17ß-estradiol protects human eyelid-derived adipose stem cells against cytotoxicity and increases transplanted cell survival in spinal cord injury.

Stem cell transplantation represents a promising strategy for the repair of spinal cord injury (SCI). However, the low survival rate of the grafted cells is a major obstacle hindering clinical success because of ongoing secondary injury processes, which includes excitotoxicity, inflammation and oxidative stress. Previous studies have shown that 17b-estradiol (E2) protects several cell types against cytotoxicity. Thus, we examined the effects of E2 on the viability of human eyelid adipose-derived stem cells (hEASCs) in vitro with hydrogen peroxide (H2O2)-induced cell model and in vivo within a rat SCI model. Our results showed that E2 protected hEASCs against H2O2-induced cell death in vitro, and enhanced the survival of grafted hEASCs in vivo by reducing apoptosis. Additionally, E2 also enhanced the secretion of growth factors by hEASCs, thereby making the local microenvironment more conducive for tissue regeneration. Overall, E2 administration enhanced the therapeutic efficacy of hEASCs transplantation and facilitated motor function recovery after SCI. Hence, E2 administration may be an intervention of choice for enhancing survival of transplanted hEASCs after SCI.

Human eyelid meibomian glands and tarsal muscle are recognized by autoantibodies from patients affected by a new variant of endemic pemphigus foliaceus in El-Bagre, Colombia, South America.

BACKGROUND: Previously, we described a new variant of endemic pemphigus foliaceus (EPF) in Colombia, South America (El Bagre-EPF). OBJECTIVE: Continuing our characterization of this variant of EPF, we now focus on one of our previously reported clinical findings: the presence of ocular lesions. These ocular lesions are seen in patients having extensive skin involvement, as measured by the Lund and Browder scale, which is generally used for patients with skin burns. METHODS: We specifically searched for evidence of autoreactivity to various eyelid structures in these patients and correlated our immunologic data with the clinical findings. We performed indirect immunofluorescence studies using normal-appearing human eyelid skin from routine blepharoplasties as substrate tissue. We tested sera from 12 patients with El Bagre-EPF and ocular lesions, 5 patients with sporadic (nonendemic) pemphigus foliaceus, and 20 healthy control subjects (10 from the El Bagre-EPF endemic area and 10 from nonendemic areas). We used fluorescein isothiocyanate conjugated goat antiserum to human total IgG/IgA/IgM as a secondary antibody. In addition, we used fluorescein isothiocyanate conjugated antibodies to human fibrinogen, albumin, IgG, IgE, C1q, and C3, Texas Red (Rockland Immunochemicals, Inc, Gilbertsville, PA), Alexa Fluor 555, or Alexa Fluor 594 (Invitrogen, Carlsbad, CA). Ki-67 (a cell proliferation marker) was used to determine the cell proliferation rate, and nuclear counterstaining was performed with either 4', 6-diamidino-2-phenylindole or Topro III (Invitrogen, Carlsbad, CA). RESULTS: We observed autoreactivity to multiple eyelid structures, including meibomian glands and tarsal muscle bundles at different levels, and some areas of the epidermis and the dermis close to the isthmus of the eyelids. Tarsal plate autoreactivity was seen in 10 of 12 of the El Bagre-EPF sera and in one control with pemphigus erythematosus. Furthermore, immunoprecipitation using an eyelid sample as a substrate with 1 mmol/L of sodium orthovanodate showed autoreactivity to several antigens, including some of possible lipid origin. LIMITATIONS: The main limitation of this study is the fact that the antigen or antigens remain unknown. CONCLUSION: We identified for the first time to our knowledge autoantibodies to meibomian glands and tarsal muscle in El Bagre-EPF. Our findings suggest that the autoantibodies to the ocular structures cause the clinical and histopathological findings in the ocular lesions in El Bagre-EPF.

Dermatologic and allergic conditions of the eyelid.

This article reviews the dermatologic and allergic conditions of the eyelid. Topics include various eyelid dermatitis, inflammatory lesions, infections, benign and malignant tumor, urticaria, vascular lesions, and others. Treatment considerations for these conditions of the eyelid are also discussed.

Ocular Surface and Tear Film Characteristics in a Sclerodermatous Chronic Graft-Versus-Host Disease Mouse Model.

PURPOSE: To report the characteristics of the ocular surface in a previously established sclerodermatous chronic graft-versus-host disease (cGVHD) mouse model. METHODS: The ocular surface features and tear film parameters of the mouse model were assessed by histopathology, immunohistochemistry, electron microscopy, quantitative polymerase chain reaction, and flow cytometry. RESULTS: The mice exhibited loss of body weight and decreased tear secretion (P < 0.001), mimicking the clinical features of patients with cGVHD. Ocular examination demonstrated significant corneal epithelial staining, conjunctival (P < 0.001), and eyelid (P = 0.015) fibrosis compared with the control mice. The density of both goblet cells (P = 0.043) and microvilli was lower (P < 0.001), and the microvilli were shorter (P = 0.007) in the conjunctiva of cGVHD mice than those of the controls. The immunohistochemical studies demonstrated greater expression of CD45, CD4, and CD8 cells in the conjunctiva and eyelid tissues compared with the controls (P < 0.05 for all). In addition, reduced Forkhead box P3 (Foxp3)+ cells were found in both the peripheral blood (P < 0.001) and conjunctiva (P = 0.042) of cGVHD mice compared with the controls. CONCLUSIONS: The constellation of these findings suggests that the sclerodermatous cGVHD mouse model well recapitulates ocular manifestations of cGVHD in humans. This model can be used to study the mechanisms involved in the pathogenesis and treatment of chronic ocular graft-versus-host disease.

Ocular disease induced in mice by anterior chamber inoculation of herpes simplex virus.

Herpes simplex virus type 1 (HSV-1) inoculated directly into the anterior chamber of the mouse eye induced an acute inflammatory process in both the injected eye and its uninjected (contralateral) counterpart. In the former, a rapid intense inflammatory reaction developed in the anterior segment (cornea, anterior chamber) and anterior portion of the uveal tract (iris and ciliary body). The retina of the injected eye was spared. In contrast, in the uninjected eyes, a delayed massive destructive reaction also developed, but was limited almost exclusively to the posterior segment ( vitreous, retina and choroid); retinas of the uninjected eyes were destroyed completely. When HSV-1 was inoculated bilaterally into both anterior chambers, destructive inflammatory responses developed in both corneas and anterior segments, but the retinas were spared bilaterally. These results indicate that (1) a unique and interesting pattern of bilateral ocular disease occurs after uniocular anterior chamber injection of HSV-1 in mice; (2) the distribution of the destructive lesions differs between the injected eye and its uninjected counterpart; and (3) local factors, perhaps produced within the eye itself, modify the progression of the virus-induced reaction within the globe.

Conjunctival basophil hypersensitivity lesions in guinea pigs. Analysis of upper tarsal epithelium.

The upper tarsal conjunctival epithelium was analyzed for inflammatory cell profile and accompanying morphological changes in a guinea pig system with histopathology resembling two human ocular diseases: vernal conjunctivitis and contact lens-associated giant papillary conjunctivitis (GPC). Female Hartley strain guinea pigs were immunized intradermally on day 0 with 200 micrograms keyhole limpet hemocyanin (KLH) and challenged on day 6 with varying doses of KLH by injection beneath the conjunctival epithelium of one lid and phosphate-buffered saline in the contralateral lid. Tissues containing the reaction site were examined by light microscopy. The 50 micrograms dose of KLH elicited the maximal accumulation of basophils and eosinophils. These values were significantly higher than in the PBS-injected control. Injection of KLH, PBS, or insertion of a sterile needle into unsensitized animals, and uninjected tissue served as additional controls. Neutrophils were significantly higher in the epithelium of the traumatized tissue (repeated needle insertions) than in the uninjected control. Basophils and mast cells were rarely found in the epithelium of unsensitized animals. Epithelial thickening, quantified by a Zeiss Videoplan2 Image Analysis system (Zeiss, West Germany), was greatest in the traumatized tissue, followed by the KLH-challenged tissue of sensitized animals. These values were significantly greater than that of the PBS-injected lid or of naive animals, uninjected or KLH-injected. These results indicate that epithelial changes can be induced by both antigen and trauma. Such epithelial changes may have a role in both vernal conjunctivitis and giant papillary conjunctivitis.

Mucosal defense of the outer eye.

A combination of mechanical, anatomical, immunological, and microbiological factors prevent infection of the outer eye. Mechanical and anatomical factors include the intact epithelium of the conjunctiva and cornea and the constant blinking action of the eyelids. Tear components that play a role in eye defense include lysozyme, immunoglobulins, lactoferrin, and betalysin. The normal bacterial flora of the conjunctiva may also have an inhibitory effect on the survival of more pathogenic species. The eye is linked to the common mucosal immune system, thus gaining the benefits of a system of microbial defense which is primed in the gastrointestinal tract, where a continuing large antigen load is capable of stimulating ongoing immune protection. The relative roles of the various factors contributing to prevention of eye infection remain to be fully defined.

Peripheral territory and neuropeptides of the trigeminal ganglion neurons centrally projecting through the oculomotor nerve demonstrated by fluorescent retrograde double-labeling combined with immunocytochemistry.

The peripheral territories of sheep trigeminal neurons which send their central process to the brainstem through the oculomotor nerve were investigated by the use of fluorescent tracers in double-labeling experiments. For this purpose Diamidino yellow (DY) injection into the oculomotor nerve was combined with Fast blue (FB) injection either into the extraocular muscles (EOMs), or the cornea, or the superior eyelid. Double-labeled DY + FB cells were found in the ophthalmic region of the trigeminal ganglion in addition to single-labeled DY or FB cells. The DY and DY + FB-labeled trigeminal cells were analysed immunocytochemically for their content of substance P (SP)-, calcitonin gene-related peptide (CGRP)-, and cholecystokinin-8 (CCK-8)-like. All single-labeled DY cells showed SP-, CGRP- or CCK-8-like immunoreactivity. Double-labeled DY + FB neurons innervating the EOMs were immunoreactive for each of the three peptides, whereas double-labeled neurons supplying the cornea were only CGRP-like positive. The findings suggest that, in the sheep, trigeminal neurons which send their process centrally through the oculomotor nerve supply the EOMs, the cornea, and the superior eyelid and contain neuropeptides which are usually associated with pain sensation.

Tissue, tear, and serum IgE concentrations in vernal conjunctivitis.

Abnormal upper tarsal tissue from four patients with vernal conjunctivitis stained immunofluorescently contained abundant IgA-, IgD-, and IgE-forming plasma cells in two of four patients. These were absent in normal upper tarsal conjunctiva. The IgE levels of paired tear-serum samples of 11 patients with vernal conjunctivitis and ten control subjects were 130 ng/ml in patients that did nor differ significantly from the normal tear IgE value of 61 ng/ml. The serum IgE means of 1,031 ng/ml for patients and 201 ng/ml for normal subjects differed significantly (P less than .05). Tear and serum IgE values correlated significantly for both groups (P less than .01 for patients, P less than .05 for normal subjects). These findings are consistent with a hyperplasia of IgA, IgD, and IgE antibody-forming cells in tarsal conjunctiva of some patients with vernal conjunctivitis, and tear IgE levels being a function of serum IgE levels.

Clinical, morphologic, immunophenotypic, and molecular genetic analysis of bilateral ocular adnexal lymphoid neoplasms in 17 patients.

We investigated the clinical, morphologic, immunologic, and molecular genetic characteristics of the lymphoid tumors occurring in 17 patients with bilateral ocular adnexal lymphoid neoplasia. We found no obvious differences in the age, sex, clinical appearance, or ophthalmic findings between patients with unilateral and those with bilateral ocular adnexal lymphoid neoplasms. Five orbital and two conjunctival lymphoid neoplasms removed from five patients were polyclonal pseudolymphomas, while nine orbital, eight conjunctival, and two eyelid lymphoid neoplasms removed from 12 patients were monoclonal B cell non-Hodgkin's lymphomas. Each pair of simultaneously bilateral lesions occurring in a single individual was morphologically and immunologically identical and exhibited identical immunoglobulin gene DNA rearrangement patterns. None of the five patients with polyclonal pseudolymphomas developed nonocular non-Hodgkin's lymphoma, but one developed a contralateral conjunctival monoclonal B cell non-Hodgkin's lymphoma. Six of the 12 patients with ocular adnexal monoclonal B cell non-Hodgkin's lymphomas developed nonocular non-Hodgkin's lymphoma. The incidence of previous or subsequent systemic nonocular non-Hodgkin's lymphoma in patients with bilateral ocular adnexal lymphoid neoplasms is comparable to that of patients with unilateral disease. In both patient groups, the development of nonocular non-Hodgkin's lymphoma is most commonly associated with ocular adnexal lymphoid neoplasms displaying follicular or diffuse small cleaved cell (poorly differentiated lymphocytic) morphologic characteristics.

Microbial and immunological investigations of chronic non-ulcerative blepharitis and meibomianitis.

Concentrations of tear lysozyme, lactoferrin, ceruloplasmin, IgG, and IgA have been measured by enzyme linked immunosorbent assay (ELISA) in patients with chronic non-ulcerative blepharitis and meibomianitis at the same time as the lid and conjunctivae were cultured for bacteria and fungi by a semiquantitative method. A group of normal controls aged 20 to 80 were similarly sampled, when strains of Staphylococcus epidermidis from their eyes and the patients' eyes were biotyped according to Baird-Parker's scheme. 5% of blepharitis cases had increased numbers of Staph. aureus present on the lids, compared with only a scanty growth obtained from 5% of normals. 7% of blepharitis cases had increased numbers of Staph. epidermidis type VI (coagulase-negative, mannitol-fermenting) present compared with a scanty growth obtained from 6% of normals. Isolation rates of other types of Staph. epidermidis did not differ from those in normals; no types were associated with meibomianitis. Tear protein profiles were normal in most patients, and there was no increase in tear IgA or IgG, which is expected with chronic infection. Overall our evidence suggests that in 88% of cases these lid conditions have an inflammatory aetiology not associated with infection. Staphylococcal isolates often found in the eye usually represent a normal commensal rather than pathogenic flora.

CD4 and CD8 antigen coexpression: a flow cytometric study of peripheral blood, bone marrow, body fluid, and solid lymphoreticular specimens.

OBJECTIVE: CD4 and CD8 antigen coexpression occurs not only on blastic T-cell malignancies, but also on a small subset of mature lymphocytes. The aim of this study was to determine the prevalence of this population of cells and to identify features that can be used to differentiate them from T lymphoblasts. DESIGN: All specimens submitted to the clinical flow cytometry laboratory from August 1, 1994, through July 31, 1995, were analyzed for CD4 and CD8 coexpression. MAIN OUTCOME MEASURE: Percentage of lymphocytes coexpressing the CD4 and CD8 antigens. RESULTS: Four percent (22/526) of all specimens contained a population of CD4/CD8 coexpressing cells. Five cases represented CD4 and CD8 antigen expression on neoplastic cells. In 17 cases, the CD4/CD8 coexpressing cells appeared to represent a population of mature lymphocytes with a normal phenotype. The immature cells of T-cell acute lymphocytic leukemia and lymphoblastic lymphoma represented a dominant uniform population of cells demonstrating strong staining with both the CD4 and CD8 antigens. Cases containing a mature population of CD4/CD8 coexpressing cells were characterized by fewer coexpressing cells and variable expression of CD8. There were cases where distinction of this population of mature CD4/CD8 coexpressing lymphoid cells from a blastic malignancy was not possible using immunophenotyping alone. CONCLUSION: Correlation of clinical, morphologic, and immunophenotypic data is recommended to prevent the misdiagnosis of subtle involvement by a blastic T-cell malignancy.

Eye and eyelid abnormalities are common in patients with Hashimoto's thyroiditis.

BACKGROUND: Overt ophthalmopathy is presumed to be uncommon in patients with Hashimoto's thyroiditis compared to Graves' disease, where significant eye changes are found in approximately 40% of patients. On the other hand, when observing, more subtle eye changes, particularly upper eyelid retraction (UER) and mild inflammatory signs, may be common in patients with Hashimoto's thyroiditis. METHODS: We have determined the prevalence and characteristics of eye signs in recently diagnosed patients with Hashimoto's thyroiditis studied prospectively since 2004 till date in Sydney (Australia). We measured serum orbital antibodies in 20 of the patients in enzyme-linked immunosorbent assay. RESULTS: The overall prevalence of eye signs in patients with Hashimoto's thyroiditis was 34%, of whom about a quarter had chronic UER, determined as a margin-reflex distance of >5 mm, as the main sign. There was no correlation between eye signs and cigarette smoking. Overall, there was only a modest correlation between eye signs and positive antibody tests, and 40% of patients with no eye signs at the time of study were antibody positive. CONCLUSION: Eye changes, in particular UER, are common in patients with Hashimoto's thyroiditis. Since thyroid stimulating hormone-receptor antibodies are not usually associated with Hashimoto's thyroiditis, autoimmune mediated damage of the levator palpebrae superioris (eyelid) muscle cannot be due to these antibodies. Although eyelid abnormalities may be a minor problem for most patients, for some there are major cosmetic implications requiring surgical management.

Ocular defense mechanisms with special reference to the demonstration and functional morphology of the conjunctiva-associated lymphoid tissue in Japanese monkeys.

In order to define the histological components of ocular defense, the conjunctiva in Japanese monkeys was studied using a whole mount method, light microscopy, and electron microscopy. We investigated the distribution of the conjunctiva-associated lymphoid tissue (CALT) using stereoscopic observations of the conjunctiva immunostained with human leukocyte antigen (HLA)-DR antibody and /or stained with alcian-blue. The outer surface of the conjunctival fornix was lined by sheets of mucus secreting goblet cells, with small epithelial patches without goblet cells, scattered among them. The patches, termed CALT, consisted of flattened epithelial cells, intraepithelial lymphocytes and dendritic cells, and lymphoid follicles with a germinal center. The CALT in Japanese monkeys was fundamentally similar in structure to those found in other animal species. CALT patches ranged in size ranging from 200 microm to 300 microm in diameter. The number of patches varied from 20 to 40 in the superior eyelid and 10 to 20 in the inferior eyelid. Latex microspheres administrated as eye drops were selectively taken up first by flattened associated epithelial cells covering the surfaces of CALT patches and then by intraepithelial dendritic cells of the CALT. These morphological findings show that CALT patches in the eyelids of primates are focal sites for particulate uptake and contact with lymphoid constituents, indicating that they are inductive sites for the common mucosal immune system as well as important components in ocular defense.