Assay concept for detecting anti-drug IgM in human serum samples by using a novel recombinant human IgM positive control.

Aim: Development and qualification of an easy-to-use ELISA for detection of IgM anti-drug antibodies (ADA) and its use in a clinical Phase I trial. Results & methodology: During the assay development two positive control (PC) approaches, the preparation of a chemically conjugated and a recombinant PC, were pursued. With both PCs, the assay was developed and successfully qualified considering the regulatory guidelines. For a case study, the IgM ADA isotyping assay with the recombinant PC was selected. Different courses and intensities of immune response regarding IgM signals were demonstrated. Conclusion: The easy-to-use ELISA allowed IgM-ADA detection in clinical samples. Conjugated and recombinant IgM PCs were comparable regarding assay sensitivity, precision and suitability.

Randomized, Double-Blind, Placebo-Controlled Trial of the Effect of Vitamin D3 on the Interferon Signature in Patients With Systemic Lupus Erythematosus.

OBJECTIVE: Vitamin D modulates the immune response and blocks induction of an interferon (IFN) signature by systemic lupus erythematosus (SLE) sera. This study was undertaken to investigate the effects of vitamin D supplementation on the IFN signature in patients with SLE. METHODS: SLE patients (n = 57) with stable, inactive disease, a serum 25-hydroxyvitamin D (25[OH]D) level ≤20 ng/ml, an elevated anti-double-stranded DNA antibody level, and an IFN signature (as determined by measuring the expression levels of 3 IFN response genes) were randomized into a 12-week double-blind, placebo-controlled trial of vitamin D3 at doses of 2,000 IU or 4,000 IU. An IFN signature response was defined as a 50% reduction in the expression of 1 of the 3 genes or a 25% reduction in the expression of 2 of the 3 genes. Disease activity, adverse events, and endocrine effects were assessed. RESULTS: Baseline characteristics of the patients in the 3 treatment groups (placebo, low-dose vitamin D3 , or high-dose vitamin D3 ) were similar. Repletion of 25(OH)D (i.e., levels ≥30 ng/ml) was not observed in any of the patients who were receiving placebo, while repletion was observed in 16 of 33 patients receiving vitamin D3 . The percentage of patients with an IFN signature response did not differ among the treatment groups. Moreover, there was no difference in the percentage of patients with an IFN signature response between those who remained vitamin D deficient and those who demonstrated repletion of vitamin D. Modular microarray analysis of a subset of patients (n = 40) did not reveal changes from baseline in any modules (including the IFN-inducible module) in any of the treatment groups, and no differences in expression were found between patients who demonstrated vitamin D repletion and patients who were persistently vitamin D deficient. Vitamin D3 was well tolerated, and there were no safety concerns. CONCLUSION: Vitamin D3 supplementation up to 4,000 IU daily was safe and well-tolerated but failed to diminish the IFN signature in vitamin D-deficient SLE patients. Higher 25(OH)D levels sustained for a longer duration may be required to affect immunologic outcomes.

Anti-M antibodies as a cause of intrauterine fetal death and neonatal hyperbilirubinaemia.

A preterm male infant (35 weeks), appropriate for gestational age with birth weight of 2.20 kg was born to a 28-year G2 P0 mother. The mother's blood group was A positive and the father's was B positive. Her first pregnancy was an intrauterine fetal death due to immune hydrops. The mother's blood was positive for indirect Coomb's test with 1:32 dilution and anti-M antibodies. This pregnancy was induced at 35 weeks of gestation. Investigations from the cord blood revealed A positive blood group, positive direct Coomb's test, haematocrit of 41.4%, cord reticulocyte count of 5.3% and total serum bilirubin (TSB) of 2.7 mg/dL. Phototherapy was started at 27 h of life for visible jaundice. In view of progressive pallor and a sudden rise of bilirubin, the infant was subjected to exchange transfusion on day 5 of life. The transfusion was given with O negative and anti-M antibodies negative donor blood. Total serum bilirubin (TSB) prior to exchange transfusion was 28 mg/dL and packed cell volume (PCV) was 21%. Phototherapy was continued for a total duration of 8 days.

[Effect of ningdong granule on stereotyped behaviors in Tourette syndrome model rats of different Chinese medical syndromes].

OBJECTIVE: To observe the effect of Ningdong Granule (NDG) on stereotyped behaviors in Tourette's syndrome (TS) model rats of different Chinese medical syndromes. METHODS: Thirty-two Wistar rats were used to establish TS models of different Chinese medical syndromes (n =8) induced by TS children patients' sera of 4 syndromes, i.e., Xin-Gan deficiency syndrome (XGDS), Gan-Shen yin deficiency syndrome (GSYDS), sputum-turbid blocking aperture syndrome (STBAS), and Gan hyperactivity Pi deficiency syndrome (GHPDS). Corresponding sera was micro-infused to them while administering NDG (120 mg/kg each time, thrice daily, for 3 successive weeks). Besides, another normal control group (n =8) was set up by injecting sera from healthy children plus intragastric perfusion of normal saline. Stereotyped behaviors were recorded on the 1st, 7th, 14th, and 21st day after administration of NDG. RESULTS: The anti-neural antibody serum concentration in TS children was significantly higher than that in healthy control [(1.28 +/- 0.36) UL vs. (0.52 +/- 0.24) U/L, P < 0.01 ]. It was (1.34 +/- 0.41) U/L in the XGDS group, (1.19 +/- 0.51) U/L in the GSYDS group, (1.29 +/- 0.61) U/L in the STBAS group, and (1. 17 +/- 0.45) U/L in the GHPDS group, showing no statistical difference (P > 0.05). There was no statistical difference in stereotypic behaviors of rats after treatment among the four different Chinese medical syndromes (P > 0.05). At day 7, 14, and 21 after treatment by NDG, the times of stereotyped behaviors were significantly less in the XGDS group than in the other three groups at the same time points except in the GHPDS group at day 14 (P < 0.05, P < 0.01). Meanwhile, the total numbers of stereotyped behaviors in the XGDS group [(42.8 +/- 12.6)] was obviously superior to that in the GSYDS group [(29.3 +/- 13.7)], the STBAS group [(21.9 +/- 10.4)], and the GHPDS group [(30.6 +/- 9.6)], showing statistical difference (P < 0.01, P < 0.05) after treatment by NDG at day 21. CONCLUSIONS: The anti-neural antibody serum concentration in TS children was significantly higher than that in healthy children. Stereotyped behaviors could be induced in rats after intrastriatal micro-infusion of TS sera rich in anti-neural antibody. TS model rats of XGDS were better improved than rats in the other 3 groups after treatment by NDG.

Amplified electrochemiluminescence of luminol based on hybridization chain reaction and in situ generate co-reactant for highly sensitive immunoassay.

In this work, we described a simple and highly sensitive electrochemiluminescence (ECL) strategy for IgG detection. Firstly, L-cysteine functionalized reduced graphene oxide composite (L-cys-rGO) was decorated on the glassy carbon electrode (GCE) surface. Then anti-IgG was immobilized on the modified electrode surface through the interaction between the carboxylic groups of the L-cys-rGO and the amine groups in anti-IgG. And then biotinylated anti-IgG (bio-anti-IgG) was assembled onto the electrode surface based on the sandwich-type immunoreactions. By the conjunction of biotin and streptavidin (SA), SA was immobilized, which in turn, combined with the biotin labeled initiator strand (S1). In the presence of two single DNA strands of glucose oxidase labeled S2 (GOD-S2) and complementary strand (S3), S1 could trigger the hybridization chain reaction (HCR) among S1, GOD-S2 and S3. Herein, due to HCR, numerous GOD was efficiently immobilizated on the sensing surface and exhibited excellent catalysis towards glucose to in situ generate amounts of hydrogen peroxide (H2O2), which acted as luminol's co-reactant to significantly enhance the ECL signal. The proposed ECL immunosensor presented predominate stability and high sensibility for determination of IgG in the range from 0.1 pg mL(-1) to 100 ng mL(-1) with a detection limit of 33 fg mL(-1) (S/N=3). Additionally, the designed ECL immunosensor exhibited a promising application for other protein detection.

A rapid assay for on-site monitoring of infliximab trough levels: a feasibility study.

Monitoring levels of biologicals against tumor necrosis factor (TNF) has been suggested to improve therapeutic outcomes in inflammatory bowel diseases (IBDs). This pilot study describes a rapid lateral flow (LF)-based assay for on-site monitoring of serum trough levels of humanized monoclonal antibody infliximab (IFX). The applied chromatographic method utilizes sequential flows of diluted serum, wash buffer, and an immunoglobulin generic label on LF strips with a Test line comprised of TNF-α. The successive flows permitted enrichment of IFX at the Test line before the label was applied. The label, luminescent upconverting phosphor (UCP) particles coated with protein-A, emits a 550-nm visible light upon excitation with 980-nm infrared light. IFX concentrations were determined through measurement of UCP fluorescence at the Test line. The assay was optimized to detect IFX levels as low as 0.17 µg/mL in serum. For patients with IBD, this limit is appropriate to detect levels associated with loss of response (0.5 µg IFX/mL). The assay was evaluated with clinical samples from patients with Crohn's disease and correlated well within the physiologically relevant range from 0.17 to 10 µg/mL with an IFX-specific ELISA. Performance of the assay was further successfully validated with samples from blood donors, IFX negative IBD patients, and rheumatoid arthritis patients that had developed anti-IFX antibodies. Because of its generic nature, the assay is suited for detecting most therapeutic anti-TNF-α monoclonal antibodies.

Restoration of regulatory and effector T cell balance and B cell homeostasis in systemic lupus erythematosus patients through vitamin D supplementation.

INTRODUCTION: Systemic lupus erythematosus (SLE) is a T and B cell-dependent autoimmune disease characterized by the appearance of autoantibodies, a global regulatory T cells (Tregs) depletion and an increase in Th17 cells. Recent studies have shown the multifaceted immunomodulatory effects of vitamin D, notably the expansion of Tregs and the decrease of Th1 and Th17 cells. A significant correlation between higher disease activity and lower serum 25-hydroxyvitamin D levels [25(OH)D] was also shown. METHODS: In this prospective study, we evaluated the safety and the immunological effects of vitamin D supplementation (100,000 IU of cholecalciferol per week for 4 weeks, followed by 100,000 IU of cholecalciferol per month for 6 months.) in 20 SLE patients with hypovitaminosis D. RESULTS: Serum 25(OH)D levels dramatically increased under vitamin D supplementation from 18.7±6.7 at day 0 to 51.4±14.1 (p<0.001) at 2 months and 41.5±10.1 ng/mL (p<0.001) at 6 months. Vitamin D was well tolerated and induced a preferential increase of naïve CD4+ T cells, an increase of regulatory T cells and a decrease of effector Th1 and Th17 cells. Vitamin D also induced a decrease of memory B cells and anti-DNA antibodies. No modification of the prednisone dosage or initiation of new immunosuppressant agents was needed in all patients. We did not observe SLE flare during the 6 months follow-up period. CONCLUSIONS: This preliminary study suggests the beneficial role of vitamin D in SLE patients and needs to be confirmed in randomized controlled trials.

Hypothermia, hypotension, hypersomnia, and obesity associated with hypothalamic lesions in a patient positive for the anti-aquaporin 4 antibody: a case report and literature review.

OBJECTIVE: To describe a patient positive for the anti-aquaporin 4 antibody with hypothalamic lesions showing hypothermia, hypotension, hypersomnia, and obesity. DESIGN: Case report. SETTING: University hospital. PATIENT: We describe a 21-year-old woman who was positive for anti-aquaporin 4 antibody and presented with hypothermia, hypotension, and hypersomnia owing to bilateral hypothalamic lesions as the only abnormal clinical finding. RESULTS: Immediate steroid administration resulted in significant improvement of the patient's vital signs and imaging findings; however, her cognitive impairment and sleepiness persisted, and she subsequently developed obesity. Decreased cerebrospinal fluid orexin levels and sleep studies confirmed the diagnosis of narcolepsy due to medical condition. Physicians should be aware that neuromyelitis optica spectrum disorders can initially involve the hypothalamus. CONCLUSIONS: We emphasize that measurement of anti-aquaporin 4 antibody is of clinical importance in the differential diagnosis of hypothalamic lesions.

Autologous whole blood and autologous serum injections are equally effective as placebo injections in reducing disease activity in patients with chronic spontaneous urticaria: a placebo controlled, randomized, single-blind study.

BACKGROUND: Recent demonstration of circulating anti-IgG antibodies towards IgE and its receptor (FcϵRI) has led to an interest in inducing tolerance to circulating histamine-releasing factors with autologous blood injections as a treatment option in chronic spontaneous urticaria (CU). The aim of the study was to assess the efficacy of autologous whole blood (AWB) and autologous serum (AS) injections in patients with CU compared to placebo. METHODS: A total of 88 CU patients with (+) autologous serum skin test (ASST) (59) and (-) ASST (29) were randomized into three parallel subgroups and were treated with weekly injections of AWB, AS or placebo for 10 weeks. Clinical assessments included urticaria activity score (UAS) and dermatology life quality index. RESULTS: In ASST (+) patients, the percentages of patients with >30% improvement in UAS and DLQI were 85% and 90% in AWB group, 65% and 65% in AS group and 79% and 90% in placebo group, respectively. In ASST (-) patients, these figures were 67% and 89% in the AWB group, 80% and 80% in the AS group and 60% and 70% in the placebo group. The intergroup difference for complete subsidence was not statistically significant. CONCLUSIONS: Even though we could not show a better efficacy than placebo, autohemotherapy resulted in a marked decrease in disease activity and improvement in quality of life scores in CU patients.

Inhibitory effects of the root extract of Dipsacus asperoides C.Y. Cheng et al T.M.Ai on collagen-induced arthritis in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Dipsaci radix, the dried root of Dipsacus asperoides C.Y. Cheng et al T.M.Ai is used as a medicinal plant in oriental clinics for the treatment of bone diseases and functions by strengthening bone and healing bone fractures. AIM OF THE STUDY: This study investigated the therapeutic efficacy of Dipsaci radix in treating rheumatoid arthritis using a type II collagen (CII)-induced arthritis (CIA) mouse model. MATERIALS AND METHODS: Arthritis was induced in male DBA/1 mice by immunization with CII. Dipsaci radix water (DR-W) extract at 50mg/kg and 100mg/kg was orally administered from days to after the induction of arthritis. Arthritic score, serum levels of anti-CII IgG2a, the inflammatory mediator prostaglandin E(2) (PGE(2)), and inflammatory cytokines (TNF-α, IL-1ß and IL-6), and histological changes in the ankle joint were analyzed in CIA mice. RESULTS: Arthritic induction increased the arthritic score, as well as serum levels of anti-CII IgG2a antibody, PGE(2), TNF-α, IL-1ß and IL-6 in mice. However, administration of DR-W extract in CIA mice significantly reduced arthritic scores and serum levels of anti-CII IgG2a antibody, PGE(2), TNF-α, IL-1ß and IL-6 compared with those in vehicle-treated CIA mice. Furthermore, histopathological improvement in joint architecture was also observed in DR-W extract-treated CIA mice. CONCLUSIONS: DR-W extract has anti-inflammatory and anti-arthritic effects in arthritic mice. This suggests that Dipsaci radix might be used as a therapeutic agent for the treatment of human arthritis.

Effects of dietary soy intake on maternal thyroid functions and serum anti-thyroperoxidase antibody level during early pregnancy.

Soy and its isoflavones have been suggested to suppress thyroperoxidase (TPO), induce goiter, inhibit deiodinase, and modulate immune functions. This study initially investigated the effects of dietary soy consumption on maternal thyroid functions and anti-TPO antibody (TPOAb) production during early pregnancy. Data were collected through questionnaire from 505 women enrolled during early pregnancy by random sampling in Shenyang, China. Based on soy intake frequency, the subjects were divided into three groups (frequent [three or more times per week], conventional [more than twice per month but less than three times per week], and occasional [two or fewer times per month]). Serum thyrotropin (TSH), free thyroxine (FT(4)), and TPOAb were measured by chemiluminescence immunoassay. Additionally, the concentrations of two primary isoflavones (daidzein and genistein) and creatinine were assessed in the spot urine samples from representative subjects (about 20%) randomly selected from the three groups. The percentages of frequent, conventional, and occasional consumers were 18.6%, 62.6%, and 18.8%, respectively. No difference was found in age, medical records, family history of thyroid diseases, serum FT(4), TSH, and TPOAb levels, TPOAb-positive percentages, or prevalence of thyroid dysfunctions among the groups. Both urinary daidzein and genistein levels were significantly higher in the frequent consumers compared with the other two groups. No correlations were found between urinary isoflavone levels and serum FT(4) or TSH. Urinary isoflavone levels were not significantly different between TPOAb-positive and -negative women among the randomly selected representative subjects. On the whole, our findings suggest dietary soy consumption during early pregnancy is not associated with the development of thyroid dysfunction or autoimmunity.

Cycloartenyl ferulate, a component of rice bran oil-derived gamma-oryzanol, attenuates mast cell degranulation.

IgE-targeting therapy could provide significant progress in the treatment of allergic inflammation. In this study, we examined the effect of cycloartenyl ferulate (cycloartenol ferulic acid ester; CAF), a natural product from rice bran oil-derived gamma-oryzanol, on allergic reaction. When CAF and gamma-oryzanol were injected intradermally with anti-DNP IgE into the dorsal skin of rats, the passive cutaneous anaphylaxis reaction induced by DNP-HSA was attenuated. CAF and gamma-oryzanol also inhibited the degranulation of DNP-IgE sensitized RBL-2H3 mast cells stimulated with anti-DNP-HSA. IgE conjugated with CAF could not be detected by anti-IgE antibody in the ELISA analysis. Although incubation of IgE with CAF did not decrease the amount of IgE, it was possible to precipitate IgE by centrifugation. These results demonstrate that CAF captures IgE, prevents it from binding to FcepsilonRI, and attenuates mast cell degranulation.

Maternal IgG anti-A and anti-B titres predict outcome in ABO-incompatibility in the neonate.

AIM: To evaluate predictors for risk of severe hyperbilirubinaemia and kernicterus in ABO-incompatible neonates with emphasize on maternal IgG anti-A/-B titres. METHODS: Blood group O women in labour at Oslo University Hospital, Ullevål, were included in the years 2004-2006. Offspring with blood group A or B had direct antiglobulin test performed and IgG anti-A/-B levels measured in maternal plasma. Blood group A or B infants developing severe hyperbilirubinaemia, received in addition to phototherapy, immunoglobulin treatment and/or exchange transfusion (EXT). RESULTS: Of 253 neonates, 61.3% had blood group O, 29.6% blood group A and 9.1% blood group B. Twenty neonates with blood group A or B received at least one immunoglobulin treatment. In multivariate analysis, maternal antibody-titres were the only significant predictors for immunoglobulin treatment (p < 0.0001), EXTs (p < 0.05) and duration of phototherapy (p < 0.0001). The need for invasive treatment increased sharply for antibody titres > or =512. Receiver operating characteristic analyses demonstrated that titres > or =512 had a sensitivity of 90% and a specificity of 72% for predicting immunoglobulin treatment and thus severe hyperbilirubinaemia. CONCLUSION: Maternal IgG anti-A/-B titres contribute to the prediction of risk of severe hyperbilirubinaemia in ABO-incompatible neonates, in addition to blood-grouping and direct antiglobulin-testing, especially following early discharge after delivery.

Serum levels of anti-cyclic citrullinated peptide antibodies are associated with a beneficial response to traditional herbal medicine (Kampo) in rheumatoid arthritis.

This study investigated the usefulness of biomarkers indicating beneficial response to traditional herbal medicine (THM) among patients with rheumatoid arthritis (RA). We assessed 34 RA patients who received keishinieppiittokaryojutsubu (KER), one of the representative THM. The observational term was 12 months, and we calculated the disease activity score of 28 joints every 3 months and evaluated the response to KER using European League Against Rheumatism (EULAR) response criteria. Additionally, serum levels of anti-cyclic citrullinated peptide antibody (ACPA) were measured by enzyme-linked immunosorbent assay at the baseline and after 6 and 12 months of the treatment with KER. As a result, 14 (41.2%) of the 34 patients were defined as responders, 13 as non-responders and 7 as out of assessment after 6 months, respectively. Pretreatment levels of serum ACPA were lower in KER responders than in non-responders (P = 0.042), although other univariate analysis did not show any significant differences in baseline clinical measures between the two groups. Furthermore, responders to KER showed a significant decrease in the serum levels of ACPA. These findings suggest that pretreatment serum levels of ACPA are a useful predictor of a good response to treatment with KER. Furthermore, a decrease in serum levels of ACPA may be an adjunctive indicator in predicting the efficacy of this kind of treatment.

[Foetomaternal erythrocyte incompatibilities: from immunohaematologic surveillance of pregnant women to haemolytic disease of the newborn]. / Incompatibilités foetomaternelles érythrocytaires (IFME): de la surveillance immunohématologique des femmes enceintes à la maladie hémolytique du nouveau-né (MHNN).

Despite the generalization of prevention measures against foetomaternal alloimmunization with anti-D immunoprophylaxis since 1970s, retrospectively 30 years later, its complications (new-born child's severe haemolytic disease, foetal death by anemia or nuclear icterus by bilirubin encephalopathy) have not disappeared. At the same time, alloimmunizations against antigens other than D increase with no possible prevention. As part of the set up in France of regional files analysing and making an inventory of serious foetomaternal incompatibilities requiring in utero or neonatal transfusion, we felt the need to synthesize current data, biological profiles (early screening of erythrocytic alloimmunization and its follow up during pregnancy, father's immunohaematologic status, evaluation of in utero immune haemolysis and impact of new non invasive techniques of diagnosis-RH1 foetal genotypage from ADN foetal of RH1--mothers' maternal plasma), clinical and paraclinical data (evaluation of foetal haemolysis by echography, recording of foetal movements and foetal cardiac rhythm), therapeutic indicators (in utero foetal transfusions or exsanguinotransfusions, neo and postnatal transfusions or exsanguinotransfusions, induced premature labour, newborn's intensive continue phototherapy and Rhesus immunoprophylaxis) in order to enable medical and paramedical professionals to carry out the specific supervision of pregnancies with foetomaternal incompatibility, the in utero, neo- and postnatal treatment of child and the efficient therapeutic prevention of anti-D alloimmunization, in a cooperative way.

Taxanes in Taxus baccata pollen: cardiotoxicity and/or allergenicity?

Diterpenic alkaloids belonging to taxine (yield: 0.18 % dry weight) and taxoids (paclitaxel, baccatine III and 10-deacetylbaccatine III, cumulated yield: 0.004 % dry weight) were isolated from Taxus baccata L. pollen. Moreover, taxoids conjugated to macromolecules were also detected by ELISA. According to these data and to semi-quantitative measurements of pollen production, a hundred-year-old yew during its flowering time releases several grams of taxanes contained in 16 to 20 kg of pollen. Toxicity assays on mice indicated that any risk of acute toxicity resulting from Taxus pollen inhalation and subsequent taxine absorption is unlikely. On the other hand, anti-paclitaxel IgG were above all detected in sera of subjects displaying hypersensitivity reactions during the pollination periods of Taxus sp. and Betula sp. This natural anti-paclitaxel IgG acquisition by individuals living in the distribution areas of these trees could be at the origin of atopic manifestations. We also postulate that such a natural sensitization could have a marked influence on the tolerance to anticancer taxoids.

IgE immune response to Ginkgo biloba pollen.

BACKGROUND: The ginkgo (Ginkgo biloba L.) continues to be planted as a shade tree in preference to other species in Seoul, Korea. The proportion of ginkgo to total shade trees was 43.2% in 1998, but the allergenic characteristics of ginkgo pollen has not been elucidated. OBJECTIVES: This study was undertaken to obtain information regarding the skin reactivity rate to ginkgo pollen in a population of Korean subjects with respiratory allergy. Possible ginkgo pollen allergens and the cross-reactivity of ginkgo pollen with other prevalent pollens were also examined. METHODS: Four hundred and forty-seven patients with asthma and/or allergic rhinitis were skin prick tested with extract of ginkgo pollen (1:20 wt/vol). Of these patients, positive skin responders (A/H ratio > or =2+) were selected for ELISA and immunoblot experiments. RESULTS: A total of 21 patients (4.7%) showed skin reactivity (A/H ratio > or =2+) to ginkgo pollen in the skin prick test. They were also cosensitized to many other tree, grass, and weed pollens. Sixteen (76%) of the 21 positive skin responders showed specific IgE responses to ginkgo pollen in ELISA. In inhibitory ELISA, IgE binding to ginkgo pollen was inhibited by more than 80% by oak, ryegrass, mugwort, and ragweed; and 34% by hop Japanese; and 10% by rBet v 2 at 10 microg/mL. In immunoblot, 10 out of 21 sera (48%) reacted to the 15-kD protein of ginkgo pollen, 9 (43%) to 33-35 kD, and 8 (38%) to 36-38 kD. In inhibitory immunoblot, IgE binding to ginkgo pollen proteins was almost completely inhibited by oak, ryegrass, mugwort and ragweed, but only partially by hop Japanese and rBet v 2. CONCLUSION: The skin reactivity rate to ginkgo pollen is approximately 4.7% in a population of Korean subjects with respiratory allergy. Since ginkgo pollen has a high cross-reactivity with other prevalent pollens, it could cause clinical symptoms during its pollen season by cross-reacting with the IgE produced in response to other pollens in patients sensitized to multiple pollens.

In vitro demonstration of IgE antibody to folate-albumin in anaphylaxis from folic acid.

BACKGROUND: Folic acid (the synthetic form of folate B vitamins in foods) is widely used in vitamin supplements. Anaphylaxis from ingestion or injection of folic acid suggests IgE antibody-mediated mechanisms, but this has not been demonstrated previously in vitro. OBJECTIVE: This study was conducted to better define the mechanism of folic acid hypersensitivity and cross-reactivity among folic acid congeners. METHODS: Skin testing was performed with folic acid congeners in a woman who developed anaphylaxis after ingestion of 2 different multivitamin preparations containing folic acid. In vitro immunologic serum studies were conducted using a folate-human serum albumin (HSA) conjugate prepared by a novel application of carbodiimide condensation. RESULTS: The patient had positive immediate-type skin test reactions to folic acid and several folate analogues including leucovorin (folinic acid). Urticaria developed during graded oral test dosing with leucovorin. Using a dot immunoblot assay or an ELISA for IgE antibody to folate-HSA, results of the patient's serum testing were positive, whereas results of sera from normal control subjects were negative, the first in vitro demonstration of IgE to a folic acid-protein conjugate. By ELISA, the positive result of the patient's serum was inhibited significantly by serum coincubation with folate-HSA, but not HSA or folic acid. CONCLUSIONS: Immediate hypersensitivity to folic acid and possibly other vitamins can be mediated by IgE antibody to conjugates formed between vitamins and self-proteins or polypeptides. Leucovorin can have clinically important immunologic cross-reactivity with folic acid. A diet rich in natural folates (pteroylpolyglutamates) appears useful as a management strategy for providing adequate nutrition to patients with folic acid hypersensitivity.