bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular cloning and characterization of a new Japanese cedar pollen allergen homologous to plant isoflavone reductase family
Authors: Kawamoto S. , Fujimura T. , Nishida M. , Tanaka T. , Aki T. , Masubuchi M. , Hayashi T. , Suzuki O. , Shigeta S. , Ono K. 
Published: March 2003

SummaryBackground Japanese cedar (Cryptomeria japonica) pollen is a major cause of seasonal pollinosis, and more than 10% of Japanese people suffer from this allergic disorder. However, only two major pollen allergens, Cry j 1 and Cry j 2, have been identified and exclusively characterized.Objective The aim of this study was to explore and identify important Japanese cedar pollen allergens other than Cry j 1 or Cry j 2.Methods C. japonica
cDNA library was immunoscreened by rabbit antiserum raised against a partially purified cedar pollen allergen fraction. An isolated cDNA clone was inserted into a glutathione S‐transferase (GST)‐tagged
Escherichia coli expression vector to obtain recombinant GST fusion protein. Non‐fusion recombinant protein was purified by glutathione Sepharose affinity chromatography in conjunction with factor Xa cleavage of the GST moiety. IgE‐binding ability of the recombinant protein was then evaluated by western blot analysis and enzyme‐linked immunosorbent assay (ELISA).Results The cDNA encodes 306 amino acids with significant sequence similarity to those of plant isoflavone reductase‐like proteins, which include a recently identified birch pollen allergen Bet v 5. Western blot analysis demonstrated that recombinant protein was recognized by cedar pollinosis patient IgE. In contrast to Bet v 5 being reported as a minor allergen, the recombinant protein exhibited 76% IgE binding frequency (19/25) against pollinosis patients.Conclusion Here we identified the third member of Japanese cedar pollen allergen homologous to isoflavone reductase. Its high IgE‐binding frequency implicates that the isoflavone reductase homologue might be an additional major pollen allergen in C. japonica.
Suplatast tosilate inhibits thymus‐ and activation‐regulated chemokine production by antigen‐specific human Th2 cells
Authors: Oda N. , Minoguchi K. , Tanaka A. , Yokoe T. , Minoguchi H. , Matsuo H. , Nakashima M. , Tasaki T. , Adachi M. 
Published: March 2003

SummaryBackground Suplatast tosilate is an anti‐allergic agent that suppresses cytokine production by human Th2 cells.Objective We investigated the effects of suplatast tosilate on the production of thymus‐ and activation‐regulated chemokine (TARC) by T cells from allergic patients with asthma.Methods Purified protein derivative (PPD)‐specific Th1 cell lines and Dermatophagoides farinae (Der f)‐specific Th2 cell lines were established from nine patients with house dust mite‐allergic asthma. The effects of suplatast tosilate on mRNA expression of TARC and protein production of TARC from antigen‐specific Th1 or Th2 cell lines were investigated after stimulation with relevant antigens or phytohemagglutinin (PHA). In addition, the effects of IL‐4, IL‐10, and IFN‐γ on TARC production by Der f‐specific Th2 cell lines in the presence or absence of suplatast tosilate were studied.Results Although PPD‐specific Th1 cell lines did not produce TARC after stimulation with PPD antigen or PHA, stimulation of Der f‐specific Th2 cell lines with Der f antigen or PHA increased production of TARC. Suplatast tosilate significantly and dose‐dependently inhibited production of TARC by Der f‐specific Th2 cell lines stimulated with either Der f antigen (76.5% inhibition at 100 µg/mL, P < 0.01) or PHA (81.9% inhibition at 100 µg/mL, P < 0.01). TARC production by Der f‐specific Th2 cell lines was significantly increased only by activation with IL‐4 but not with IL‐10 or IFN‐γ; this increase in TARC production was significantly inhibited by suplatast tosilate (97.5% inhibition at 100 µg/mL, P < 0.01).Conclusion Suplatast tosilate inhibits TARC production by human Th2 cells. Therefore, this agent inhibits both Th2 cytokine and Th2 chemokine and may be a useful anti‐allergic agent.
Increased aeroallergen‐specific interleukin‐4‐producing T cells in asthmatic adults
Authors: Pala P. , Message S. D. , Johnston S. L. , Openshaw P. J. M. 
Published: March 2003

SummaryBackground Asthma, atopy and some forms of respiratory syncytial virus (RSV) disease are thought to be caused by T cells making IL‐4 (Th2 cells). However, not all patients with similar patterns of clinical disease have the same underlying pathogenesis and the ability to detect immunopathogenic T cells by examination of the peripheral blood remains in doubt. With the prospect of specific immunotherapy for diseases caused by T cell subsets, it is important to determine whether peripheral blood mononuclear cell (PBMC) reactivity can be used to establish the presence of immunopathogenic responses and therefore to predict therapeutic effects.Objective To detect IL‐4 and IFN‐γ production as markers of Th1 and Th2 responses in the peripheral blood of atopic and asthmatic adults.Methods PBMC from 22 adult asthmatics (18 of whom were atopic) and 21 non‐asthmatic volunteers (ten of whom were atopic) were stimulated with cat, birch and house dust mite allergens, human rhinovirus, RSV and recombinant chimaeric F/G protein from RSV in vitro. ELISPOT assays were used to enumerate cells producing IL‐4 and IFN‐γ.Results Asthmatics had a sixfold increase in frequencies of IL‐4‐producing cells to cat and birch allergen (median values: 37 vs. 7 per million PBMC, P < 0.01 and 20 vs. 3 per million PBMC, P < 0.04, respectively) compared to non‐asthmatics. By contrast, non‐asthmatic atopics showed no specific increase in antigen‐specific IL‐4 responses and there was no evident correlation between skin prick test reactivity and ELISPOT results. Atopics had significantly more IFN‐ γ‐producing cells specific for FG than nonatopics. while IFN‐γ and IL‐4 responses to other antigens were not significantly different.Conclusion Enhanced IL‐4 responses to non‐viral aeroallergens are seen in adults with asthma, while enhanced IFN‐γ responses to viral antigen FG were seen in atopics. In practical terms, ELISPOT assays for specific cytokines may provide a method that could be used to monitor antigen‐specific T cell responses in peripheral blood.
Comparison of four allergen‐sampling methods in conventional and allergy prevention classrooms
Authors: Karlsson A.‐S. , Renström A. , Hedrén M. , Larsson K. 
Published: March 2003

SummaryBackground Several allergen‐sampling methods are used to assess level of personal or indirect exposure to cat in homes, schools and other public buildings and working environments.Objective To compare four different allergen‐sampling methods (dust collectors, Petri dishes, person‐carried pumps and intranasal samplers) by simultaneous sampling in classrooms and to compare the cat allergen levels between conventional classrooms and allergy prevention classrooms. Another aim was to relate the results to self‐reported frequency of allergy and asthma symptoms among the children, to their perception of the school environment.Methods Among all compulsory schools (n = 257) in the Stockholm suburban area, 35 classrooms (five with implemented allergy prevention measures, seven with additional cleaning and 23 with normal cleaning routines) were chosen for allergen‐sampling. Dust collectors (two models), Petri dishes, person‐carried pumps and intranasal samplers were used simultaneously. All children (n = 829) received a self‐administered questionnaire which included questions about home and school environment, allergic disease, asthma symptoms and pet contact.Results The correlation between sampling methods was generally poor. Furthermore, there was no significant difference in allergen levels between allergy prevention and allergen avoidance classes compared to conventional classes. Median levels were generally, but not significantly, lower in classes with few cat owners, compared to classes with many cat owners. Children in allergy prevention classes were more satisfied with the indoor air quality and cleaning than children attending classes with fewer or no allergy prevention measures (P < 0.0001). Nine per cent of all children reported allergic symptoms while at school.Conclusion The lack of correlation between sampling methods used simultaneously demonstrates the difficulty in assessing allergen levels in schools and similar environments. The implemented intervention measures (allergy prevention/allergen avoidance) did not influence cat allergen levels at school.
Skin test with a timothy grass (Phleum pratense) pollen extract vs. IgE to a timothy extract vs. IgE to rPhl p 1, rPhl p 2, nPhl p 4, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, and rPhl p 12: epidemiological and diagnostic data
Authors: Mari A. 
Published: March 2003

SummaryBackground The diagnostic approach to grass pollen allergy is now possible by detecting specific IgE to its allergenic components.Objective To compare the IgE reactivity to a timothy grass pollen extract with the IgE reactivity to eight allergenic components from the same source (Phl p 1, 2, 4, 5, 6, 7, 11, 12). Both were compared with the skin test reactivity to a timothy grass extract.Methods A population survey was carried out by means of the skin test to identify grass‐allergic subjects, and to characterize them in terms of demographic and allergological parameters. Seven hundred and forty‐nine sera were available for IgE detection to a timothy extract, to the recombinant Phl p 1, 2, 5, 6, 7, 11, 12, and to native Phl p 4 and bromelain. Results were stratified by means of demographic and allergy parameters.Results Ninety‐five per cent of the sera had detectable IgE to the timothy extract. Prevalence of IgE reactivity increased from 86.8% to 93.3% as the number of combined reactive molecules rose from 2 to 8. Adjusted prevalences for each allergen were: rPhl p 1 = 83%, rPhl p 2 = 55%, nPhl p 4 = 70%, rPhl p 5 = 50%, rPhl p 6 = 44%, rPhl p 7 = 7%, rPhl p11 = 43%, rPhl p 12 = 15%. Isolated reactivity to rPhl p 1 was 6%, whereas it was negligible for the remaining molecules. IgE reactivity prevalence and mean values differed when patients were stratified on the basis of their associated pollen reactivity and their skin test reactivity grade. No differences were found when age, symptom type and duration were considered. Up to eight‐fold higher IgE concentrations were found when the sum of IgE to molecules was compared with IgE to the extract. Testing for the IgE reactivity to the glycan of the native Phl p 4 allergen showed a possible interference with prevalence and value estimation. Higher prevalence values were found in previously immunotherapy‐treated patients.Conclusions The use of a complete panel of grass allergenic molecules can mimic the current use of allergenic extracts, but new relevant information, such as individual pattern of reactivity, adjusted prevalence, correct specific IgE concentration, can be achieved only by means of discrete allergenic molecules.
HIV-induced IL-6/IL-10 dysregulation of CD4 cells is associated with defective B cell help and autoantibody formation against CD4 cells
Authors: Weimer R , Zipperle S , Daniel V , Zimmermann R , Schimpf K , Opelz G 
Published: March 2003

AbstractTo analyse CD4 cell cytokine secretion and helper/suppressor function at a clonal level we established 446 CD4+ T cell clones (TCC) in four healthy controls, three HIV− haemophilia patients, four CDC II,III and four CDC IV patients. Spontaneous TCC secretion of Th1 cytokines (IL-2, interferon-gamma (IFN-γ)) and Th2 cytokines (IL-4, IL-6, IL-10) was determined by ELISA. TCC helper and suppressor functions were tested in a pokeweed mitogen (PWM)-stimulated allogeneic co-culture system using a reverse haemolytic plaque assay for assessment of B cell responses. There was a significant association of TCC surface marker expression (Leu-8, CD45RA) with TCC IL-6 secretion in healthy controls (P &lt; 0.01), HIV− patients (P 0.001) and CDC II,III patients (P 0.01) but not in CDC IV patients. Likewise, TCC expression of Leu-8 and CD45RA was significantly associated with TCC suppressor function in healthy controls (P 0.0005) but not in HIV-infected patients. A reduced TCC helper frequency (10% of TCC) and an enhanced TCC suppressor frequency (&gt; 80% of TCC) were detected only in those HIV-infected patients who showed an excessively increased TCC IL-6 secretion (&gt; 70% of TCC) together with a significantly diminished TCC IL-10 secretion (10% of TCC). CD4 cell autoantibodies also were found only in patients with this type of cytokine dysregulation. These data indicate that CD4 cell surface markers lose their functional relevance in HIV-infected patients. HIV-induced IL-6/IL-10 dysregulation of CD4+ T cells, i.e. the up-regulation of spontaneous IL-6 and down-regulation of spontaneous IL-10 secretion, appears to be involved in inducing CD4 helper defects and may promote autoantibody formation against CD4 cells.
Complement-regulatory protein expression and activation of complement cascade on erythrocytes from patients with rheumatoid arthritis (RA)
Authors: Arora M , Kumar A , Das S N , Srivastava L M 
Published: March 2003

AbstractIt has been previously reported that the expression of the complement receptor, CR1, on erythrocytes is reduced in patients with RA and that the reduced expression of CR1 is related to disease activity. In this study we investigate the role of other regulatory proteins, i.e. decay-accelerating factor (DAF) and CD59 (membrane inhibitor of reactive lysis), in the pathogenesis of RA by checking the expression of DAF and CD59 on erythrocytes of RA patients to establish whether reduced expression of DAF and CD59 on erythrocytes could be related to increased ability of erythrocytes to activate complement in RA. Flow cytometry was used to measure the expression of DAF and CD59 on erythrocytes from RA patients as well as the deposition of C3 fragments occurring in vivo or after in vitro complement activation. Significantly reduced expression of DAF and CD59 was observed on erythrocytes of RA patients. A significant inverse relationship was observed between DAF expression and in vitro complement activation, whereas no significant relationship between CD59 and complement activation was observed. Finally, we demonstrated an inverse relationship between CH50 activity and DAF expression. Thus, determination of DAF on erythrocytes can emerge as an additional tool in the assessment of extent of complement activation in RA.
Meningococcal disease and polymorphism of FcγRIIa (CD32) in late complement component-deficient individuals
Authors: Platonov A E , Kuijper E J , Vershinina I V , Shipulin G A , Westerdaal N , Fijen C A P , Van De Winkel J G J 
Published: March 2003

AbstractLate complement component-deficient (LCCD) individuals lack plasma bactericidal activity and are highly susceptible to meningococcal disease. Phagocytosis plays a significant role in immune defence against meningococci and involves FcγRIIa (CD32) on leucocytes. Two allotypic forms are currently recognized: FcγRIIa-R131 and RIIa-H131. Neutrophils with the IIa-H/H131 allotype are more effective in phagocytosis than IIa-R/R131. We studied the distributions of IIa-R131 and IIa-H131 allotypes among 29 Russian LCCD patients who had suffered from recurrent episodes of meningococcal disease. The distribution of IIa-R/R131 to heterozygous IIa-R/H131 to homozygous IIa-H/H131 genotypes was 0.14:0.29:0.57 for LCCD patients who developed the first episode of disease before 10 years of age. The distribution was 0.21:0.64:0.14 for patients who experienced meningococcal disease above the age of 10 years (χ2 = 6, P &lt; 0.05, odds ratio for IIa H/H131 versus R/R131 = 8). Meningococcal disease had a ‘grave’ course in 14 of 31 disease episodes in patients with IIa-R/R131 and IIa-R/H131 allotypes, in contrast to 1 of 18 episodes in patients with IIa-H/H131 allotype (χ2 = 7, P &lt; 0.01, odds ratio = 14). We conclude that IIa-H/H131 individuals appear to have a higher acquired antibody-mediated phagocytosis-dependent resistance to meningococcal disease above the age of 10 years. Additionally, effective CD32-mediated phagocytosis may restrict the severity of meningococcal disease in LCCD patients with IIa-H/H131 phenotype.
Pentoxifylline decreasesin vivoandin vitrotumour necrosis factor-alpha (TNF-α) production in lepromatous leprosy patients with erythema nodosum leprosum (ENL)
Authors: Sampaio E P , Moraes M O , Nery J A C , Santos A R , Matos H C , Sarno E N 
Published: March 2003

SUMMARYIncreasing evidence has implicated TNF-α as a pivotal molecule involved in the systemic inflammatory manifestations of ENL, an acute inflammatory complication that may occur in the chronic course of leprosy. In the present study, the mechanism of action of pentoxifylline (PTX) as an alternative therapy for management of leprosy reactions has been evaluated. The effect of PTX on TNF-α production was examined in leprosy patients at the protein level and at the transcriptional level as well. Treatment of ENL patients with PTX (1200 mg daily) ameliorated the systemic symptoms and favoured the evolution of reactional leprosy lesions. Serum TNF-α was assayed before and during treatment with PTX in 15 patients. The increased TNF-α levels seen in the circulation during the reaction were dramatically reduced within 3–7 days of therapy. No significant effect on serum IL-6 was noted. In vitro TNF-α production was assayed upon culture stimulation with Mycobacterium leprae. A reduction of inducible TNF-α in peripheral blood mononuclear cells (PBMC) was seen after 1–2 weeks of in vivo administration of PTX. Furthermore, no effect of the drug on IL-10 secretion was detected in these cultures. A kinetic analysis of the expression of TNF-α and IL-6 mRNA at the site of leprosy lesion was performed in six reactional patients by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The amount of TNF-α mRNA was increased in the tissue during ENL compared with before the reaction, and decreased thereafter following treatment for reaction (either PTX or thalidomide). These data suggest that PTX inhibits TNF-α production in ENL patients both in vivo and in vitro, and it may be useful in the treatment of leprosy patients undergoing ENL.
Essential pathogenic role for endogenous interferon-gamma (IFN-γ) during disease onset phase of murine experimental autoimmune orchitis. I.In vivostudies
Authors: Itoh M , Yano A , Xie Q , Iwahashi K , Takeuchi Y , Meroni P L , Nicoletti F 
Published: March 2003

SUMMARYWe previously found that immunization of CH3/He male mice with syngeneic testicular germ cells (TGC) without the aid of any adjuvants was sufficient to induce DTH to TGC and experimental autoimmune orchitis (EAO). To evaluate the role of endogenous IFN-γ in this model, C3H/He mice immunized subcutaneously with TGC on days 0 and 14 received a single injection of anti-murine IFN-γ MoAb on day 15, 20 or 25. On day 45, DTH to TGC was tested, testis specimens were collected for histological examination, and blood samples collected for IFN-γ measurement. The results showed that whilst MoAb treatment on day 15 or 25 did not influence DTH responses, EAO development, and appearance of IFN-γ in the circulation, treatment on day 20 significantly suppressed all of them. Thus, a single injection with anti-IFN-γ MoAb may successfully down-regulate testicular autoimmunity, provided that the treatment is given at an optimal time point during disease development.
Serum levels of soluble CD30 are increased in ulcerative colitis (UC) but not in Crohn's disease (CD)
Authors: Giacomelli R , Passacantando A , Parzanese I , Vernia P , Klidara N , Cucinelli F , Lattanzio R , Santori E , Cipriani P , Caprilli R , Tonietti G 
Published: March 2003

SUMMARYImbalance in Th1 and Th2 subsets and their derived cytokines seems to be involved in the immune abnormalities underlying UC and CD. CD30 is a member of the tumour necrosis factor/nerve growth receptor superfamily expressed on T cells producing Th2 cytokines and released as a soluble form. In this study high levels of soluble CD30 were found in sera of UC patients independently of disease activity. Furthermore, increased titres of soluble CD30 molecule were shown, in the same patients, by mitogen-stimulated cultures of peripheral blood mononuclear cells. Our data seem to indicate that an activation of Th2 immune response is involved in the pathogenesis of UC, but not of CD. Furthermore, this finding indicates that serum soluble CD30 measurement may be helpful for differentiating these two forms of inflammatory bowel disease.
Expression of low-density lipoprotein receptors in peripheral blood and tonsil B lymphocytes
Authors: De Sanctis J B , Blanca I , Rivera H , Bianco N E 
Published: March 2003

SUMMARYB lymphocytes, purified from peripheral leucocytes from young normolipaemic humans, expressed and internalized low-density lipoprotein receptors (LDLR). The expression was assessed by a monoclonal anti-LDLR. The internalization of LDL was assessed by LDL labelled with 125I (125I-LDL) and 1,1′-dioctadecyl-3,3,3′,3′ tetramethyl-indocarboxycyanine perchlorate (LDL-DiI). The expression of LDLR, assessed by anti-LDLR, was: 38 ± 8% (n = 5) for fresh purified cells, 60 ± 10% (n = 12) for non-stimulated cells, 79 ± 5% (n = 10) for IL-2 (100 U/ml)-stimulated cells and 95 ± 5% (n = 8) for pokeweed mitogen (PWM) (1:200 dilution)-stimulated cells. The optimal concentrations of agonist were 100 U/ml of IL-2, and 1:200 dilution of PWM. IL-2 and PWM increased the internalization of LDL-DiI by 1.5-fold. The internalization of LDL-DiI was maximal at 60 μg of protein/ml (48 ± 8%). Scatchard analysis revealed a Kd of 3.2 ± 0.22 × 10−8 m and 2180 ± 190 binding sites in non-stimulated cells, a Kd of 7.73 ± 0.36 × 10−9 m and 12 500 ± 430 binding sites for IL-2 (100 U/ml)-stimulated cells, and a Kd of 7.2 ± 0.43 × 10−9 m and 13 250 ± 450 binding sites for PWM (1:200 dilution)-stimulated cells. Lineweaver–Burk analysis of LDL binding (LDL-DiI) revealed that the apparent Kd for non-stimulated cells was 1.3 ± 0.11 × 10−8 m, and 9.2 ± 0.2 × 10−9 m and 7.5 ± 0.25 × 10−9 m for IL-2- and PWM-stimulated cells, respectively. B lymphocytes from tonsils also showed a high expression of LDLR assessed with anti-LDLR (70 ± 6%). The high expression of LDLR and the avid internalization of LDL suggest that LDL may be important for B cell physiological responses.