Cytological and immunohistochemical study of the limbal form of vernal keratoconjunctivitis by the replica technique.

The cellular composition of the inflammatory infiltrate present in 13 patients with the limbal form of vernal keratoconjunctivitis was examined in the conjunctival scrapings and the limbic replicas by means of Giemsa stain and immunohistochemistry. Conjunctival scrapings showed the presence of mast cells, lymphocytes, plasma cells, polymorphonuclear leucocytes, and very few basophils in all the specimens. Eosinophils were present in only four scrapings. The superficial epithelium of the limbic lesion and the adjacent cornea and conjunctiva was studied by the replica technique. The limbic lesion area showed the presence of necrotic epithelial cells mixed with inflammatory cells, including eosinophils, mast cells, lymphocytes, plasma cells, and polymorphonuclear leucocytes and very few basophils. Most of the inflammatory cells were HLA-DR+. Many OKT6+ cells were present, indicating the presence of Langerhans cells. T-lymphocytes including a few helper/inducer cells and many suppressor/cytotoxic cells, were detected in the infiltrate. In addition many B-lymphocytes were observed. These findings suggest that other immune mechanisms in addition to type 1 reaction are involved in the pathogenesis of the disease.

Immunopathology of trachomatous conjunctivitis.

Upper palpebral conjunctival biopsy specimens obtained from eight patients with active trachoma were examined by routine histological and immunohistochemical methods. The epithelium expressed class I major histocompatibility complex (MHC) products throughout and class II MHC products in the superficial layers. The epithelial inflammatory infiltrate consisted of polymorphonuclear leucocytes, macrophages, T lymphocytes, and dendritic cells. In the underlying stroma the inflammatory infiltrate was organised as B lymphoid follicles, and there was also a diffuse infiltrate consisting of plasma cells and scattered B lymphoid cells, dendritic cells, T cells, macrophages, and polymorphonuclear leucocytes. Each type of cell has its special location in the tissue. Plasma cells were located on a subepithelial band and as a dense infiltrate round the acini of accessory lacrimal glands. IgA+ plasma cells outnumbered IgG+ cells, whereas IgM+ and IgE+ cells were few. Our data provide good evidence for the presence of both humoral and cell mediated immune responses and a possible role for autoimmune mechanisms in the conjunctival tissues of trachoma patients.

Recombinant interferon-gamma induces HLA-DR expression on human corneal epithelial and endothelial cells in vitro: a preliminary report.

The effect of interferon-gamma on the expression in situ of major histocompatibility complex (MHC) products in human corneas was studied in vitro. Incubation for four days with 5 or 50 mg/l of Escherichia coli-derived recombinant human interferon-gamma resulted in the appearance de novo of MHC class II or HLA-DR antigens on variable numbers of corneal epithelial cells as well as on corneal endothelium, whereas it had no effect on the expression of MHC class I or HLA-ABC antigens. These results may help to explain the mechanism underlying the expression of HLA-DR antigens on corneal and limbal epithelium in various inflammatory eye diseases.

Developmental anomalies in the organogenesis of the eyeball. Bilateral diophthalmos.

A case is described of a 4-month-old boy presenting with a bilateral microphthalmos with cyst. Detailed examination of one eye and cyst suggested that the cyst was atypical and represented an attempt at the formation of an extra eye on the same optic stalk. Such a developmental anomaly has not been described before and the term 'diophthalmos' is proposed as an appropriate name.

Expression of MHC class II antigens and immunoglobulin M by the corneal epithelial cells in herpetic keratitis.

The corneal buttons obtained from 4 patients with active epithelial and stromal herpetic keratitis were studied with routine microscopy and immunohistochemistry. We used an immunoperoxidase technique with monoclonal antibodies directed against Langerhans cells, lymphocyte subsets, MHC products and immunoglobulins A, G, M and D. The epithelium and stroma contained an inflammatory infiltrate composed of polymorpho-nuclear leukocytes, dendritic cells, B-lymphocytes and T-lymphocytes (helper/inducer and suppressor/cytotoxic subsets). The epithelial cells of all the corneal buttons expressed MHC class II antigens. IgM was bound to the membrane of the epithelial cells in 3 specimens. HSV antigenic material was localized in the epithelial cells and in the stromal keratocytes by a direct immunofluorescence technique. Our data suggest that cell-mediated as well as antibody-mediated immune responses are involved, with a possible role for an autoimmune mechanism in the pathogenesis of this condition.

Experimental chlamydial keratitis in rabbits. Correlation with chlamydia infected McCoy tissue culture cells.

Rabbit corneas were inoculated three times at weekly intervals with the agent of chlamydia trachomatis using the scratch method. Specimens of the corneal epithelium were obtained using the replica technique on the 1st, 2nd, 3rd and 4th day after each inoculation and at two weeks after the last inoculation. The development of chlamydial inclusions and the inflammatory cell response were monitored using Giemsa stain, acridine orange stain and direct immunofluorescent technique. Primary inoculation produced mild clinical disease associated cytologically with polymorphonuclear leucocytic cellular inflammatory response. Repeated inoculations produced more severe disease associated clinically with pannus formation and cytologically with the presence of lymphocytes and Leber cells in addition to polymorphonuclear leucocytes. Halberstaedter Prowazek inclusion bodies were detected in all the specimens. Additional intracytoplasmic and intranuclear inclusions of different morphological appearances were present. The cytological findings detected in the corneal epithelium of rabbits were correlated with the findings in McCoy tissue culture cells inoculated with chlamydia trachomatis.

Basic biochemical parameters of one hundred cataractous lenses from Egyptian patients.

Cataractous lenses from an extended population of 100 Egyptian patients whose age, sex and case history had been recorded, were collected and classified morphologically. After determination of the lens wet weights, lenses were microsectioned and basic parameters such as wet weight, dry weight, water content and contents of water-soluble and water-insoluble crystallins of the samples were determined. The data obtained document moderate as well as drastic changes of the investigated parameters along with aging and cataractogenesis, and extend our knowledge from small numbers of hitherto investigated human lenses to a considerably high number of cases.

Distribution of water-soluble crystallins in microsectioned cataractous lenses from one hundred Egyptian patients.

Morphologically classified lenses from 100 cataract patients from the Mansoura Eye Hospital (Egypt) were microdissected and the crystallin composition and distribution were analyzed by thin-layer isoelectric focusing (IEF). The IEF profiles of cataractous lenses were compared with each other, with those of sclerotic lenses and with a normal lens profile. The alterations in the composition of high-molecular-weight (HMW)-, alpha-, beta H-, beta L-, beta s-, and gamma-crystallins along with normal aging, are superimposed by pronounced cataract-related changes which are different for the various types of cataracts. The general feature includes a continuous loss of gamma-, beta s- and beta L-crystallins of higher IEPs and an increase of HMW material. This is highly pronounced in the nucleus of nuclear cataractous lenses. In cortical cataractous lenses, changes start in the cortical layers. No differences could be observed between alterations in Caucasian lenses and this extended Egyptian lens population.

Immunocytological study of phlyctenular eye disease.

Scrapings from phlyctens and conjunctiva of 12 patients with phlyctenular keratoconjunctivitis were studied using OKT4-Leu3a, OKT8, B1, BA1, S-100 and HLA-DR monoclonal antibodies. T-lymphocytes were present in both conjunctival and phlyctenular scrapings. OKT4-Leu3a positive cells outnumbered the OKT8 positive cells in both conjunctival and phlyctenular scrapings. B1 and BA1 positive cells were absent from the conjunctival scrapings, but were present in the phlyctenular scrapings. S-100 positive cells were present in both conjunctival and phlyctenular scrapings. However, they were very few in the conjunctival scrapings. Most of the cells in both conjunctival and phlyctenular scrapings were HLA-DR positive. These findings support the hypothesis that cell mediated immunity is responsible for the pathogenesis of phlyctenular eye disease.

Phenotypic characterization of inflammatory cells in phlyctenular eye disease.

Phlyctenular conjunctival biopsy specimens obtained from seven patients presenting with ulcerated limbal phlyctens were examined with routine histology and immunohistochemistry using a panel of monoclonal and polyclonal antibodies and immunoperoxidase techniques. Large numbers of mononuclear phagocytes, dendritic Langerhans cells and polymorphonuclear leukocytes and moderate numbers of T-lymphocytes were observed in the epithelium. This infiltrate was most intense at the basal epithelial layers which expressed HLA-DR antigens. The underlying stromal inflammatory infiltrate was organized as perivascular cuffs and a scattered subepithelial infiltrate, and consisted of many mononuclear cells and poly-morphonuclear leukocytes. Among the mononuclear cells, monocyte derived cells dominated and included monocytes macrophages and dendritic cells. T-lymphocytes were present in moderate numbers, whereas B-lymphocytes and plasma cells, mostly of IgA class, were infrequent. These findings provided in situ immunohistochemical evidence that phlyctenular eye disease is the result of delayed type hypersensitivity immune response in which monocytes and monocyte derived cells play a central role.

Immunopathological study of vernal keratoconjunctivitis.

The in situ immune reaction occurring in the conjunctival tissues of patients with vernal keratoconjunctivitis was studied using immunohistochemical techniques and a panel of monoclonal and polyclonal antibodies. Our data point to a complex immunopathogenesis of the disease. The anaphylactic response was demonstrated by the presence of many eosinophils and mast cells showing membranous IgE staining in the epithelial and stromal inflammatory infiltrate. IgE+ plasma cells were rare or absent in the conjunctival tissues and around the acini of accessory lacrimal glands, suggesting that IgE is not produced locally in the conjunctival tissues. A humoral immune response was evidenced by the presence of small B-lymphoid follicles, scattered B-lymphocytes, and many plasma cells in the stromal inflammatory infiltrate. IgA+ and IgG+ plasma cells were of more or less equal numbers and outnumbered IgM+ plasma cells. The majority of plasma cells around the acini of accessory lacrimal glands were IgA+. A role played by cell-mediated immunity was evident from the presence of scattered stromal T-lymphocytes and many dendritic cells in the epithelium and stroma. Some stromal dendritic cells carried IgE on their surfaces.