Total IgE In Sera Research Articles

An Antibody Persistent and Protective Two rSsCLP-Based Subunit Cocktail Vaccine against Sarcoptes scabiei in a Rabbit Model.

Scabies is a highly contagious disease caused by Sarcoptes scabiei which burrows into stratum corneum of host’s skin. In this study, after optimizing vaccination schedule, a vaccination trial is comprised of three test groups of rabbits (n = 10/group) by immunization with (1) rSsCLP5; (2) rSsCLP12; or (3) a mixture of rSsCLP5 and rSsCLP12, three biological replicates groups (n = 10/group) and three control groups (n = 10/group). Levels of specific IgG, total IgE and cytokines in sera were detected and histopathologically analyzed as indicators of vaccine effects. The results showed that 85% (17/20) of rabbits exhibited no detectable skin lesions of S. scabiei infestation in mixed protein groups compared to single protein groups with 75% (15/20) and 70% (14/20), respectively. Moreover, the deworming rates of mixed groups are increased by 10%–20% compared with that of single groups. Each of six groups immunized with rSsCLP displayed significant increases of specific IgG, total IgE, IL-10, and TNF-α. The degree of skin damage in test groups also significantly lower than that of control groups. Thus, purified rSsCLP5 and rSsCLP12 subunit cocktail vaccine induced robust immune protection and could significantly decrease mite populations to reduce the direct transmission between rabbits.

Considerations for Use of Acupuncture as Supplemental Therapy for Patients with Allergic Asthma

This study examines the clinical and immunomodulatory effects of acupuncture in the treatment of patients with allergic asthma. The acupuncture points GV14, BL12, and BL13 were selected based on the theory of traditional Chinese medicine in treating asthma. Manual acupuncture was performed once every other day (three times per week) for 5 weeks. The needles were twisted approximately 360° evenly at the rate of 60 times/min for 20 s, manipulated every 10 min and withdrawn after 30 min. Concentrations of sIgA and total IgA in secretions were determined by the combination of sucrose density gradient ultracentrifugation and RIA. Levels of cortisol in the plasma were measured by RIA. Total IgE in the sera was examined by ELISA. Flow cytometry was used to detect the numbers of CD3+, CD4+, CD8+, and IL-2R + T lymphocytes in the peripheral blood. The absolute and differential numbers of eosinophils in peripheral blood were counted with eosin staining. The total efficacy of the acupuncture treatment in patients with allergic asthma at the end of one course of treatment was 85 %. After treatment, the concentrations of sIgA and total IgA in the saliva (P<0.01, P<0.02) and nasal secretions (P<0.02, P<0.02) were significantly decreased in patients with allergic asthma. The levels of total IgE in sera (P<0.001), the counts of IL-2R + T lymphocytes (P<0.001), and the absolute and differential numbers of eosinophils (P<0.01, P<0.01) in the peripheral blood were also significantly decreased. The numbers of CD3+, CD4+, and CD8+ T lymphocytes in the peripheral blood were significantly increased in the allergic asthmatics treated by acupuncture (P<0.001, P<0.01, and P<0.001, respectively). The concentration of cortisol in the plasma of asthmatic patients did not change significantly after the acupuncture treatment (P>0.05). Acupuncture has regulatory effects on mucosal and cellular immunity in patients with allergic asthma and may be an adjunctive therapy for allergic asthma.

Generation of therapeutic antibody responses against IgE in dogs, an animal species with exceptionally high plasma IgE levels

Allergic diseases are common in dogs and the involvement of IgE in the pathogenesis of canine atopic dermatitis (CAD) and flea allergy dermatitis (FAD) is documented. However, many dogs do not achieve sufficient control of their allergic disease, and there is a great need for new treatment strategies. In order to address this issue we first needed to obtain a better picture of IgE-levels in dogs and how plasma IgE-levels are affected by breed, age and health status. IgE is normally present at diminutive concentrations in sera and detection by diagnostic methods has been a technical challenge. Here, we present a new in vitro assay for determining absolute levels of total IgE in sera from dogs. In a panel of 76 adult dogs diagnosed either as healthy or suffering from atopic dermatitis, autoimmunity or infection by skin parasites, we show that levels of IgE range from 1 to 41 μg/ml. This is almost 100 times the level observed in non-atopic humans. However, these exceptionally high IgE-levels in the dogs could not be correlated to either breed or health status. To address the issue of novel treatment strategies, the possibility of reducing the IgE-levels in nine Beagle dogs by immunization with a new therapeutic vaccine was investigated. High levels of anti-IgE antibodies were induced in all dogs, and the IgE-levels were subsequently decreased by a mean of 65%. This shows that the allergy vaccine is potent enough to break the tolerance against IgE, even when the initial IgE-levels are as high as those observed in dogs. Thus, the vaccination treatment may have the potential to serve as a future therapy for dogs with atopic diseases.

Presence of staphylococcal exfoliative toxin A in sera of patients with atopic dermatitis.

It has been reported that the toxins that Staphylococcus aureus produces are associated with the exacerbation of atopic dermatitis (AD). It has been shown in many studies that staphylococcal enterotoxin (SE) A and SEB contribute to AD by humoral immunity through IgE production as a superantigen. On the other hand, little attention has been paid to the relationship between AD and exfoliative toxin x (ETx). We investigated the toxins that are frequently detected from the skin of patients and how these toxins affect AD. S. aureus, isolated from the skin of 100 patients with mild to severe AD, were examined for the producibility of toxins by polymerase chain reaction. Serum samples were obtained from 21 patients with mild and moderate AD. The levels of SEB, ETA, total IgE, specific IgE, and specific IgG in sera were measured by ELISA. SEB was most frequently detected from S. aureus on the skin of these patients as previously reported. And ETx, to which little attention has been paid so far, was frequently detected next to SEB. Furthermore, ETA was detected from the sera of almost all the AD patients. SEB was not detected at all. Although the level of ETA in the AD group was significantly higher than that of controls, ETA-specific IgE was not detected from their sera. High levels of ETA tended to be detected from infantile patients. Although there were no significant differences in the levels of ETA-IgG between AD and the controls, its prevalence was more than twice as high as the controls in AD. These results suggest that many AD patients were exposed to ETx. We conclude that ETx may contribute to exacerbation of AD, particularly in infants, by a mechanism that is not through specific IgE production, unlike SEB.

Heterologous antigen extract in ELISA for the detection of human IgE anti-Strongyloides stercoralis.

Strongyloides ratti larval extract was used for the standardization of ELISA to detect genus-specific IgE in human strongyloidiasis. Forty serum samples from monoinfected patients shedding S. stercoralis larvae (Group I), 40 from patients with other intestinal parasites (Group II), and 40 from copronegative healthy subjects (Group III) were analyzed. Genus-specific IgE levels (ELISA Index: EI) were significantly higher in the group I (EI = 1.43) than groups II (EI = 0.70) and III (EI = 0.71), showing positivity rates of 55%, 2.5% and 0%, respectively. Similarly, sera from copropositive patients had significantly higher levels of total IgE (866 IU/mL) as compared to those from group II (302 IU/mL) and III (143 IU/mL). A significant positive correlation was found between levels of Strongyloides specific-IgE and total IgE in sera from patients with strongyloidiasis. In conclusion, S. ratti heterologous extract showed to be a useful tool for detecting genus-specific IgE by ELISA, contributing for a better characterization of the immune response profile in human strongyloidiasis.

Piezoelectric quartz crystal based label-free analysis for allergy disease

This work presents a piezoelectric (Pz) quartz crystal based label-free quantification of total IgE and allergen-specific IgE in human sera for allergy testing. An evaluation of the different brands of crystals was first initiated with respect to variability in mass sensitivity, frequency measurement reliability and stability, and surface roughness. Thereafter, for total IgE quantification, a direct assay format was adopted. By means of thioctic acid (TA) and coupling reagents, anti-human IgE antibodies were immobilized on AT-cut Pz crystals (10 MHz). The modified crystals could detect serum IgE directly corresponding to a downward frequency shift. The results showed that silver-coated crystals as compared with their gold-coated counterparts provided approximately 1.5 times higher mass detection sensitivity for total IgE in the range of 5–300 IU/ml with a linear regression line, y=1.8957 x+1.5603, R 2=0.995. For the detection of allergen-specific IgE, a sandwiched assay format was used. As the allergen-modified sensor surface captured various classes of associated antibodies (IgE, IgG, etc) and interfering serum proteins as well, the initial frequency shift downwards caused by sera sample incubation would not be proportional to specific IgE levels. Thus, following sample incubation, a second incubation step with secondary anti-human IgE was added to recognize IgE from other bound substances. The frequency shift after secondary antibody binding reflected the amount of allergen-specific IgE proportionally. Compared with 10 MHz crystals, the 20 MHz counterparts provided approximately four times higher mass detection sensitivity for allergen specific IgE in the range of 0.15–17.5 IU/ml with a linear regression line, y=50.525 x+107.777, R 2=0.954. Total IgE and allergen specific IgE assay results of real patients’ sera using the Pz sensors agreed well with those obtained by commercially available test kits with correlation coefficient 0.96–0.98. The possibility of regenerating the quartz crystals for further re-use was also dealt with.

The effect of feeding carrots on immunoglobulin E production and anaphylactic response in mice.

Carrot juice was administered orally to BALB/c mice immunized intraperitoneally with dinitrophenylated (DNP)-OVA for about 1 month. The titers of DNP-specific IgE, DNP-specific IgG, and the levels of total IgE in mouse sera were determined. The DNP-specific IgE production by mice fed carrot juice was significantly inhibited. On the other hand, the DNP-specific IgG production and the level of total IgE in mice fed carrot juice were not significantly different from those in control mice. We also examined the effect of feeding carrots on immediate-type hypersensitivity. One hour after antigen stimulation, the ears of mice fed carrots swelled less than those of control mice. Furthermore, the rise in serum histamine in the mice fed carrots under active systemic anaphylaxis was lower than in controls. We then examined the pattern of cytokine production by spleen cells from mice followed by restimulation with DNP-OVA in vitro. The spleen cells from the mice fed carrots produced more interferon-gamma than those from the control group. In contrast, the spleen cells from the mice fed carrots produced less interleukin-4 than those from the control group. Furthermore, the interleukin-12 production of the spleen cells from mice fed carrots was also higher than that of the control group. These findings suggest that feeding carrots improves the helper T cell (Th)1/Th2 balance, inhibiting specific IgE production and antigen-induced anaphylactic response.

Inhalation of diesel exhaust enhances antigen-specific IgE antibody production in mice

To examine the effects of diesel exhaust (DE) inhalation on IgE antibody production, BALB/c mice were exposed to 0 (control), 3.0 and 6.0 mg/m3 DE inhalation for 3 weeks. Intranasal sensitization with ovalbumin (OA) three times at intervals of 3 weeks was conducted immediately before, immediately after and 3 weeks after DE inhalation. Body weight and thymus weight for the DE-exposed and control mice were essentially the same but spleen weight in mice exposed to 6 mg/m3 significantly increased. Anti-OA IgE antibody titers in the sera of mice exposed to 6 mg/m3 was significantly higher than the control. Total IgE and anti-OA IgG in sera for DE-exposed and control mice remained basically the same. To investigate cytokine production in mice exposed to 6 mg/m3, spleen cells from DE-exposed and control mice were stimulated with OA in vitro and cytokine production in the culture supernatants was measured by ELISA. In vitro antigen-stimulated interleukin-4 (IL-4) and -10 (IL-10) production in spleen cells of exposed mice significantly increased compared to the control. In vitro interferon (IFN)-γ production in spleen cells of exposed mice markedly decreased. DE inhalation is thus shown to have adverse effect on antigen-specific IgE antibody production in mice through alteration of the cytokine network.

Detection of allergen-specific IgE in tears of grass pollen-allergic patients with allergic rhinoconjunctivitis.

Allergic conjunctivitis is a common symptom amongst Type I (IgE-mediated) allergic diseases; and most frequently seen as rhinoconjunctivitis. However, the site of production and the significance of allergen specific IgE needs further elucidation. We investigated whether the presence of IgE in tears of grass pollen allergic patients correlated with disease and clinical symptoms, whether the IgE binding pattern to the different grass pollen antigens was different in sera and tears, and whether IgA antibodies to grass pollen allergens were present in tears. Finally, we looked whether specific IgE was produced locally or was exudated from serum. Sera from 44 grass pollen allergic patients suffering from either allergic rhinitis (n = 11) or allergic rhinoconjunctivitis (n = 33) and from healthy controls (n = 7) were used for the experiments. Binding of specific IgE and IgA antibodies to the different groups of grass pollen allergens (Phleum pratense) was evaluated by means of immunoblotting. Specific IgE was detected in sera as well as in tears of allergic patients, whereby tear-derived allergen-specific IgE exerted similar specificities to the corresponding IgE from serum. The correlation between symptoms of ocular allergy and the presence of allergen-specific IgE in tears was highly significant (P < 0.0001). In contrast, only a poor correlation was found between specific and/or total IgE in sera and the manifestation of ocular allergy (P = 0.73). Allergen-specific IgE antibodies in tears seem to be produced locally rather than exudated from serum. IgE in tears seems to be responsible for allergic conjunctivitis. IgA in tears cannot exert a protective function since the IgA antibodies recognize different antigens in a grass pollen (Phleum pratense) extract than IgE antibodies. The highly significant correlation between allergic conjunctivitis and the presence of specific tear IgE emphasizes the diagnostic value of immunoblots with tear IgE, especially in cases in which serum provides inconclusive results.