摘要:

Rabbit uterine collagenase was purified from the medium of involuting uterus (1-2 days postpartum) in culture using ammonium sulfate fractionation, DEAE-cellulose, hepari n-af f inity, and high performance liquid chromatography. The enzyme was purified more than 1600 fold. Hybridoma cell - 1 ines produci ng monoclonal anti bodi es were prepared by fusing the spleen cells of mice immunized with the purified enzyme with mouse myeloma cells (Sp2/0-Ag14). The hybridoma cells were selected with HAT medium, cloned, and screened by ELISA. Antibody-producing ascites were prepared by injecting hybridoma cell-lines into the peritoneal cavities of mice. Western-blot analysis indicated that the antibodies recognized a polypeptide having a molecular weight of 52,000. The IgG isolated from the ascites inhibited the enzyme. Indirect immunofluorescent staining demonstrated that polymorphonuclear leukocytes (PMNs) in the superficial layer of alkali- burned corneas contained collagenase, whereas stronal cells and PMNs within the stroma were not stained by the antibodies. Our results suggest that collagenases produced by rabbit PMNs are different from those produced by fibroblasts from cornea. We hypothesize that PMNs in alkali-burned corneas secrete all or most of their collagenases by degranulation at the anterior surface of the cornea, and then continue to migrate into the deeper portion of the stroma.

ISSN:0271-3683    

DOI:10.3109/02713688609029231

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

We performed a phase I study to assess the therapeutic efficacy of an anti-T cell monoclonal antibody conjugated with ricin A-chain in a corneal graft rejection model. Corneal allografts were exchanged between Dutch and New Zealand rabbits. Rejections occurred within 16-22 days in untreated animals. Graft rejection was delayed by topical or retrobulbar cyclosporine, but not by subconjunctival injections of a murine anti-rabbit T cell monoclonal antibody, nor by either subconjunctival or intravenous F(ab')2ricin conjugate.

ISSN:0271-3683    

DOI:10.3109/02713688609029232

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

Individual mucus samples were collected from normal individuals and from patients with ocular cicatricial pemphigoid (CP), Stevens-Johnson syndrome (SJS), and various types of conjunctival inflammation (rosacea, meibomianitis, atopy, keratoconjunctivitis sicca, etc.). The mucus samples were dissolved in sample buffer containing 8M urea, 2% SDS and 5% 2-mercaptoethanol and were electrophoresed on gradient 2-16% polyacrylamide gels. Four glycoproteins with molecular weights greater than 200,000 daltons were consistently observed in both individuals with normal conjunctiva and patients with CP, SJS, and other diseases exhibiting conjunctival inflammation. The amounts of each glycoprotein appeared to vary from one individual to another; however, the presence or absence of specific glycoproteins could not be correlated with the different ocular diseases. The techniques described for mucus analysis offer advantages over previously published techniques since improved resolution of the mucous glycoproteins can be achieved by electrophoresis on 2-16% gradient gels, and individual samples can be analyzed. Our results suggest that substantial amounts of ocular mucous glycoprotein are present in the eyes of patients with CP and SJS, diseases which have been previously described as mucin-deficient dry eye syndromes.

ISSN:0271-3683    

DOI:10.3109/02713688609029233

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

We have developed a polyclonal anti-Histoplasma capsulatum antibody to detect II. capsulatum antigens in ocular tissue. Antibodies were specific for II. capsulatum as determined by enzyme linked immunosorbant assay and immunoprecipitation. A total of 21 choroidal histoplasmic lesions in 4 primate eyes (Macaca speciosa and Macaca mulatta). taken at various times from 14 to 60 days after the internal carotid artery injection of yeast phase organisms, were evaluated. Using an indirect immunofluorescent technique, these antibodies stained yeast phase organisms within acute choroidal lesions at 14 days after infection. By 60 days intact organisms were no longer detected; occasional cells, however, contained intracellular inclusions that stained with these antibodies. Although yeast phase organisms are rapidly cleared from the primate choroid, these data indicate that residual H. capsulatum antigens may remain in choroidal lesions after the acute infectious stage. These findings are consistent with the hypothesis that H. capsulatum antigens play a role in the immunologic reactivation of atrophic choroidal scars in ocular histoplasmosis.

ISSN:0271-3683    

DOI:10.3109/02713688609029234

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

A fast and sensitive high-performance liquid chromatographic (HPLC) method for the determination of vitamin A (all-trans retinol) in 50μ1 human tear fluid is described. After deproteinization with ethanol and extraction with n-hexane, retinol is separated from the extract on a straight-phase HPLC column and detected fluorometrically. Vitamin A can be determined in concentrations as low as 0.4 ng/ml. A single analysis can be completed in 12 min while the analysis of a series of 60 samples takes about 8 h. The within-assay and between-assay coefficients of variation were 3.1 and 4.2 % resp. The between-assay analytical recovery of retinol added to tear fluid was 97.4±6.0 % (mean±SD). Retinol was determined in tear fluid of nine adult volunteers. The amounts observed ranged from<0.4 - 10.6 ng/ml.

ISSN:0271-3683    

DOI:10.3109/02713688609029235

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

This paper reports the development of a non-contact corneal irrigation chamber (NC-CIC) which enables non-invasive collection of epithelial cells from the corneal surface of human subjects. Cells were viewed by fluorescence microscopy following vital staining with acridine orange (AO). Staining characteristics revealed two corneal epithelial cell types: cells with (i) green and (ii) orange-red cytoplasmic staining. The green cytoplasmic stain appeared to indicate a more viable cell. Multi-cell aggregates were regularly collected from the corneal epithelial surface. Groups of up to seven epithelial cells were obtained. Quantitative studies of corneal epithelial cell sloughing, using isotonic NaCl (305 mOsm/kg) and isotonic“basic tear solution”(BTS, 305 mOsm/kg) as irrigating solutions, involving hourly irrigations between 8 a.m. and 4 p.m. were conducted. Consistently higher cell counts were obtained with NaCl. Using BTS, data scatter was reduced sufficiently to reveal significant differences in sloughing rate as a function of time of day. Instillation of one drop of 0.5% proparacaine caused a significant, but gradual, increase in epithelial cell sloughing rate over a period of hours, as indicated by subsequent BTS irrigations of the cornea. Since the NC-CIC technique is able to discriminate these effects, it may be an appropriate system for i_n vivo studies of the relationship between corneal epithelial cell mitosis and sloughing.

ISSN:0271-3683    

DOI:10.3109/02713688609029236

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

Inflammation was induced in the eye by active and passive Arthus reactions using a non-ocular antigen and was examined by histologic and immunopathologic techniques. T cells were seen to infiltrate the eye when the inflammation occured in the active Arthus reaction and were not seen after passive immunization. The findings were compared to those of experimental autoimmune uveoretinitis.

ISSN:0271-3683    

DOI:10.3109/02713688609029237

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

Inhibitor studies with peptide substrates demonstrate that bovine lens neutral proteinase comprises three distinct activities. Diisopropylfluoro-phosphate distinguishes the activity hydrolyzing carbobenzoxy-Gly-Gly-Leu-p-nitroanilide (inhibited) from that hydrolyzing carbobenzoxy-Leu-Leu-Glu-2-naphthylamide (not inhibited). Leupeptin inhibits hydrolysis of the substrate carbobenzoxy-Leu-Leu-Arg-2-naphthylamide, but not hydrolysis of car-bobenzoxy-Gly-Gly-Leu-p-nitroanilide or carbobenz-oxy-Leu-Leu-Glu-2-naphthylamide, demonstrating the presence of the third activity. Inhibition of the three activities by thiol reagents suggests that each activity may be dependent on an active-site cysteine residue.

ISSN:0271-3683    

DOI:10.3109/02713688609029238

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

Uveitis can be induced by systemic or intravitreal administration of endotoxin (lipopolysaccharide, LPS). In this study we correlated the expression of class II antigens (la in rat) of the Major Histocompatibility Complex (MHC), with this experimental model of uveitis, la antigen was detected by immunohistochemistry using the Avidin-Biotin-Peroxidase Complex (ABC) method and the monoclonal antibody OX6. la antigen was not expressed in normal eyes. However, la was expressed in the anterior uvea epithelial cells in all eyes with LPS induced uveitis.This study demonstrates that the ocular la expression is a localized process in the anterior uvea in response to systemic or intravitreal LPS. This response appears to be distinct from the action of LPS on macrophage la expression, where LPS has been shown to inhibit the induction of la antigen in macrophages by gamma interferon.

ISSN:0271-3683    

DOI:10.3109/02713688609029239

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor

摘要:

The corneal uptake and penetration of14C-labelled 0.15% amphotericin B and 5% natamycin were studied in Dutch-belted rabbits. Corneal levels of natamycin were substantially higher than amphotericin B. For both drugs, these levels were influenced by corneal contact time. In corneas debrided of epithelium, both agents entered the corneal stroma and levels were detected in aqueous in the therapeutic range. However, in corneas with intact epithelium, penetration was negligible for amphotericin B (0.23μg/gm at 2 mins). By contrast, although penetration of natamycin was greatly reduced, 7.Oμg/gm were present at 2 mins.

ISSN:0271-3683    

DOI:10.3109/02713688609029240

出版商:Taylor&Francis    

年代:1986

数据来源: Taylor