperitoneal route with two different antigens (latex proteins and ovalbumin) with or without cornstarch powder. Airway responsiveness after specific bronchial provocation was evaluated and specific IgG and IgG1 levels were determined by enzyme‐linked immunosorbant assay (ELISA). Controls were treated with cornstarch powder or saline alone.

Results Animals sensitized with latex proteins (n = 7 in each group) showed significant bronchoconstriction (P < 0.03) and higher anti‐latex antibody levels than the controls (P < 0.005). Guinea‐pigs sensitized with latex‐contaminated cornstarch had higher levels of specific antibodies than those sensitized with latex alone (P < 0.05). Animals sensitized to latex mixed with cornstarch showed higher bronchospasm than those treated with latex alone (P < 0.003). Animals sensitized to ovalbumin mixed with cornstarch also showed higher antibody and bronchoconstriction levels (P < 0.05) than those immunized with ovalbumin alone but antibody titres were significantly lower than those of the animals treated with ovalbumin and Freund's complete adjuvant (P < 0.01; n = 5 in each group).

Conclusion Our findings show that cornstarch powder increases antigen‐induced bronchoconstriction and antibody production. This role of immunoadjuvant is not antigen‐specific. The cornstarch powder used as donning agent in latex gloves is an allergen carrier and it can enhance latex‐induced hypersensitivity.
Elicitation of the allergic reaction in β‐lactoglobulin‐sensitized Balb/c mice: biochemical and clinical manifestations differ according to the structure of the allergen used for challenge
Authors: Adel‐Patient K. , Nahori M.‐A. , Proust B. , Lapa e Silva J. R. , Créminon C. , Wal J.‐M. , Vargaftig B. B.
Published: March 2003

Background Mouse models of allergy are used to study the mechanisms of induction and perpetuation of bronchopulmonary hyper‐reactivity (BHR) as related to eosinophils and specific IgE.

Objective Our aim was to adapt the current model for the study of bovine β‐lactoglobulin (BLG), a major cow's milk allergen, and to further analyse the mechanisms of the acute and late allergic reaction.

Methods Female Balb/c mice were sensitized intraperitoneally with BLG and the influence of the adjuvant and of the BLG dose on the IgE response was analysed, IgE and IgG1 epitopes being characterized. Once optimized, this model was applied to the study of the active phase of allergy in the respiratory tract after a single airway challenge using native or denatured BLG, which contains only linear epitopes.

Results An immediate allergic reaction was characterized by the rapid release of histamine into the bronchoalveolar lavage fluids. Prostaglandin (PG)D2 was only present when the standard histamine‐releasing agent compound 48/80 or denatured BLG were used as triggers, whereas native BLG induced leukotriene release. Twenty‐four hours after challenge, BHR, eosinophil influx, IL‐4 and IL‐5 production, plasma exudation and mucus production were very much increased, differently depending on the allergen structure, and indicated the occurrence of the late allergic reaction. Our results show that the murine model can be used to study the mechanisms of allergy to clinically relevant antigens, such as those contained in cow's milk. The acute allergic reaction, which depends on the structural feature of the allergen, is composed of two distinct pathways characterized by peptido‐leukotrienes or PGD2 production, which may result from distinct activation intensities of mast cells, leading to distinct late reactions.

Conclusion This study thus demonstrates a clear link between the structural feature of a protein, and the physiopathology of the experimental asthmatic reaction.
Psoriasis, psoriatic arthropathy and relapsing orbital myositis
Authors: Ajitsaria R. , Dale R. , Ferguson V. , Mayou S. , Cavanagh N.
Published: March 2003

Orbital myositis is an inflammatory disorder of the orbital muscles causing orbital pain and restriction of eye movements. Although rare in children, it is most frequently seen after orbital trauma or as a post-infectious process. We describe a child with chronic relapsing psoriasis, juvenile psoriatic arthritis and relapsing bilateral orbital myositis.
Early increase of CD4+ CD45RA+ and CD4+ CD95− cells with conserved repertoire induced by anti-retroviral therapy in HIV-infected patients
Authors: Silvestri G , Munoz-Calleja C , Bagnarelli P , Piedimonte G , Clementi M , Montroni M
Published: March 2003

Administration of anti-retroviral drugs induces a decrease of viral load associated with increase of CD4+ cell count in most HIV-infected patients. To investigate the early changes in CD4+ cell phenotype induced by anti-retroviral therapy, six patients with CD4+ cell count > 100/mm3 and never treated with anti-HIV therapy were enrolled and blood samples collected several times within 14 days from the initiation of therapy with Zidovudine plus Didanosine. CD4+ cell count and HIV viraemia were investigated at each time point, as well as the expression of CD45RA, CD45RO and CD95/Fas molecules on CD4+ cells, and the T cell receptor (TCR) Vβ repertoire of CD4+ cells. All patients showed a rapid and dramatic decrease in viral load with a corresponding increase of CD4+ cell count. The main remodelling of CD4+ cell subpopulations took place in the first 14 days of therapy, and consisted of: (i) increased CD4+ CD45RA+/CD4+  CD45RO+ ratio; (ii) decrease of CD95/Fas expression. The rise in absolute number of CD4+ CD45RA+ cells was paralleled by an increase of CD4+ CD95/Fas− cells and accounted for most of the early increment of CD4+ cell count. The TCR Vβ repertoire of CD4+ cells was conserved after anti-HIV therapy, with the exception of two patients with expanded CD4+ Vβ12+ cells, which also tested CD45RA+ and CD95/Fas−. These experiments show that newcomer CD4+ lymphocytes are CD45RA+ CD95/Fas− cells, suggesting that blocking HIV replication causes an early and antigen-independent proliferation of possibly ‘naive’ cells unprimed for CD95/Fas-mediated apoptosis. These cells expressed a conserved and widespread TCR repertoire, suggesting that their capability for antigenic recognition is intact.
A study of the epitopes on steroid 21-hydroxylase recognized by autoantibodies in patients with or without Addison's disease
Authors: Volpato M , Prentice L , Chen D , Betterle C , Rees Smith B , Furmaniak J
Published: March 2003

Steroid 21-hydroxylase (21-OH) autoantibodies are found in patients with autoimmune Addison's disease (AAD), either isolated or associated with autoimmune polyglandular syndrome (APS) type I and II and in adrenal-cortex autoantibody (ACA)-positive patients without AAD. In order to assess any differences in the 21-OH autoantibodies in these different patient groups, we have studied their reactivity with different epitopes on 21-OH using full length and modified 35S-labelled 21-OH proteins produced in an in vitro transcription/translation system. There were no major differences in the pattern of autoantibody reactivity with the different modified 21-OH proteins in patients with isolated AAD or with APS types I and II, and in 21-OH autoantibody-positive patients with clinical AAD, subclinical AAD and those maintaining a normal adrenal function. Our studies also indicate that the main epitopes for 21-OH autoantibodies in patients with different forms of autoimmune adrenal disease are located in the C-terminal end and in a central region of 21-OH.
Phenothiazine-induced increase in thyroid autoantigens and costimulatory molecules on thyroid cells: a pathophysiological mechanism for drug-induced autoimmunity?
Authors: Takorabet L , Ropars A , Stasiuk L , Raby C , Charreire J
Published: March 2003

We have previously demonstrated (J Immunol 1995; 154:3593) that MHC class II antigens can be induced on thyroid epithelial cells (TEC) by alimemazine, a member of the phenothiazine group. Although this expression of MHC class II antigens on TEC confers the theoretical ability to behave as antigen-presenting cells (APC), the simultaneous expression of self antigens and co-receptor(s) must also occur for efficient presentation of self antigens. Therefore, we investigated whether alimemazine applied at pharmacologic doses would modify the expression of thyroid antigens, and simultaneously, the expression of intercellular adhesion molecule-1 (ICAM-1), B7, and LFA-1 co-receptors in human TEC in culture. Using polymerase chain reaction (PCR) amplification and Northern blot analysis, we showed that alimemazine induces increases in thyroglobulin (Tg) and thyroid-stimulating hormone receptor (TSH-R) cDNA, within the first 2 h following its addition. This phenomenon is followed 48 h later by an increase of Tg and TSH-R protein expression on the surface of TEC. Furthermore, increases in the expression of ICAM-1 and B7 co-receptors were concomitantly observed. These results suggest that alimemazine, a drug currently used in paediatrics, could play a role in the induction and perpetuation of thyroid autoimmune disorders by transforming TEC into functional APC.
Tenidap decreases IL-8 and monocyte chemotactic peptide-1 (MCP-1) mRNA expression in the synovial tissue of rabbits with antigen arthritis and in cultured synovial cells
Authors: Palacios I , Lopez-Armada M J , Hernandez P , Sanchez-Pernaute O , Gutierrez S , Miguelez R , Martinez J , Egido J , Herrero-Beaumont G
Published: March 2003

Since IL-8 and MCP-1 are chemoattractant proteins that participate in the recruitment of inflammatory cells into the arthritic joint, we examined the effects of tenidap, a new anti-inflammatory drug of the oxindole family, on IL-8 and MCP-1 expression in the joints of rabbits with acute antigen arthritis. The model was induced by injecting 5 mg/ml ovalbumin into the knees of 20 preimmunized rabbits. Animals were randomized into two groups: treated with tenidap (15 mg/kg per 12 h), or untreated. The effect of tenidap treatment was evaluated on chemokine production in synovial membranes of rabbits with arthritis and in cultured monocytic and synovial cells (SC). By immunoperoxidase staining, chemokines were localized in the synovial tissue. Chemokine messenger RNA levels in the synovial membranes and in cultured cells were analysed by reverse transcription-polymerase chain reaction (RT-PCR). At the end of the study, tenidap significantly reduced neutrophil infiltration into the joint cavity (27 ± 4 × 106 cells/ml versus 45 ± 6 × 106 cells/ml in untreated; P < 0.05), and synovial effusion (134 ± 15 μl versus 236 ± 19 μl in untreated; P < 0.005). Untreated rabbits showed synovial membrane up-regulation in mRNA expression of IL-8 and MCP-1 (11- and seven-fold versus healthy rabbits, respectively) that was markedly decreased by tenidap (two- and three-fold versus healthy rabbits, respectively). IL-8 and MCP-1 were localized in the synovial tissue in a perivascular pattern and areas of the interstitium and lining, mostly coinciding with cell infiltration. Tenidap also reduced the accumulation of IL-8 and MCP-1 proteins. In cultured synovial and monocytic cells, tumour necrosis factor-alpha (TNF-α) elicited an increase in gene expression of IL-8 (four- and nine-fold, respectively) and MCP-1 (nine- and four-fold, respectively) that was significantly reversed in both cell types by 10 μm tenidap. These results suggest that the beneficial effect of tenidap in acute antigen arthritis could be related to the down-regulation in gene expression and synthesis of IL-8 and MCP-1, two key chemokines involved in the recruitment of inflammatory cells.
Oxidative stress suppresses transcription factor activities in stimulated lymphocytes
Authors: Flescher E , Tripoli H , Salnikow K , Burns F J
Published: March 2003

Effects of oxidative stress on stimulation-dependent signal transduction, leading to IL-2 expression, were studied. Purified quiescent human blood T lymphocytes were subjected to: (i) acute exposure to hydrogen peroxide; (ii) chronic exposure to hydrogen peroxide; and (iii) acute exposure to ionizing radiation. The cells were then stimulated for 6 h. DNA-binding activities (determined by the electrophoretic mobility shift assay) of three transcription factors: NFκB, AP-1 and NFAT, were abolished in the lymphocytes by all three modes of oxidative stress. The lymphocytes exhibited lipid peroxidation only upon exposure to the lowest level of hydrogen peroxide used (20 μm). All three modes of oxidative stress induced catalase activity in the lymphocytes. The only exception was hydrogen peroxide at 20 μm, which did not induce catalase activity. We conclude that: (i) suppression of specific transcription factor functions can potentially serve as a marker of exposure to oxidative stress and its effects on human lymphocytes; (ii) lipid peroxidation is only detectable in human lymphocytes upon exposure to weak oxidative stress which does not induce catalase activity; (iii) therefore, transcription factor DNA-binding activities are more sensitive to oxidative stress than lipid peroxidation.
Changes to the cytokine microenvironment in the genital tract mucosa of HIV+ women
Authors: Olaitan A , Johnson M A , Reid W M , Poulter L W
Published: March 2003

As previous studies have indicated that genital tract mucosal T cell function may be impaired in HIV infection, we investigated the T cell cytokine mRNA in the genital tract mucosa of HIV-infected women to determine if there are alterations in the cytokine profile which may explain the T cell impairment. The in situ hybridization technique was used to investigate the T helper-1 (Th1: IL-2, interferon-gamma (IFN-γ)) and Th2 cytokine (IL-4, IL-5, IL-10) mRNA profile in cervical biopsies from 10 HIV+ and 10 HIV− subjects. Cervical intraepithelial neoplasia (CIN) and genital infection had previously been excluded and the distribution of immunocompetent cells within the cervical mucosa was known for each subject. Non-parametric tests were used to compare the optical density (OD) of cytokine mRNA in the HIV+ and HIV− groups. Comparisons were also made between peripheral CD4 lymphocyte counts, cervical CD4/CD8 T lymphocyte ratios and cytokine mRNA OD in HIV+ subjects. The HIV+ women had significantly higher mRNA OD for the Th2 cytokines IL-4, IL-5 and IL-10 than HIV− women. There was also significantly lower IL-2 mRNA OD in the former group. HIV+ women had lower IFN-γ mRNA than HIV− women, but the difference was not statistically significant. There was no correlation between cytokine mRNA OD and peripheral CD4 count or cervical CD4/CD8 ratio. The predominance of Th2 cytokines, which are immuno-inhibitory, in the cervical mucosa of HIV+ women may underlie the impaired cytotoxic potential observed in the CD8+ T lymphocytes and may contribute to the susceptibility of HIV-infected women to recurrent genital tract infections and cervical neoplasia.
Binding of anticardiolipin antibodies to protein C via β2-glycoprotein I (β2-GPI): a possible mechanism in the inhibitory effect of antiphospholipid antibodies on the protein C system
Authors: Atsumi T , Khamashta M A , Amengual O , Donohoe S , Mackie I , Ichikawa K , Koike T , Hughes G R V
Published: March 2003

It is known that antiphospholipid antibodies (aPL) hamper the anticoagulant activity of the protein C system, but the mechanism is still obscure. In this study, we demonstrate that anticardiolipin antibodies (not anti-protein C autoantibodies) can bind protein C via β2-GPI, which bears their binding epitope, in a fashion dependent on negatively charged phospholipids. We studied the binding of IgG from aPL to protein C in the presence of β2-GPI by ELISA (anti-‘protein C’ antibody ELISA), and compared their binding with those obtained in the absence of β2-GPI. In the anti-‘protein C’ antibody ELISA system, 47% of 78 aPL+ patients had a positive titre in the presence of cardiolipin (CL) and β2-GPI, but binding was not found in the absence of β2-GPI. Highly significant correlations were found between the titre of anti-‘protein C’ antibody in the presence of β2-GPI and that of anti-β2-GPI antibody (r = 0·802, P = 0·0001). We further analysed the interaction between protein C, phospholipids, β2-GPI and human aCL MoAbs established from patients with antiphospholipid syndrome. In a first set of experiments, the binding of β2-GPI to protein C and its phospholipid dependency were investigated. β2-GPI bound to protein C in the presence of CL or phosphatidylserine, but not in the presence of phosphatidylcholine or phosphatidylethanolamine. In a second group of experiments, the binding of three human monoclonal aCL recognizing the cryptic epitope of β2-GPI (virtually anti-β2-GPI antibodies) was evaluated in the presence of cardiolipin and β2-GPI. All three human monoclonal aCL bound to protein C in the presence of CL and β2-GPI, whereas they did not in the absence of either β2-GPI or CL. These data suggest that protein C could be a target of aCL by making a complex with CL and β2-GPI, leading to protein C dysfunction.
Detection of Fcγ receptors on human endothelial cells stimulated with cytokines tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ)
Authors: Pan L-F , Kreisle R A , Shi Y-D
Published: March 2003

This investigation was conducted to detect Fcγ receptors (FcγR) on cytokine-stimulated human endothelial cells (EC) by measuring anti-FcγR MoAb binding with an ELISA. TNF-α and IFN-γ significantly increased the expression of FcγR type II (FcγRII) and type III (FcγRIII) on aortic EC. Simultaneous treatment with both cytokines had a synergistic effect and pretreatment of EC with IFN-γ augmented the effect of TNF-α. The greatest effect was the increase (up to four-to-six-fold) in expression of FcγRII found by the simultaneous treatment of aortic EC with both cytokines. The receptors were expressed on the cell surface and showed receptor capping after incubation at 37°C. This study showed that the inflammatory cytokines TNF-α and IFN-γ enhanced low-affinity FcγR expression on human EC in vitro. The expression of FcγR may contribute to the specific localization of circulating immune complexes on blood vessels in areas of vasculitis.
T lymphocyte responses to anti-neutrophil cytoplasmic autoantibody (ANCA) antigens are present in patients with ANCA-associated systemic vasculitis and persist during disease remission
Authors: King W J , Brooks C J , Holder R , Hughes P , Adu D , Savage C O S
Published: March 2003

ANCA with specificity for myeloperoxidase (MPO) and proteinase 3 (PR3) are present in patients with systemic vasculitis. The aim of this work was to determine whether such patients have T cell responses to these antigens and whether these responses are related to disease activity. Peripheral blood lymphocytes from 45 patients and 19 controls were cultured with ANCA antigens and proliferation measured. The antigens used were heat-inactivated (HI) MPO, HI PR3, native (non-HI) PR3, HI whole α-granules, and 25 overlapping peptides covering the entire PR3 sequence. Significant responses to both whole PR3 preparations were seen from patient and control groups, and to the α-granules from the patient group. Patients responded at all stages of disease: active, remitting, treated or untreated. Only two patients responded significantly to MPO. Responses were significantly higher with the patient group than the control group to all four whole ANCA antigens. Responses to those PR3 peptides containing epitopes known to be recognized by ANCA were detected from one patient. Thus, these studies demonstrate that T cells from vasculitis patients can proliferate to PR3 and occasionally to associated ANCA antigens. Further, responses may persist even after disease remission has been achieved.
Reduced IL-1 production in adolescents with mite antigen asthma in remission
Authors: Noma T , Ichikawa K , Yoshizawa I , Aoki K , Kawano Y , Baba M
Published: March 2003

To determine the immunological mechanisms associated with outgrowing mite antigen-induced bronchial asthma during adolescence, we studied the relationship between clinical status and Dermatophagoides farinae (Df) antigen-induced peripheral cell activation by measuring IL-1α and IL-lβ production in patients with bronchial asthma. After antigen-driven restimulation in vitro, there was increased IL-1α, IL-1β production by peripheral blood mononuclear cells (PBMC) from patients with active bronchial asthma, while cellular IL-1α, IL-1β production was reduced in patients with asthma in remission. IL-1α and IL-1β production by PBMC (possibly reflecting airway inflammation) after exposure to Df antigen might be down-regulated in patients outgrowing mite antigen-induced asthma, because lipopolysaccharide-induced IL-1α, IL-1β production (seen in both normal individuals and patients with active asthma) was also reduced when patients were in remission.
Interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α) are necessary in the early stages of induction of CD4 and CD8 cytotoxic T cells byMycobacterium lepraeheat shock protein (hsp) 65 kD
Authors: Del C Sasiain M , DE LA Barrera S , Fink S , Finiasz M , Alemán M , Fariña M H , Pizzariello G , Valdez R
Published: March 2003

Cytotoxic T cells (CTL) may play an important role in host defence against mycobacterial infections. CD4 CTL are preferentially induced by mycobacteria, but both CD4 and CD8 CTL may be necessary components of a protective immune response. The 65-kD mycobacterium heat shock protein (hsp65) is a poor inducer of CTL in multibacillary leprosy (MB) patients. In this study we evaluate the possible role of cytokines in modulating the cytotoxic activity of CTL from leprosy patients and normal individuals (N) against autologous macrophages presenting Mycobacterium leprae hsp65. Our results show that hsp65-specific CTL were generated from both CD4 and CD8 lymphocytes. In N, individual cytokines as well as the combination of them were able to modify the hsp65-induced cytotoxic activity. The effect of cytokines on leprosy patients' lymphocytes was different in MB and paucibacillary (PB) patients. Thus, IL-6, IL-2, IFN-γ or TNF-α did not modify the generation of hsp65-CTL from either MB (with or without an erythema nodosum episode (ENL)) or PB. In all the patients the simultaneous addition of two cytokines was required in order to increase CTL generation. In MB, IL-6 plus IFN-γ or IL-2 increased both CD4 and CD8 CTL, while TNF-α plus IFN-γ up-regulated only CD4 CTL. In PB, CD8 CTL were prominent with IL-6 plus IFN-γ, while the increase was significant in CD4 CTL with IL-6 plus IL-2. Down-regulation of CTL was observed by addition of IL-4, IL-10, anti-IFN-γ or anti-TNF-α in N controls. Our data demonstrate that IFN-γ and TNF-α must be present for at least the first 60 h of the induction stage in order to generate full hsp65 CTL. Hence, IFN-γ and TNF-α would be key factors in the generation of hsp65 CTL.