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PubMed1992

Restricted tissue reactivity of autoantibodies to a 64-kDa eye muscle membrane antigen in thyroid-associated ophthalmopathy.

We studied the tissue specificity of eye muscle (EM) membrane-reactive autoantibodies detected in the serum of patients with thyroid-associated ophthalmopathy (TAO). In preliminary studies, such antibodies were shown to react, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, with human thyroid (THY) and other human skeletal muscle (HSM) membrane antigens. We carried out absorption with human EM (HEM), THY, and HSM membranes of sera from patients with TAO and autoimmune thyroid disease without ophthalmopathy which reacted with one or more of 55-, 64-, and 95-kDa antigens in pig eye muscle (PEM) membrane in immunoblotting, the majority of which were also cytotoxic to HEM cells in an antibody-dependent cell-mediated cytotoxicity assay. In Western blotting, serum antibodies reactive with PEM membrane antigens of 55, 64, and 95 kDa were cross-absorbed by HEM, THY, and HSM but not by spleen or brain membranes and showed some species specificity, being absorbed by pig and human, but not bovine, EM membranes. When incubated with cultured HEM, THY, and HSM cells in vitro, autoantibodies in TAO sera immunoprecipitated a 64-kDa antigen from the first two tissues, but not from HSM, suggesting a specific binding to autoantigenic epitopes in HEM and THY. Sera from patients with TAO as well as those from patients with thyroid autoimmunity without ophthalmopathy immunoprecipitated a approximately 66-kDa protein, shown to be distinct from the 64-kDa antigen. The restricted immunological cross-reactivity of antibodies to a THY and HEM 64-kDa membrane antigen is discussed in the context of the association of ophthalmopathy with thyroid autoimmunity. Further experiments are needed to show whether autoantibodies to the 64-kDa eye muscle and thyroid shared antigen are cytotoxic, and thus likely to play a major role in the pathogenesis of the eye disease, or just markers of the orbital autoimmune process.

Adult↗
PubMed1998

Prevalence and significance of antibodies reactive with eye muscle membrane antigens in sera from patients with Graves' ophthalmopathy and other thyroid and nonthyroid diseases.

We have investigated the antibody responses made to porcine eye muscle antigens by sera from patients with Graves' disease and ophthalmopathy (GO) on Western blotting in a retrospective cross sectional study. Reactivity to several different eye muscle antigens was observed; however, antibodies to two antigens of 64 and 95 kd were found predominantly in sera with a prevalence of 63.6% (14/22) and 50.0% (11/22), respectively. In Graves' patients without GO the levels dropped to 18.2% (6/33) and 9.1% (3/33), and to 10.0% (2/20) and 20.0% (4/20) in patients with other thyroid diseases, and 0% (0/29) and 20.7% (6/29) in patients with nonthyroid diseases. Healthy control subjects gave positive responses in 8.0% (2/25) and 12.0% (3/25) of sera tested against the two antigens. The prevalence of the responses in patients with GO was significantly different when compared with normals and other patient groups (p < 0.05 and p < 0.01). All patients with ophthalmopathy graded class 3 or 4 and most of the patients with GO and a suppressed thyrotropin (TSH) value at the time of investigation reacted with one or other of the two antigens. Similar prevalences of response were found to the 64 and 95 antigens in those patients with GO whether they were thyrotropin receptor antibodies (TSHR-Ab) positive or negative (p > 0.05). However, the prevalence of the response to the 64 kd antigen in the TSHR-Ab positive group of patients without GO (31.3%) was significantly higher than that found in the TSHR-Ab negative (5.9%, p < 0.05) group. 81.8% (9/11) of patients with GO and a suppressed TSH recognized the 64 kd antigen compared with 45.5% with a normal TSH value (p < 0.05) and 16.7% without GO and a normal TSH (p < 0.01). In additional studies we have shown that a mouse monoclonal antibody (3B12) raised to the extracellular domain of the human TSH receptor also reacted with a 55 kd antigen in pig eye muscle but was not inhibited from its binding to either pig eye antigen or human TSH receptor antigen (3A1) by any of our patients' sera. These data suggest that serum antibodies reactive with porcine eye muscle antigens are potentially useful markers of ophthalmopathy in Graves' patients and should be further evaluated in longitudinal studies to see if they can predict the onset or regression of Graves' ophthalmopathy.

Adult↗
PubMed1998

Possible involvement of Fas-mediated apoptosis in eye muscle tissue from patients with thyroid-associated ophthalmopathy.

In order to investigate the possible involvement of apoptosis in the pathogenesis of thyroid-associated ophthalmopathy (TAO), we studied the expression of Fas, Fas ligand (FasL), and Bcl-2 in extraocular muscle tissues from patients with TAO by immunohistochemistry using monoclonal antibodies against Fas, FasL, and Bcl-2. Apoptosis was detected by in situ end-labeling of fragmented DNA. Apoptosis was detected in extraocular muscle tissues from 17 of 19 patients with TAO and, to a smaller degree in 4 of 7 normal control subjects. The mean percentages of apoptotic nuclei were 12.1% in TAO eye muscle fibers and 16.4% in TAO interstitial cells, compared with 2.2% and 0.4% in normal eye muscle fibers and interstitial cells, respectively. The percentage of apoptotic nuclei in eye muscle fibers correlated with the enlargement of eye muscle tissue (r = 0.47, p < 0.05). Fas was demonstrated on the surfaces of extraocular muscle fibers in 11 of 18 patients with TAO, but not in normal extraocular muscle tissues. Neither TAO nor normal extraocular muscle tissues expressed Bcl-2. FasL was detected in infiltrating mononuclear cells in extraocular muscle tissue from a patient with TAO. These data suggest that Fas-mediated apoptosis may occur in extraocular muscle tissue from patients with TAO and may be involved in the late stage of TAO.

Adult↗
PubMed1992

Significance of antibodies reactive with a 64 kDa eye muscle membrane antigen in patients with thyroid autoimmunity.

SDS-polyacrylamide gel electrophoresis and Western blotting for antibodies reactive with a 64 kDa protein in pig eye muscle membrane was carried out in patients with lid lag and retraction, but no other signs of ophthalmopathy, associated with thyroid disease or nonimmunologic goiter and in patients with Graves' hyperthyroidism without ophthalmopathy who were studied prospectively to determine the relationship of eye muscle antibodies to clinical features of the ophthalmopathy as they appeared in this group of predisposed patients. Seventy-one percent of euthyroid patients with lid lag and retraction but no established ophthalmopathy had detectable serum antibodies to a 64 kDa eye muscle membrane protein. Much smaller proportions had antibodies to proteins of other MW. In normal subjects with previously detectable antibodies to a 64 kDa protein, serum titers, determined by carrying out immunoblotting at serum dilutions of 1:25-1:6400, were low (< or = 1:100) in all cases tested. On the other hand, titers were higher (1:200-1:6400) in 16 of 22 patients with established ophthalmopathy and in 5 of 7 patients with lid lag and retraction tested. Titers tended to be lower in patients with ophthalmopathy of > or = 3 years duration than in those of < or = 1 year duration. Antibody titers were low (1:25) in 6 of 7 patients with Graves' hyperthyroidism without evident eye disease tested. Antibodies to a 64 kDa eye muscle membrane protein were predictive of the development of ophthalmopathy in patients with Graves' hyperthyroidism studied prospectively for periods of 8-42 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Evaluation of an enzyme-linked immunosorbent assay for the measurement of autoantibodies against eye muscle membrane antigens in Graves' ophthalmopathy.

We have tested for antibodies against human and pig eye muscle membrane antigens in the serum of patients with Graves' ophthalmopathy using an enzyme-linked immunosorbent assay (ELISA). Several different membrane preparations were used as source of putative antigen including a 100,000 X g pellet, a pellet depleted of the 100,000 X g (microsome) fraction, and solubilized membranes. With eye muscle membrane pellets there were no significant differences for either serum or immunoglobulins between patients with ophthalmopathy, those with autoimmune thyroid disorders without eye disease, and normal subjects for either human or pig membranes, although tests were positive determined from the upper limit of normal in a few patients with or without eye disease. This was the case regardless of the enzyme-antibody conjugate used, the membrane protein concentration or serum or immunoglobulin dilution. Pre-absorption of tissue fractions, serum, or immunoglobulins, with red blood cells or liver powder, eye muscle membranes or skeletal muscle membranes did not significantly reduce back-ground binding which was often very high, or enhance the difference between patients with ophthalmopathy and normal subjects. It was found that non-specific binding to the plastic surface of the microplates and/or tissue proteins, the presence, in human tissues, of blood-derived immunoglobulins which gave strong reactions in the ELISA, and variable fixation of membrane pellets to the plates were factors which made ELISA unsatisfactory when crude membrane pellets were used as antigen.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed2004

[The combination of eye muscle surgery with other interventions on or in the eye].

Strabismus and motility disorders of the eye can be linked in many different ways to intraocular diseases and disorders of the lid and orbit. Different functional aspects have to be considered before performing combined eye muscle and intraocular or adnexal surgery. This allows to avoid unnecessary operations, to choose the right dosage of the operation, to limit the risk of diplopia, and finally to achieve a satisfying result for the patient.

Combined Modality Therapy↗
PubMed1985

[Does proprioception of the extrinsic eye muscles participate in equilibrium, vision and oculomotor action?].

Extrinsic eye muscles respond to both proprioceptive and visual impulses. Histology of ocular proprioceptive receptors shows them to be qualitatively and original, neuromuscular bundles are found in primates, and so called "palisade sensory nerve endings" in felines. Golgi's osteotendinous receptors are apparently absent. The number of muscle receptors is high and their proportion in relation to muscle weight even more so. The afferent pathway for this sensory system initially accompanies the motor fibers of oculomotor III, IV, ans VI nerves. Some fibers continue with these nerves to the brain stem while others join the ophthalmic branch of the trigeminal nerve (probably in the cavernous sinus). Cell bodies of proprioceptive fibers are located either in the mesencephalic root of the trigeminal nerve and in the Gasserian ganglion. The superior colliculus, center of eye-head coordination integration, also receives proprioceptive data. Proprioceptive afferent signals also project on two cerebellar cortex regions, one in the VI th and VII th lobules of the posterior lobe vermis, the other in the flocculus, all major centers for control and interaction of visuocervicovestibular activity. Extraocular proprioceptive impulses play an important role as do visual influx, in the maturation of the visual cortex in mammalia. Moreover, maintenance of binocular integration appears to require both coherent binocular vision and balanced bilateral, extraocular afferent impulses. Clinical consequences are multiple. Proprioceptive imbalance due to oculomotor dysfunction could explain the attendant visual deficit, which is reversible after correction of they motor disorder. Forced excentration of gaze provokes equilibrium disturbances in normal subjects. In cases of oculomotor palsy, the accompanying vertigo often observed could thus be due à lack of coordination between the intention of movement and its execution. Many oculomotor signs are dependent on the position of gaze alone, in the absence of any visual guide: end-position nystagmus gaze nystagmus, rebound nystagmus, gaze drift, periodic alternating and vertical superior nystagmus, ocular flutter and opsoclonus, nystagmus retractorius, etc. Proprioceptive impulses from extrinsic eye muscles are probably necessary for assessment of orbital coordinates of the ocular globe position during target fixation: this emphasizes the importance of this proprioceptive factor in the origin of ocular jerks. Proprioceptive impulses may play a crucial role in the triggering of the rapid phase of nystagmus, thus conditioning the variations in amplitude seen in provoked nystagmus tests.

Afferent Pathways↗
PubMed1991

Contribution of eye muscle proprioception to velocity-response characteristics of eye movements: involvement of the cerebellar flocculus.

The vestibulo-ocular reflex (VOR) and optokinetic nystagmus (OKN) were examined in alert pigmented rabbits following interruption of proprioceptive afferents from the extraocular muscles in one eye by surgical section of the ophthalmic branch of the trigeminal nerve (V1 nerve). Deficits were mainly produced in movement dynamics of the ipsilateral eye including reduction of (1) the VOR gain at a high frequency of head rotation, (2) the OKN gain and (3) the velocity of quick eye movements in the OKN. In some of the rabbits examined, the cerebellar flocculus was lesioned by local injection of kainic acid before severance of the V1 nerve. No significant additional reductions of VOR or OKN gains were produced by V1 nerve section in the flocculus-lesioned rabbits. These results suggest that proprioceptive signals from eye muscles act to improve VOR and OKN dynamics through the neuronal mechanisms involving the cerebellar flocculus.

Animals↗
PubMed1993

Use of eye muscle antibody measurements to monitor response to plasmapheresis in patients with thyroid-associated ophthalmopathy.

We have measured eye muscle antibodies, in immunoblotting, in the serum from five patients with severe ophthalmopathy associated with Graves' hyperthyroidism who underwent plasmapheresis, correlating their levels with clinical features of the eye disorder and response to treatment. Blood taken before each plasma exchange was tested in SDS-polyacrylamide gel electrophoresis and Western blotting for antibodies reactive with pig eye muscle membrane (PEMM) antigens and in a 51Cr release assay for antibodies which are cytotoxic to human eye muscle cells in antibody-dependent cell-mediated cytotoxicity (ADCC). Antibodies reactive with a 64 kDa PEMM antigen were detected in three patients who had eye disease of less than six months duration, but not in the two with more chronic disease. Antibodies against a 95 kDa PEMM antigen were detected in one patient in whom anti-64 kDa antibodies were also demonstrated. All five patients showed significant improvement in their eye disease following plasmapheresis exchange and titres of the anti-64 kDa protein antibody decreased in the three patients with detectable levels before treatment. TSH receptor stimulating antibodies were detected in all five patients before treatment, falling during plasmapheresis in four and becoming undetectable in three by the end of treatment. There was no close correlation between levels of TSH receptor antibodies and titres of anti-64 kDa protein antibodies although both tended to fall during and following plasmapheresis. ADCC tests were negative in all five patients before plasmapheresis but, surprisingly, transiently positive in three following treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1979

Cell-mediated lysis of lipid vesicles containing eye muscle protein: implications regarding pathogenesis of Graves ophthalmopathy.

We prepared artificial vesicles that are lysed upon cell-mediated immunological attack by human lymphocytes. These vesicles are made from a mixture of dimyristoyl lecithin, dipalmitoyl lecithin, and cholesterol, have eye muscle membrane protein (EMP) inserted into the bilayer wall, and contain intravesicular (99m)Tc marker. Injury to the vesicular membrane was assessed by measurement of (99m)Tc release. Thyroglobulin (Tg) and Tg-anti-Tg complex (TgA) bind to EMP-vesicles to an extent equal to or greater than to native eye muscle membranes in vitro; this binding requires the presence of normal human IgG. The role of Tg, TgA, IgG, and peripheral blood lymphocytes in altering membrane permeability was analyzed. Incubation of vesicles for up to 3 hr alone, with added IgG alone, or with further addition of Tg or TgA did not result in (99m)Tc release. Addition of lymphocytes from normal donors to the above four preparations showed release in the presence of TgA. Lymphocytes from each of eight patients with Graves ophthalmopathy caused release not only in the presence of TgA, but also in the presence of Tg. Separation of a patient's lymphocytes into high- and low-affinity rosette-formers (T and K cells, respectively) showed that cell-mediated vesicle lysis in the presence of TgA was greater with K cells than with T cells, while vesicle lysis in the presence of Tg was greater with T cells than with K cells. Vesicles made with inserted Tg but lacking EMP were not lysed by such T cells. Lymphocytes failed to induce permeability changes in vesicles containing other inserted proteins obtained from human nonextraocular muscle, liver, spleen, or adrenal, even if Tg or TgA were present. The results support the concept that muscle cell damage in Graves ophthalmopathy is immunological, cell-mediated, and of two types: (i) K lymphocytes reacting to immune complex, TgA, on the eye muscle cell surface (i.e., antibody-dependent cytotoxicity) and (ii) sensitized T lymphocytes reacting to Tg on the eye muscle cell surface. An antigenic role for EMP is possible, but has not been unequivocally proven.

Antigen-Antibody Complex↗
PubMed1991

Prevalence of antibodies reactive with a 64 kDa eye muscle membrane antigen in thyroid-associated ophthalmopathy.

We have determined the prevalences of antibodies reactive with certain pig eye muscle membrane antigens, as determined from SDS-polyacrylamide gel electrophoresis (PAGE) and western blotting, in patients with autoimmune thyroid disorders with or without associated ophthalmopathy. The most frequently detected antibody was that directed against a protein of 64 kDa. Antibodies against this antigen were detected in 33% of unselected patients with ophthalmopathy and in 75% of those with severe, active, orbital inflammation of less than 12 months duration. Such antibodies were detected also in 33% of patients with Graves' hyperthyroidism with no apparent eye disease and in 17% of patients with Hashimoto's thyroiditis with no eye disease but were not detected in patients with nonautoimmune thyroid disorders in 25 normal subjects tested. In the majority of sera showing reactivity with this protein, reactivity with membrane antigens of 55 and 95 kDa also were observed, suggesting that the three proteins were conformationally associated. Cytotoxic antibodies against human eye muscle cells, measured in antibody-dependent cell-mediated cytotoxicity (ADCC), were demonstrated in 37% of patients with ophthalmopathy. There was no significant correlation, in patients with eye disease, between positive ADCC tests and antibodies to the 64 kDa protein, the 64 kDa reactive antibody being found in 50% of patients with positive ADCC and in 35% of those with negative tests. In this study, a clear association has been shown between thyroid-associated ophthalmopathy and the detection, in immunoblotting, of antibodies reactive with a 64 kDa eye muscle membrane antigen.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1976

Somatotopic localization of the eye muscle afferents in the semilunar ganglion.

In the medial dorsolateral portion of the semilunar ganglion of curarized and anaesthetized lambs a cellular pool has been identified which contains the perikarya of the first-order neurons of the eye muscle proprioception. Responses to moderate manual stretch of individual eye muscles were recorded by means of tungsten microelectrodes, from single units of the ganglion. They were of the type induced by muscle spindle excitation. Such responses showed a somatotopic localization. The superior rectus and the superior oblique muscles were represented in the most dorsal layers of the ganglion, while the inferior rectus and the inferior oblique muscles projected on the most ventral portion of the pool. The medial and the lateral recti were represented in the medial and lateral parts and occasionally wedged themselves between the cells innervating the superior and the inferior muscles. Thus a somatotopic arrangement of the eye muscle proprioception has been demonstrated for the first time in the semilunar ganglion.

Animals↗
PubMedDate not supplied

Influence of rectus eye muscles tenotomy on the aqueous humor composition in rabbits.

INTRODUCTION: Anterior segment ischaemia manifesting in aqueous malformation may follow tenotomy of the extraocular muscles. MATERIAL AND METHODS: Tenotomy of 2 or 4 rectus eye muscles was performed on the right eyes of 23 adult rabbits. The aqueous humor was withdrawn from the anterior chamber of both eyes in 7 rabbits on the 11th, on the 22nd in 8 and on the 32nd postoperative day in 8 rabbits. Aqueous natrium, potassium and glucose concentrations and protein electrophoresis were examined. RESULTS: The only significant change was lower mean aqueous potassium concentration in the operated eyes as compared to controls on the 11th postoperative day. CONCLUSION: Tenotomy of rectus eye muscles in healthy rabbits evoked a discrete blood-aqueous barrier break-down.

Albumins↗
PubMed2006

Study of serum antibodies against three eye muscle antigens and the connective tissue antigen collagen XIII in patients with Graves' disease with and without ophthalmopathy: correlation with clinical features.

OBJECTIVE: The extraocular muscles are one of the primary tissues implicated in the autoimmune-mediated inflammation of thyroid-associated ophthalmopathy (TAO). Our aim was to determine the prevalence and level of antibodies against three candidate eye muscle antigens and the orbital fibroblast membrane antigen collagen XIII, in well-characterized patient groups. STUDY DESIGN AND PATIENTS: The study cohort consisted of patients with Graves' hyperthyroidism with and without ophthalmopathy, controls patients with other thyroid or other autoimmune disorders and healthy subjects. The presence of eye muscle antibodies was determined using an optimized and standardized enzyme-linked immunosorbent assay. We measured antibodies against (i) the 67-kDa flavoprotein (Fp) subunit of the mitochondrial enzyme succinate dehydrogenase; (ii) G2s, a 141 amino acid fragment of the winged-helix transcription factor FOXP1; (iii) calsequestrin, a 63-kDa calcium-binding protein; and (iv) collagen XIII, a connective tissue protein that is closely linked to the congestive ophthalmopathy subtype of TAO. Eye muscle antibody levels were correlated with clinical diagnosis and presence or not of ophthalmopathy. RESULTS: Prevalences of positive antibody tests to calsequestrin (75.0%) and collagen XIII (43.8%) were significantly greater in Graves' disease (GD) patients with ophthalmopathy than in healthy subjects, whereas modest significance was demonstrated with antibodies against Fp, but not G2s. Significantly greater serum levels of antibodies against calsequestrin, G2s, and collagen XIII, but not Fp, were found in GD patients with ophthalmopathy compared to control patients without eye disease and healthy subjects. CONCLUSIONS: Calsequestrin and collagen XIII antibodies are the most specific to TAO, whereas antibodies against G2s, and to a lesser extent Fp, are also markers of ophthalmopathy, but less reliable. These results are unique in that it is the first time the significance of a panel of three candidate eye muscle antibodies and a connective tissue antibody have been evaluated in the same patients with ophthalmopathy.

Adult↗