bluuebunny/crossref_metadata_2025 huggingface dataset:
A Correlation of the Uterine and Ovarian Blood Flows with Parity of Nonpregnant Women Having a History of Recurrent Spontaneous Abortions
Authors: Jirous J. , Diejomaoh M. , Al-Othman S. , Al-Abdulhadi F. , Al-Marzouk N. , Sugathan T.
Published: April 2003

The aim of this prospective study was to explore the relationship between the incidence of recurrent spontaneous abortions (RSA) and values of Doppler indices characterizing the uterine and ovarian flows in nonpregnant women with a history of primary and secondary RSA using color, power and pulsed Doppler transvaginal ultrasonography. A total of 19 healthy women as a control group A and 81 women with a history of RSA as a study group B were examined on the 21st day of a regular menstrual cycle. Our study demonstrated the presence of significantly decreased uterine and ovarian blood flows in women with primary and secondary RSA if more than 3 abortions had occurred. The obtained values were less favorable in the primary RSA compared with the secondary RSA cases.
Transient Increase in the Levels of T-Helper 1 Cytokines in Postmenopausal Women and the Effects of Hormone Replacement Therapy
Authors: Kamada Masaharu , Irahara Minoru , Maegawa Masahiko , Ohmoto Yasukazu , Murata Kaoru , Yasui Toshiyuki , Yamano Shuji , Aono Toshihiro
Published: April 2003

The aim of this study was to determine, at least in part, T-cell function in postmenopausal women and the effects of hormone replacement therapy (HRT). Levels of T-helper 1 (Th1) cytokines (IL-2 and IFN-γ) and T-helper 2 (Th2) cytokines (IL-4 and IL-10) produced by phytohemagglutinin-stimulated whole blood cells from 72 untreated and 44 HRT-treated women were measured by ELISA. Thirteen of the 44 HRT-treated women were examined before and during HRT. The production of IL-2 increased gradually with advance of the postmenopausal period. The levels of IL-2 in women in the early (≤10 years) and mid (>10 and <30 years) postmenopausal stages were significantly higher than those in women in their second, third and fourth decades. The level in women in the late (≧30 years) postmenopausal stage, however, was significantly lower than those in women in the early and mid postmenopausal stages. The level of IFN-γ was highest in women in the mid postmenopausal stage. On the other hand, the levels of Th2 cytokines did not change with age or after menopause until the mid postmenopausal period but were significantly lower in women in the late postmenopausal stage. IFN-γ levels in women on HRT were significantly lower than those in untreated postmenopausal women at all postmenopausal stages. HRT induced a significant decrease in the production of IL-2 and IL-4. In conclusion, production of Th1 cytokines is augmented in women after menopause. HRT prevents this increase, thereby improving the aberration of Th1/Th2 balance that is implicated in an inadequate immune response and pathological conditions.
NK Cell Activity and Subsets in Women with a History of Spontaneous Abortion
Authors: Morikawa Mamoru , Yamada Hideto , Kato Emi H. , Shimada Shigeki , Ebina Yasuhiko , Yamada Takahiro , Sagawa Tadashi , Kobashi Gen , Fujimoto Seiichiro
Published: April 2003

Objective: The aim of this study was to assess the role of NK cells in nonpregnant women with a history of spontaneous abortion. Study Design: 113 nonpregnant women with a history of spontaneous abortion were assessed for peripheral NK cell activity and percentage of NK cell subsets, in relation to the cause of abortions, the number of spontaneous abortions, and subsequent pregnancy outcome (n = 56). Results: Neither NK cell activity nor subsets showed a significant difference in relation to the cause or number of spontaneous abortions. NK cell activity in nonpregnant women who later experienced subsequent abortion with normal chromosomes (n = 10) (mean ± SD: 42.8 ± 15.8%) was relatively higher than that in women with subsequent live birth (control, n = 39) (32.1 ± 13.7%) (p = 0.099). NK cell activity in women who later experienced subsequent abortion with abnormal chromosomes (n = 7) (28.7 ± 21.4%) was the same as the level in the control. Conclusion: Peripheral NK cell activity or subsets during nonpregnant status were not related to the cause or number of previous spontaneous abortions. A relation between preconceptional NK cell activity and later experiencing abortion with normal chromosomes should be further studied.
Risk Factors for Breast Cancer among Pre- or Post-Menopausal Women in Belo Horizonte, Brazil
Authors: Gomes Ana Lúcia R.R. , Guimarães Mark D.C. , Gomes Carlos C. , Chaves Indelécio G. , Gobbi Helenice , Camargos Aroldo F.
Published: April 2003

Background: Much controversy has been generated about pre- and post-menopausal breast cancer patients and investigators have sought to identify whether risk factors differ between these two groups. In Brazil, breast cancer is an important cause of death among women and there are few analytical studies concerning pre- or post-menopausal comparisons. Methods: A case-control study was carried out at the Federal University Hospital, Belo Horizonte, Brazil, to determine if selected socio-economic and reproductive risk factors for breast cancer differed between pre-menopausal and post-menopausal women. Cases were 300 women with breast carcinoma and controls were 600 women with other benign diseases matched for age and date of diagnosis, admitted to the same hospital during the same period (1978–1987). Univariate and multivariate conditional logistic regression analyses were performed. Results: Multivariate analysis showed no differences in breast cancer risk in pre- and post-menopausal women (risk factors were similar in direction and magnitude). Occupation, irregular menstrual cycles, parity, history of breast cancer in at least one first-degree female relative, and oral contraceptive use had similar associations in both groups. Conclusions: The present study indicates that breast cancer diagnosed before and after menopause has a similar risk profile.
Induction of Labor in Grand Multiparous Women and Previous Cesarean Section: Is It Safe?
Authors: Abu-Heija Adel T. , Ali Abdulhai Mohammada
Published: April 2003

Objective: To compare the outcome of induced and spontaneous labor in grand multiparous women with one previous lower segment cesarean section, so that the safety of labor induction could be assessed. Method: In 56 women labor was induced and their outcomes were compared with those of 177 women with spontaneous labor. All women were multiparous and had had one previous cesarean section. Results: There were no significant differences in the incidences of 1- and 5-min Apgar scores, congenital malformation, cesarean section rates and uterine scar dehiscence or uterine rupture. There were 4 cases of intrauterine fetal death in the induction group, but no case of intrapartum or early neonatal death in the 2 groups. In the study group, 80.4% of the women delivered vaginally compared with 84.3% in the control group. Conclusion: In this moderate-sized study, we may conclude that when there is no absolute indication for repeating cesarean section, induction of labor may be a safe option in these high-risk women.
Testing Quantitative Traits for Association and Linkage in the Presence or Absence of Parental Data
Authors: Zhu Xiaofeng , Elston Robert C. , Cooper Richard S.
Published: April 2003

Zhu and Elston developed a transmission disequilibrium test for quantitative traits by defining a linear transformation to condition out founder information. The method tests the null hypothesis of no linkage or association and can be applied to general pedigree structures. However, this method requires both genotype and phenotype parental information, which may be difficult to obtain. In this paper, we describe parametric and non-parametric methods to relax this requirement when only nuclear families are sampled. We show that neither method is affected by population stratification in the absence of linkage. The statistical power and validity of the tests are investigated by simulation. A simple simulation method to calculate the power of the nonparametric method is also discussed. In practice, the data may have some families with parental phenotype and genotype information available and some without. We briefly discuss how all the data may be analyzed jointly.
Population Genetic Analysis of the Genes APOE, APOB (3′VNTR) and ACE in Some Black and Amerindian Communities from Colombia
Authors: Jaramillo-Correa J.P. , Keyeux G. , Ruiz-Garcia M. , Rodas C. , Bernal J.
Published: April 2003

A population genetic study was carried out with the APOE, APOB and ACE loci in 17 Colombian human populations. Ten of them were Amerindian communities coming from the northeastern part of Colombia, Pacific region, Eastern Plains and Amazonia. Six were black populations from Providence Island, Caribbean and Pacific coasts. Finally, the Mestizo population of Bogota was studied as well. The APOE and ACE loci were in Hardy-Weinberg equilibrium, whereas the APOB locus was not studied in all populations. The genetic heterogeneity was substantially greater among the Amerindian populations (GST = 0.059) than in the Afrocolombian populations (GST = 0.009). Also the gene flow population pair estimates were so much higher among the Afrocolombian populations (Nm = 49.08 ± 43.07) than among Amerindian populations (Nm = 9.66 ± 18.04). Different phylogenetic and multivariant analyses showed that the Amerindian populations analyzed were clustered in three different arrays: one constituted by the Colombian northeastern and Pacific populations, the second one by the two Amazon populations (Coreguaje and Nukak) and the last one by the Yuco (the unique Caribbe-speaking population among those studied). The latter population was highly divergent from a genetic point of view from the remainder Amerindian populations studied. By using the Mantel test, the existence of a positive and significant correlation between the genetic and geographical distances found among Amerindian populations was demonstrated. This fact was not observed among the Afrocolombian populations. Nevertheless, an isolation-by-distance Slatkin analysis test did not show a significant clear structure of this special pattern among the Indian tribes studied.
Estimation of the Heritability of Latent Variables Which Are Included in a Structural Model for Metabolic Syndrome
Authors: Koch Rainer , Julius Ulrich , Jaross Werner , Schröder Hans-Egbert
Published: April 2003

In a study looking for risk factors of atherosclerosis in families with combined hyperlipidemia and hypertension, clinical and biochemical data of 1,149 persons were analyzed to develop two hypothetical multivariate scores concerning the degree to which a patient is affected by the metabolic syndrome. The scores are based on a structural model for low-density cholesterol (LDL) and high-density cholesterol (HDL), triglycerides, uric acid, creatinine, glucose, insulin, systolic blood pressure and waist-to-hip ratio. Age, gender and body mass index were used for adjusting all variables. In segregation analyses of 42 pedigrees without using genotype information, estimations of the heritabilities and environmentally caused variance and covariance components were computed for the individual score values of the two latent factors. The first score shows a heritability of 42%; the environment component disappeared. The score mainly reflects the HDL, LDL and triglyceride levels. The second score shows a heritability of 16% with an environment component of 7%. It includes mainly insulin, uric acid and creatinine. In the search for genetic causes, both scores could be a basis for further phenotypic classification of the metabolic syndrome.
Altered Expression of CD11b and CD62L after Cross-Linking of CD45 Isoforms on Human Eosinophils
Authors: Matsumoto Kenji , Bochner Bruce , Wakiguchi Hiroshi , Kurashige Takanobu
Published: April 2003

Background: Phosphotyrosine phosphatase CD45 is exclusively found on nucleated hematopoietic cells. CD45 is an essential component of signaling of mast cel degranulation and of lymphocyte activation, but little is known about its role in eosinophils. In the present study, we have determined the expression and function of CD45 isoforms on human eosinophils. Methods: Expression of CD45 isoforms on purified peripheral blood eosinophils was examined using indirect immunofluorescence and flow cytometry. Eosinophils were cultured with anti-CD45 isoform monoclonal antibodies (mAbs) for up to 24 h. Eosinophil activation was evaluated by measuring surface expression of CD11b or CD62L by flow cytometry. Results: Fresh eosinophils express CD45, CD45RB, CD45RO epitopes but not CD45RA. Incubation with anti-CD45 isoform mAb for 30 min did not alter the expression of either CD11b or CD62L on eosinophils. In contrast, the expression of CD11b was significantly enhanced after 24 h of incubation with mAbs against CD45, CD45RB, or CD45RO. In addition, the expression of CD62L was also significantly reduced with anti-CD45RB and with anti-CD45RO mAbs. Conclusions: Cross-linking of surface CD45 isoforms for 24 h significantly induced upregulation of CD11b expression and CD62L shedding, consistent with activation of eosinophils. Our data suggest that CD45 isoforms are functionally expressed on human eosinophils, and are capable of modulating eosinophil function and might participate in allergic inflammation.
Suppressive Effects of SP-A on Ionomycin-Induced IL-8 Production and Release by Eosinophils
Authors: Cheng Gang , Ueda Takashi , Nakajima Hirokazu , Nakajima Akiko , Kinjyo Syunichi , Motojima Shinji , Fukuda Takeshi
Published: April 2003

Recent studies have demonstrated that pulmonary surfactant protein (SP)-A plays a potential role in modifying inflammation and immune function. To see whether SP-A could modify IL-8 production and release by eosinophils stimulated with ionomycin, SP-A purified from surfactant recovered from patients with alveolar proteinosis was added to eosinophils isolated by the negative-selection method with immunomagnetic beads, and cultured for 24 h. The concentrations of IL-8 in the cell-free supernatants and cell lysates were then measured by ELISA. SP-A attenuated the production of IL-8 by eosinophils in a concentration-dependent manner. SP-A also attenuated the release of IL-8 from the eosinophils. The addition of SP-A antibody (PE10) reversed these effects of SP-A completely. These data suggest that SP-A may have the potential to modify allergic inflammation by inhibiting the release and production of IL-8 by eosinophils.
Regulatory Mechanisms of Eosinophil Adhesion to and Transmigration across Endothelial Cells by α4 and β2 Integrins
Authors: Yamamoto Hideaki , Nagata Makoto
Published: April 2003

To participate in allergic airway inflammation, it is necessary for blood eosinophils (Eos) to adhere to and migrate across pulmonary vascular endothelial cells (EC). Accumulating evidence has established that Eos adhesion molecules, such as α4 and β2 integrins, play central roles in these processes. We have reported that Eos spontaneously adhere to recombinant human (rh)-VCAM-1, while adhesion to rh-ICAM-1 requires a second stimulus such as GM-CSF. Furthermore, our study employing human pulmonary microvascular endothelial cells (HPMEC) revealed that although the α4 integrin/VCAM-1 pathway is crucial for the firm adhesion of Eos, subsequent transendothelial migration occurred dependent on the β2 integrin/ICAM-1 pathway. We also discuss secretagogues that would affect Eos recruitment to the airways via regulation of α4 and β2 integrins.
Matrix Metalloproteinase-9 in Peripheral Blood Eosinophils
Authors: Fujisawa Takao , Kato Yoshiko , Terada Akihiko , Iguchi Kosei , Kamiya Hitoshi
Published: April 2003

Background: Matrix metalloproteinases (MMPs) are major contributors to tumor invasion, remodeling of connective tissue and infiltration of inflammatory cells and may be important mediators in developing allergic inflammation. Overexpression of MMP-9 mRNA by eosinophils in the asthmatic airways has been reported. To clarify the relative significance of MMP as an inflammatory mediator from eosinophils, we determined the content of MMP-9 in the peripheral blood eosinophils and compared it with the other leukocyte fractions. Methods: Peripheral blood eosinophils, neutrophils, and mononuclear cells were purified from normal and allergic donors with Percoll gradient centrifugation and CD16 negative selection. Cell lysate and culture supernatants stimulated with IL-5, PAF, and PMA were tested for MMP-9 with gelatin zymography and ELISA. Results: The amount of MMP-9 in highly purified eosinophils, neutrophils, and mononuclear cells was 2.5 ± 0.9, 4,073 ± 581, and 7.6 ± 1.4 ng/5 × 106 cells, respectively. There was no difference in MMP-9 content of eosinophils between normal donors and patients with asthma. Culture of peripheral blood eosinophils with IL-5 for 4 days did not induce MMP-9 production. The stimulation of eosinophils with PMA and other secretogogues caused only small amounts of MMP-9 secretion as compared with neutrophils. Conclusions: These findings suggest that circulating eosinophils normally have only small amounts of MMP-9 and that eosinophils may need complex activation signals to produce significant amounts of MMP as seen in tissues of allergic inflammation.
Localization of Sialyl Glycoconjugates in Eosinophil-Specific Granules after Degranulation Stimuli
Authors: Ohmori Jun , Nawa Yukifumi
Published: April 2003

Sialyl glycoconjugates, a group of the acidic glycoconjugates, are expressed in murine eosinophil-specific granules through their maturational stages. To clarify whether the sialyl glycoconjugates work as functional molecules in the process of eosinophil degranulation, we examined the localization of sialyl residues after degranulation stimulated with calcium ionophore A23187 or interleukin-5. Although sialyl residues were localized in the specific granules before degranulation, they were dissociated from the granules after the stimuli. These results suggest that sialyl glycoconjugates participate extensively in the process of eosinophil degranulation
Sequence Polymorphisms and Antibody Binding to the Group 2 Dust Mite Allergens
Authors: Smith Alisa M. , Benjamin David C. , Derewenda Urszula , Smith Wendy A. , Thomas Wayne R. , Chapman Martin D.
Published: April 2003

Background: The group 2 allergens Der p 2, Der f 2 and Eur m 2 are 14-kD proteins with >80% sequence identity. Isoforms within each genus have been identified which differ by 3–4 amino acids. The aim of this study was to investigate the importance of these substitutions to antibody binding. Methods: Recombinant allergens were expressed and purified from Escherichia coli. ELISA and skin testing were used to evaluate antibody binding. Molecular modeling of the tertiary structure was preformed to examine the location of substitutions. Results: The three Der f 2 isoforms and two of three of the Der p 2 isoforms reacted with all monoclonal antibodies (mAb). Der p 2.0101, the isoform with aspartate at position 114, bound all mAb except 1D8. Substitution of asparagine for aspartate restored binding of rDer p 2.0101 to mAb 1D8 and increased the correlation coefficient for IgE binding from 0.72 to 0.77. The three Der p 2 isoforms showed comparable skin test reactivity to nDer p 2 and commercial extract. rEur m 2.0101 bound to all mAb except 7A1 and when compared with rDer p 2 for IgE binding, r2 = of 0.58 (n = 72). Lep d 2 did not react with mAb or with Dermatophagoides spp. allergic sera. Modeling revealed that Eur m 2, Lep d 2 and Tyr p 2 retain the tertiary fold of Der p 2 and the substitutions are on the surface. Conclusions: mAb could distinguish isoform substitutions. IgE binding showed a good correlation among all isoforms, thus the recombinant allergens are useful for diagnosis.
Lipid Transfer Protein: A Pan-Allergen in Plant-Derived Foods That Is Highly Resistant to Pepsin Digestion
Authors: Asero Riccardo , Mistrello Gianni , Roncarolo Daniela , de Vries Sacco C. , Gautier Marie-Françoise , Ciurana Caroline L.F. , Verbeek Eddy , Mohammadi Tamimount , Knul-Brettlova Vlasta , Akkerdaas Jaap H. , Bulder Ingrid , Aalberse Rob C. , van Ree Ronald
Published: April 2003

Background: Lipid transfer proteins (LTPs) are stable and highly conserved proteins of around 10 kD. They have recently been identified as allergens in fruits of the Rosaceae family. Objective: The aim of this study was to investigate whether the highly conserved structure of LTPs justifies a designation as a true pan-allergen, and to study the role of protein stability in allergenicity. Methods: Thirty-eight patients with a positive skin prick test to Rosaceae fruit extracts were characterized by interviews and skin prick tests. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified natural and recombinant LTPs. To address the role of protein stability in the allergenicity of LTP, fruit extracts and LTPs were digested with pepsin. Results: IgE antibodies to Rosaceae LTPs cross-reacted with a broad range of non-Rosaceae vegetable foods. Inhibition studies with purified natural and recombinant LTPs confirmed the role of LTP in this cross-reactivity. Many of the patients with this type of cross-reactive IgE antibodies had a clinical food allergy. In contrast to the typical birch Rosaceae cross-reactive patients, the oral allergy syndrome was frequently accompanied by more severe and systemic reactions. IgE reactivity to LTP was shown to be resistant to pepsin treatment of the allergen. Conclusion: LTP is a true pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.
The IgE-Binding Regions of the Major Allergen Pen a 1: Multiple Epitopes or Intramolecular Cross-Reactivity?
Authors: Reese G. , Ayuso R. , Leong-Kee S.M. , Plante M. , Lehrer S.B.
Published: April 2003

The muscle protein tropomyosin is the major allergen (Pen a 1) in brown shrimp, Penaeus aztecus, lobster, crab, mollusks and other invertebrates such as house dust mite, and cockroach. Pen a 1 contains five major IgE-binding regions, and sequences that are rarely or never recognized by IgE antibodies from shrimp-allergic subjects. The molecular structure of tropomyosins is simple and repetitive; it is made up of multiple heptad repeats. The analysis of sequence identities and similarities of major, minor and non-IgE-binding regions indicate that intramolecular cross-reactivity to the multiple IgE-binding regions of Pen a 1 is not due to repetitive, identical or similar, cross-reacting sequences but due to multiple, independent epitopes. However, an allergenic motif may explain the reactivities to the five identified major IgE-binding regions.
Secretion of Proinflammatory Eicosanoid-Like Substances Precedes Allergen Release from Pollen Grains in the Initiation of Allergic Sensitization
Authors: Behrendt H. , Kasche A. , Ebner von Eschenbach C. , Risse U. , Huss-Marp J. , Ring J.
Published: April 2003

It is commonly believed that allergic sensitization starts when an allergen contacts the surface of an antigen-presenting cell in mucosal or skin epithelia. Most studies dealing with this aspect use allergen extracts as stimulus. Under natural exposure conditions, however, the bioavailability of allergen depends on allergen liberation from internal binding sites within the allergen carrier, e.g. pollen grains. In comparing total protein and major allergen release from timothy grass (Phleum pratense L.) pollen freshly collected on rural meadows or near high-traffic roads, there was a striking difference between the pollen, with higher allergen release rates from rural meadow pollen grains. Thus, allergen release does not explain the higher prevalence rates of atopic sensitization and disease observed in many epidemiological studies in children exposed to automobile exhaust. Therefore, other possible effectors from pollen grains were investigated. Pollen grains incubated in protein- free buffer were found to secrete significant amounts of eicosanoid-like substances, namely leukotriene (LT) B4-like and prostaglandin E2-like substances, in a pH-, time- and temperature-dependent fashion. The highest values of eicosanoid secretion were found in birch, grass and mugwort pollen, while pine (Pinus sylvestris L.) pollen showed only marginal eicosanoid-like secretion. Additionally, the release of these substances was significantly higher from pollen which had been collected near roads with heavy traffic, indicating a stronger proinflammatory activity of these pollen grains. In order to investigate the effects of air pollutants, native pollen grains were exposed in a dose- and time-dependent fashion in a fluidized bed reactor to traffic-related pollutants, e.g. volatile organic compounds (toluene, m-xylene), leading again to a significant increase in the secretion of LTB4-like immunoreactivity, in contrast to exposure with sulfur dioxide. This finding opens a new dimension of understanding of the early events in allergic sensitization, indicating that proinflammatory effects of the allergen carrier, e.g. the pollen grain itself, can lead to activation of the mucosal membrane. These findings might help to also explain the higher prevalence rates of pollen allergy in areas with high automobile exhaust emissions. Furthermore, the allergenic ‘potency’ of various allergens has to be redefined at the allergen carrier level with regard to different stages of allergen and mediator release prior to the contact with the host’s immune system.
Mechanisms of T Cell Peptide Epitope-Dependent Late Asthmatic Reactions
Authors: Larché Mark , Haselden Brigitte M. , Oldfield William L.G. , Shirley Karen , North Janet , Meng Qiu , Robinson Douglas S. , Ying Sun , Kay A. Barry
Published: April 2003

Short peptide sequences corresponding to T cell epitopes have been identified in the major cat allergen Fel d 1. In order to directly activate allergen-specific T cells in cat-allergic asthmatic individuals, peptides were administered by intradermal injection. Subsequently, a proportion of subjects experienced a delayed reduction of airway calibre manifested as a decrease in FEV1. Changes in lung function occurred approximately 3 h after peptide injection, peaked at 6 h and resembled an isolated late asthmatic reaction (LAR). Using molecular tissue typing techniques, it was determined that many of the individuals experiencing isolated LAR expressed particular HLA-DR molecules. These molecules were shown in subsequent experiments to bind individual peptides within the preparation and thus to activate T cells in a major histocompatibility complex (MHC)-restricted fashion. The precise mechanisms whereby MHC-restricted activation of allergen-specific T cells gives rise to bronchoconstriction are currently under investigation.
Nerve Growth Factor Is Produced by Human Fibroblasts and Modulates Them: Role of Nerve Growth Factor in Tissue Repair
Authors: Levi-Schaffer F. , Micera A. , Maquart F.X. , Aloe L.
Published: April 2003

Some recent findings suggest the contribution of nerve growth factor (NGF) in tissue repair and fibrosis because of changed levels of this neurotrophin in these conditions that often follow an inflammatory insult. Data on the potential fibrogenicity of mast cells and eosinophils in fibrosis with different etiopathologies and in the one resulting from some allergic conditions are increasing. Both these cells produce NGF and their functions are modulated by this factor. However, the direct effects of NGF on fibroblasts, which represent the main effector and target cells of fibrosis, have been poorly investigated. This minireview reports on recent data concerning the involvement of NGF in allergic inflammation and in tissue repair/remodeling.
Activation of the Plasma Kinin Forming Cascade along Cell Surfaces
Authors: Kaplan Allen P. , Joseph Kusumam , Shibayama Yoji , Reddigari Sesha , Ghebrehiwet Berhane
Published: April 2003

Proteins of the plasma kinin-forming cascade bind to endothelial cells and activation of the cascade can be initiated along the surface. The light chain of high molecular weight kininogen (HK) (domain 5) and factor XII bind to gC1qR, the heavy chain of HK (domain 3) binds to cytokeratin 1 and the interactions are zinc dependent. Prekallikrein binds to domain 6 of HK. Antisera to gC1qR and cytokeratin 1 inhibit binding and activation. Incubation of normal plasma with endothelial cells leads to gradual conversion of prekallikrein to kallikrein, while plasma deficient in factor XII or HK are inactive within a 2-hour time frame. Thus factor XII is critical for activation to proceed. Augmentation of these reactions may occur when C1 inhibitor is functionally deficient or with ACE inhibitors which also inhibit kininases.
Allergic Manifestations as the Results of a Conditional Autoimmune Response
Authors: Stadler Beda M. , Miescher Sylvia , Horn Michael , Pachlopnik Jana , Stadler Michael , Kricek Franz , Vogel Monique
Published: April 2003

By means of repertoire cloning we have isolated human anti-IgE antibodies as well as human anti-FcΕRI antibodies. Whether the naturally occurring anti-IgE autoantibodies play a pathophysiological role may be disputed, but the beneficial role of recombinant anti-IgE antibodies as a therapeutic agent has been shown. On the other hand, the natural antibodies isolated from an antibody library of a nonallergic individual against the FcΕRI α-chain are anaphylactogenic, if FcΕRI was not occupied. Thus, anti-FcΕRI autoantibodies may be part of a conditional autoimmune reaction, leading to urticaria if local IgE is consumed, e.g. after an immediate reaction. Thus, anti-FcΕRI antibodies may represent an amplification arm of the late reaction. The normal occurrence of anti-IgE and anti-IgE receptor autoantibodies may suggest that it might also be feasible to induce such autoantibodies by vaccination. In a monkey model using a mimotope of human IgE it was possible to induce a beneficial anti- IgE autoimmune response. The actual epitope of the IgE molecule was then also molecularly reconstructed by generating recombinant anti-idiotypic antibodies. These antibodies also induced effectively an anti-IgE response in monkeys, suggesting that not only mimotopes but also anti-idiotypic antibodies may be used to generate an autoimmune response. Both of our projects suggest that active immunization may be a new form of immunomodulation for the treatment of allergic disease.
Chemokines, Chemokine Receptors and Allergy
Authors: Kaplan Allen P.
Published: April 2003

Chemokines are a group of cytokines that are responsible for the influx of blood cells, including T and B lymphocytes, monocytes, neutrophils, eosinophils and basophils, in allergic and other inflammatory conditions. They function as G protein-coupled chemotactic factors which also activate the cells with which they interact. Certain chemokines function within the afferent arm of the immune system, in which antigen is processed and antibody formation initiated, and others are active within the effector pathways of cellular immunity and late-phase allergic reactions. Th2 lymphocytes, which are critical for allergy, employ the CC chemokine receptors CCR4 and CCR8 with the ligands thymus- and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC) and I-309, respectively. The chemokine receptor CCR3 and ligands monocyte chemoattractant protein (MCP)-3, MCP-4, regulated upon activation normal T cell expressed and secreted (RANTES) and eotaxins I and II are of particular relevance for the recruitment and activation of eosinophils. Th1 reactions depend upon interferon γ-induced CXC chemokines interferon- inducible protein (IP)-10, interferon-inducible T cell-α chemoattractant (iTAC) and monokine induced by interferon-γ (MiG), which bind to chemokine receptor CXCR3.
Regulation of Surface FcεRI Expression on Human Eosinophils by IL-4 and IgE
Authors: Iikura Motoyasu , Yamaguchi Masao , Hirai Koichi , Miyamasu Misato , Yamada Hirokazu , Nakajima Toshiharu , Fujisawa Takao , Ra Chisei , Morita Yutaka , Yamamoto Kazuhiko
Published: April 2003

Background: Recent studies have demonstrated that eosinophils from allergic patients express low levels of FcΕRI on their surface, but the regulatory mechanisms of eosinophil surface FcΕRI expression are not fully understood. We investigated whether IL-4 and IgE, which are reported to regulate surface FcΕRI expression on human mast cells, are able to affect surface FcΕRI expression in normal human eosinophils. Methods: Eosinophils purified from peripheral blood were cultured with IL-5 and with or without IL-4 and/or IgE, and surface FcΕRI expression was analyzed by flow cytometry using an anti-FcΕRI mAb, CRA-1. Results: Apparent FcΕRI expression (∼1% of mast cell FcΕRI levels) was observed in eosinophils cultured with both IL-4 and IgE. A combination of IL-4 (≧1 ng/ml) and IgE (≧ 0.5 µg/ml) was necessary for the maximal induction of surface FcΕRI expression. In the presence of IL-4 and IgE, eosinophils cultured for 2 days demonstrated low but statistically significant levels of surface FcΕRI, which reached a plateau after 7 days of culture. However, cross-linkage of surface FcΕRI molecules by CRA-1 or anti-IgE did not induce any eosinophil activation. Conclusions: IL-4 and IgE can affect the levels of surface FcΕRI on normal human eosinophils. FcΕRI expression on eosinophils may be regulated by a mechanism similar to that in mast cells.
Does IL-4 Play a Role in the Expansion of Vβ8a T Cell Receptor-Bearing Cells?
Authors: Liebers V. , Gellert B. , Raulf-Heimsoth M. , Baur X.
Published: April 2003

Background: Peripheral blood mononuclear cells (PBMC) of subjects allergic to the insect-derived allergen Chi t 1–9 are characterized by an allergen-induced pronounced proliferation and increased expression of activation markers (CD25, HLA-DR, CD23). T cell lines showed an elevated percentage of Vβ8a-positive cells following stimulation by Chi t 1–9. Objective: The aim of the present study was to investigate whether Vβ8a dominance plays an important role in PBMC short-term cultures (24 h) as well. The role of exogenous added cytokines, especially IL-4, has been determined. Methods: The T cell receptor repertoire was measured with 16 monoclonal antibodies to epitopes on the variable region of the β chain by flow cytometry. Patients allergic to Chi t 1–9 were compared to nonallergic subjects as well as to subjects with other occupational allergies. In addition, cytokines were determined intracellulary by flow cytometry. Studies were performed with PBMC cultured for 24 h. Results: After cultivation for 24 h without or with different stimuli (cytokines, allergen, phytohaemagglutinin), changes in the T cell receptor profile and the cytokine profile were measurable compared to the baseline value (without cultivation). Stimulation with IL-4 revealed increased percentages of Vβ8a-expressing cells in Chi t 1–9-sensitized patients. This IL-4-induced Vβ8a increase did not occur in PBMC from the two control subject groups (non-allergic and allergic to other allergens than Chi t 1–9). Conclusion: In conclusion, the dominance of certain T cell receptor types seems to arise due to the exposure to specific allergens and cytokine production. Some T cell receptors are often affected, for example Vβ8a, whereas others only show minor variations. Vβ8a expression obviously plays an important role in Chi t 1–9 allergy.
Recombinant Dirofilaria immitis-Derived Antigen Can Suppress Passive Cutaneous Anaphylaxis Reactions
Authors: Furuhashi Yuko , Imai Shinjiro , Tezuka Hiroyuki , Fujita Koichiro
Published: April 2003

Background: High levels of antigen-nonspecific IgE are produced in animals infected with helminth parasites. Generally, the increase in IgE is thought to exacerbate allergic reactions. However, high levels of antigen-nonspecific IgE may alter some features of anaphylactic reactions. To investigate the molecular mechanisms of antigen-nonspecific IgE production induced during filarial infections, we previously constructed rDiAg (recombinant Dirofilaria immitis-derived antigen) in Escherichia coli. In the present study, we examined the effect of rDiAg on the production of antigen-nonspecific IgE and on allergic cutaneous reactions in rats. Methods: Osmotic pumps filled with 200 µg of rDiAg or with 200 µl of PBS (control) were subcutaneously implanted in Wistar rats, and plasma samples were collected weekly thereafter. IgE levels were determined by ELISA. Homologous passive cutaneous anaphylaxis (PCA) reactions with anti-DNP-As IgE were examined 21 days after implantation. 125I-IgE binding assays were examined on peritoneal mast cells from rDiAg-infused rats and control rats. Results: Antigen-nonspecific IgE production was induced in rDiAg-infused rats. PCA reactions were suppressed in rDiAg-infused rats in spite of high levels of IgE and a markedly increased expression of FcΕRI. 125I-IgE did not bind to mast cells derived from rDiAg-infused rats, but it bound dose dependently to mast cells derived from control rats. Conclusion: The present data support the hypothesis that antigen-nonspecific IgE might protect against antigen-specific IgE by means of competition for mast cell receptors. rDiAg is an essential factor to induce antigen-nonspecific IgE in helminth infections.
Expression and Purification of Recombinant Rat Eosinophil-Associated Ribonucleases, Homologues of Human Eosinophil Cationic Protein and Eosinophil-Derived Neurotoxin, and Their Characterization
Authors: Nakajima Masahiro , Hirakata Mikito , Nittoh Takeaki , Ishihara Kenji , Ohuchi Kazuo
Published: April 2003

Background: Human eosinophils contain two eosinophil ribonucleases, eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). In rats, 8 homologues of human ECP and EDN have been identified. To clarify the biological activity of rat eosinophil ribonucleases, we cloned rat eosinophil-associated ribonuclease (EAR)-1/rat ribonuclease 7 and rat EAR-2/rat ribonuclease 4, and produced recombinant rat pre-EAR-1 and pre-EAR-2 in a bacterial expression system. Methods: As we have already cloned the complete nucleotide sequence for rat EAR-1, we determined that for rat EAR-2 cDNA by the rapid amplification of cDNA ends procedure. Recombinant rat pre-EAR-1 and pre-EAR-2 were expressed in Escherichia coli as N-terminal 6 × histidine-tagged proteins, isolated from the insoluble fraction of the cell lysate and purified by a single-step method using an Ni-NTA resin column after solubilization with a 6 M guanidine solution. Results: The deduced amino acid sequence revealed that the molecular weight of EAR-2 containing the signal peptide is 17.3 kD and the isoelectric point is 8.59. The homology in amino acid sequence between rat pre-EAR-2, and human pre-ECP and human pre-EDN is 51 and 53%, respectively. The purified and refolded recombinant rat pre-EAR-1 and pre-EAR-2 showed bactericidal activity against E. coli and Staphylococcus aureus. Conclusions: These findings suggest that rat EAR-1 and EAR-2 act as host defense factors against bacterial infection in rats.
Cupressaceae Pollinosis: Identification, Purification and Cloning of Relevant Allergens
Authors: Di Felice Gabriella , Barletta Bianca , Tinghino Raffaella , Pini Carlo
Published: April 2003

Allergy to Cupressaceae pollen is a worldwide pollinosis caused by several species. Pollen extracts prepared from allergenic species belonging to this family are characterised by low protein and high carbohydrate content. The allergenic components represented in the pollen extracts from different species of the Cupressaceae family show high levels of cross-reactivity when probed with human IgE from allergic subjects and share a number of common epitopes also identified by polyclonal rabbit antisera and monoclonal antibodies. A close relationship has also been described with the Taxodiaceae and Podocarpaceae families. Although both proteic and carbohydrate epitopes appear to be involved in IgE recognition and allergenic cross-reactivity, a large portion of the IgE reactivity of Cupressaceae-allergic patients seems to be associated with sugar moieties present on the relevant allergenic molecules. From this point of view, Cupressaceae/Taxodiaceae allergens constitute a particularly useful model to study IgE cross-reactivity, as they have been shown to display different levels of homology. Moreover, the availability of the purified allergens, together with their recombinant counterparts, may shed light on the actual role played by carbohydrate in allergic sensitisation, IgE recognition and allergenic cross-reactivity.
Dietary Cholesterol Enhances Pulmonary Eosinophilic Inflammation in a Murine Model of Asthma
Authors: Yeh Ya-Fan , Huang Song-Lih
Published: April 2003

Background: Epidemiological studies have suggested that the dietary pattern may be associated with the prevalence of asthma. We previously reported that an increased intake of foods of animal origin was associated with the occurrence of allergic rhinitis and asthma in adolescents. Here we examined the effect of dietary cholesterol in a murine model of allergic pulmonary inflammation. Methods: Weanling C57BL/6 mice were fed a control diet containing 0.02% cholesterol or a diet supplemented with 1% or 2% cholesterol. Four weeks later the mice were sensitized with intraperitoneal ovalbumin (OVA) followed by OVA or saline inhalation 2 weeks later. OVA aerosol-induced inflammation was significantly enhanced by dietary supplementation of 1% or 2% cholesterol. Results: Among OVA-challenged mice, leukocyte numbers, particularly those of eosinophils, in the bronchoalveolar space increased by 3- to 5-fold with the cholesterol supplement. Among OVA aerosol-challenged mice, the levels of interleukin-5 and cysteinyl leukotrienes in the bronchoalveolar lavage fluid were significantly higher in those fed the 2% cholesterol diet compared with mice on the control diet. Conclusions: Dietary cholesterol may enhance pulmonary allergic inflammation.
Effect of Roxithromycin on Eotaxin-Primed Reactive Oxygen Species from Eosinophils
Authors: Cui Chang-hao , Honda Kohei , Saito Norihiro , Yamada Yoshiyuki , Sannohe Satoshi , Ueki Shigeharu , Hamada Kazuyuki , Yamaguchi Kazutoshi , Kobayashi Yoshimi , Adachi Tetsuya , Kayaba Hiroyuki , Chihara Junichi
Published: April 2003

Background: The CC chemokine eotaxin not only attracts eosinophils to inflamed sites but also promotes adhesion, degranulation and reactive oxygen species production of eosinophils. Reactive oxygen species released from eosinophils are believed to injure epithelial cells at inflamed sites, resulting in airway hyperresponsiveness. Roxithromycin has been reported to have antiasthmatic effects, although its mechanism of action is not thoroughly understood. Therefore, the effect of roxithromycin on eotaxin-primed reactive oxygen species production from eosinophils was studied. Methods: Reactive oxygen species production by eosinophils cultured with or without roxithromycin was evaluted using luminol-dependnet chemiluminescence. Results: Roxithromycin inhibited the release of reactive oxygen species from eosinophils evoked with the calcium ionophore A23187, regardless of pretreatment with or without eotaxin. Conclusion: Roxithromycin may protect epithelial cells at inflamed sites, at least partly by inhibiting the release of reactive oxygen species from eosinophils.
cDNA Cloning and Immunological Characterization of a Newly Identified Enolase Allergen from Penicillium citrinum and Aspergillus fumigatus
Authors: Lai Hsiu-Yu , Tam Ming F. , Tang Ren-Bin , Chou Hong , Chang Ching-Yun , Tsai Jaw-Ji , Shen Horng-Der
Published: April 2003

Background:Penicillium citrinum and Aspergillus fumigatus are prevalent indoor airborne fungal species that have been implicated in human respiratory allergic disorders. It is important to understand the allergenic profile of these fungal species. The purpose of the present study is to characterize a newly identified enolase allergen from P. citrinum and A. fumigatus. Methods: Fungal proteins were separated by two-dimensional (2D) gel electrophoresis and blotted onto polyvinylidene difluoride membranes. Protein spots that reacted with IgE antibodies in serum samples from asthmatic patients were identified and the N-terminal amino acid sequences were determined by Edman degradation. The peptide sequences obtained were utilized in cloning the cDNA of the allergen genes by reverse transcriptase-polymerase chain reaction and the 5′- and 3′-rapid amplification cDNA end reactions. Results: Our results from 2D immunoblotting identified a 47-kD IgE-reactive component in the extracts of P. citrinum and A. fumigatus. The N-terminal amino acid sequences of the 47-kD proteins are homologous to those of fungal enolases. The corresponding enolase cDNA from P. citrinum contains 1,552 bp and encodes a protein of 438 residues. In A. fumigatus, the isolated enolase cDNA has 1,649 bp and contains a 438-amino acid open reading frame. The deduced amino acid sequences of these two enolases have 94% identity. These enolases from P. citrinum and A. fumigatus were expressed in Escherichia coli as a His-tagged protein and designated as rPen c 22 and rAsp f 22, respectively. Sera from 7 (30%) of the 23 Penicillium-sensitized asthmatic patients showed IgE binding to the 47-kD P. citrinum component (Pen c 22) and rPen c 22. In addition, six of seven Pen c 22-positive serum samples have IgE immunoblot reactivity to the 47-kD A. fumigatus component (Asp f 22) and rAsp f 22. A polyclonal rabbit antiserum generated against the N-terminal peptide of Pen c 22 can react with Pen c 22, rPen c 22, Asp f 22 and rAsp f 22. In addition, the presence of IgE cross-reactivity between rPen c 22 and rAsp f 22 and between enolases from A. fumigatus and Alternaria alternata was also detected by immunoblot inhibition. Conclusions: These results demonstrated that a novel enolase allergen from P. citrinum (Pen c 22) and A. fumigatus (Asp f 22) was identified. In addition, IgE cross-reactivity between enolase allergens from A. fumigatus and P. citrinum and between enolases from A. fumigatus and A. alternata was also detected. Results obtained provide more information on fungal enolase allergens.
Human Cytomegalovirus Reactivation in Bone-Marrow-Derived Granulocyte/Monocyte Progenitor Cells and Mature Monocytes
Authors: Prösch Susanna , Döcke Wolf-Dietrich , Reinke Petra , Volk Hans-Dieter , Krüger Detlev H.
Published: April 2003

Monocyte/granulocyte progenitor cells of the bone marrow are a major site of human cytomegalovirus (HCMV) latency. The mechanisms of establishment and maintenance of HCMV latency are still unknown. Reactivation of the latent virus in bone-marrow-derived progenitor cells has been demonstrated in vitro and suggested to occur also in vivo. Clinical studies have shown that reactivation is a rather frequent event not only in immunosuppressed but also in nonimmunosuppressed patients and in healthy blood donors. At least three independent mechanisms of virus reactivation are discussed: systemic inflammation connected with strong tumor necrosis factor alpha release; application of cAMP-elevating drugs, and highly stressful events associated with increased plasma catecholamine levels.
Molecular Mimicry by Cytomegaloviruses
Authors: Vink Cornelis , Beisser Patrick S. , Bruggeman Cathrien A.
Published: April 2003

Cytomegaloviruses (CMVs) are well known for their high prevalence rate within host populations as well as their ability to induce lifelong infections. To maintain a persistent and stable relationship with their host, CMVs have evolved various molecular mechanisms to both control host cell metabolism and evade immune surveillance. Among the viral gene products that are likely to be involved in these processes are homologues of cellular G protein-coupled receptors, MHC class I molecules and chemokines. The viral genes encoding these homologues have probably been pirated by the viruses during a long pathogen/host coevolution. In this report, we will discuss the possible functions of these homologues in the pathogenesis of CMV infections.