bluuebunny/crossref_metadata_2025 huggingface dataset:
The Influence of CpG Motifs on a Protein or DNA-Based Th2-Type Immune Response against Major Pollen Allergens Bet v 1a, Phl p 2 and Escherichia coli-Derived β-Galactosidase
Authors: Hochreiter Romana , Hartl Arnulf , Freund Johann , Valenta Rudolf , Ferreira Fatima , Thalhamer Josef
Published: April 2003

Background: DNA immunization and protein immunization with CpG motifs as adjuvants represent promising approaches in allergen-specific immunotherapy. Objective: We investigated the effect of coinjection or prepriming with CpG-ODN on Th2-type responses induced by gene gun and protein immunization. Methods: BALB/c mice were immunized with the gene gun using plasmid DNA containing the cDNAs coding for the genes of Bet v 1a, Phl p 2 and β-galactosidase or with the purified Al(OH)3-adsorbed proteins. In addition, CpG-ODN were applied by coinjection or by prepriming treatment. Antibody and cytokine responses were measured by ELISA, proliferative and cytotoxic responses were determined by standard labeling procedures. Furthermore, the allergenic activity of sera was measured by passive cutaneous anaphylaxis. Results: Gene gun immunization and protein immunization induced a clear Th2-type response for all antigens. The Th1-promoting effect of CpG-ODN coinjection together with gene gun immunization was restricted to β-galactosidase as indicated by the increase of IgG2a and a marked expression of IFN-γ. CpG motifs also increased the specific cytotoxic response against β-galactosidase. Prepriming with CpG-ODN and gene gun or protein immunization with Bet v 1a exhibited no significant difference to the non-CpG control group. However, sera from mice preprimed with CpG-ODN induced no anaphylaxis with gene gun immunization, but with protein immunization. Conclusions: The effect of CpG motifs in vivo depends on a variety of parameters like the nature of the antigen and the immunization modality. Furthermore, our studies indicate that a combination of CpG + DNA immunization may be more effective in antagonizing Th2 responses than the combination of CpG + protein immunization.
New Developments in the Diagnosis and Treatment of Hymenoptera Venom Allergy
Authors: Müller Ulrich R.
Published: April 2003

According to most textbooks, diagnostic tests with Hymenoptera venoms are reliable, and immunotherapy with these venoms in Hymenoptera-venom-allergic patients leads in near to 100% to full protection. Careful analysis of the literature shows however that the specificity of diagnostic tests is far from perfect and that both efficacy and tolerance, especially in patients receiving honeybee venom immunotherapy, are still suboptimal. The major allergens of honeybee and vespid venoms are now available in recombinant form. Preliminary trials analyzing diagnostic tests with recombinant allergens in honeybee venom allergy are promising: the specificity is clearly increased in both skin testing and in determining venom-specific IgE antibodies when compared to natural venom allergens. An important recent finding is the frequent association of severe Hymenoptera venom allergy and elevated basal serum levels of the mast-cell-specific enzyme tryptase. Elevated levels are found in up to 30% of the patients with a history of severe shock reactions following Hymenoptera stings. The current findings indicate that basal tryptase levels indicating an increased mast cell load are much more frequent than previously thought and are a risk factor for severe or even fatal sting reactions. Premedication with antihistamines in the initial phase of venom immunotherapy reduced both local and systemic allergic side effects in several controlled studies. In a retrospective analysis of one of these trials it was found that reexposure during immunotherapy resulted in significantly more systemic allergic reactions in patients on placebo than on antihistamine premedication, suggesting that initial antihistamine premedication might increase the efficacy of venom immunotherapy. Different ways of allergen modification for venom immunotherapy have been proposed. While the results with chemical modifications were not convincing, recent studies with T-cell epitope peptides from the major bee venom allergen phospholipase A2 look promising. Patient-tailored cocktails of recombinant venom allergens or isoforms thereof may be another possibility in the future. A number of prospective studies analyzing the duration of venom immunotherapy required for long-term protection have been published in the last decade. While most patients are still fully protected 1 year after discontinuation of therapy, relapses may occur in up to 20% of patients reexposed many years after treatment. Various risk factors for such relapses have been identified.
The Impact of the Route and Frequency of Antigen Exposure on the IgE Response in Allergy
Authors: Nelde A. , Teufel M. , Hahn C. , Duschl A. , Sebald W. , Bröcker E.B. , Grunewald S.M.
Published: April 2003

Background: Knowledge of the factors which control IgE production is essential in order to understand the pathogenesis of immediate hypersensitivity reactions. We have studied the extent to which the route and frequency of antigen application as well as different antigen amounts may influence IgE synthesis. Methods: We established sensitisation protocols in BALB/c mice, in which various doses of ovalbumin (Ova) were applied via intranasal, epicutaneous or intraperitoneal routes. Ova-specific antibodies were measured by ELISA. After 6 weeks of sensitisation, anaphylactic shock was measured following intravenous challenge with Ova. In addition, bronchoalveolar lavages were performed in intranasally sensitised mice. Results: We were able to show that the most efficient IgE production was achieved by long-term antigen application via the airways, leading to local allergic airway pathology. The epicutaneous route of antigen application also induced very high IgE titres, while intraperitoneal sensitisation led to significantly lower IgE levels. After intraperitoneal sensitisation, IgE synthesis was best induced by increasing the frequency of antigen application, but not by increasing the amount of antigen. In all groups of mice, Ova-specific IgE antibodies were high enough to induce systemic allergic symptoms leading to anaphylactic shock. The severity of shock correlated with the amount of specific IgE. Conclusions: Taken together, our results demonstrate that antigen application via the airways or skin induces IgE synthesis more efficiently than via the intraperitoneal route. Few exposures with high-dose antigen are less efficient than multiple exposures with low doses. Our finding that both the route and the frequency of antigen application strongly influence IgE synthesis may help to understand how environmental antigens lead to allergic sensitisation.
CD4+IL13+ T Lymphocytes at Birth and the Development of Wheezing and/or Asthma during the 1st Year of Life
Authors: Spinozzi Fabrizio , Agea Elisabetta , Russano Anna , Bistoni Onelia , Minelli Liliana , Bologni Donatella , Bertotto Alberto , de Benedictis Fernando M.
Published: April 2003

Background: Despite our knowledge that maternal inheritance influences the development of asthma in childhood, attempts to identify a clear-cut Th2-oriented cytokine production by T lymphocytes at birth have given conflicting results. The prognostic significance of these cells for asthma development later in life remains to be determined. Methods: We evaluated at the single cell level Th1- and Th2-type cytokines in 208 randomly selected cord blood mononuclear cell (CBMC) samples obtained from pregnant women (group A, n = 68 with diagnosed respiratory allergic disease; group B, n = 140, with no evidence of atopy), and prospectively followed newborns for 1 year. Results: There was no difference in IFN-γ, IL-4 and IL-5 production at birth between both groups, whereas a correlation between CD4+IL13+ lymphocytes from CBMC samples derived from atopic mothers and the occurrence of wheezing and/or asthma during the 1st year of life was found. Conclusions: Our observations suggest that the intracellular cytokine profile of cord blood CD4+ cells, in terms of IL-13 production, could be considered a useful tool for a more accurate identification of newborns from atopic mothers who are at high risk of developing asthma.
Human Mast Cell Transcriptome Project
Authors: Saito Hirohisa , Nakajima Toshiharu , Matsumoto Kenji
Published: April 2003

After draft reading of the human genome sequence, systemic analysis of the transcriptome (the whole transcripts present in a cell) is progressing especially in commonly available cell types. Until recently, human mast cells were not commonly available. We have succeeded to generate a substantial number of human mast cells from umbilical cord blood and from adult peripheral blood progenitors. Then, we have examined messenger RNA selectively transcribed in these mast cells using high-density oligonucleotide probe arrays. Many unexpected but important transcripts were selectively expressed in human mast cells. We discuss the results obtained from transcriptome screening by introducing our data regarding mast-cell-specific genes.
Identification of Cyclophilin as an IgE-Binding Protein from Carrots
Authors: Fujita Chizuko , Moriyama Tatsuya , Ogawa Tadashi
Published: April 2003

Background: Plant food allergies have been associated with pollenosis, although most of the causative allergens are as yet undefined. It is important to elucidate the properties of plant food allergens in order to minimize a patient’s risks in food selection. The purpose of the present study was to examine and characterize the IgE-binding proteins in carrots as possible allergens by using patients’ sera. Method: IgE-binding proteins in carrot extract were screened by an immunoblot technique using sera of patients with atopic dermatitis (selected based upon a case history of food allergies). An allergenic protein was purified from carrot extract by chromatographic procedures. The N-terminal amino acid sequence of allergenic protein was determined and subjected to a computer homology search. Cross-reactivity between carrot and birch allergens was examined by immunoblotting. Results and Conclusion: A unique, approximately 20-kDa allergenic protein that reacted with about 14% of patients’ sera was isolated and characterized. The N-terminal amino acid sequence of the purified protein was found to be homologous with those of plant cyclophilins. This allergen exhibited a peptidyl-prolyl cistrans isomerase activity, which was inhibited by the conjugation of cyclosporin A. These properties of the allergenic protein isolated from carrot identified it as a cyclophilin, a possible plant food allergen. No cross-reactivity between this 20-kDa carrot allergen and Bet v 7, a birch pollen cylcophilin, was observed.
Helicobacter pylori Infection Affects Eosinophilic Cationic Protein in the Gastric Juice of Patients with Idiopathic Chronic Urticaria
Authors: Ojetti V. , Armuzzi A. , De Luca A. , Nucera E. , Franceschi F. , Candelli M. , Zannoni G.F. , Danese S. , Di Caro S. , Vastola M. , Schiavino D. , Gasbarrini G. , Patriarca G. , Pola P. , Gasbarrini A.
Published: April 2003

Background:Helicobacter pylori, the main cause of gastritis and peptic ulcer, has been associated with idiopathic chronic urticaria (ICU), an immunological skin disorder of unknown origin. Eosinophilic cationic protein (ECP) is a cytotoxic molecule secreted by the activated eosinophils involved in the pathogenesis of ICU. We assessed serum/gastric juice ECP levels and gastric mucosal eosinophil infiltration in ICU patients infected or not with H. pylori and evaluated the modification after bacterium eradication. Methods: 33 patients with ICU and 25 dyspeptic controls underwent upper gastrointestinal endoscopy for histological evaluation and assessment of H. pylori infection. One-week triple therapy was given to H. pylori-positive patients. Serum and gastric juice ECP levels, eosinophil infiltration from gastric mucosal sections and urticaria symptoms were evaluated in all patients at enrolment and 8 weeks after eradication. Results: 19 of 33 (57%) ICU patients and 16 of 25 (64%) controls were found to be infected with H. pylori. Serum ECP was significantly higher in ICU patients compared to controls, regardless of infectious status. Gastric juice ECP and gastric eosinophil infiltration were significantly higher in infected ICU patients when compared both to uninfected ICU patients and controls. H. pylori eradication determined a significant decrease in gastric juice ECP and gastric eosinophil infiltration only in ICU patients. Moreover, a total or partial remission of urticaria symptoms was observed only in ICU patients in whom the bacterium was eradicated. Conclusions: Although H. pylori infection affects gastric juice ECP and eosinophil infiltration of ICU patients, the role of the bacterium in the pathogenesis of this skin disorder still remains uncertain.
Recombinant Allergens for Skin Testing
Authors: Schmid-Grendelmeier Peter , Crameri Reto
Published: April 2003

Skin testing is a basic diagnostic procedure widely used to explore immediate-type reactions to allergen preparations in vivo. Despite their reliability, if standardized extracts are used, skin tests suffer from limited reproducibility due to difficulties in preparing consistently standardized extracts from natural raw material. Starting from allergen-encoding cDNAs, large amounts of highly pure allergens with a high batch-to-batch consistency satisfying the quality requirements of medicinal products manufactured by recombinant DNA technology can be produced. These reagents are expected to be qualitatively superior to the commercially available allergen preparations used for the in vitro and in vivo diagnosis of allergic conditions. In this article the current literature available on skin testing with such recombinant allergens (rAllergens) is reviewed and critically analyzed. To date many different rAllergens of various pollens, moulds, mites, bee venom, latex and celery have been used in skin testing in more than 1,600 allergic and control individuals. Skin prick tests as well as intradermal skin tests with rAllergens prove to be highly specific and safe. The diagnostic sensitivity of single rAllergens is generally lower than those obtained with allergen extracts, but can be considerably increased by using rAllergen panels covering the most important allergenic structures present in a given complex allergenic extract. Moreover, quantitative skin testing with single rAllergens allows interesting insights into correlations between the in vivo and in vitro sensitization to a given allergen. In conclusion, skin testing with rAllergens offers a highly specific and safe additional diagnostic tool to elucidate patient- and disease-specific sensitization patterns which will be needed for the development of patient-tailored immunotherapeutic treatments.
Can Immunoregulatory Lactic Acid Bacteria Be Used as Dietary Supplements to Limit Allergies?
Authors: Cross M.L. , Gill H.S.
Published: April 2003

Studies in gnotobiotic animals have suggested that the intestinal bacterial flora may play an important role in priming the immune system during ontogeny to limit dysfunctional responses, including allergy. Prospective clinical studies have identified a higher incidence of allergy expression in early childhood among children who have low enteric populations of lactic acid bacteria (LAB), such as lactobacilli and bifidobacteria, further supporting a role for gut-colonizing bacteria in regulating immunological atopy. There is some evidence to suggest that supplementing the human diet with probiotic LAB might combat both allergy development and expression of atopy in allergy sufferers; however, definitive information, in the form of controlled intervention trials, remains scant. Recent immunological evidence has indicated that certain strains of LAB can stimulate the production of type I and II interferons and pro-interferon monokines (IL-12 and IL-18), following contact with the immune system; therefore, probiotic forms of immunoregulatory LAB could be used as dietary supplements to modify the gut microflora and provide pro-T helper cell 1 (Th1) STAT-activating signals sufficient to deviate the immune phenotype and correct the Th2-type bias which promotes allergy. This review outlines the clinical and laboratory evidence of a role for LAB in combating allergies, and attempts to explain this phenomenon in terms of our current understanding of immunoregulatory signals produced by gut-colonizing microbes.
The Spectrum of Olive Pollen Allergens
Authors: Rodríguez Rosalía , Villalba Mayte , Monsalve Rafael I. , Batanero Eva
Published: April 2003

Olive pollen is one of the most important causes of seasonal respiratory allergy in Mediterranean countries, where this tree is intensely cultivated. Among the high number of protein allergens detected in this pollen, 8 – Ole e 1 to Ole e 8 – have been isolated and characterized. Ole e 1 is the most frequent sensitizing agent, affecting more than 70% of the patients suffering of olive pollinosis, although others, such as Ole e 4 and Ole e 7, have also been shown to be major allergens. In this context, the prevalence of many olive pollen allergens seems to be dependent on the geographical area where the sensitized patients live. Some of the olive allergens have been revealed as members of known protein families: profilin (Ole e 2), Ca2+-binding proteins (Ole e 3 and Ole e 8), superoxide dismutase (Ole e 5) and lipid transfer protein (Ole e 7). No biological function has been demonstrated for Ole e 1, whereas Ole e 4 and Ole e 6 are new proteins without homology to known sequences from databases. cDNAs encoding for Ole e 1, Ole e 3 and Ole e 8 have been overproduced in heterologous systems. The recombinant products were correctly folded and exhibited the functional activities of the natural allergens. In addition to the Oleaceae family, other species, such as Gramineae or Betulaceae, contain pollen allergens structurally or immunologically related to those of the olive tree. This fact allows to detect and evaluate antigenic cross-reactivities involving olive allergens. The aim of this research is the development of new diagnostic tools for olive pollinosis and new approaches to improve the classical immunotherapy.
γδ T Cells as Regulators of Airway Hyperresponsiveness
Authors: Lahn Michael , Kanehiro Arihiko , Takeda Katsuyuki , Konowal Anatole , O’Brien Rebecca L. , Gelfand Erwin W. , Born Willi K.
Published: April 2003

Airway responsiveness (AR) is determined by complex mechanisms reflecting lung responses to airborne stimuli. Murine studies have identified a number of potential factors modulating AR and thus have contributed to the current understanding of these mechanisms. In allergic inflammation, immune cells, in particular αβ T cells, have emerged as important contributors to increased AR. We have found that in contrast to αβ T cells, γδ T cells can have a negative regulatory effect on AR. Here, we review the current studies on γδ T cells in allergic inflammation and discuss their role in modulating AR. We propose that γδ T cells exhibit different immune properties depending on the type of stimulus and inflammation. These differential immune properties appear to be associated with specific γδ T cell subsets, which control AR to airborne stimuli. In particular, our recent data indicate that the Vγ4+ T cell subset acts as an important negative regulator of AR and contributes to maintaining normal lung function in mice.
Sera from Patients with Kawasaki Disease Induce Intercellular Adhesion Molecule-1 but Not Fas in Human Endothelial Cells
Authors: Inoue Yoshinari , Kimura Hirokazu , Kato Masahiko , Okada Yasunori , Morikawa Akihiro
Published: April 2003

Background: Kawasaki disease (KD) is an acute vasculitis of unknown etiology occurring in childhood, characterized by abnormalities of the immune system including elevations of proinflammatory cytokines in the serum. We investigated the effect of serum from patients with KD on the expression of intercellular adhesion molecule-1 (ICAM-1) and Fas by human umbilical vein endothelial cells (HUVEC). Methods: Confluent monolayers of HUVEC were incubated with sera from patients in the acute or convalescent phase of KD. Expression of ICAM-1 and Fas by HUVEC was assessed by flow cytometry. Concentrations of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in sera from patients with KD were measured by an immunoradiometric assay and an enzyme-linked immunosorbent assay, respectively. Results: Sera from patients in the acute phase of KD produced significantly greater ICAM-1 expression by HUVEC than sera from patients in the convalescent phase. In contrast, KD sera did not induce Fas expression. While the mean serum concentration of TNF-α in patients in the acute phase of KD was significantly higher than in those in the convalescent phase, IL-1β concentrations did not differ between the acute and convalescent phases. Exposure of HUVEC to recombinant human TNF-α increased the expression of both ICAM-1 and Fas, but a much lower concentration was required for an effect upon ICAM-1. Exogenous TNF-α did not induce apoptosis in HUVEC. Conclusions: These results suggest that increased expression of ICAM-1 by endothelial cells might be involved in the pathogenesis of acute KD, and that TNF-α might induce ICAM-1 expression.
Antitumor Activity of Expanded Human Tumor-Infiltrating γδ T Lymphocytes
Authors: Chen Juan , Niu Haitao , He Wei , Ba Denian
Published: April 2003

Background: The objective of this study was to investigate the antitumor activity of selectively expanded γδ T cells in tumor-infiltrating lymphocytes (γδTILs) or tumor ascites lymphocytes (γδTALs) from patients with colorectal and ovarian epithelial carcinoma (OEC) in vitro and in vivo. Methods: γδTILs/TALs were expanded by the solid-phase antibody method; their cytolytic and proliferative activities in vitro were detected by the MTT method and 3H-TdR incorporation and their effect in vivo was evaluated by the nude mice model. Results: Expanded γδTILs from colorectal tumors demonstrated marked cytotoxicities to allogeneic human colon adenocarcinoma HR8348 and lymphoma Daudi cells, as well as xenogeneic murine thymoma EL-4 cell lines. Cytokines, including IL-2, IL-4, IL-12, IL-15, TNF-α and INF-γ, could promote the cytotoxicities of γδTILs to tumor cells, whereas IL-10, GM-CSF and TFG-β had no effect on such killing activities. Rested γδTILs could proliferate strongly in response to mitomycin C-treated Daudi and EL-4 tumor cells, but not to HR8348 tumor cells, suggesting that the latter might possess only cytotoxicity-related antigen recognized by γδTILs. Either αβTILs or γδTILs from patients with OEC displayed cytotoxicities to allogeneic or autologous OEC cell lines at a similar strength in vitro. Transferring γδTILs into Daudi cell-bearing BALB/c nude mice with an injection of IL-2 was able to maintain a high survival rate of the mice for 30 days, when compared with mice treated with αβTILs or without any treatment (p < 0.05). Without coinjection of IL-2, after 3 months of Daudi tumor inoculation, a high survival rate was observed in γδTIL-treated mice. Similarly, adoptive γδTALs from the ascites of patients with OEC transferred into nude mice displayed a stronger antitumor response to OEC SKOV3 cells than αβTALs in vivo. Tumor volumes in γδTAL-treated mice were smaller than in αβTAL-treated or non-TAL-treated mice within the period from day 23 to day 50 after tumor inoculation (p < 0.05). Fifty days after SKOV3 tumor inoculation, a decreasing trend of carcinogenic rate was observed in γδTAL-treated nude mice. Conclusion: Taken together, our results suggest that γδT cells could be a new candidate for adoptive immunotherapy in the future treatment of patients with cancer.
Chemotaxis of Human CD4+ Eosinophils
Authors: Okubo Yoshio , Tsukadaira Akihiro , Takashi Shuji , Kubo Keishi , Koyama Sekiya
Published: April 2003

We reviewed the biological functions of CD4+ eosinophils, which are observed in peripheral blood, sputum and bronchoalveolar lavage fluid of various diseases. We have shown that CD4 molecules on human eosinophils are induced by tumor necrosis factor-alpha (TNF-α) stimulation. Interleukin-16 (IL-16) has been reported to bind a natural soluble ligand for the CD4 molecule. We reported that TNF-α-stimulated eosinophils migrate in a time- and dose-dependent manner against IL-16. Theophylline and dexamethasone significantly inhibited CD4 expression. Theophylline inhibited CD4+ eosinophil chemotaxis, but dexamethasone did not. Theophylline may prevent airway inflammation by downregulating the expression of CD4 molecule and CD4+ eosinophil migration. However, dexamethasone may inhibit airway inflammation through the downregulation of CD4 expression.
A Study of Clinical Features of Cough Variant Asthma
Authors: Okada Chiharu , Horiba Masahide , Matsumoto Hiroshi , Torigoe Rikako , Mizuuchi Hidetsugu , Murao Masaharu , Soda Ryo , Takahashi Kiyoshi , Kimura Goro , Tanimoto Yasushi
Published: April 2003

Patients with cough variant asthma (CVA) and classic asthma are frequently among subjects who present at clinics complaining of a chronic persistent cough. To reveal the features of CVA, we examined the differences in the clinical appearance between CVA and classic asthma. Ten CVA subjects and 11 classic asthmatics were enrolled in the study; they were recruited among patients who presented at the National Minamiokayama Hospital complaining of a chronic cough. The number of eosinophils in peripheral blood was 256 ± 45.8/µl in CVA and 400 ± 123/µl in classic asthma. Eosinophils represented 67% of the cells of sputum in CVA and 82% in classic asthma. Bronchial responsiveness to methacholine was Dmin 1.37 ± 0.56 U in CVA and 0.71 ± 0.46 U in classic asthma. There was no significant difference in these three parameters. There was only a significant difference in v̇25 between CVA and classic asthma, 80.0 ± 6.9 and 52.2 ± 10.0%, respectively. Eosinophil inflammation was almost the same in both CVA and classic asthma.
T Cell Vaccination Eliminates Antigen-Specific T Cells and Prevents Antigen-Induced Eosinophil Recruitment into the Tissue
Authors: Kasai Masaaki , Kurasawa Kazuhiro , Nakajima Hiroshi , Iwamoto Itsuo
Published: April 2003

Asthma is characterized by airway inflammation with prominent eosinophil infiltrates. In a murine model of asthma, antigen-induced eosinophil recruitment into the airways of sensitized mice is mediated by CD4+ T cells and their cytokines, especially IL-5. In the present study, using ovalbumin-specific T cell receptor transgenic mice, we found that T cell vaccination, which was the administration of preactivated and attenuated antigen-specific T cells by the intraperitoneal route, prevented antigen-induced eosinophil recruitment into the airways. This effect was antigen-specific because ovalbumin-specific T cell vaccination did not affect BSA-induced eosinophil recruitment into the airways. We also found that antigen-specific IgE production as well as antigen-induced proliferation and cytokine production of splenocytes were diminished by T cell vaccination. Moreover, flow-cytometric analyses revealed that T cell vaccination eliminated antigen-specific T cells in the periphery. Together, these results indicate that T cell vaccination prevents antigen-induced eosinophil recruitment into the airways presumably by eliminating antigen-specific T cells.
Influence of Maternal Murine Immunization with Dermatophagoides pteronyssinus Extract on the Type I Hypersensitivity Response in Offspring
Authors: Fusaro A.E. , Maciel M. , Victor J.R. , Oliveira C.R. , Duarte A.J.S. , Sato M.N.
Published: April 2003

Background: Maternal exposure to environmental ubiquitous allergens could exert an influence on the newborn’s immune repertoire and the later development of allergy. The aim of this study was to investigate the effects of maternal immunization with Dermatophagoides pteronyssinus (Dp) on the hypersensitivity response and IgG subclass production in offspring using a murine model. Methods: A/Sn mice were immunized with Dp before mating with normal A/Sn males. Diaplacental serum samples were collected from newborn mice delivered by cesarean section, and maternal milk samples were extracted from the stomachs of newborn mice. Groups of offspring 25 or 45 days old were Dp immunized and boosted on the 10th day after sensitization. The animals were bled 7 days after the booster. Results: High levels of anti-Dp IgG subclasses – mainly IgG1, but also IgG2a and IgG2b – were transmitted by immunized mice via the placenta to the offspring. In the milk from immunized mothers, significant levels of anti-Dp IgG subclasses and anti-Dp IgM and IgA antibodies were detected. Moreover, the increase in total IgA antibodies in the milk of the immunized females correlated with a significantly increased level of TGF-β1. TGF-β2 levels were markedly higher than the β1 isoform in the milk, although no difference was observed between the groups. When offspring from immunized mothers were sensitized at 25 days, a significant decrease in total and anti-Dp IgE antibodies as well as total and anti-Dp IgG1, IgG2a and IgG2b subclasses was observed compared to normal female offspring, whereas when offspring were sensitized at 45 days, both offspring groups showed similar levels of IgE and IgG subclasses. Conclusions: Our study showed that maternal immunization with Dp promotes the transference of specific antibodies and/or TGF-β, which can negatively modulate the allergic response in offspring, and suggests that maternal preexposure to allergen before mating can protect mice during the early phase.
A Candidate Vaccine against Influenza Virus Intensively Improved the Immunogenicity of a Neutralizing Epitope
Authors: Lu Yun , Ding Jian , Liu Wanli , Chen Ying-Hua
Published: April 2003

Background: The hemagglutinin (HA) of influenza viruses is one of the major targets of the humoral response. The role of serum antibody to HA in the protection against infection has been demonstrated by long-standing observation. In previous studies, we suggested that an epitope vaccine might be a new strategy against the virus. Methods: HA sequences of 491 H3 subtype strains from the influenza sequence database were compared and analyzed. To acquire information on the immunogenicity of the F3 epitope, F3-epitope-specific antibody levels in 81 patient sera infected with influenza virus were tested by ELISA. Based on the theory of the epitope vaccine, we designed an epitope peptide F3 (C-KAYSNCYPYDVPDY-G-KAYSNCYPYDVPDY), which contains the repeated F3 epitope KAYSNCYPYDVPDY (aa92–105) on HA (H3N2). The specificity and the titer of the antibodies induced by the epitope vaccine were determined by ELISA. The neutralizing activities of these anti-F3 antibodies were shown by inhibiting influenza virus infection of MDCK cells. Results and Conclusion: Comparison of HA sequences of 491 H3 subtype strains indicates that this epitope is highly conservative. Analysis of the sera from influenza virus-infected patients revealed a very low level of F3 epitope-specific antibodies, suggesting the poor immunogenicity of the F3 epitope on influenza virus. The epitope vaccine based on the F3 epitope induced high levels of F3 epitope-specific antibodies recognizing the epitope peptide F3 (antibody titer in antisera up to 1:25,600). Besides, the antisera could also recognize the natural HA in Western blotting. Interestingly, these antisera induced by the epitope vaccine could inhibit infection of MDCK cells by influenza virus (strain A/Wuhan/359/95) in the neutralization assay. These results suggest that the epitope vaccine can intensively increase the immunogenicity of neutralizing epitopes and may provide a new way to develop an effective vaccine against influenza virus.
In utero Hematopoietic Stem Cell Transplantation: A Caprine Model for Prenatal Therapy in Inherited Metabolic Diseases
Authors: Lovell Kathryn L. , Kraemer Stacey A. , Leipprandt Jeffrey R. , Sprecher David J. , Ames N. Kent , Nichols-Torrez Jodi , Carter Kawanaa D. , Rahmani Darush K. , Jones Margaret Z.
Published: April 2003

Objectives: We explored the feasibility and efficacy of in utero hematopoietic stem cell transplantation in the caprine animal model system with the objectives of determining procedures for transplantation and establishing methods for detecting engraftment. Methods: Male fetal liver hematopoietic stem cells were injected into female fetuses during the immunotolerant period, using either hysterotomy or ultrasound-guided injections. Results: The rate of fetal death was much lower for the ultrasound-guided injections. Donor cells were observed in the peritoneal fluid of 4 fetuses 3 days after injection, but no donor cells were detected in tissues at longer time periods. Conclusions: Ultrasound-guided injection of hematopoietic stem cells into the abdomen of a developing fetus is safe and feasible. The parameters required for successful engraftment have not yet been identified.
Congenital Parvovirus B19 Infection: Experience of a Recent Epidemic
Authors: Kailasam C. , Brennand J. , Cameron A.D.
Published: April 2003

Objective: We report the management and outcome of 6 cases of non-immune fetal hydrops secondary to parvovirus B19 infection presenting over a 5-month period. Methods: The Queen Mothers Hospital is a tertiary referral centre for fetal medicine. All cases were suspected on the basis of ultrasound evidence of hydrops. Two cases were managed conservatively owing to the presence of an active fetus with evidence of resolving hydrops. Fetal blood sampling intra-uterine transfusion and drainage of ascitic fluid were performed in 3 cases. The 6th case unfortunately resulted in an intra-uterine death prior to fetal blood sampling. Results: Maternal parvovirus specific B19 was identified in all cases. Fetal parvovirus B19 IgM was identified in the 3 cases in whom fetal blood sampling was performed. A single intra-uterine transfusion was performed in these 3 cases; fetal hydrops resolved in 2 of these pregnancies progressing to the birth of a healthy baby at term, whereas 1 case was complicated by intra-uterine death. Fetal hydrops resolved in both cases managed conservatively, leading to the birth of a healthy baby at term. Conclusions: Parvovirus B19 infection should always be suspected in cases of non-immune hydrops. Conservative management will be appropriate in some cases and should involve weekly ultrasonography. The outlook for pregnancies presenting with gross hydrops remains guarded, even if intra-uterine transfusion is performed successfully.
A Case of Fetal Complete Heart Block Recorded by Magnetocardiography, Ultrasonography and Direct Fetal Electrocardiography
Authors: Hosono Takayoshi , Chiba Yoshihide , Shinto Mika , Miyashita Susumu , Muramaki Kazunori , Kandori Akihiko , Tsukada Keiji
Published: April 2003

Fetal magnetocardiograms (FMCGs) were recorded in a case of fetal complete heart block (CHB) from the 30th to the 37th week of gestation using the multichannel SQUID system (Hitachi, Japan). M-mode ultrasonography and direct fetal electrocardiography using needle electrodes revealed fetal CHB. We identified independent fetal P-waves and QRS complexes in the FMCG recorded in the 32nd week of gestation when the fetal atriums were close to the FMCG sensor. We also recorded FMCG P-waves in the 37th week of gestation when the fetal heart was larger. Fetal heart position and size are important for obtaining a useful FMCG. To establish FMCG as a diagnostic tool of fetal arrhythmia, comparative studies with FECG are needed.
Applicability of Four New Antibodies for the Detection of Fetal Nucleated Cells Out of Maternal Blood by FISH Analysis
Authors: Hohmann Harald , Michel Susanne , Reiber Wolfgang , Günther Michael , Claussen Uwe , von Eggeling Ferdinand
Published: April 2003

The availability of fetal cells from the maternal peripheral blood opens up the possibility for a noninvasive prenatal diagnosis. Until now this prospect has been hampered by the fact that there are no highly specific antibodies to fetal cells. In the present study, four as yet untested monoclonal antibodies – 2G5, 3F9, EPO-R and Flk1 – were analyzed in view of their effectiveness to stain fetal nucleated cells. Therefore, 20 ml of peripheral blood from 40 women carrying a male fetus were collected in the 16th to 28th weeks of pregnancy. Mononucleated cells, enriched by density gradient, were stained with one of the four antibodies and isolated with magnetic cell sorting. Subsequently, fluorescence in situ hybridization with chromosome X and Y centromere probes was performed which allowed us to analyze the maternal or fetal origin of the enriched cells. For evaluation, 500 nuclei per case were examined. The antibody 3F9 detected 0.2 cells on average (n = 5), 2G5 1.2 (n = 11), EPO-R 1.6 (n = 10) and Flk1 1.8 cells (n = 8) with fetal specific Y-positive hybridization signals. The antibody against CD71 (n = 6) used as control gave no Y-specific signals. Considering these results, the tested antibodies, especially Flk1, appear to be more successful in detecting fetal cells than the commonly used CD71 antibody.
Safety of Chronic Fetal Vascular Access in the Sheep Model
Authors: Paek Bettina W. , Lopoo John B. , Jennings Russell W. , Farmer Diana L. , Albanese Craig T. , Harrison Michael R.
Published: April 2003

Objective: Long-term access to the fetal circulation has the potential to open up new perspectives in the treatment of numerous fetal anomalies. The purpose of this study was to investigate the safety of long-term catheterization of fetal placental vessels. Methods: A midline laparotomy was performed in 4 time-mated pregnant ewes at 125 days’ gestation (term 145 days). Placental vessels were exposed by a small uterine incision. A specially designed catheter was inserted into a placental vessel over a length of 3 cm, the distal end of the catheter was tunneled underneath the maternal skin and attached to a subcutaneous port implanted in the maternal flank. All pregnancies were allowed to go to term. Results: Ewes and fetuses tolerated the placement of the catheter and port without complications. The catheter remained patent in all cases. All lambs were delivered vaginally at term and did not require resuscitation after birth. No fetal anomalies or growth restriction were noted. Conclusion: The ovine placental vessel can be accessed long term without complication.
Knowledge on Periconceptional Use of Folic Acid in Women of British Columbia
Authors: Morin Valérie I. , Mondor Myrto , Wilson R. Douglas
Published: April 2003

Objective: Our purpose was to evaluate the knowledge of folic acid and its use preconceptionally in women of British Columbia. Methods: The study was conducted at British Columbia Women’s Hospital in Vancouver, Canada, between April 15 and June 15, 1999. Pregnant women and women in the postpartum period were asked to complete a survey on folic acid. Results: In total, 1,004 women completed the questionnaire during the study period. Seventy-one percent of the women knew that vitamins could help prevent birth defects. Of those, 76.3% identified folic acid as the one vitamin specifically associated with reduction of birth defects. It was identified that 49.4% of all women took vitamins prior to pregnancy. Conclusions: Women in the population studied were relatively well informed about the benefits of folic acid, but less than 50% of them took vitamins prior to conception.
Meconium Staining and Meconium Aspiration Syndrome
Authors: Blackwell Sean C. , Carreno Carlos A. , Hassan Sonia S. , Moldenhauer Julie , Wolfe Honor M. , Berry Stanley M. , Kruger Michael , Sorokin Yoram
Published: April 2003

Objective: To determine whether the incidence of pregnancies complicated by meconium-stained amniotic fluid (MSAF) or meconium aspiration syndrome (MAS) differs with seasonal changes. Methods: An established perinatal database was used to identify all term (≧37 weeks) singleton gestations resulting in a live birth from January 1, 1997 to December 31, 1999. Patients were divided into groups based on the season of delivery: winter (December-February), spring (March-May), summer (June-August), and fall (September-November). Rates of MSAF (%MSAF/total deliveries) and MAS (%MAS/total deliveries) were calculated and compared among seasons. Local climatic data (average monthly temperature and monthly precipitation) were obtained from the National Weather Service. Multiple logistic regression analysis was performed to control for the effects of confounding variables and odds ratio (OR) with 95% confidence intervals (CI) were calculated. p < 0.05 was considered significant. Results: Over the 3-year study period there were a total of 14,888 deliveries meeting the criteria. MSAF occurred in 3,206 (21.5%) deliveries and MAS developed in 92 (0.6% of total, 2.9% of MSAF). There were no differences in the rate of MSAF (p = 0.2) or MAS (p = 0.6) between seasons. By logistic regression neither season, temperature, nor precipitation were associated with MSAF or MAS. Conclusions: Our findings suggest that over the period examined there were no significant seasonal variations in the incidence of MSAF or MAS.
Pathogenesis of Twin-Twin Transfusion Syndrome: The Renin-Angiotensin System Hypothesis
Authors: Mahieu-Caputo Dominique , Muller Françoise , Joly Dominique , Gubler Marie-Claire , Lebidois Jérome , Fermont Laurent , Dumez Yves , Dommergues Marc
Published: April 2003

In spite of active perinatal management, twin-twin transfusion syndrome (TTTS) remains a severe disease with a high risk of neonatal mortality and morbidity. TTTS initially results from an unbalanced blood flow from a donor to a recipient twin. However, its pathogenesis remains unclear, although cardiovascular disturbances and regulation of fetal volemia and diuresis seem central in this syndrome. Previously, we demonstrated that the renin-angiotensin system (RAS) was up-regulated in donor twins as a consequence of hypovolemia, and down-regulated in recipients. This was the first evidence of the implication of the RAS in TTTS. We hypothesize that the RAS plays a key role in the pathogenesis of TTTS. In the donor, RAS up-regulation aggravates oligohydramnios and may increase arterial resistance, which could contribute to placental dysfunction leading to intrauterine growth restriction. In the recipient, paradoxical RAS activation, due to transfer of effectors such as angiotensin II through placental shunts, could explain fetal vascular disturbances and cardiomyopathy. According to our hypothesis, TTTS would appear similar to the classical model of hypertension referred to as ‘2 kidneys-1 clip’ with a donor twin, comparable to the clipped kidney, intoxicating its cotwin, comparable to the normal kidney.
Bilateral Diaphragmatic Hernia Followed by Fetal Ultrasonography
Authors: Kamata Shinkichi , Sawai Toshio , Usui Noriaki , Nose Keisuke , Kitayama Yasuhiro , Nakai Hiroshi , Okuyama Hirohmi , Okada Akira , Kawaguchi Hideki , Chiba Yoshihide
Published: April 2003

Two cases of bilateral congenital diaphragmatic hernia (CDH) followed by fetal ultrasonography were described. Although many cases of CDH are diagnosed by fetal ultrasonography, it is difficult to diagnose bilateral CDH in utero, which is a relatively rare and fatal condition. Two fetuses were diagnosed as having left CDH associated with severe anomalies. However, a retrospective review of fetal ultrasonography indicated elevation of the liver in the right posterior chest. Both patients died shortly after surgical repair for left CDH despite the use of extracorporeal membrane oxygenation. Diagnosis of bilateral CDH by fetal ultrasonography and the evaluation of its prognosis were discussed.
Validity of Sonographic Formulas for Estimating Fetal Weight below 1,250 g: A Series of 119 Cases
Authors: Jouannic Jean-Marie , Grangé Gilles , Goffinet François , Benachi Alexandra , Cabrol Dominique
Published: April 2003

Objective: The aim of this study was to determine the accuracy of sonographic methods for estimating fetal weight <1,250 g on the basis of ten published formulas falling into two principal categories: general formulas applied to all fetuses, and formulas specifically developed for very-low-weight fetuses. Methods: Recent biometric data (obtained less than 7 days before birth) on 119 fetuses weighing <1,250 g were used retrospectively. Estimated fetal weights derived from ten published formulas were compared to actual weights. For each equation, the errors in predicting fetal weight were used to calculate mean percentage error and standard deviation of the mean error. The t test was used to determine whether the mean errors were significatively different from zero. The F test was used to determine if there were significant differences in the standard deviation of the mean errors. Results: The mean birth weight of infants was 956 ± (SD) 183 g at a gestational age of 29 ± (SD) 2.3 weeks. The best three formulas were the Hadlock, Sabbagha and Mielke which generated a mean error of –0.25, 2.81 and 0.29 not statistically different from zero with standard deviations of 13.02, 9.14 and 11.96, which were not statistically different. Conclusion: In our population of very-low-birth-weight infants, the use of specific formulas targeted to those fetuses does not provide a more accurate estimation of fetal weight.
First-Trimester Diagnosis of Robinow Syndrome
Authors: Percin E. Ferda , Guvenal Tevfik , Cetin Ali , Percin Sıtkı , Goze Fahrettin , Arici Sema
Published: April 2003

We present 2 cases with Robinow syndrome in a nonconsanguineous Turkish couple. The first case, second living child of the family, has all of the cardinal features of this syndrome including short stature, mesomelic shortening of forearms, frontal bossing, hypertelorism, anteverted nares, triangular mouth, hypoplastic genitalia and vertebral and costal anomalies. The second case was diagnosed with first-trimester ultrasonographic findings such as shortening of extremities and increased nuchal translucency thickness at 12 + 4 weeks of gestation, and the family wished to terminate this pregnancy. After abortion, we obtained findings such as typical face features, shortening of forearms, ambiguous genitalia suggesting Robinow syndrome with autopsy examination.
Mild Ventriculomegaly as a Counselling Challenge
Authors: Greco Pantaleo , Vimercati Antonella , De Cosmo Lucrezia , Laforgia Nicola , Mautone Angelina , Selvaggi Luigi
Published: April 2003

Objective: Our purpose was to evaluate the outcome of a group of fetuses with mild ventriculomegaly. Methods: We retrospectively collected all cases of antenatally diagnosed mild enlargement of the lateral cerebral ventricles (tranverse diameter of the atrium between 10 and 15 mm) between 1992 and 1997. Cases were included in the study if no other ultrasonic anomalies (including soft markers) were found. TORCH screening, karyotyping, search for associated anomalies and neurological examination (including imaging) were performed. Outcome information was available for all cases up to 30 months. Results: Fourteen cases were examined. TORCH was always negative, 1 case with a microdeletion of chromosome 1 was identified. During the course of pregnancy, 2 fetuses showed progression to frank hydrocephalus. Five cases of agenesis of the corpus callosum, 1 of lissencephaly and 1 of heterotopia were eventually diagnosed. After birth 2 syndromes were identified. Three babies died during the follow-up period, among the survivors 4 had severe neurological damage, 2 of them had no associated dysmorphic findings. Discussion: A heterogeneous group of central nervous system disorders may manifest antenatally as mild ventriculomegaly. The current management may not be able to identify dangerous conditions that present subtle dysmorphic features, or be able to predict abnormal outcome in most of the cases.