Component-resolved diagnostics in vernal conjunctivitis.

PURPOSE OF REVIEW: Conventional diagnostic tests in allergy are insufficient to clarify the cause of vernal conjunctivitis. Component-resolved diagnostic (CRD) by microarray allergen assay may be useful in detecting allergens that might be involved in the inflammatory process. RECENT FINDINGS: In a recent trial in patients suffered from eosinophilic esophagitis, after 2 years of the CRD-guided exclusion diet and specific immunotherapy, significant clinical improvement was observed, and 68% of patients were discharged (cure based on negative biopsy, no symptoms, and no medication intake). Our new objective was to evaluate IgE-mediated hypersensitivity by CRD in tears and serum from patients with vernal conjunctivitis and treat patients with identified triggering allergens by specific immunotherapy. Twenty-five patients with vernal conjunctivitis were evaluated. The identified triggering allergens were n Lol p 1 (11 cases), n Cyn d 1 (eight cases), group 4 and 6 grasses (six cases) and group 5 of grasses (five cases). Prick test and pollen IgE were positive in one case. Clinical improvement was observed in 13/25 vernal conjunctivitis patients after 1-year specific immunotherapy. SUMMARY: CRD seems to be a more sensitive diagnostic tool compared with prick test and IgE detection. Specific CRD-led immunotherapy may achieve clinical improvements in vernal conjunctivitis patients.

Nasal and ocular responses after specific and nonspecific nasal challenges in seasonal allergic rhinitis.

BACKGROUND: Different nasal challenges induce neural and immune response leading to nasal and ocular symptoms in patients with seasonal allergic rhinitis (SAR). The release of neural mediators from nasal mucosa and conjunctiva after no-specific challenges in patients with SAR remains unknown. OBJECTIVES: To compare the release of mediators from the nose and conjunctiva with symptoms after different nasal challenges in patients with SAR. METHODS: Three types of consecutive nasal challenges were performed outside the pollen season in 25 patients with SAR. Challenges consisted of 500 biological units (BU) of allergen, 80 µg of histamine, and 1 mL of 2% hypertonic saline per nostril, within 24-hour and 72-hour intervals, respectively. Before and 15 minutes after challenges, evaluation of symptoms was performed with a visual analog scale. Concentrations of tryptase, eosinophil cationic protein in nasal lavages after 15 minutes, and substance P in tears after 5 minutes were measured with enzyme immunoassays. RESULTS: Concentrations of substance P in tears were significantly higher after nonspecific challenges. Substance P concentration in tears significantly correlated with eye itchiness after histamine and hypertonic saline and with tearing after allergen. Ocular symptoms correlated significantly with tryptase concentration in nasal lavage collected 15 minutes after allergen challenge. There is a significant correlation in tear volume comparing different nasal challenges. CONCLUSIONS: Nasal challenges with allergen, histamine, or irritants outside the pollen season induce a significant increase in nasal and ocular symptoms in patients with SAR. Interaction of the early-phase response and neurogenic inflammation define the pattern and severity of eye symptoms.

Component-resolved diagnostics in vernal conjunctivitis.

BACKGROUND: Conventional diagnostic tests in allergy are insufficient to clarify the origin of vernal conjunctivitis (VC). OBJECTIVES: To evaluate IgE-mediated hypersensitivity by component-resolved diagnosis (CRD) in tears and serum from patients with VC and to evaluate how to treat patients with identified triggering allergens by specific immunotherapy. METHODS: Patients were divided into 3 groups: (1) patients with VC (25 patients), (2) patients allergic to grass pollen with seasonal allergic conjunctivitis (AC) (50 patients), and (3) healthy blood donors (50 patients). If triggering allergens were detected, specific conventional immunotherapy was administered for 1 year. RESULTS: Twenty-five patients with VC were evaluated. The identified triggering allergens were n Lol p 1 (11 patients), n Cyn d 1 (8 patients), group 4 and 6 grasses (6 patients), and group 5 grasses (5 patients). Prick test and pollen IgE test results were positive in one patient. Clinical improvement was observed in 13 of the 25 patients with VC after 1 year of specific immunotherapy. CONCLUSION: CRD seems to be a more sensitive diagnostic tool compared with prick test and IgE detection. Specific CRD-led immunotherapy may achieve clinical improvements in patients with VC.

Conjunctival provocation with airborne allergen in patients with atopic keratoconjunctivitis.

BACKGROUND: Atopic keratoconjunctivitis (AKC) is a chronic eye disease with periods of exacerbations. Many patients experience no obvious seasonal variation, although a majority of patients are allergic to common airborne allergens. OBJECTIVE: To investigate the allergic reaction, to conjunctival provocation with airborne allergens, in patients with AKC. METHODS: Eleven patients with AKC and birch and/or grass pollen allergy participated in the study, which was performed outside the pollen season. Five patients with seasonal allergic conjunctivitis (SAC) and five healthy subjects were included for validation purposes. The challenge was performed in one eye with the allergen, to which the patient was reactive, and with dilution buffer in the other eye. Signs and symptoms from both eyes were graded at baseline and at 10 min, 8 and 48 h after provocation. Tear fluid was collected from both eyes for cytokine analyses at baseline and at 8 and 48 h. RESULTS: A significant change in clinical symptoms and signs, (redness and chemosis) was evident 10 min after provocation compared with baseline (P = 0.005) and compared with the unprovoked eye (P = 0.005) in AKC subjects. These parameters were normalized after 8 and 48 h. A significant increase for IFN-γ (P = 0.021) and IL-6 (P = 0.015), and a near significant increase for IL-10 (P = 0.066) were seen in the tear fluid of the challenged eye at 48 h after provocation vs. baseline and vs. the control eye for IFN-γ (P = 0.005), IL-6 (P = 0.028) and IL-10 (P = 0.008) in AKC subjects. CONCLUSION AND CLINICAL RELEVANCE: In this single dose allergen provocation study, AKC patients responded with a typical IgE-mediated allergic reaction. An increase in cytokines at 48 h after the challenge was demonstrated and might, with further studies, give us a better understanding of the nature of inflammation in AKC.

Specific tear IgE in patients with moderate-to-severe autumnal allergic conjunctivitis.

BACKGROUND: Recently, a new commercial test for specific IgE based on immunochromatography (Immfast Check J1®) has been developed. We previously reported on the use of this kit for tear fluid samples during spring. In this study, we compared the level of specific IgE in tear fluid among patients with seasonal allergic conjunctivitis during spring or autumn and control subjects. METHODS: A nonrandomized, cross-sectional study was conducted in patients with moderate-to-severe allergic conjunctivitis occurring in spring (n = 56, spring group) or autumn (n = 52, autumn group), as well as in age- and sex-matched healthy control subjects (n = 50, control group). Levels of specific IgE for cedar pollen, cat epithelium and Dermatophagoides pteronyssinus were measured in tear fluid with the Immfast Check J1. RESULTS: Specific IgE scores for cedar pollen and D. pteronyssinus were significantly higher in the spring and autumn groups than in the control group (p < 0.0001). In the autumn group, the highest detection rate for a specific allergen was 96.2% for D. pteronyssinus, followed by cedar pollen (86.5%) and cat epithelium (25.5%). In the spring group, the highest detection rate was 98.3% for cedar pollen, followed by D. pteronyssinus (51.7%) and cat epithelium (19.0%). CONCLUSIONS: These results suggest that both house dust mite allergen and outdoor allergens such as cedar pollen can be causes of allergic conjunctivitis during both spring and autumn in Japan. The Immfast Check J1 provides rapid measurement of specific IgE in tear fluid, which allows easy diagnosis of allergic conjunctivitis in an outpatient setting.

Rapid immunochromatographic measurement of specific tear immunoglobulin E in moderate to severe cases of allergic conjunctivitis with Immfast Check J1 in the spring.

PURPOSE: To evaluate whether Immfast Check J1, a new commercial immunochromatographic test for the measurement of specific immunoglobulin (Ig) E, could be used as a screening test for the diagnosis of allergic conjunctivitis. METHODS: A prospective, nonrandomized, cross-sectional study was conducted in 64 moderate to severe cases of allergic conjunctivitis (allergic group) and 44 age- and sex-matched healthy control subjects (control group). Specific IgE levels for cedar pollen, cat epithelium, and Dermatophagoides pteronyssinus were measured in tear fluid with the Immfast Check J1. RESULTS: Specific IgE levels in tear fluid could be assayed within 20 minutes in all subjects. The positive rate of specific IgE was significantly higher in the allergic group than in the control group (cedar pollen, 96.9% vs. 2.3%, P < 0.00001; cat epithelium, 23.4% vs. 0.0%, P = 0.00018; and D. pteronyssinus, 53.1% vs. 2.3%, P < 0.00001). Specific IgE scores were also significantly higher in the allergic group than in the control group (cedar pollen, P < 0.00001; cat epithelium, P = 0.03907; and D. pteronyssinus, P < 0.00001). CONCLUSIONS: These results suggest that Immfast Check J1 is a rapid and easy test for detection of allergen-specific IgE in tear fluid samples from patients with allergic conjunctivitis. The test is reliable and easy to perform on an outpatient basis.

Effects of multiple dexamethasone treatments on aggravation of allergic conjunctivitis associated with mast cell hyperplasia.

In a Japanese cedar pollen-induced allergic conjunctivitis model in guinea pigs, symptoms were aggravated by repeated pollen challenges. In addition, the number of mast cells in the conjunctiva was increased by multiple challenges. The amount of a mast cell mediator, histamine in ophthalmic lavage fluid was also increased by multiple challenges. In the present study, we evaluated the effects of multiple dexamethasone treatments to assess the relationship between the aggravation of symptoms and mast cell hyperplasia. Sensitized guinea pigs were challenged by dropping a pollen suspension onto their eye surface once a week until the 15th challenge. Dexamethasone (10 mg/kg, p.o.) was administered once 3 h before the 15th challenge or 3 h before every 1st--15th challenge. Mast cells in the conjunctival tissue were detected by toluidine blue staining. Histamine was fluorometrically assayed by high-performance liquid chromatography. Serum Cry j 1-specific IgE titer was measured by an enzyme-linked immunosorbent assay. The results indicated that a single treatment with dexamethasone did not affect the 15th challenge-induced symptoms; however, multiple treatments with the corticosteroid suppressed not only conjunctivitis symptoms after every challenge but also the mast cell hyperplasia and the increase in histamine in the lavage fluid. Conversely, the increase in the IgE titer in the serum was not affected by multiple treatments with dexamethasone. In conclusion, increased numbers of mast cells in the conjunctival tissue may be associated with the aggravation of allergic conjunctivitis symptoms.

Quantitative analysis of antigen specific IgE in tears in comparison to serum samples.

We determined pollen specific IgE in tears and compared these results to the concentration of specific IgE in serum samples. We obtained tears (using Schirmer strips) and serum samples from subjects with Japanese cedar (Cryptomeria japonica) pollinosis, and tested for C. japonica pollen specific IgE using a quantitative ELISA. Time kinetic analyses through the pollen season showed that specific IgE levels in tears were found to increase earlier than those in sera and reached their maximum at the end of or after the pollen season, from March to early June. In the C. japonica pollen free season, July to December, the specific IgE levels in tears decreased, although the serum levels remained relatively high. These results indicate that the quantitative assay for specific IgE in tears might be useful to identify specific eye allergens.

Brain-derived neurotrophic factor selectively enhances allergen-specific IgE production.

We studied the effect of brain-derived neurotrophic factor (BDNF) on in vitro Japanese cedar pollen (JCP)-specific IgE production by mononuclear cells from atopic keratoconjunctivitis patients with JCP allergy. BDNF enhanced JCP-specific IgE production in a dose-dependent fashion in cultures of mononuclear cells stimulated with JCP, and maximal enhancement was achieved at 10 ng/ml. In contrast, BDNF had no effect on JCP-specific IgA or IgG4 production. On the other hand, other neurotrophins, NGF, NT-3, or NGF failed to enhance JCP-specific IgE production. Moreover, anti-BDNF mAb specifically blocked BDNF-induced enhancement of JCP-specific IgE production. Study for cytokine production revealed that BDNF decreased production of Th1 cytokines, IFN-gamma and IL-12, while it had no effect on production of TH2 cytokines, IL-4, IL-10 and IL-13, in cultures of mononuclear cells stimulated with JCP. These results indicate that BDNF relatively skews cytokine pattern toward Th2 type. Collectively, BDNF may increase allergen-specific IgE production, which may in turn aggravate allergic symptoms.

The staphylococcal superantigen-like protein 7 binds IgA and complement C5 and inhibits IgA-Fc alpha RI binding and serum killing of bacteria.

The staphylococcal superantigen-like proteins (SSLs) are close relatives of the superantigens but are coded for by a separate gene cluster within a 19-kb region of the pathogenicity island SaPIn2. rSSL7 (formally known as SET1) bound with high affinity (K(D), 1.1 nM) to the monomeric form of human IgA1 and IgA2 plus serum IgA from primate, pig, rat, and horse. SSL7 also bound the secretory form of IgA found in milk from human, cow, and sheep, and inhibited IgA binding to cell surface FcalphaRI (CD89) and to a soluble form of the FcalphaRI protein. In addition to IgA, SSL7 bound complement factor C5 from human (K(D), 18 nM), primate, sheep, pig, and rabbit serum, and inhibited complement-mediated hemolysis and serum killing of a Gram-negative organism Escherichia coli. SSL7 is a superantigen-like protein secreted from Staphylococcus aureus that blocks IgA-FcR interactions and inhibits complement, leading to increased survival of a sensitive bacterium in blood.

Dissociation of allergen-specific IgE and IgA responses in sera and tears of pollen-allergic patients: a study performed with purified recombinant pollen allergens.

BACKGROUND: Trees and grass pollen allergens represent potent elicitors of allergic rhinoconjunctivitis and asthma. Little is known regarding the presence of allergen-specific IgA antibodies in sera and tears and their association with IgE responses in patients with allergic conjunctivitis. OBJECTIVE: The purpose of this study was to compare the specificities of IgE and IgA antibodies in sera and tears of pollen-allergic patients with conjunctivitis by using purified recombinant pollen allergens. METHODS: Sera and tears collected from 23 pollen-allergic and from 23 nonatopic individuals were analyzed for IgE and IgA reactivity to nitrocellulose-blotted birch and timothy grass pollen extracts. In addition, we determined the specificities of IgE, IgG(1-4), and IgA antibodies with use of a panel of purified recombinant pollen allergens (timothy grass: rPhl p 1, rPhl p 2, rPhl p 5; birch: rBet v 1, rBet v 2) in serum and tear samples by immunoblotting and ELISA. Statistical analyses of data were performed by t test and Mann Whitney U test. RESULTS: Serum and tears of many of the pollen-allergic individuals with conjunctivitis exhibited specificity for the very same pollen allergens. No allergen-specific IgE antibodies were detected in tears of nonatopic individuals. IgA antibodies in sera and tears of patients with allergic conjunctivitis were mainly directed against nonallergenic moieties and showed specificities that were significantly different from those of IgE antibodies. CONCLUSION: The dissociation of IgE and IgA responses and the lack of allergen-specific IgA antibodies in mucosal secretions (eg, tears) may contribute to allergic manifestations in target organs of atopy. Induction of allergen-specific IgA antibodies may hence be considered as a promising strategy for the treatment of mucosal forms of atopy.

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

BACKGROUND: 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. OBJECTIVE: 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. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

Effects of certain antiallergic drugs on experimental conjunctivitis in guinea pigs.

Effects of certain antiallergic drugs on experimental conjunctivitis were studied with guinea pigs. Chlorpheniramine, ketotifen and levocabastine were effective in inhibiting histamine- and antigen-induced conjunctivitis in guinea pigs. By contrast, amlexanox was only effective in inhibiting antigen-induced conjunctivitis. Topical application of antigen released 46.5 +/- 3.8% of histamine from the conjunctiva in sensitized guinea pigs. Both levocabastine and amlexanox were effective in inhibiting conjunctival histamine release induced by antigen application. Lacrimal histamine contents were also increased after challenge. The increase in the histamine content of tears was inhibited by pretreatment with levocabastine and amlexanox, but no significant effect was observed with chlorpheniramine and ketotifen. From these findings, it is concluded that certain antiallergic drugs, but not amlexanox, exhibited potent inhibition on experimental conjunctivitis in guinea pigs. In addition, it has been established that measurement of histamine in the conjunctiva and tears as well as observation of conjunctivitis syndromes are useful for evaluating the effectiveness of antiallergic drugs on various kinds of allergic conjunctivitis in clinical situations.

Secretory IgA response in oral immunotherapy. Investigation in birch pollinosis.

A rationale for oral immunotherapy (OIT) might be founded on two potential mechanisms: induction of a mucosal secretory IgA response, or induction of systemic hyporesponsiveness (oral tolerance). Previous studies have shown clinically that there is a beneficial effect of OIT in birch pollinosis, in both children and adults. During OIT, birch pollen antigens in enterocoated capsules were given to 20 adults (participating in a double-blind, placebo-controlled trial) and 10 children, all suffering from birch pollinosis. Saliva and tears (only adults) samples were collected before, during, and after OIT. Each sample was assayed for both IgA antibodies against birch pollen antigens and total IgA by enzyme-linked immunosorbent assay. IgA antibody levels were also expressed in relation to total IgA concentrations, to correct for variations in secretion and flow rate between subjects and at different times. Changes in birch-specific secretory IgA antibodies in saliva and tears could not explain the beneficial effect of OIT in birch pollinosis. Further studies in this field are warranted.

Epidemiologic factors and isotype-specific antibody responses in serum and mucosal secretions of dairy calves with bovine coronavirus respiratory tract and enteric tract infections.

Blood, feces, and nasal swabs specimens were collected 12 to 24 hours after birth and then 3 times/week (blood only once per week) from one group of 10 calves until they were 10 weeks old and from a second group of 10 calves until they were 10 to 20 weeks old. Colostrum was collected from all calves' dams and tears from 5 randomly selected calves in the first group. All fecal and nasal specimens were assayed for bovine coronavirus (BCV) antigens by ELISA. Nasal epithelial cells were examined for BCV antigens by direct immunofluorescence. Isotype antibody titers to BCV in all samples from 5 calves in group 1 were evaluated by ELISA. Zinc sulfate turbidity (ZST) values were determined on the first serum samples taken from all calves in group 1. To determine whether any correlation existed between ZST values, isotype antibody titers to BCV (12 to 24 hours after birth), number of respiratory sick days, number of enteric sick days, or days to first shedding of virus, a Spearman rank order correlation coefficient was done. Bovine coronavirus respiratory tract and enteric tract infections were common on this farm. Most initial infections developed when calves were 1 to 3 weeks old; however, there were also multiple incidences of shedding of viral antigens or seroconversions at later times during the study. Persistence of infection or reinfection of the upper respiratory tract with BCV was common. Colostral antibody titers to BCV (IgG1) were in all cows at moderate amounts; however, calf serum antibody titers and ZST values (12 to 24 hours after birth) were highly variable.(ABSTRACT TRUNCATED AT 250 WORDS)

Mucosal and systemic antibody responses to bovine coronavirus structural proteins in experimentally challenge-exposed calves fed low or high amounts of colostral antibodies.

Ten colostrum-deprived calves were assigned to 1 of 2 treatment groups (5 calves/group), and fed colostrum that had either low (naturally infected cows) or high (immunized cows) antibody titers to bovine coronavirus (BCV). All calves were inoculated orally and intranasally with virulent BCV when they were 24 to 48 hours old and challenge exposed 21 days later. Blood, feces, nasal secretions, tears, saliva, and bronchoalveolar lavage (BAL) fluids were collected weekly from each calf for 5 weeks after inoculation. The titers to whole BCV or the relative amounts of isotype-specific antibodies to BCV structural proteins were evaluated in these samples by ELISA or immunoblotting, respectively. Both pools of colostrum contained primarily IgG1, IgG2, and IgA antibodies to the E2 and E3 BCV proteins. Calves fed the high-titer colostrum had correspondingly higher amounts of passive IgG1 and IgA antibodies to whole BCV and to the E2 and E3 BCV proteins in serum, feces, and BAL fluid at postinoculation week 1 than those calves fed low-titer colostrum. Active IgG1, IgA, and IgM antibody responses in serum and active IgA and IgM antibody responses in most mucosal secretions to whole BCV and to the E2 and E3 proteins were lower or delayed in calves fed high-titer colostrum, compared with responses in calves fed low-titer colostrum. In contrast, increased responses to the BCV N protein were observed in all samples (except in serum and BAL fluid) in the calves fed high-titer colostrum, compared with calves fed low-titer colostrum. Upon challenge exposure, responses to E2 and E3 BCV proteins in serum and BAL fluid were lower in the group fed high-titer colostrum, compared with those in the group fed low-titer colostrum. Our findings indicate that the level of passive immunity in calves at the time of BCV inoculation can influence the development of active antibody responses in serum, feces, and mucosal secretions to whole BCV and to some BCV proteins individually.

Protective secretory immunoglobulin A antibodies in humans following oral immunization with Streptococcus mutans.

Ingestion of a vaccine containing killed Streptococcus mutans, originally isolated from each volunteer, daily for 10 consecutive days induced increased levels of specific secretory immunoglobulin A (sIgA) antibodies to S. mutans cells and two cell surface proteins, glucosyltransferase and surface antigen I/II, in parotid saliva and tears of four healthy males and in parotid saliva, tears, colostrum, and milk of a pregnant woman. In addition, these antibodies inhibited glucosyltransferase activity. Both IgA1 and IgA2 antibodies were induced. The levels of IgA antibodies in all secretions remained significantly above preimmunization levels for more than 50 days after oral administration of antigen. A second series of immunizations for 7 consecutive days resulted in even higher levels of sIgA antibodies, which peaked earlier and persisted longer than those observed after the primary immunizations. No increase in levels of antibodies in serum were detected in any subject. Antibodies reactive with human heart and kidney antigens could not be detected in saliva, tears, colostrum, milk, or serum samples collected at any time during the immunization regimen. The numbers of viable S. mutans organisms in dental plaque and whole saliva decreased after each series of immunizations, which correlated with increased levels of IgA antibodies in saliva, suggesting that IgA antibodies in saliva were responsible for the reduced adherence of this bacterium. These results indicate that ingested S. mutans antigen induces secretion of specific IgA1 and IgA2 antibodies in saliva, tears, colostrum, and milk, providing further evidence for the existence of a common mucosal immune system.