bluuebunny/crossref_metadata_2025 huggingface dataset:
Power to Detect Linkage Based on Multiple Sets of Data in the Presence of Locus Heterogeneity: Comparative Evaluation of Model-Based Linkage Methods for Affected Sib Pair Data
Authors: Vieland Veronica J. , Wang Kai , Huang Jian
Published: April 2003

The development of rigorous methods for evaluating the overall strength of evidence for genetic linkage based on multiple sets of data is becoming increasingly important in connection with genomic screens for complex disorders. We consider here what happens when we attempt to increase power to detect linkage by pooling multiple independently collected sets of families under conditions of variable levels of locus heterogeneity across samples. We show that power can be substantially reduced in pooled samples when compared to the most informative constituent subsamples considered alone, in spite of the increased sample size afforded by pooling. We demonstrate that for affected sib pair data, a simple adaptation of the lod score (which we call the compound lod), which allows for intersample admixture differences can afford appreciably higher power than the ordinary heterogeneity lod; and also, that a statistic we have proposed elsewhere, the posterior probability of linkage, performs at least as well as the compound lod while having considerable computational advantages. The companion paper (this issue, pp 217–225) shows further that in application to multiple data sets, familiar model-free methods are in some sense equivalent to ordinary lod scores based on data pooling, and that they therefore will also suffer dramatic losses in power for pooled data in the presence of locus heterogeneity and other complicating factors.
Offspring Risk and Sibling Risk for Multilocus Traits
Authors: Koivisto Mikko , Mannila Heikki
Published: April 2003

The recurrence risk of a trait in a relative of type R is the probability that an individual who is in relationship of type R to an affected proband has the trait. It is intuitively clear that closer relationships lead to higher recurrence risks. However, no exact analysis of this phenomenon has been presented for multilocus traits. We prove a theorem that shows how recurrence risks are influenced by the degree of closeness of the relationship R. For example, our theorem implies that sibling risk is always higher than offspring risk. The loci influencing the trait are assumed to be autosomal and unlinked, but arbitrary epistasis between the loci is allowed. We give a detailed proof of the theorem by using stochastic matrices. A shorter proof based on the additive and dominance genetic variances is also sketched. Additionally, we also give some empirical results and discuss generalizations of the theorem.
Comparison of ‘Model-Free’ and ‘Model-Based’ Linkage Statistics in the Presence of Locus Heterogeneity: Single Data Set and Multiple Data Set Applications
Authors: Huang Jian , Vieland Veronica J.
Published: April 2003

Earlier work [Knapp et al.: Hum Hered 1994;44:44–51] focusing on affected sib pair (ASP) data established the equivalence between the mean test and a test based on a simple recessive lod score, as well as equivalences between certain forms of the maximum likelihood score (MLS) statistic [Risch: Am J Hum Genet 1990;46:242–253] and particular forms of the lod score. Here we extend the results of Knapp et al. [1994] by reconsidering these equivalences for ASP data, but in the presence of locus heterogeneity. We show that Risch’s MLS statistic under the possible triangle constraints [Holmans: Am J Hum Genet 1993;52:362–374] is locally equivalent to the ordinary heterogeneity lod score assuming a simple recessive model (HLOD/R); while the one-parameter MLS assuming no dominance variance is locally equivalent to the (homogeneity) recessive lod. The companion paper (this issue, pp 199–208) showed that when considering multiple data sets in the presence of locus heterogeneity, the HLOD can suffer appreciable losses in power. We show here that in ASP data, these equivalences ensure that this same loss in power is incurred by both forms of the MLS statistic as well. The companion paper also introduced an adaptation of the lod, the compound lod score (HLOD/C). We confirm that the HLOD/C maintains higher power than these ‘model-free’ methods when applied to multiple heterogeneous data sets, even when it is calculated assuming the wrong genetic model.
Gene-Environment Interaction and Affected Sib Pair Linkage Analysis
Authors: Gauderman W. James , Siegmund Kimberly D.
Published: April 2003

Objectives: Gene-environment (G×E) interaction influences risk for many complex disease traits. However, genome screens using affected sib pair linkage techniques are typically conducted without regard for G×E interaction. We propose a simple extension of the commonly used mean test and evaluate its power for several forms of G×E interaction. Methods: We compute expected IBD sharing by sibling exposure profile, that is by whether two sibs are exposed (EE), unexposed (UU), or are discordant for exposure (EU). We describe a simple extension of the mean test, the ‘mean-interaction’ test that utilizes heterogeneity in IBD sharing across EE, EU, and UU sib pairs in a test for linkage. Results: The mean-interaction test provides greater power than the mean test for detecting linkage in the presence of moderate or strong G×E interaction, typically when the interaction relative risk (Rge) exceeds 3 or is less than 1/3. In the presence of strong interaction (Rge = 10), the required number of affected sib pairs to achieve 80% power for detecting linkage is approximately 30% higher when the environmental factor is ignored in the mean test, than when it is utilized in the mean-interaction test. Conclusion: Linkage methods that incorporate environmental data and allow for interaction can lead to increased power for localizing a disease gene involved in a G×E interaction.
Consanguinity and Prereproductive Mortality in the Utah Mormon Population
Authors: Jorde L.B.
Published: April 2003

To test the effects of parental consanguinity on mortality among offspring, inbreeding coefficients were estimated for 303,675 members of the Utah Mormon population who were born between 1847 and 1945. Although consanguinity has been relatively rare in this population, the large sample size permitted the identification of more than 3,500 inbred offspring. Among the offspring of unrelated parents, 13.2% died before the age of 16. Significant elevations in prereproductive mortality were seen among the offspring of first-cousin marriages (22%) and among the offspring of closer unions (32%). The cor- responding relative risks are 1.70 (95% confidence limits = 1.52, 1.91) and 2.41 (95% confidence limits = 1.59, 3.41), respectively. Other categories of relationship did not produce significant elevations in offspring mortality. Similar results were obtained when a case-control approach was used to remove the effects of socioeconomic variation. Consistent with many other studies of populations with low consanguinity rates, this population experienced a relatively high absolute increase in mortality among the offspring of first-cousin marriages (9%). Preliminary evidence is offered for the hypothesis that mortality differentials are larger in populations with low inbreeding and low mortality because nongenetic causes of death do not obscure the effects of consanguinity.
Association Tests in Nuclear Families
Authors: Siegmund Kimberly D. , Gauderman W. James
Published: April 2003

We present a conditional likelihood approach for testing linkage disequilibrium in nuclear families having multiple affected offspring. The likelihood, conditioned on the identity-by-descent (IBD) structure of the sibling genotypes, is unaffected by familial correlation in disease status that arises from linkage between a marker locus and the unobserved trait locus. Two such conditional likelihoods are compared: one that conditions on IBD and phase of the transmitted alleles and a second which conditions only on IBD of the transmitted alleles. Under the log-additive model, the first likelihood is equivalent to the allele-counting methods proposed in the literature. The second likelihood is valid under the added assumption of equal male and female recombination fractions. In a simulation study, we demonstrated that in sibships having two or three affected siblings the score test from each likelihood had the correct test size for testing disequilibrium. They also led to equivalent power to detect linkage disequilibrium at the 5% significance level.
Statistical Considerations for Genome-Wide Scans: Design and Application of a Novel Software Package POLYMORPHISM
Authors: Niu Tianhua , Struk Berthold , Lindpaintner Klaus
Published: April 2003

Objective: Given the cost and complexity of genome-wide scans, optimization of study design is of critical importance. Available algorithms only partially satisfy this need. We designed a software package called ‘POLYMORPHISM’ to meet these needs. Methods: The program is designed to calculate linkage parameters for both ‘single-point’ and ‘two-point’ settings that are applicable also to incompletely informative microsatellite markers. In single-point analysis, the heterozygosity, polymorphism information content (PIC) and linkage information content (LIC) statistics based on marker allele frequencies are provided. In two-point analysis, joint PIC values for two markers, the conditional probability of detecting linkage phase, the frequency of double heterozygotes and the expected number of informative meioses are calculated. Results: Results were obtained using S.A.G.E./DESPAIR (Design of Linkage Studies Based on Pairs of Relatives) in addition to applying this program to a Centre d’Etude du Polymorphisme pedigree-derived genotyping data set, which estimated critical parameters used in a two-stage genome scan. A single nucleotide polymorphism (SNP)-based one-stage genomic screen strategy is also considered. Conclusions: LIC values are crucial for getting accurate estimates on those parameters that are important for a two-stage genome screening study. Optimization of the cost-effectiveness of an SNP-based genomic screen strategy is possible by modeling a balance between marker information content and marker density.
Haemochromatosis Gene Mutations in Finns, Swedes and Swedish Saamis
Authors: Beckman L.E. , Sjöberg K. , Eriksson S. , Beckman L.
Published: April 2003

Frequencies of three different mutant haemochromatosis (HFE) alleles (282Tyr, 63Asp and 65Cys) were studied in three northern European populations, i.e. Finns, Swedes and Swedish Saamis. In Finns and Swedes the allele frequencies were within the range found in other populations from northern and western Europe. The Saamis differed from the Swedes with respect to all mutant alleles. Lower frequencies compared to Swedes were found for the 282Tyr (p = 0.0046) and 63Asp (p = 0.034) alleles, whereas the frequency of the 65Cys allele was higher (p = 0.046) in the Saamis. The total distribution of HFE alleles in Saamis showed a highly significant difference from that in Swedes (χ2 = 16.7, 3 d.f., p = 0.0008). These results further underline the genetic uniqueness of the Saamis.
Sample Size Considerations in Genetic Polymorphism Studies
Authors: B-Rao Chandrika
Published: April 2003

Objectives: Molecular studies for genetic polymorphisms are being carried out for a number of different applications, such as genetic disorders in different populations, pharmacogenomics, genetic identification of ethnic groups for forensic and legal applications, genetic identification of breed/stock in animals and plants for commercial applications and conservation of germ plasm. In this paper, for a random sampling scheme, we address two questions: (A) What should be the minimum size of the sample so that, with a prespecified probability, all alleles at a given locus (or haplotypes at a given set of loci) are detected? (B) What should be the sample size so that the allele frequency distribution at a given locus (or haplotype frequency distribution at a given set of loci) is estimated reliably within permissible error limits? Methods: We have used combinatorial probabilistic arguments and Monte Carlo simulations to answer these questions. Results: We found that the minimum sample size required in case A depends mainly on the prespecified probability of detecting all alleles, while in case B, it varies greatly depending on the permissible error in estimation (which will vary with the application). We have obtained the minimum sample sizes for different degrees of polymorphism at a locus under high stringency, as well as a relaxed level of permissible error. We present a detailed sampling procedure for estimating allele frequencies at a given locus, which will be of use in practical applications. Conclusion: Since the sample size required for reliable estimation of allele frequency distribution increases with the number of alleles at the locus, there is a strong case for using biallelic markers (like single nucleotide polymorphisms) when the available sample size is about 800 or less.
IL-5-Producing T Cells that Induce Airway Eosinophilia and Hyperresponsiveness Are Suppressed by Dexamethasone and Cyclosporin A in Mice
Authors: Wada Kazuteru , Kaminuma Osamu , Mori Akio , Nakata Aya , Ogawa Koji , Kikkawa Hideo , Ikezawa Katsuo , Suko Matsunobu , Okudaira Hirokazu
Published: April 2003

We have recently demonstrated that airway eosinophilic inflammation can be transferred to unprimed mice by infusion of IL-5-producing T cell clones. In this study, we investigated the effects of dexamethasone and cyclosporin A on the airway eosinophilic inflammation in mice transferred with T cell clones. An ovalbumin-reactive T cell clone, KW29, produced IL-5 as well as IL-2 and IL-4 upon stimulation with relevant antigen. Dexamethasone and cyclosporin A dose-dependently suppressed the production of these cytokines in vitro. The number of eosinophils recovered in the bronchoalveolar lavage fluid and the airway responsiveness to acetylcholine were increased in KW29-transferred mice after antigen provocation. Both responses were dose-dependently suppressed by the administration of dexamethasone or cyclosporin A in vivo. We concluded that airway eosinophilic inflammation can be controlled by agents capable of downregulating IL-5 production in T cells.
Heterogeneous Expression of Tumor Necrosis Factor-α Receptors I and II on Human Peripheral Eosinophils
Authors: Matsuyama Go , Ochiai Kenichi , Ishihara Chie , Kagami Masaru , Tomioka Hisao , Koya Niroku
Published: April 2003

Background: Expression of the tumor necrosis factor-α receptor by human eosinophils has not been determined. We examined the surface expression and mRNA for tumor necrosis factor-α receptors I and II (TNF-αRI and TNF-αRII) on peripheral eosinophils. Methods: Eosinophils were obtained from 17 asthma patients and 3 healthy volunteers. Flow cytometry was used to examine receptor expression, and the reverse transcriptase-polymerase chain reaction was used to examine receptor mRNA expression. Results: Flow cytometry revealed that 16 out of 20 subjects had eosinophils expressing TNF-αRI and 18 subjects had eosinophils expressing TNF-αRII. All eosinophils expressed at least one receptor. The mRNA for TNF-αRII was detected in all eosinophils examined, but the signals for TNF-αRI mRNA were only found in cells expressing this receptor. Conclusions: Human peripheral eosinophils show heterogeneous expression of TNF-αRI and TNF-αRII.
Eosinophil Degranulation in the Presence of Lung Fibroblasts
Authors: Takafuji Shigeru , Shoji Shunsuke , Ito Koji , Yamamoto Kazuhiko , Nakagawa Takemasa
Published: April 2003

Background: Although eosinophils (Eos) and fibroblasts (Fb) are closely approximated in the bronchial submucosae of asthmatics, and are believed to play important roles in the pathogenesis of bronchial asthma, the interaction between Eos and Fb has not been thoroughly elucidated. In this study, we have examined eosinophil cationic protein (ECP) release from human Eos cultured in the presence of human lung Fb. Methods: Eos from healthy donors were cultured with or without C5a for 16 h in the presence of human fetal lung Fb which had previously been incubated with or without some cytokines for 4 h. ECP in supernatants was measured by RIA. Results: ECP release was potentiated only when both Eos and Fb were activated by C5a and TNF, respectively, while it was not significantly potentiated when either Eos or Fb were activated. ECP release from Eos activated by C5a was also potentiated when Fb were stimulated by IL-1β. The enhancement of ECP release in cocultured Eos and Fb with stimulation was partly inhibited by monoclonal antibodies against GM-CSF and was accompanied by the enhancement of adhesion of Eos to Fb. Conclusions: This study shows that the stimulation of both Eos and Fb increases ECP release. It is suggested that Fb may influence Eos degranulation and play a role in the pathogenesis of bronchial asthma.
Human Eotaxin Induces Eosinophil-Derived Neurotoxin Release from Normal Human Eosinophils
Authors: El-Shazly Amr , Masuyama Keisuke , Nakano Koji , Eura Masao , Samejima Yasuhiro , Ishikawa Takeru
Published: April 2003

Background: Eosinophil granule proteins deposition at the site of allergic inflammation contributes to the late-phase reaction of hypersensitivity diseases. In the present communication, we describe the effect of human eotaxin on normal human eosinophil exocytosis measured as degranulation of eosinophil-derived neurotoxin (EDN). Methods: Purified eosinophils were obtained from normal healthy volunteers with the CD16-negative procedure. Purified eosinophils were stimulated with various concentrations of eotaxin and the amount of EDN released was analysed by radioimmunoassay. Flow cytometry was used to examine the surface expression of adhesion molecules on eosinophils. Results: Eotaxin significantly induced EDN release in a dose-dependent manner. The potency of eotaxin in this effect was equal to that of RANTES, and comparable to that of platelet-activating factor. Eotaxin-induced EDN release was blocked by cytochalasin B in a dose-dependent manner. The surface expression of CD11a, CD11b, CD18 and VLA-4 adhesion molecules on normal human eosinophils were not modulated by eotaxin stimulation. Conclusions: These results indicate that eotaxin may play an important role not only as a selective chemotaxin for the cell type but also as a secretagogue. Furthermore, they demonstrate a degranulation mechanism(s) involving cytoskeletal changes which is probably independent of the quantitative expression of adhesion molecules.
Signal Transduction in Activation of Human Eosinophils: G Protein-Dependent and -Independent Pathways
Authors: Kato Masahiko , Kita Hirohito , Tokuyama Kenichi , Morikawa Akihiro
Published: April 2003

Degranulation of eosinophils and subsequent release of toxic granule proteins play a key role in allergic diseases such as bronchial asthma. We have previously shown that stimulation of eosinophils with immobilized secretory immunoglobulin A (sIgA) induced the phosphorylation of several proteins including 51-, 65-, 73-, 78-, 100-, 105- and 113-kD proteins. Pervanadate, a protein tyrosine phosphatase inhibitor, also induced at least 7 tyrosine phosphorylated proteins including those observed with immobilized sIgA. Pervanadate also induced inositol phosphate (IP) production and degranulation of eosinophils in a concentration-dependent manner. Eosinophil production of IP and degranulation as well as tyrosine phosphorylation of proteins induced by sIgA were completely inhibited by a tyrosine kinase inhibitor, genistein, and pertussis toxin (PTX), suggesting the involvement of both the PTX-sensitive guanine nucleotide-binding (G) protein and protein tyrosine kinases (PTK) in sIgA-induced activation of eosinophils. In contrast, PTX did not affect tyrosine phosphorylation induced by sIgA or pervanadate. Furthermore, pervanadate-induced IP production was partially inhibited by PTX. Finally, a phospholipase C-γ2 isoform was tyrosine phosphorylated by pervanadate, but not by sIgA. These findings suggest that at least two different pathways, i.e. PTK-mediated G protein-dependent or -independent PLC activation, are involved in the activation of human eosinophils.
Cross-Linking of the β2 Integrin, CD11b/CD18, on Human Eosinophils Induces Protein Tyrosine Phosphorylation and Cellular Degranulation
Authors: Kato Masahiko , Kita Hirohito , Tokuyama Kenichi , Morikawa Akihiro
Published: April 2003

Adhesion molecules, including integrins, play an important role in the selective recruitment of eosinophils. It has recently been shown that integrins also modulate the functions of eosinophils. Here, we tested the hypothesis that cross-linking of the β2 integrin, αMβ2Mac-/, leads to intracellular signaling events such as activation of protein tyrosine kinases leading to eosinophil degranulation. Cross-of cell surface CD11b/with anti-tibody and goat anti-G immobilized onto the plate triggered tyrosine phosphorylation of several intracellular proteins, including the one with a 115-ular mass (pp115).The same stimulus also provoked degranulation of eosinophils. These findings suggest that engagement of β integrin on eosinophils triggers the activation of intracellular signaling cascade which leads to cellular degranulation.
Ecalectin as a T Cell-Derived Eosinophil Chemoattractant
Authors: Hirashima Mitsuomi
Published: April 2003

In our previous papers, we have shown that human T cells produce a unique eosinophil chemotactic factor (ECF), termed Ecalectin, with a molecular weight of about 30–50 kD during interaction with BALL-1 (a B cell line) extracts, antigen or mitogen. A 1.6-kB cDNA was isolated from a human T cell-derived expression library that encodes Ecalectin. Ecalectin is a 36-kD protein consisting of 323 amino acids based on its deduced amino acid sequence. Ecalectin was found to be a variant of human galectin-9, a member of the growing family of animal lectins exhibiting affinity for β-galactosides. Recombinant Ecalectin, expressed in COS cells, exhibited potent ECF activity in vitro and in vivo in a dose-dependent manner but not chemotactic activity for neutrophils, lymphocytes, or monocytes. The finding that the Ecalectin transcript is present in various lymphatic tissues and that its expression increases substantially in antigen-activated peripheral blood mononuclear cells suggests that Ecalectin is an important T cell-derived regulator of eosinophil recruitment in allergic reaction sites.
Analysis of Biological Activities of Recombinant Rat Interleukin-5
Authors: Ishihara Kenji , Nittoh Takeaki , Satoh Ikuko , Ohuchi Kazuo
Published: April 2003

We analyzed the biological activities of recombinant rat interleukin-5 (IL-5). Purified recombinant rat IL-5 promoted the proliferation of T88-M cells in a concentration-dependent manner, and its effect was inhibited by an anti-murine IL-5-neutralizing polyclonal antibody. When bone marrow cells from normal rats were incubated with recombinant rat IL-5 in medium containing methylcellulose, the colony formation by eosinophilic cells was induced. Furthermore, when rat peritoneal eosinophils were incubated with recombinant rat IL-5, the spontaneous decrease in eosinophil viability was inhibited in a time- and concentration-dependent manner. In addition, the recombinant rat IL-5-induced eosinophil survival was inhibited by an anti-murine IL-5-neutralizing polyclonal antibody. These findings suggest that rat IL-5 acts as B cell growth factor II, eosinophil differentiation factor, and eosinophil survival-enhancing factor.
Interferon-γ Increases CD62L Expression on Human Eosinophils
Authors: Momose T. , Okubo Y. , Horie S. , Takashi S. , Tsukadaira A. , Suzuki J. , Isobe M. , Sekiguchi M.
Published: April 2003

Background: L-selectin (CD62L) and Mac-1 (CD11b/CD18) play a crucial role in the infiltration of eosinophils. However, changes in CD62L and CD11b expression on eosinophils after stimulation with cytokines were little studied. Methods: Eosinophils in peripheral blood of healthy volunteers were purified and cultured with interleukin-5 (IL-5), granulocyte/macrophage colony-stimulating factor (GM-CSF) or interferon-γ (IFN-γ). After up to 24 h incubation, CD62L and CD11b expression were examined using flow cytometry. The effects of dexamethasone (Dex), cycloheximide (CHX) or theophylline on CD62L expression on IFN-γ-stimulated eosinophils were also studied. Results: IL-5 or GM-CSF downregulated CD62L and upregulated CD11b expression on eosinophils after 30 min stimulation. Conversely, IFN-γ upregulated CD62L after 12 h stimulation in a time- and dose-dependent manner, and had no effect on CD11b expression. Dex, CHX or theophylline dose-dependently decreased CD62L expression on IFN-γ-stimulated eosinophils. Conclusions: IFN-γ is a particular cytokine which can increase CD62L expression on circulating or infiltrated human eosinophils. It is suggested that protein synthesis and intracellular cAMP participate in the increase in L-selectin expression on IFN-γ-stimulated eosinophils.
Adhesion to Fibronectin Regulates Expression of Intercellular Adhesion Molecule-1 on Eosinophilic Cells
Authors: Higashimoto Ikkou , Chihara Junichi , Kawabata Masaharu , Nakajima Shigenori , Osame Mitsuhiro
Published: April 2003

Background: Adhesion molecules may play an important role in eosinophilic activation as well as adherence of extracellular matrix (ECM) including fibronectin (FN) in the inflamed focus. Tissue eosinophils expressed inter-cellular adhesion molecule-1 (ICAM-1, CD54), whereas peripheral blood eosinophils did not. The molecular mechanisms of ICAM-1 expression on eosinophils are not clear. We immunologically examined the effect of adherence to FN on the expression of adhesion molecules in an eosinophilic cell line (EoL-1). Methods: EoL-1 cells, suspended at a concentration of 2 × 106 cells/ml, were cultured at 37°C in BSA or FN-coated 24-well plates. After 4 h, the cells were removed by pipetting, and the expression of adhesion molecules was evaluated by immunofluorescence. Results: Adherence of EoL-1 to FN enhanced ICAM-1 (CD54) expression on EoL-1 (FN vs. BSA: 84.76 ± 17.90 vs. 43.73 ± 7.60, mean fluorescent intensity). Regarding other adhesion molecules on EoL-1 (CD11a, CD18, CD49d), the expression was not significantly augmented by adherence to FN. Conclusions: We concluded that the signal via adhesion of FN regulates the expression of ICAM-1 on eosinophilic cells. This study suggests that eosinophilic adhesion to ECM via adhesion molecules plays an important role in the pathogenesis of allergic inflammation through eosinophilic functional changes, including regulation of expressive adhesion molecules such as ICAM-1.
CCR3 mRNA Expression in Bronchial Epithelial Cells and Various Cells in Allergic Inflammation
Authors: Oyamada Hajime , Kamada Yumiko , Kuwasaki Tomoe , Yamada Yoshiyuki , Kobayashi Yoshimi , Cui Chang-Hao , Honda Kohei , Kayaba Hiroyuki , Saito Norihiro , Chihara Junichi
Published: April 2003

Background: RANTES and eotaxin are important chemokines involved in the activation and migration of eosinophils and are considered to play a major role in allergic inflammation. Methods: In this study, we used RT-PCR to investigate the kinds of cells that express mRNA for CCR3, a common receptor of these chemokines, and eotaxin, a ligand for CCR3. Results: CCR3 mRNA was expressed in eosinophils, peripheral mononuclear cells, an eosinophilic cell line (EoL-1), a bronchial epithelial cell line (NCI-H292), human endothelial cells and nasal washings from patients with allergic rhinitis.  Conclusion: These results suggest that the CCR3-eotaxin system plays an important role in generating inflammation, since these substances are expressed not only in cells implicated in activation or migration of eosinophils but also in various other cells involved in allergic inflammation.
Mechanisms of Eosinophil Cationic Protein Release in the Serum: Role of Adhesion Molecules
Authors: Kato Yoshiko , Fujisawa Takao , Terada Akihiko , Iguchi Kosei , Kamiya Hitoshi
Published: April 2003

Background: Measurement of eosinophil cationic protein (ECP) in serum has been utilized as a marker for allergic inflammation. The serum level of ECP represents the level found in vivo plus additional proteins released in vitro from peripheral blood eosinophils during the coagulation period. The mechanisms of release, however, are unclear. We investigated a possible involvement of adhesion molecules in the ECP release. Materials and Methods: Venous blood was drawn in the presence of EDTA from allergic donors. The blood was incubated with neutralizing monoclonal antibodies to CD18, CD11a, CD11b, CD29, CD49d, CD54, α4β7, or isotype-matched control antibodies, respectively, at 4°C for 30 min. Calcium gluconate (calcium) was then added to induce coagulation. The blood was further incubated for 90 min and centrifuged to obtain the serum. ECP in the serum was measured with RIA. In some experiments, purified eosinophils were incubated with plasma and calcium, then ECP in the supernatants was assayed. Results: ECP in the samples with calcium was significantly higher than in those without calcium. Purified eosinophils released ECP upon plasma coagulation. Anti-CD18, CD49d, and α4β7 antibodies significantly suppressed ECP levels in the serum. Conclusions: These results suggest that ECP release in the serum is calcium and plasma coagulation-dependent and that cell adhesion through αLβ2, αMβ2, α4β1 and α4β7 integrins is at least in part responsible for ECP release.
Difference in Apoptotic Function between Eosinophils from Peripheral Blood and Bronchoalveolar Lavage in Chronic Eosinophilic Pneumonia
Authors: Saita Naoki , Yamanaka Tohru , Kohrogi Hirotsugu , Ando Masayuki , Hirashima Mitsuomi
Published: April 2003

We compared apoptosis in eosinophils from bronchoalveolar lavage (BAL-Eos) with that in eosinophils from peripheral blood (PB-Eos) of 4 patients with chronic eosinophilic pneumonia (CEP). The survival rate of the BAL-Eos on the 3rd day of the culture was significantly higher than that of the PB-Eos (39.1 vs. 1.3%). The percentage of apoptotic cells in the PB-Eos after a 24-hour incubation was higher than that in the BAL-Eos (21.7 vs. 10.6%) according to an analysis with annexin V. We further found that ECF-PI9, an eosinophil chemotactic factor (ECF) derived from an established T cell line (STO-2), significantly suppressed the apoptosis of both PB-Eos and BAL-Eos and prolonged their survival. The expression of Fas on PB-Eos was significantly suppressed by ECF-PI9 (18.5 to 7.37%, p < 0.05), whereas ECF-PI9 failed to suppress the Fas expression on BAL-Eos (3.3 to 3.6%). In addition, an ECF with similar physicochemical properties and biological functions was isolated from the BAL fluid of patients with CEP. These data demonstrate differences between PB-Eos and BAL-Eos, and indicate that ECF-PI9 is involved in the pathogenesis of CEP.
Ecalectin/Galectin-9, a Novel Eosinophil Chemoattractant: Its Function and Production
Authors: Hirashima Mitsuomi
Published: April 2003

We have recently succeeded in purifying and cloning a novel eosinophil chemoattractant, ecalectin/galectin-9, which belong to a rapidly growing galectin family. The eosinophil chemoattractant activity of ecalectin is potent and selective for eosinophils. The cellular source of ecalectin has been thought to be antigen-stimulated T cells. Herein, we will review the significance of its divalent galactoside-binding activity in ecalectin-induced eosinophil chemoattraction and the regulation of ecalectin expression in immune cells.
Eosinophil-Adhesion-Inducing Activity Produced by Antigen-Stimulated Mononuclear Cells Involves GM-CSF
Authors: Nagata Makoto , Yamamoto Hideaki , Tabe Kazuaki , Sakamoto Yoshio , Matsuo Hiroshi
Published: April 2003

Background: The initial step of eosinophil accumulation in allergic inflammation is adhesion of circulating eosinophils to vascular endothelial cells (EC). There is evidence that the adhesive property of circulating eosinophils is upregulated following antigen exposure. Although the exact mechanism remains to be established, cytokine(s) produced by antigen-stimulated mononuclear cells is (are) likely key factor(s). Objective: The objective of this study was to examine the factor(s) responsible for eosinophil adhesion and migration induced by the antigen-stimulated mononuclear cells obtained from atopic asthmatics. Methods: Peripheral blood mononuclear cells (PBMC) isolated from house-dust-mite-sensitive bronchial asthmatics were cultured for 96 h in the presence or absence of 1 µg/ml Dermatophagoides farinae (Df) antigen. Eosinophils were isolated from peripheral blood of healthy subjects. Eosinophil-adhesion-inducing activity in the culture supernatants of PBMC was examined by the ability to modify the adhesion of eosinophils to human pulmonary microvascular endothelial cells (HPMEC) in the presence or absence of anti-cytokine/chemokine antibodies. Eosinophil migration induced by the supernatants was also examined. Results: Eosinophil adhesion to HPMEC was significantly augmented by the supernatants of Df-stimulated PBMC, which was significantly inhibited by anti-GM-CSF, but not by anti-IL-5, anti-RANTES, or isotype-matched controls. On the other hand, eosinophil migration induced by the supernatants was inhibited by anti-GM-CSF and partly by anti-RANTES. Conclusion: Both eosinophil adhesion and migration induced by the antigen-stimulated PBMC involve GM-CSF. In contrast, RANTES is involved only in the eosinophil migration. These molecules may participate in the development of eosinophil accumulation at the allergic inflammation sites.
Regulation of Human Eotaxin Generation by Th1-/Th2-Derived Cytokines
Authors: Miyamasu Misato , Misaki Yoshikata , Yamaguchi Masao , Yamamoto Kazuhiko , Morita Yutaka , Matsushima Kouji , Nakajima Toshiharu , Hirai Koichi
Published: April 2003

Background: Accumulating evidence indicates that eotaxin is the primary mediator of tissue eosinophilia. In the present study, we analyzed the mechanisms of eotaxin generation by Th1-/Th2-derived cytokines in vitro. Methods: Human dermal fibroblasts, human umbilical vein endothelial cells and A549 human bronchial epithelial cell line cells were stimulated with TNF-α, IL-4, IFN-γ or TNF-α in combination with IL-4 or IFN-γ and the amount of eotaxin production was analyzed. Results: Fibroblasts produced nanogram/milliliter quantities of eotaxin. Proinflammatory cytokine TNF-α and Th2-type cytokine IL-4 each induced eotaxin production, and combination of them caused a marked synergistic increase in that production. On the other hand, Th1-type cytokine IFN-γ inhibited eotaxin generation at the protein/mRNA levels. Conclusion: The Th2-derived cytokine upregulated while the Th1-derived cytokine inhibited eotaxin production by fibroblasts. In view of the Th1/Th2 paradigm, these results indicate that (1) eotaxin regulates eosinophil accumulation in the Th2-dominant state such as allergic disease, and (2) direct suppression of eotaxin production by IFN-γ is one of the major mechanisms by which IFN-γ suppresses eosinophilic inflammation.
Increased Sensitivity to IL-4 in Patients with Allergic Bronchopulmonary Aspergillosis
Authors: Khan Seema , McClellan Jim S. , Knutsen Alan P.
Published: April 2003

Background: Allergic bronchopulmonary aspergillosis (ABPA) is characterized by a heightened Th2 CD4+ T cell response to Aspergillus fumigatus allergens and a hyper-IgE state compared to atopic asthmatic and cystic fibrosis patients without ABPA. We hypothesized that one reason for this response is increased sensitivity to IL-4 in ABPA, resulting in increased expression of CD23 and CD86, leading to a positive amplification mechanism which increases Th2 CD4+ T cell responses. Methods: Peripheral blood mononuclear cells isolated from 10 ABPA, 9 atopic, and 8 nonatopic subjects and stimulated for 48 h with varying concentrations of rIL-4 ranging from 0.1 to 50 ng/ml. The percentages of CD23+ and CD86+ B cells and the number of CD23+ molecules on CD20+ and CD86+CD20+ B cells were quantified by flow cytometry. Results: Total serum IgE levels were elevated in ABPA patients compared to atopic and nonatopic controls. At day 0 prior to culture, CD23 molecules per CD20+ B cell were significantly elevated in ABPA patients compared to atopic and to nonatopic patients. CD23 molecules per CD20+ B cell in ABPA and atopic patients decreased after 48 h in culture without IL-4 added and were similar. With IL-4 stimulation, ABPA patients had significantly increased rates of CD23 expression per B cell compared to atopic and nonatopic subjects (p < 0.001). Furthermore, ABPA had significantly increased numbers of CD23+ molecules per B cell and CD86+ B cell following IL-4 stimulation compared to atopic and nonatopic patients. Both ABPA and atopic patients at day 0 prior to culture had increased expression of CD86+ and CD23+CD86+ B cells compared to nonatopic patients. After 48 h in culture without IL-4, the percentages of CD86+ and CD23+CD86+ B cells decreased in ABPA and atopic patients. After stimulation with IL-4, ABPA patients had significant upregulation of CD23+CD86+ B cells compared to atopic and nonatopic patients. Similarly, the number of CD23 molecules per CD86+CD20+ B cell was significantly upregulated following IL-4 stimulation in ABPA patients compared to atopic and to nonatopic subjects. Conclusions: This is the first study to demonstrate that ABPA patients have increased sensitivity to IL-4 stimulation compared to other atopic individuals, such that ABPA > atopic >> nonatopic patients. The B cells from ABPA patients were significantly more sensitive to IL-4 stimulation compared to atopic and nonatopic patients with upregulation of CD23 and CD86 expression. ABPA subjects had increased CD86+ and CD23+CD86+ B cell expression on day 0 prior to culture and with upregulation of CD23+ molecules on CD86+CD20+ B cells. IL-4 also stimulated upregulated CD86+ expression on B cells in atopic patients with little effect on nonatopic patients. This study supports the premise that IL-4, IL-4Rα and CD86 are central targets in the treatment of ABPA and atopic disease.
Immunoprotective Properties of Peptidoglycan Monomer Linked with Zinc in Cholestatic Jaundice
Authors: Ravlić-Gulan Jagoda , Radošević-Stašić Biserka , Gulan Gordan , Štimac Davor , Pavelić Krešimir , Rukavina Daniel
Published: April 2003

Background: Previously it was shown that a new immunostimulator, peptidoglycan monomer linked with zinc (PGM-Zn), might have immunocorrective and hepatotropic effects. Owing to this in the present study we investigated its effects on jaundice-induced immunodysfunction, which might be responsible for serious peri- and postoperative complications in biliary obstruction. Methods: In vivo effects of PGM-Zn were analyzed in mice subjected to common bile duct ligation (CBDL), where we estimated phenotypic profile and cell cycle of thymocytes, splenocytes and phagocytic function of peritoneal macrophages. In vitro effects of PGM-Zn were evaluated on blastogenesis of human peripheral blood mononuclear cells (PBMNC), obtained from healthy donors and stimulated with anti-CD3 monoclonal antibody and/or PMA, in the presence or absence of jaundice serum obtained from patients with biliary calculosis. Results and Discussion: Jaundice induced marked disarrangement of lymphatic homeostasis, which at several points might be blocked by PGM-Zn. In mice it delayed the CBDL-induced decline of CD4+ CD8+ thymocytes, decreased the proportion of CD8+ T cells, and increased the percentage of CD4– CD8– thymocytes, augmenting simultaneously the proportion of thymic cells in S and G2 + M phase of cycle. Similar hyperplastic reaction with increased percentage of CD4+, Ig+ and CD5+ cells was noticed in the spleen, together with the enhanced phagocytic ability of peritoneal macrophages. In human PBMNC jaundice reduced the percentages of CD3, CD5, CD4, CD8 and HLA-DR-expressing cells and increased the proportion of CD25 and perforin-positive lymphocytes. PGM-Zn given in vitro was able to abrogate the antiproliferative activity of jaundice serum on PMA and anti-CD3 + PMA-induced blastogenesis.
Inhibition of Passive Cutaneous Anaphylaxis-Associated Scratching Behavior by μ-Opioid Receptor Antagonists in ICR Mice
Authors: Inagaki Naoki , Nakamura Nobuaki , Nagao Masafumi , Kawasaki Hirokazu , Nagai Hiroichi
Published: April 2003

Background: Itching in humans is attenuated by µ-opioid receptor antagonists. ICR mice display increases in scratching behavior upon induction of IgE-mediated passive cutaneous anaphylaxis (PCA), or intradermal injection of compound 48/80 or histamine. Methods: Cutaneous reactions were induced in ICR mice by IgE-mediated PCA, compound 48/80 and histamine, and the scratching behavior associated with the cutaneous reactions was evaluated. Results: Naloxone and nalmefene reduced the incidence of scratching behavior associated with PCA. Naloxone also inhibited the induction of scratching behavior caused by compound 48/80 and histamine. Naloxone did not affect the increase in vascular permeability caused by PCA and injection of compound 48/80. Conclusion: Scratching behavior in mice may be induced by a sensation or a mechanism similar to itching in humans and should become a useful model for examining itching in humans.
The Monocyte/IgE Connection: May Polymorphisms in the CD14 Gene Teach Us about IgE Regulation?
Authors: Vercelli Donata , Baldini Mauro , Martinez Fernando
Published: April 2003

Background: Total IgE levels are known to be under genetic control. Linkage studies have indicated that one or more loci on chromosome 5q may control total IgE, as well as asthma and bronchial hyperresponsiveness to nonspecific stimuli. Our group has undertaken a systematic analysis of chromosome 5q, and has recently characterized five single nucleotide polymorphisms at position –1619, –1359, –1145, –809 and –159 in the promoter of the gene encoding CD14, the myeloid pattern recognition receptor that is critical for efficient innate immune response to lipopolysaccharide (LPS) and bacterial ligands. Methods: Carriers of the major CD14 haplotypes were analyzed for serum levels of IgE and soluble CD14. In vitro IgE synthesis was assessed in peripheral blood mononuclear cells stimulated with IL-4 in the presence or absence of LPS or anti-CD14 monoclonal antibodies. Results: An inverse correlation was found between serum levels of IgE and soluble CD14. On the other hand, in vitro IL-4-dependent IgE synthesis was strongly upregulated by LPS, but suppressed by anti-CD14 monoclonal antibodies. Conclusions: Our results highlight the complex role played by monocytes in IgE regulation.
Sensitisation to the Lipid-Binding Apolipophorin Allergen Der p 14 and the Peptide Mag-1
Authors: Epton M.J. , Dilworth R.J. , Smith W. , Thomas W.R.
Published: April 2003

Background: The IgE-binding peptides Mag 1 and Mag 3 and the high-molecular-weight protein M-177 have been identified as parts of the apolipophorin-like group 14 house dust mite allergen. By analogy with the homologous insect proteins, apolipophorins are hydrophobic proteins found in lipid bodies and lipid transport particles. This explains why they degrade and are poorly represented in extracts. Methods: We have examined the T cell stimulation induced by a 341-residue recombinant Der p 14 peptide equivalent to the Mag 1 polypeptide examined by others. Results: Peripheral blood mononuclear cells from both allergic and non-allergic donors responded strongly in in vitro proliferation assays to the Der p 14 peptide to induce markedly more 3H-thymidine incorporation than Der p 2 and the release of Th2 cytokines. Conclusions: The apolipophorin-like group 14 allergens, despite their predicted hydrophobicity and lipid-binding activity, can induce high IgE responses and T cell stimulation. They appear to be important mite allergens unsuited to representation by aqueous extracts of mites.
The Role of Particulate Matter in Exacerbation of Atopic Asthma
Authors: Gavett Stephen H. , Koren Hillel S.
Published: April 2003

Increasing evidence shows that elevated levels of particulate matter (PM) can exacerbate existing asthma, while evidence that PM can promote the induction of asthma is limited. PM in ambient air has been associated with increased emergency room visits and medication use by asthmatics. Controlled human exposure studies of acid aerosols suggest increased responses among adolescent asthmatics. Increased ambient and indoor levels of bioaerosols (e.g., house dust mite, fungal spores, endotoxin) have been associated with exacerbation of asthma. Environmental Protection Agency (EPA) studies focus on the effects of exposing humans and animal models to a combination of various PM samples (e.g., diesel exhaust particles, oil fly ash) and allergens (e.g., house dust mite, ovalbumin). These research efforts to understand the mechanisms by which PM exposure can promote allergic sensitization and exacerbate existing asthma concentrate on the role of transition metals. Exposure of animal models to combined PM and allergen promotes allergic sensitization and increases allergic inflammation and airway hyperresponsiveness. Exposure of healthy human volunteers to emission source PM samples promotes inflammation and increased indices of oxidant formation correlating with the quantity of transition metals in the samples. Results of these studies suggest that transition metals in ambient PM promote the formation of reactive oxygen species and subsequent lung injury, inflammation, and airway hyperresponsiveness leading to airflow limitation and symptoms of asthma.
SDF-1 Induces IL-8 Production and Transendothelial Migration of Human Cord Blood-Derived Mast Cells
Authors: Lin T.-J. , Issekutz T.B. , Marshall Jean S.
Published: April 2003

Background: Mast cell numbers and expression of chemokines are known to increase in the context of angiogenesis and inflammation, but the mechanisms by which this occurs are not understood. Stromal-derived factor-1 (SDF-1) is an important chemokine in angiogenesis and cell migration. The effects of SDF-1 on human mast cells were examined. Methods: Expression of the SDF-1 receptor CXC chemokine receptor 4 (CXCR4) on mast cells was examined by RT-PCR and flow cytometry. The ability of labeled cord blood-derived mast cells to migrate across HUVEC monolayers in response to SDF-1 was determined. The cytokine and chemokine responses of cord blood-derived mast cells to SDF-1 treatment over 24 h were examined by ELISA. Results: Cord blood-derived human mast cells expressed the CXCR4 receptor for SDF-1 and migrated across HUVEC monolayers in response to this chemokine. Treatment of cord blood-derived mast cells with SDF-1 did not induce degranulation or the production of several cytokines but did induce a highly selective IL-8 response. Conclusion: Human mast cells can both migrate across vascular endothelium and produce the pro-angiogenic chemokine IL-8 in response to SDF-1. These responses may be important in angiogenic processes.