ovalbumin and Eye-Diseases

Ocular allergy leads to acute and chronic inflammation that may deteriorate the conjunctiva and other ocular tissue. The conjunctiva is covered with abundant lymphatic vessels but how the conjunctival lymphatic system patriciates in the development of allergic eye disease (AED) remains to be elucidated.. By using ovalbumin (OVA)+pertussis toxin (PTX) as a sensitizer followed by daily OVA challenges, we induced optimized AED manifestations in mice. We show that conjunctival lymphatics underwent significant expansion after 28 days of chronic OVA challenge, and this process can be prevented by inducible genetic ablation of lymphatic Vegfr3. Through transcriptomic profile analysis in combination with histopathological examinations, we found that pro-lymphangiogenic VEGFR3 signal promoted allergy-induced activation of T helper 2 (Th2) type immune responses, including antigen presentation, and Th2 cells, B cells and mast cell-related pathways in the conjunctiva, thereby critically contributing to the immunoglobulin E (IgE) production and AED manifestations. As a result, ocular allergy can be alleviated by genetic inhibition of lymphatic Vegfr3. Interestingly, pro-lymphangiogenic VEGFR3 signal did not appear to affect the obstruction of meibomian glands (MGs) or the activation of Th17 type and neutrophil pathways that are associated with AED.. These data reveal the key role of pro-lymphangiogenic VEGFR3 signaling in the development of AED and provide experimental evidence that VEGFR3 inhibition may be useful in treating ocular allergy in patients.

Topics: Animals; Disease Models, Animal; Eye Diseases; Hypersensitivity; Lymphangiogenesis; Mice; Ovalbumin; Vascular Endothelial Growth Factor Receptor-3

Severe, chronic eye allergy is an understudied, vision-threatening condition. Treatments remain limited. We used a mouse model of severe allergic eye disease (AED) to determine whether topical application of the pro-resolution mediator Resolvin D1 (RvD1) terminates the response. AED was induced by injection of ovalbumin (OVA) followed by topical challenge of OVA daily. RvD1 was applied topically prior to OVA. Clinical symptoms were scored. Eye washes were assayed for MUC5AC. After 7 days, eyes were removed and the number of goblet cells, T helper cell responses and presence of immune cells in draining lymph nodes and conjunctiva determined. Topical RvD1 treatment significantly reduced symptoms of AED. RvD1 did not alter the systemic type 2 immune response in the lymph nodes. AED increased the total amount of goblet cell mucin secretion, but not the number of goblet cells. RvD1 prevented this increase, but did not alter goblet cell number. Absolute numbers of CD4 + T cells, total CD11b + myeloid cells, eosinophils, neutrophils, and monocytes, but not macrophages increased in AED versus RvD1-treated mice. We conclude that topical application of RvD1 reduced the ocular allergic response by local actions in conjunctival immune response and a decrease in goblet cell mucin secretion.

Topics: Allergens; Animals; Cells, Cultured; Chronic Disease; Disease Models, Animal; Docosahexaenoic Acids; Eye Diseases; Goblet Cells; Humans; Hypersensitivity; Immunity, Cellular; Mice; Mice, Inbred C57BL; Mucin 5AC; Ovalbumin

Pruritus is the major symptom of ocular allergy but currently available treatments are often ineffective. Previous studies demonstrated that subpopulations of primary sensory neurons express Fc receptors and may contribute to antigen-specific pain. We investigated the role of neuronal Fc-epsilon Receptor I (FcεRI) in allergic ocular pruritus. Ovalbumin (OVA) was used as allergen together with alum adjuvant (OVA+alum) to produce a mouse model of ocular allergy with a significant elevation in the serum levels of both antigen-specific IgE and IgG. Mice sensitized by OVA without alum only induced elevation of serum IgG but not IgE. Scratching behavior toward the eyes with the hindlimb was used as an indicator of ocular itch. Topical OVA challenging to the eye dose-dependently induced scratching toward the eye in the OVA+alum sensitized mice, but not those sensitized by OVA only. The antigen-induced scratching was largely abolished by topical application of the blocking antibody to FcεRIα, but was only partially alleviated by pretreatment of mast cell stabilizer or histamine I receptor antagonist. The expression of FcεRI was detected in subpopulations of trigeminal ganglion (TG) neurons including those expressing pruriceptive markers and innervating the conjunctiva in the naïve mice. Moreover, FcεRI was found significantly upregulated in small-sized TG neurons in the OVA+alum sensitized mice. In acutely dissociated TG neurons, IgE-immune complex (IC), but not the antibody or antigen alone, induced intracellular calcium increase. The neuronal responses to IgE-IC could be specifically blocked by pre-application of a siRNA for FcεRIα. Our results indicate that FcεRI expressed on peripheral nociceptive neurons in the TG may be directly activated by IgE-IC and contribute to allergic ocular pruritus. This study may suggest a novel mechanism for the development of pathological itch in allergic diseases.

Topics: Alum Compounds; Animals; Disease Models, Animal; Eye Diseases; Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Neurons; Ovalbumin; Pruritus; Receptors, IgE

Allergic eye disease, as in most forms of atopy, ranges in severity among individuals from immediate hypersensitivity to a severe and debilitating chronic disease. Dendritic cells play a key role in stimulating pathogenic T cells in allergen re-exposure, or secondary responses. However, molecular cues by dendritic cells underpinning allergic T cell response levels and the impact that this control has on consequent severity of allergic disease are poorly understood. Here, we show that a deficiency in thrombospondin-1, a matricellular protein known to affect immune function, has subsequent effects on downstream T cell responses during allergy, as revealed in an established mouse model of allergic eye disease. More specifically, we demonstrate that a thrombospondin-1 deficiency specific to dendritic cells leads to heightened secondary T cell responses and consequent clinical disease. Interestingly, whereas thrombospondin-1-deficient dendritic cells augmented activity of allergen-primed T cells, this increase was not recapitulated with naïve T cells in vitro. The role of dendritic cell-derived thrombospondin-1 in regulating secondary allergic T cell responses was confirmed in vivo, as local transfer of thrombospondin-1-sufficient dendritic cells to the ocular mucosa of thrombospondin-1 null hosts prevented the development of augmented secondary T cell responses and heightened allergic eye disease clinical responses. Finally, we demonstrate that topical instillation of thrombospondin-1-derived peptide reduces T cell activity and clinical progression of allergic eye disease. Taken together, this study reveals an important modulatory role of dendritic cell-derived thrombospondin-1 on secondary allergic T cell responses and suggests the possible dysregulation of dendritic cell-derived thrombospondin-1 expression as a factor in allergic eye disease severity.

Topics: Allergens; Animals; Dendritic Cells; Eye Diseases; Hypersensitivity; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; T-Lymphocytes; Thrombospondin 1

We have investigated whether CD95-CD95 ligand interactions are important in anterior chamber-associated immune deviation (ACAID) induced by soluble protein antigen, and if so, to identify the participating cells on which these molecules are expressed. Peritoneal exudate cells as antigen-presenting cells (APC) obtained from B6.lpr/lpr, B6.gld/gld and C57BL/6 mice were cultured with ovalbumin (OVA) and transforming growth factor-beta2 (TGF-beta2) overnight, then injected intravenously into C57BL/6 or B6.lpr/lpr recipients. Some B6.lpr/lpr mice were reconstituted with naive T cells from wild-type C57BL/6 donors. In other experiments, B6. lpr/lpr and B6.gld/gld mice received an anterior chamber injection of OVA followed 7 days later by subcutaneous immunization with OVA plus adjuvant. Delayed hypersensitivity (DH) was assessed with an ear swelling assay. T cells activated in vitro with OVA-pulsed, TGF-beta-treated APC were tested in vivo for their capacity to suppress DH expression in a local adoptive transfer assay. The results indicate that when ACAID was induced by in-vitro generated ACAID-inducing cells, the APC expressed CD95L, and recipient T cells expressed CD95. The capacity of in vitro generated regulatory T cells to suppress DH expression to OVA in vivo was not governed by CD95-CD95L interactions. When OVA was injected into the anterior chamber of naive mice, CD95 expression was required for ACAID induction, although ACAID was readily induced in CD95L-deficient mice. We conclude that CD95-CD95L interactions are required in ACAID for the initial stage of APC presentation of eye-derived antigens to T cells, and that CD95-CD95L interactions participate at one or more additional step in the process by which ACAID is induced by soluble protein antigens.

Topics: Animals; Anterior Chamber; Eye Diseases; Fas Ligand Protein; fas Receptor; Hypersensitivity, Delayed; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Mutant Strains; Ovalbumin; Protein Binding; T-Lymphocytes

The development of tolerance induced by subcutaneous or intraocular injections of lipopolysaccharide (LPS, Escherichia coli) in rat eyes has been studied. In addition, the ocular inflammatory responses to the reversed passive Arthus (RPA) reaction in the tolerant eyes were investigated. The tolerance in the eye after a single injection of LPS persisted for at least 42 days. Up to 42 days, vasodilatation, disruption of the blood-aqueous barrier, and leukocyte accumulation in the anterior chamber after a second injection of LPS were inhibited. Unilateral intraocular injection of LPS produced local tolerance, which was not observed in the contralateral eyes. The inflammatory reactions in response to RPA in the LPS-tolerant eyes were also significantly attenuated. It was also found that inflammatory reactions induced by RPA or 16,16-dimethyl prostaglandin E2 had no inhibitory effect on the responses to subsequent RPA or LPS administration, which indicated that initial inflammatory reactions do not render the tissues refractory to the response to a second stimuli. The results of this study suggest that some, as yet unknown, local changes in the ocular tissues caused by LPS may be involved in the development of tolerance.

Topics: 16,16-Dimethylprostaglandin E2; Animals; Drug Tolerance; Escherichia coli; Eye Diseases; Immune Sera; Inflammation; Iris; Leukocytes; Lipopolysaccharides; Male; Ovalbumin; Proteins; Rats; Rats, Inbred Strains; Vasodilation

A model of local ocular anaphylaxis has been developed in the rat. Erythema, oedema, and enhanced retention of radioiodinated rat serum albumin ([125I]-RSA) were noted in ocular adnexal tissues of immunized rats within 5 min of injection of antigen; these changes reached a maximum 15 min after antigen injection. Erythema, oedema, and retention of [125I]-RSA subsided to baseline levels 1--6 hr after challenge. A significant increase in weight of ocular adnexal tissues was seen within 15 min after challenge. The weight increase reached a maximum at 45 min and persisted through 6 hr. Weight approached baseline values by 24 hr. Although antigen was injected into the ocular adnexa and not directly into the globe, the globes of the antigen-injected eyes of immunized rats underwent anaphylaxis, possibly because of absorption of antigen through the sclera. In addition, the adnexa and globes of the contralateral eyes, which did not receive antigen, also underwent anaphylactic changes. These changes were not as marked as those observed in the antigen-injected tissues, but followed the same time-course of development. We conclude that anaphylaxis can be locally induced in ocular tissues, that the onset of anaphylaxis is within minutes, and the effects last for at least 24 hr.

Topics: Anaphylaxis; Animals; Antigens; Disease Models, Animal; Eye Diseases; Ovalbumin; Rats; Rats, Inbred Strains; Serum Albumin, Radio-Iodinated

Topics: Cats; Coloring Agents; Eye Diseases; Humans; Hypersensitivity; Microscopy; Ovalbumin; Pathology; Research; Retinal Vessels; Staining and Labeling; Vitreous Body