Allergic Diseases and Chronic Adenotonsillar Diseases: A Mendelian Randomization Study.

OBJECTIVE: The existing epidemiological evidence regarding the intricate relationship between allergic diseases and chronic adenotonsillar diseases (CATD) remains inconclusive. Herein, the objective of our study is to explore the causal association using Mendelian randomization (MR). METHODS: Employing data from large genome-wide association studies, a comprehensive two-sample bidirectional MR study was conducted. The studied traits encompassed allergic rhinitis (cases n = 9707, controls n = 331173), allergic asthma (cases n = 8525, controls n = 193857), allergic conjunctivitis (cases n = 18321, controls n = 324178), atopic dermatitis (cases n = 11964, controls n = 306909), and CATD (cases n = 38983, controls n = 258553). All the patients were of European descent and participants in cohort studies. The primary analysis was executed using inverse-variance-weighted MR. Furthermore, six additional MR methods (MR-Egger, weighted median, simple mode, weighted mode, MR pleiotropy residual sum and outlier, MR robust adjusted profile score) were employed to ensure the reliability and detect potential horizontal pleiotropy within the results. The estimates obtained from the MR analysis were factored into the overall effect calculation. RESULTS: Genetically anticipated outcomes demonstrated a significant association between CATD risk and allergic rhinitis (OR = 1.141, p = 6.30E-06), allergic asthma (OR = 1.115, p = 8.31E-05), allergic conjunctivitis (OR = 1.197, p = 8.69E-07), and a suggestive association with atopic dermatitis (OR = 1.053, p = 0.040). However, no substantial correlation was observed in the reverse direction. CONCLUSIONS: Findings of our study provide evidence supporting a causal role of allergic diseases in the development of CATD, whereas the converse relationship does not appear to hold true. LEVEL OF EVIDENCE: 3 Laryngoscope, 134:2653-2658, 2024.

The Protective Effects of KAT5 Inhibition on Ocular Inflammation by Mediating the PI3K/AKT Pathway in a Murine Model of Allergic Conjunctivitis.

Purpose: We aimed to explore the effect of lysine acetyltransferase KAT5 on allergic conjunctivitis (AC). Methods: The effect of KAT5 on inflammatory response during AC progression was analyzed in the experimental allergic conjunctivitis (EAC) mouse model. Results: The clinical score, permeability, total IgE, ovalbumin (OVA)-specific IgE, and IgG1/IgG2a were induced in the EAC mice, in which the overexpression of KAT5 could further enhance but KAT5 inhibitor NU9056 reduce the phenotypes. The eosinophilic infiltration was induced in EAC mice, in which the overexpression of KAT5 was able to further promote but NU9056 attenuate the phenotype. The expression of Eotaxin and RANTES and the inflammatory factors were upregulated in EAC mice and KAT5 overexpression increased, but NU9056 decreased the expression in the model. Significantly, the CD11c+ dendritic cells and CD4+ T cells infiltration in the conjunctiva was enhanced in EAC mice, whereas KAT5 overexpression induced but NU9056 suppressed the effect in the model. Mechanically, the phosphorylation of PI3K and Akt and the levels of histone H3 lysine 27 acetylation (H3K27ac) were enhanced in EAC mice, whereas the overexpression of KAT5 promoted and NU9056 repressed the phenotype in the mice. The enrichment of KAT5 and H3K27ac on PI3K promoter was increased in EAC mice, and the overexpression of KAT5 further enhanced the enrichment in the mice. Significantly, we observed similar results in the KAT5 knockout mice as well. Moreover, PI3K/AKT signaling inhibitor LY294002 reversed KAT5 overexpression-mediated phenotypes and inflammatory response after induction AC in vivo. Conclusions: Therefore we concluded that KAT5 inhibition protected against ocular inflammation by mediating the PI3K/AKT pathway in EAC mouse model.

Effects of Docosahexaenoic Acid on Chemokine Expression in Human Conjunctival Fibroblasts.

Purpose: We assessed the production of chemokines by human conjunctival fibroblasts in response to inflammation and the effects of omega (ω)-3 fatty acids on chemokine expression.Methods: Primary cultures of human conjunctival fibroblasts were incubated with interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-α). The expression of eotaxin-1 and RANTES in response to pretreatment with docosahexaenoic acid (DHA) was investigated. Moreover, western blotting was used to evaluate the effects of DHA on the activation of nuclear factor (NF)-κB and signal transducer and activator of transcription 6 (STAT6).Results: The expression of eotaxin-1 mRNA was significantly suppressed by pretreatment with DHA with IL-4 and TNF-α costimulation. RANTES expression was similarly suppressed, but the difference was not significant. The secretion of eotaxin-1 and RANTES was significantly lower in DHA-pretreated cells than in vehicle-treated cells. Western blotting for NF-κB and STAT6 showed that these proteins were downregulated in the DHA pretreatment group compared with those in the vehicle control group.Conclusion: The results of this study suggested that DHA could have applications in the management of allergic inflammation.

Self-Reported Allergic Rhinitis and/or Allergic Conjunctivitis Associate with IL13 rs20541 Polymorphism in Finnish Adult Asthma Patients.

BACKGROUND: Our aim was to observe factors associated with IL13 rs20541 polymorphism and other factors with or without allergic comorbidities such as subject-reported allergic rhinitis (AR) and/or allergic conjunctivitis (AC) symptoms in adult asthmatics. METHODS: A population-based sample of Finnish adult asthma patients (n = 1,156) and matched controls (n = 1,792) filled in a questionnaire. Asthma was diagnosed based on a typical history of asthma symptoms and lung function tests. Skin prick tests with 17 aeroallergens and blood tests including analysis of interleukin 13 (IL13) rs20541 (G/A) genotypes were performed for a subsample (n = 193). RESULTS: The proportion of asthmatics reporting AR was 61.9% and reporting AC was 40.7%. After adjustments, the presence of the IL13 rs20541A- allele (OR 3.06, 95% CI 1.42-6.58, p = 0.004) or multisensitization (adjusted OR 4.59, 95% CI 1.48-14.26, p = 0.008) was associated with AR/AC asthma. Nasal polyps and acetylsalicylic acid-exacerbated respiratory disease was also associated with AR/AC asthma. CONCLUSIONS: Adult AR/AC asthma could putatively be a phenotype, characterized by the presence of atopic and/or eosinophilic factors and a high prevalence of the IL13 rs20541A- allele. Studies on the mechanisms behind this and in other populations are needed.

Preservation of epithelial cell barrier function and muted inflammation in resistance to allergic rhinoconjunctivitis from house dust mite challenge.

BACKGROUND: An emerging paradigm holds that resistance to the development of allergic diseases, including allergic rhinoconjunctivitis, relates to an intact epithelial/epidermal barrier during early childhood. Conceivably, the immunologic and genomic footprint of this resistance is preserved in nonatopic, nonallergic adults and is unmasked during exposure to an aeroallergen. OBJECTIVE: The aim of this study was to obtain direct support of the epithelial/epidermal barrier model for allergic rhinoconjunctivitis. METHODS: Twenty-three adults allergic to house dust mites (HDMs) (M+) and 15 nonsensitive, nonallergic (M-) participants completed 3-hour exposures to aerosolized HDM (Dermatophagoides pteronyssinus) powder on 4 consecutive days in an allergen challenge chamber. We analyzed: (1) peripheral blood leukocyte levels and immune responses; and (2) RNA sequencing-derived expression profiles of nasal cells, before and after HDM exposure. RESULTS: On HDM challenge: (1) only M+ persons developed allergic rhinoconjunctivitis symptoms; and (2) peripheral blood leukocyte levels/responses and gene expression patterns in nasal cells were largely concordant between M+ and M- participants; gross differences in these parameters were not observed at baseline (pre-exposure). Two key differences were observed. First, peripheral blood CD4+ and CD8+ T-cell activation levels initially decreased in M- participants versus increased in M+ participants. Second, in M- compared with M+ participants, genes that promoted epidermal/epithelial barrier function (eg, filament-aggregating protein [filaggrin]) versus inflammation (eg, chemokines) and innate immunity (interferon) were upregulated versus muted, respectively. CONCLUSION: An imprint of resistance to HDM challenge in nonatopic, nonallergic adults was muted T-cell activation in the peripheral blood and inflammatory response in the nasal compartment, coupled with upregulation of genes that promote epidermal/epithelial cell barrier function.

Chaperone patterns in vernal keratoconjunctivitis are distinctive of cell and Hsp type and are modified by inflammatory stimuli.

BACKGROUND: Vernal keratoconjunctivitis (VKC) is a severe ocular allergy with pathogenic mechanism poorly understood and no efficacious treatment. The aims of the study were to determine quantities and distribution of Hsp chaperones in the conjunctiva of VKC patients and assess their levels in conjunctival epithelial and fibroblast cultures exposed to inflammatory stimuli. METHODS: Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, Hsp105, and Hsp110 were determined in conjunctiva biopsies from nine patients and nine healthy age-matched normal subjects, using immunomorphology and qPCR. Conjunctival epithelial cells and fibroblasts were cultured and stimulated with IL-1ß, histamine, IL-4, TNF-α, or UV-B irradiation, and changes in Hsp levels were determined by Western blotting. RESULTS: Hsp27, Hsp40, Hsp70, and Hsp90 levels increased in the patients' conjunctiva, whereas Hsp10, Hsp60, Hsp100, and Hsp105 did not. Double immunofluorescence demonstrated colocalization of Hsp27, Hsp40, Hsp70, and Hsp90 with CD68 and tryptase. Testing of cultured conjunctival cells revealed an increase in the levels of Hsp27 in fibroblasts stimulated with IL-4; Hsp40 in epithelial cells stimulated with IL-4 and TNF-α and in fibroblasts stimulated with IL-4, TNF-α, and IL-1ß; Hsp70 in epithelial cells stimulated with histamine and IL-4; and Hsp90 in fibroblasts stimulated with IL-1ß, TNF-α, and IL-4. UV-B did not induce changes. CONCLUSIONS: VKC conjunctiva displays distinctive quantitative patterns of Hsps as compared with healthy controls. Cultured conjunctival cells respond to cytokines and inflammatory stimuli with changes in the Hsps quantitative patterns. The data suggest that interaction between the chaperoning and the immune systems drives disease progression.

Culture medium from TNF-α-stimulated mesenchymal stem cells attenuates allergic conjunctivitis through multiple antiallergic mechanisms.

BACKGROUND: The immunomodulatory and anti-inflammatory functions of mesenchymal stem cells (MSCs) have been demonstrated in several autoimmune/inflammatory diseases, but their contribution to allergic conjunctivitis and underlying antiallergic mechanisms remain elusive. OBJECTIVE: We sought to explore the clinical application of MSCs to experimental allergic conjunctivitis (EAC) and its underlying antiallergic mechanisms. METHODS: Culture medium from TNF-α-stimulated, bone marrow-derived MSCs (MSC-CMT) was administered topically to mice with EAC, and the related allergic symptoms and biological changes were evaluated. Murine spleen-derived B cells, bone marrow-derived mast cells (MCs), and lung vascular endothelial cells were cultured in vitro to investigate the antiallergic MSC-CMT mechanisms. RESULTS: Topical instillation of MSC-CMT significantly attenuated the clinical symptoms of short ragweed pollen-induced EAC, with a significant decrease in inflammatory cell frequency, nuclear factor κB p65 expression, and TNF-α and IL-4 production. In vitro MSC-CMT significantly inhibited the activation of MCs and B-cell IgE release and reduced histamine-induced vascular hyperpermeability. During EAC, MSC-CMT treatment also decreased IgE production, histamine release, enrichment and activation of MCs, and conjunctival vascular hyperpermeability. The MSC-CMT-mediated inhibition of B cells, MCs, and histamine and its antiallergic effects during EAC were abrogated when MSCs were pretreated with COX2 small interfering RNA. CONCLUSIONS: Our findings provide compelling evidence that MSC-CMT inhibits EAC through COX2-dependent multiple antiallergic mechanisms and support the use of MSC-CMT as a novel strategy for treating allergic conjunctivitis.

CCL20/MIP-3 alpha mRNA expression in the conjunctival epithelium of normal individuals and patients with vernal keratoconjunctivitis.

BACKGROUND: CCL20, the single chemokine ligand for CCR6, contributes to recruiting CCR6-expressing memory B cells, memory T cells, Th17 cells and dendritic cells, and is involved in regulating immune responses, homeostasis, and inflammation in mucosal tissues. METHODS: CCL20 messenger RNA (mRNA) expression was analyzed in the conjunctival epithelium in an in vivo study of patients with vernal keratoconjunctivitis (VKC group) and healthy volunteers (control group) using impression cytology. In vitro analysis of CCL20 mRNA was performed using cultured conjunctival epithelial cells (CECs). Real-time polymerase chain reaction was used to assess IL-8 and eotaxin-2 mRNA expression for comparison with CCL20 mRNA expression. RESULTS: In the control group, CCL20 mRNA expression was present in all conjunctival locations. However, CCL20 mRNA expression was significantly higher in the upper palpebral conjunctiva in the severe VKC group than in the mild VKC and control groups (p < 0.05, Steel test). In vitro stimulation of CECs with lipopolysaccharide (LPS) significantly increased CCL20 expression in a concentration-dependent manner that was significantly correlated with expression of IL-8 (p < 0.001, Spearman's rank correlation coefficient), but not eotaxin-2. CONCLUSION: We conclude that CCL 20 mRNA expression in the conjunctival epithelium plays a crucial role in regulating homeostasis at the ocular surface and in exacerbation of VKC.

IL3 SNP rs40401 variant is a risk factor for rhinoconjunctivitis in Japanese women: the Kyushu Okinawa maternal and child health study.

BACKGROUND: No studies have investigated the relationship between IL3 single nucleotide polymorphism (SNP) rs40401 and allergic rhinitis. We performed a case-control study to examine this issue and to assess interactions between the SNP and smoking or older siblings in young adult Japanese women. METHODS: Included were 393 women who met the criteria of the International Study of Asthma and Allergies in Childhood (ISAAC) for rhinoconjunctivitis. Controls were 767 women without rhinoconjunctivitis according to the ISAAC criteria who had not been diagnosed with allergic rhinitis by a doctor. RESULTS: Compared with women with the TT genotype of SNP rs40401, those with the CC genotype had a significantly increased risk of rhinoconjunctivitis: the adjusted OR was 1.52 (95% CI: 1.05-2.19). This positive relationship was significant under the additive model: the adjusted OR was 1.23 (95% CI: 1.02-1.47). The positive association fell just short of the significance level under the dominant or recessive model. There was no significant interaction between SNP rs40401 and smoking with respect to rhinoconjunctivitis. Compared with subjects with the TT or TC genotype of IL3 SNP rs40401 who had one or more older siblings, those with the CC genotype who had no older siblings had a 2.33-fold increased risk of rhinoconjunctivitis; nevertheless, the interaction was not significant. CONCLUSION: This is the first study to show a significant positive association between IL3 SNP rs40401 variant and the risk of rhinoconjunctivitis. We could not find evidence for interactions between SNP rs40401 and smoking or older siblings affecting rhinoconjunctivitis.

Interaction between conjunctival epithelial cells and mast cells induces CCL2 expression and piecemeal degranulation in mast cells.

PURPOSE: Intraepithelial mast cells are observed in giant papillae tissue samples obtained from patients with atopic keratoconjunctivitis (AKC)/vernal keratoconjunctivitis (VKC). We examined the roles of interaction between the conjunctival epithelial cells and mast cells. METHODS: The interaction between human mast cells and conjunctival epithelial cells (HCjE) was investigated using a coculture model. Protein array analysis, ELISA, and real-time PCR were performed to test the interaction. Tissue samples (n = 6) from giant papillae were resected for therapeutic purposes, and subjected to immunohistological analysis of CCL2 expression. Recombinant CCL2 (10 ng/mL) was reacted with the cultured human mast cells and ultrastructural analysis was performed. A ragweed (RW)-induced mouse experimental allergic conjunctivitis model was used to examine ccl2 mRNA expression and mast cell morphology. RESULTS: Protein array and real-time PCR analyses showed that CCL2 protein/mRNA expression was induced by mast cell-HCjE coculture. Upregulation of CCL2 mRNA was observed in mast cells, whereas in situ CCL2 expression was observed at the conjunctival epithelium of the giant papillae by immunohistochemistry. Ultrastructural analysis showed that recombinant CCL2 treatment induced piecemeal degranulation (PMD) in the mast cells. Ultrastructural analysis of tissues from the giant papillae showed PMD of mast cells within the conjunctival epithelial cells. The RW-induced experimental allergic conjunctivitis model showed increased ccl2 mRNA expression and PMD morphology in the conjunctivae. CONCLUSIONS: Mast cell-conjunctival epithelial cell interaction induces CCL2 expression and subsequent PMD.

Prevalence and risk factors of atopic diseases in German children and adolescents.

BACKGROUND: Atopic diseases became an important health problem in affluent Western societies. METHODS: To study the prevalence and factors associated with the risk of atopic diseases in Germany, data from the German Health Interview and Examination Survey for Children and Adolescents (KiGGS) were analysed (n = 17,450). Standardized, computer-assisted personal interviews with parents and parent-administered questionnaires provided physician diagnoses of allergic rhinoconjunctivitis, atopic dermatitis and asthma as well as data on demographic characteristics, migration background, birth order, age at the beginning of nursery, atopic diseases of parents, parents' smoking status, parents' occupation, breastfeeding and living environment. RESULTS: The life-time prevalence of atopic dermatitis was 13.2% (95% confidence limit: 12.5-13.9%), 10.7% (10.1-11.3%) for allergic rhinoconjunctivitis and 4.7% (4.3-5.1%) for asthma. At least one atopic disease in parents was the strongest factor associated with atopic diseases in the offspring, with a prevalence ratio of up to 2.6. High and middle socio-economic status (prevalence ratio, 95% confidence limit: 1.28, 1.12-1.46; 1.15, 1.01-1.32) were associated with the risk of atopic dermatitis, whereas a two-sided background of migration reduced the risk (0.76, 0.65-0.88). Factors that reduced the risk of allergic rhinoconjunctivitis were parents working as self-employed farmers (0.48, 0.30-0.76) and older siblings (0.80, 0.71-0.89), whereas the beginning of nursery school at older age was associated with an increased risk in children who were cared for outside the family before school age (1.05, 1.00-1.10). Living in mould-infested rooms (1.64, 1.23-2.19), an urban living environment (1.20, 1.02-1.42) and a smoking mother and/or father (1.20, 1.02-1.40) were associated with the risk of asthma. CONCLUSIONS: Our results are in line with the so-called 'hygiene hypothesis', which emphasizes the role of environmental factors in addition to a genetic predisposition in the development of atopic diseases. Research on factors associated with atopic diseases can facilitate decisions on preventive strategies. Further studies are needed to explore trends in prevalence and risk factors for atopic diseases.

Polymorphisms in the IL4 gene, smoking, and rhinoconjunctivitis in Japanese women: the Kyushu Okinawa Maternal and Child Health Study.

Epidemiological evidence on the relationship between IL4 single nucleotide polymorphisms (SNPs) and allergic rhinitis is limited. We conducted a case-control study to investigate this issue in young adult Japanese women. Included were 393 women who met the criteria of the International Study of Asthma and Allergies in Childhood (ISAAC) for rhinoconjunctivitis. Controls were 703 women without rhinoconjunctivitis based on the ISAAC criteria who had not been diagnosed with allergic rhinitis and/or asthma by a doctor. Compared with the TT genotype of IL4 SNP rs2227284, the GG genotype, occurring in 10.2% of control subjects, was significantly inversely associated with the risk of rhinoconjunctivitis: the adjusted OR was 0.60 (95% CI: 0.37-0.98). There were no significant relationships between SNP rs2243250, rs2070874, or rs2243290 and rhinoconjunctivitis. None of the haplotypes were significantly related to rhinoconjunctivitis. A significant inverse relationship between the combination of the TG and GG genotypes of SNP rs2227284 and rhinoconjunctivitis was observed in women who had never smoked, but not in those who had ever smoked (P for interaction=0.11). This is the first study to demonstrate a significant relationship between IL4 SNP rs2227284 and rhinoconjunctivitis. Smoking may modify the relationship between SNP rs2227284 and rhinoconjunctivitis.

Histamine H4 receptors in normal conjunctiva and in vernal keratoconjunctivitis.

BACKGROUND: While it is known that histamine is the primary mediator of ocular allergy, the presence and distribution of histamine receptors are not well documented in the human eye. Our aim was to evaluate histamine receptor expression in normal and vernal keratoconjunctivitis conjunctiva. METHODS: Mucosal biopsies were obtained from conjunctiva of healthy donors and from tarsal conjunctiva of vernal patients. Immunostaining and semi-quantitative reverse-transcriptase polymerase chain reaction for H(1), H(2), H(3), and H(4) receptors were performed. Histamine receptor expression was also evaluated in conjunctival cell cultures exposed to histamine, interleukin-4, interleukin-5, interferon-γ and tumor necrosis factor-α. RESULTS: Immunostaining for H(1) and H(2) receptors was slightly positive in normal and over-expressed in vernal tissues. H(3) receptors were rarely present in normal and inflamed conjunctiva. In striking contrast to control tissues, H(4) receptors were highly expressed in all inflamed tissues, particularly by stromal inflammatory cells. Semi-quantitative reverse-transcriptase polymerase chain reaction demonstrated an over-expression of H(1), H(2), and H(4) receptors in vernal vs control tissues. Notably, H(4) receptors were five times more expressed in vernal vs control tissues. In cell cultures, H(2) receptor expression was stimulated eight times the normal levels by interleukin-4 and three times by histamine, but the H(4) receptor was only slightly affected by stimulation with these mediators. CONCLUSIONS: Increased expression of H1, and particularly of H(2) and H(4) receptors in vernal keratoconjunctival tissues indicate their important role in the pathogenesis of this disease. H(4) receptors may be a target in the treatment of allergic inflammation.

Expression patterns of HLA-DR+ or HLA-DR- on CD4+/CD25++/CD127low regulatory T cells in patients with allergy.

BACKGROUND: Allergic rhinoconjunctivitis induced by pollen is a highly prevalent chronic inflammatory disease in Europe. Parietariajudaica is a frequent trigger in the Mediterranean area. The function of regulatory T cells (Treg cells) in allergy has recently been investigated, but further data are necessary to better understand their role and to find new strategies to treat allergic diseases such as allergic rhinoconjunctivitis. OBJECTIVE: To characterize gene expression of HLA-DR+ or HLA-DR- on peripheral CD4+/CD25++/CD127low Treg cells in patients with allergy. METHODS: Peripheral Treg cells (CD4+/CD25++/CD127low) were quantified using flow cytometry and sorted according to HLA-DR expression during the pollen season in patients with allergic rhinoconjunctivitis caused by P. judaica. The results were compared with those of nonatopic controls. Expression of associated cytokines and their receptors was measured using quantitative reverse transcription-polymerase chain reaction after extraction of mRNA in sorted populations. RESULTS: During the pollen season, no significant differences were observed between allergic patients with rhinoconjunctivitis and healthy controls in terms of the absolute number or the percentage of Treg cells in peripheral blood. All patients had a higher number/percentage of HLA-DR- Treg cells than HLA-DR+ Treg cells. In both groups we found high levels of FOXP3 mRNA expression. Despite being lower in number, HLA-DR+ Treg cells presented higher expression of CD28, PRF1, GZMB, and FASL than HLA-DR-Treg cells. CONCLUSIONS: The most relevant results obtained suggest that HLA-DR+ Treg cells tend to present higher gene expression of molecules associated with contact-dependent cell activation and cytotoxicity.

Functional role of thymic stromal lymphopoietin in chronic allergic keratoconjunctivitis.

PURPOSE: Previous reports have shown that thymic stromal lymphopoietin (TSLP) plays a role in atopic diseases. This study was undertaken to investigate the expression of TSLP in the giant papillae obtained from patients with vernal keratoconjunctivitis (VKC) or atopic keratoconjunctivitis (AKC), and its functional roles were analyzed. METHODS: TSLP mRNA expression was examined in resected conjunctival samples obtained from four patients with VKC/AKC and three control subjects by reverse transcription-polymerase chain reaction. Anti-TSLP, anti-dendritic cell-limbic system-associated membrane protein (anti-DC-LAMP), and anti-tryptase immunohistochemical staining was performed with 10 resected giant papillae. Human conjunctival epithelial (HCJE) cells were stimulated with poly I:C, with and without endosomal inhibitor, to examine TSLP mRNA expression. Cultured human mast cells were stimulated with recombinant (r)TSLP to analyze the downstream effect of TSLP. RESULTS: All four VKC/AKC samples showed TSLP mRNA expression; however, no TSLP mRNA expression was found in the control conjunctivae. Anti-TSLP immunohistochemical staining showed preferential expression in the epithelial cells and some infiltrated cells of the giant papillae, but not in the control conjunctivae. Double immunohistochemical staining with TSLP and DC-LAMP or tryptase showed the existence of activated dendritic cells and mast cells near TSLP-positive cells in the giant papillae. Real-time PCR analysis showed that poly I:C induced TSLP mRNA expression in HCJEs in an endosomal-function-dependent manner and that rTSLP could induce IL-13 mRNA expression in the mast cells synergistically with IL-33. CONCLUSIONS: The TSLP protein produced in conjunctival epithelial cells plays a role in severe ocular allergy through the activation of dendritic cells and mast cells in synergy with other cytokines.

Macrophage inflammatory protein-1alpha as a costimulatory signal for mast cell-mediated immediate hypersensitivity reactions.

Regulation of the immune response requires the cooperation of multiple signals in the activation of effector cells. For example, T cells require signals emanating from both the TCR for antigen (upon recognition of MHC/antigenic peptide) and receptors for costimulatory molecules (e.g., CD80 and CD60) for full activation. Here we show that IgE-mediated reactions in the conjunctiva also require multiple signals. Immediate hypersensitivity reactions in the conjunctiva were inhibited in mice deficient in macrophage inflammatory protein-1alpha (MIP-1alpha) despite normal numbers of tissue mast cells and no decrease in the levels of allergen-specific IgE. Treatment of sensitized animals with neutralizing antibodies with specificity for MIP-1alpha also inhibited hypersensitivity in the conjunctiva. In both cases (MIP-1alpha deficiency and antibody treatment), the degranulation of mast cells in situ was affected. In vitro sensitization assays showed that MIP-1alpha is indeed required for optimal mast cell degranulation, along with cross-linking of the high-affinity IgE receptor, FcepsilonRI. The data indicate that MIP-1alpha constitutes an important second signal for mast cell degranulation in the conjunctiva in vivo and consequently for acute-phase disease. Antagonizing the interaction of MIP-1alpha with its receptor CC chemokine receptor 1 (CCR1) or signal transduction from CCR1 may therefore prove to be effective as an antiinflammatory therapy on the ocular surface.

Cross-sectional study of allergic disorders in relation to familial factors in Japanese adolescents.

AIM: The roles of heredity and the household environment in the development of allergic disorders are not clearly established. This study examined the relationship between selected familial factors and the prevalence of symptoms of wheeze, atopic eczema and rhinoconjunctivitis in Japanese adolescents. METHODS: Study subjects were 5539 students aged 12-15 y in Suita City. A questionnaire ascertained gender, grade, number of older siblings, maternal age at childbirth, smoking in the household, domestic pets and parental history of allergy, as well as signs and symptoms of allergy in the previous 12 mo. The latter were based on diagnostic criteria from the International Study of Asthma and Allergies in Childhood. RESULTS: Male gender was independently associated with an increased prevalence of wheeze and a decreased prevalence of atopic eczema. The prevalence of rhinoconjunctivitis, but not wheeze or atopic dermatitis, significantly increased with advancing grade. A significant inverse dose-response relationship between the number of older siblings and the prevalence of rhinoconjunctivitis, but not wheeze or atopic dermatitis, was observed. Maternal age at childbirth, smoking in the household and domestic pets were not apparently related to any of the allergic disorders. A positive maternal allergic history was more evidently associated with an increased prevalence of wheeze and rhinoconjunctivitis, but not atopic eczema, than a positive paternal allergic history. CONCLUSION: The findings suggest that the factors associated with allergic disorders in Japan are largely the same as those already identified in Western populations.

Vernal keratoconjunctivitis: a model of 5q cytokine gene cluster disease.

Clinical studies of vernal keratoconjunctivitis (VKC) patients show that total IgE serum levels are increased even in the absence of IgE antibodies to common allergens. Activated eosinophils are also a constant feature of VKC at both the circulation (cytofluorimetry) and tissue (tear cytology and conjunctival scrapings) levels. Moreover, allergen challenge induces a prolonged inflammatory reaction with a prevalent participation of eosinophils, lymphocytes and possibly basophils. Immunohistochemical studies of VKC biopsies show a multicellular inflammatory infiltrate with prevalence of activated eosinophils, mast cells and CD4 lymphocytes in both epithelium and subepithelium. Mediator studies indicate that eosinophil products (eosinophil peroxidase, eosinophinal cationic protein and eosinophil-derived neurotoxin/eosinophil protein X) are increased in both serum and tears, where tryptase and interleukin (IL)-5 are also detectable in higher amounts than in controls. On the basis of these findings, we postulate that VKC can represent a phenotypic model of up-regulation of the cytokine gene cluster on chromosome 5q which through its products (IL-3, IL-4, IL-5 and granulocyte/macrophage-colony-stimulating factor) regulates Th2 prevalence, IgE production as well as mast cell and eosinophil growth and function in VKC.

Familial occurrence of atopic disease: genetic versus environmental factors.

Cumulative life prevalence of atopic disease (any of reported symptoms of asthma/wheezy bronchitis, allergic rhinitis, eczema and urticaria) was studied by means of a questionnaire in 19814 (7-, 10- and 14-year-old) Swedish school children and their parents. Maternal history was found to be twice as common as paternal history. The children of affected mothers contracted atopic disease in the same proportion as the children of affected fathers yielding twice as many affected children with affected mothers than with affected fathers. The strongest parental influence on childhood disease was seen for multiple symptoms and for congruent symptoms with both parents. The number of children with both parents affected was 1.6 times larger than expected. A possible dose-response effect in polygenic inheritance is discussed as well as assumed impact of environmental factors with a tendency to familial clustering.