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1.
J Invest Dermatol ; 97(3): 511-6, 1991 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-1875051

RESUMO

Blood mononuclear cells (MNC) from patients with psoriasis were more adherent to monolayers of endothelial cells prepared from human umbilical cord vein than otherwise similar cells from control subjects. This increase in adherence occurred in the presence (mean 37% increase; p less than 0.01) and absence (mean 47% increase; p less than 0.05) of 10% autologous serum and was not related to the disease severity of the patients. The augmented adhesiveness of the patients' cells was also apparent when using monolayers of endothelial cells isolated from human skin. The levels of immune complexes, complement, alpha 2-macroglobulin, acute phase proteins (alpha 1-acid glycoprotein, C-reactive protein and alpha 1-antitrypsin), and tumor necrosis factor alpha (TNF alpha), interleukin-1 alpha (IL-1 alpha), and interleukin-1 beta (IL-1 beta) in the patients' sera were within normal limits. When MNC were added to endothelial monolayers that had been incubated with either TNF alpha or the highest concentration of rIL-1 beta used in the study, both the patients' and control's cells exhibited a similar increase in attachment (p less than 0.01). Pretreatment of endothelium with interferon-gamma did not enhance the attachment of MNC from either group of subjects. The augmented adherence of the patient's MNC appears to be due to an abnormal adhesiveness of the lymphocytes rather than the monocytes and is not related to an enhanced expression of the cell-surface adhesion molecules CD11a/CD18. It is likely that the circulating MNC of psoriatic patients may be predisposed for extravasation into skin.


Assuntos
Endotélio Vascular/citologia , Leucócitos Mononucleares/citologia , Psoríase/sangue , Adesão Celular , Citocinas/farmacologia , Endotélio Vascular/efeitos dos fármacos , Feminino , Humanos , Microcirculação , Pele/irrigação sanguínea
2.
J Immunol Methods ; 113(2): 157-63, 1988 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-3171188

RESUMO

Porcine aortic vascular endothelial cells were grown to confluence on microporous PTFE membranes and incorporated into a two-compartment chemotaxis assembly. Human peripheral blood mononuclear cells (20-25% monocytes) were placed in the upper compartment and zymosan-activated human serum (ZAHS) as chemoattractant in the lower compartment. Over a 3 h period monocytes migrated across the endothelialized membrane and adhered to a collecting filter sited in the lower compartment. Addition of ZAHS to the cell suspension in the upper compartment virtually abolished the migration response whilst only minimal leucocyte migration was supported by the bare unendothelialized PTFE membrane. The extent of transendothelial monocyte chemotaxis was dependent upon the concentration of chemoattractant in the lower compartment and upon the cell density of the suspension containing monocytes. A confluency test of fluid flow across the endothelialized filter showed that monocyte migration could take place without disturbing endothelial barrier function. The culture system is easy to assemble and the method provides experimental versatility.


Assuntos
Quimiotaxia de Leucócito , Endotélio Vascular/citologia , Monócitos/fisiologia , Animais , Complemento C5 , Endotélio Vascular/fisiologia , Humanos , Contagem de Leucócitos , Membranas Artificiais , Monócitos/ultraestrutura , Politetrafluoretileno , Suspensões , Suínos , Zimosan
3.
J Immunol Methods ; 82(2): 189-98, 1985 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-3900214

RESUMO

Using a cell electrophoretic apparatus, which was sensitive in detecting small changes in electrophoretic mobility (EPM), the macrophage electrophoretic mobility (MEM) test was investigated as a routine method for detecting lymphokine activity. Electrophoretic analysis of guinea-pig macrophages revealed 2 main subpopulations, one with an EPM of 0.90 micron cm s-1 V-1 (fast) and the other, an EPM of 0.83 micron cm s-1 V-1 (slow). From 23 experiments the fast and slow populations were found to consist of 90% and 10% cells, respectively. When macrophages were incubated with standard guinea-pig lymphokine preparations there was a significant decrease in the fast population with a corresponding increase in the slow population. This lymphokine induced 'slowing' of the macrophages was shown to be very reproducible. Since only 50% of macrophages of high EPM were observed to respond to lymphokine activity, it is not surprising that the MEM test has failed in the past when investigators have accepted as significant a 10-15% reduction in EPM, estimated from measurements made on only 10 macrophages. Parallel bioassays indicated that there were appreciable potency differences for macrophage slowing factor (MSF) and macrophage migration inhibition factor (MIF) activities in the lymphokine preparations used which suggest that these activities may be due to different molecular entities.


Assuntos
Quimiotaxia de Leucócito/efeitos dos fármacos , Eletroforese , Fatores Inibidores da Migração de Leucócitos/farmacologia , Linfocinas/farmacologia , Macrófagos/imunologia , Animais , Resistência a Medicamentos , Estudos de Avaliação como Assunto , Cobaias , Macrófagos/efeitos dos fármacos , Macrófagos/fisiologia , Propriedades de Superfície
4.
J Immunol Methods ; 180(2): 165-80, 1995 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-7714332

RESUMO

In order to identify the factors that control the binding of blood leucocytes to cerebral blood vessels we have modified and applied the frozen section assay of Stamper and Woodruff to the study of human brain. Cryostat sections of brain tissue obtained at post mortem were overlaid with blood lymphocytes and experimental conditions were defined which permitted optimum binding of the cells to transected blood vessel walls. The maximal binding of lymphocytes to cerebral vessels occurred when 6 x 10(6) lymphocytes were overlaid onto brain sections for 30 min at 7 degrees C with gentle agitation. Only a small proportion (0.01%) of the added lymphocytes bound to exposed cerebral vessels. However, lymphocytes were far more adherent than monocytes and polymorphonuclear cells (7-fold and 11-fold respectively: p < 0.001) and activation of lymphocytes with IL-2 enhanced their binding to blood vessel walls (mean 130% increase; p < 0.03). Further analysis revealed that CD4-positive T lymphocytes were the predominant cell population binding to the blood vessels. Antibody blocking studies showed that lymphocyte binding to cerebral blood vessels was inhibited by pretreating the lymphocytes with anti-CD11a, anti-CD18 or anti-CD49d (p < or = 0.02) and immunohistochemical studies revealed the presence of the counter-receptors ICAM-1 (CD54) and VCAM-1 (CD106) for these adhesion molecules in addition to the presence of E-selectin (CD62E) and P-selectin (CD62P) on the cerebral blood vessels. The establishment of a technique in situ which measures selective binding of CD4-positive peripheral lymphocytes to sections of cerebral blood vessels will assist in the molecular characterization of factors that control the interaction of leucocytes with the blood-brain barrier in health and disease.


Assuntos
Encéfalo/irrigação sanguínea , Linfócitos/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais/imunologia , Vasos Sanguíneos/citologia , Antígenos CD18/imunologia , Linfócitos T CD4-Positivos/fisiologia , Linfócitos T CD8-Positivos/fisiologia , Adesão Celular/fisiologia , Moléculas de Adesão Celular/fisiologia , Humanos , Técnicas In Vitro , Interleucina-2/fisiologia , Leucócitos/fisiologia , Ativação Linfocitária/fisiologia , Antígeno-1 Associado à Função Linfocitária/imunologia , Pessoa de Meia-Idade
5.
J Immunol Methods ; 54(3): 317-29, 1982 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-6757327

RESUMO

The leucocyte migration inhibition test in agarose as described by Clausen (1971) was modified into a statistically designed assay of LIF activity using human polymorphonuclear leucocytes from single blood donors. Individual assays included a laboratory standard of lymphokine with LIF activity prepared from the culture supernatants of the RPMI 1788 human lymphoblastoid cell line (LCL-LK). Analysis of 157 LIF assays revealed simple criteria by which the acceptability of an individual assay could be judged before subjecting it to statistical analysis. The failure of LIF assays to meet these criteria of acceptability was particularly associated with low areas of control polymorph migration in the absence of lymphokine ('spontaneous migration'). We demonstrate that the statistically designed assay permits the measurement, with precision, of LIF activity in units/ml by reference to a working standard of LCL-LK. We illustrate the use of this assay in the measurement of LIF activity generated by tuberculin-stimulated human peripheral blood lymphocytes.


Assuntos
Fatores Inibidores da Migração de Leucócitos/análise , Linfocinas/análise , Linfocinas/normas , Animais , Linhagem Celular , Inibição de Migração Celular , Relação Dose-Resposta Imunológica , Cobaias , Humanos , Fatores Inibidores da Migração de Leucócitos/farmacologia , Ativação Linfocitária , Linfocinas/biossíntese , Neutrófilos/imunologia , Padrões de Referência , Tuberculina/imunologia
6.
J Immunol Methods ; 100(1-2): 147-52, 1987 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-3496396

RESUMO

This paper describes the use of phenyl-Sepharose CL-4B as a solid-phase hydrophobic adsorbent in the purification of S-antigen from protein extracts of bovine, porcine and human retina. Chromatographic conditions were ascertained whereby the majority of contaminating proteins were bound to the adsorbent leaving S-antigen in the liquid phase. In combination with size fractionation on Ultrogel AcA, the method conveniently yielded porcine and bovine S-antigen preparations up to 100% purity. Immunogenicity of purified S-antigens was verified by induction of experimental autoimmune uveoretinitis in albino Lewis rats. The method is preparative in scale, fast in performance and yields S-antigen in high purity and antigenic potency.


Assuntos
Antígenos/isolamento & purificação , Proteínas do Olho/isolamento & purificação , Adsorção , Animais , Antígenos/imunologia , Arrestina , Bovinos , Cromatografia , Proteínas do Olho/imunologia , Humanos , Masculino , Ratos , Ratos Endogâmicos Lew , Sefarose/análogos & derivados , Suínos
7.
Invest Ophthalmol Vis Sci ; 33(1): 30-5, 1992 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-1730546

RESUMO

Electroretinographic supernormality, affecting both the a- and b-waves of the electroretinogram (ERG), occurs consistently before the onset of experimental autoimmune uveoretinitis (EAU) in rabbits and rats. To investigate the possible role of antibody to S-antigen (S-ab) in this phenomenon, affinity-purified polyclonal rat S-ab was injected intravenously into normal rats and administered to isolated rat eyecup preparations by bolus perfusion. In both situations, ERG supernormality was observed. The effect in vivo peaked 90 min after injection, and ERG changes in vitro were observed within 15 sec. The ERG response in vivo and in vitro was dose dependent and was abolished in vivo by pretreatment with cyproheptadine (a serotonin antagonist). The ERG was not affected in either system by a control rat antibody (antiovalbumin) or by murine monoclonal or rabbit polyclonal antibodies to S-antigen. The induction of ERG supernormality in vivo and in vitro by homologous S-ab indicates the operation of species-specific mechanisms both involving and bypassing the blood-retinal barrier and highlights a significant role for humoral autoimmunity in the pathogenesis of S-antigen-induced EAU in the rat.


Assuntos
Antígenos/imunologia , Autoanticorpos/administração & dosagem , Autoantígenos/imunologia , Eletrorretinografia , Proteínas do Olho/imunologia , Retinite/fisiopatologia , Uveíte/fisiopatologia , Animais , Arrestina , Bovinos , Cromatografia de Afinidade , Ciproeptadina/administração & dosagem , Eletroforese em Gel de Poliacrilamida , Injeções Intravenosas , Coelhos , Ratos , Retinite/imunologia , Uveíte/imunologia
8.
Invest Ophthalmol Vis Sci ; 28(3): 604-7, 1987 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-3549614

RESUMO

Retinal S-antigen (S-ag) was purified by monoclonal antibody (MoAb) immunoaffinity chromatography from soluble protein extracts of bovine and human retina. Purification of S-ag was readily achieved by affinity chromatography using four different MoAb-Sepharose 4B columns. The four antibody columns gave different recoveries with material of comparable enrichment with greater than 95% purity as judged by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The use of two different MoAbs covalently bound to Sepharose 4B and known to be directed to disparate, spacially distant epitopes on S-ag led to at least a twofold increase in recovery, with the aforementioned purity. Immunoaffinity purified S-ag retained its serological and uveitogenic properties. The high recovery of S-ag associated with this one-step procedure is preferable to conventional preparatory techniques, and enables high antigen recovery when tissue availability is limited (eg human retina).


Assuntos
Anticorpos Monoclonais , Antígenos/isolamento & purificação , Proteínas do Olho/isolamento & purificação , Arrestina , Sítios de Ligação de Anticorpos , Reações Cruzadas , Técnicas Imunológicas , Imunoadsorventes
9.
Invest Ophthalmol Vis Sci ; 29(1): 12-21, 1988 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-2447030

RESUMO

Eight murine monoclonal antibodies to retinal S-antigen were found to recognize three antigenic clusters in competitive binding experiments. Variable patterns of immunohistochemical staining of retinal sections supported the competitive binding studies. Monoclonal antibodies to two of these three antigenic clusters on S-antigen reacted by Western immunoblotting with both intact S-antigen and two cyanogen bromide cleaved peptide fragments of bovine S-antigen. The binding of the monoclonal antibodies was localized to epitopes contained on two peptide fragments of 26,000 and 18,000 molecular weight. The same peptide fragments also react with polyclonal rat or rabbit antisera to bovine S-antigen. Monoclonal antibodies to the third antigenic cluster did not react with either intact or fragmented S-antigen in immunoblotting studies.


Assuntos
Anticorpos Monoclonais , Antígenos/imunologia , Epitopos , Proteínas do Olho/imunologia , Animais , Anticorpos Monoclonais/imunologia , Arrestina , Sítios de Ligação de Anticorpos , Ligação Competitiva , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos BALB C
10.
Invest Ophthalmol Vis Sci ; 38(12): 2608-18, 1997 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9375580

RESUMO

PURPOSE: To develop a sensitive and reproducible technique for measuring the adherence of blood lymphocytes to vessel walls exposed in sections of human retina and for examining the role of lymphocyte and vascular adhesion molecules in these events. METHODS: Cryostat sections of human retina were overlaid with blood lymphocytes from healthy subjects, and experimental conditions were sought by which preferential attachment of the cells occurred to blood vessel walls in the retinal sections. Adherent lymphocytes were identified by staining with methyl green-thionine, and transected blood vessels were identified by their structure and by staining of basement membranes with periodic acid-Schiff. The adherence of enriched preparations of CD4+ (T-helper) and CD8+ (T-cytotoxic) lymphocytes, of interleukin-2 (IL-2)-activated cells, and of lymphocytes from patients with ocular Behçet's disease was examined. The distribution of adhesion molecules on retinal vessel walls was determined by immunohistochemistry, and the contribution of leukocyte integrins to lymphocyte binding was studied by blocking experiments with monoclonal antibodies. RESULTS: The optimal selectivity of blood lymphocyte attachment to retinal vessel walls occurred when purified lymphocytes were suspended in culture medium with 10% fetal calf serum and overlaid onto retinal sections for 30 minutes at 23 degrees C with gentle agitation. Under these conditions, 92% of the lymphocytes that adhered to the section were confined to the retinal microvasculature, and CD4+ T cells were more adherent than CD8+ T cells (P < 0.01). Prior exposure of normal lymphocytes to IL-2 enhanced their binding to retinal blood vessels, and lymphocytes from patients with Behçet's disease showed supranormal vascular adherence (P < 0.005). Many transected vessels stained positively for CD31; PECAM (mean 62%), CD54; ICAM-1 (mean 73%), CD62E; E-selectin (mean 35%), CD62P; P-selectin (mean 61%), and CD106; VCAM-1 (mean 42%). However, these vascular adhesion molecules occupied < 20% of the area of the blood vessel walls. Lymphocyte adhesion to the retinal vessels was more dependent on CD29 (the common chain of the beta 1 integrins) expression than either CD11a/CD18 or CD49d. CONCLUSIONS: This technique allows measurements to be made of lymphocyte adherence to vascular and nonvascular structures of retina ex vivo. Extension of this approach to the study of leukocyte adherence to sections of pathologic retina may be of clinical and experimental applicability in understanding mechanisms of retinal inflammation.


Assuntos
Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Adesão Celular , Vasos Retinianos/metabolismo , Antígenos CD/metabolismo , Síndrome de Behçet/metabolismo , Moléculas de Adesão Celular/metabolismo , Humanos , Técnicas Imunoenzimáticas , Interleucina-2/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
11.
Invest Ophthalmol Vis Sci ; 38(5): 1043-8, 1997 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9113001

RESUMO

PURPOSE: To measure vitreous levels of the soluble intercellular adhesion molecule (sICAM-1) in eyes with rhegmatogenous retinal detachment (RRD) complicated or uncomplicated by proliferative vitreoretinopathy (PVR) to investigate whether levels of this molecule related to history of previous retinal surgery or to the duration and severity of PVR. METHODS: The authors measured vitreous sICAM-1 by enzyme-linked immunosorbent assay in 28 eyes with PVR and 35 eyes with uncomplicated RRD. Vitreous from 10 eyes with macular holes and from 12 cadaveric eye donors were used as control specimens. RESULTS: Vitreous sICAM-1 levels were higher in the group with RRD complicated by PVR as a whole than in the group with RRD alone or in the control groups. In patients with no previous retinal surgery, there was no difference in vitreous sICAM-1 levels between the groups with RRD alone and RRD complicated by PVR. However, in patients who had undergone previous external surgery, those with PVR showed higher levels of vitreous sICAM-1 than those with RRD alone. In PVR, raised levels of sICAM-1 were associated preferentially with a history of previous vitrectomy as well as with a longer duration of the condition, although these levels were not related to the grade of PVR. In eyes with RRD alone, the levels of sICAM-1 were not enhanced with the duration of the detachment. Despite showing high vitreous levels of sICAM-1, patients with PVR did not exhibit increased serum levels of this adhesion molecule. CONCLUSIONS: The current observations suggest that those persons in whom PVR develops may have an impairment of the mechanisms that control the inflammatory response to retinal trauma. Persistently raised vitreous levels of sICAM-1 point to the continued operation of cytokine-mediated vascular reactions at the blood-retinal barrier.


Assuntos
Molécula 1 de Adesão Intercelular/metabolismo , Vitreorretinopatia Proliferativa/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Descolamento Retiniano/complicações , Descolamento Retiniano/metabolismo , Vitreorretinopatia Proliferativa/etiologia , Corpo Vítreo/metabolismo
12.
Hum Immunol ; 4(3): 197-208, 1982 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-6181030

RESUMO

In vitro primary sensitization of human peripheral blood T-cells to hemocyanins was detected during a 12-14 day culture period with antigen (KLH or HCH)-pulsed macrophages. Primed T-cells proliferate in secondary culture (2-3 days) when restimulated with antigen presented by macrophages. The kinetics of primary sensitization and secondary responsiveness are interrelated and are dependent on the antigen doses employed. Antigen-induced proliferation of cells sensitized in vitro is identical to proliferation of T-cells from immunized donors in terms of antigen specificity and the clonal nature of the response to antigen. Through the use of thymidine suicide techniques, distinct populations of cells responding to KLH or HCH can be demonstrated using cells from actively immunized donors or cells that have been sensitized in vitro to both hemocyanins.


Assuntos
Epitopos , Hemocianinas/imunologia , Imunização/métodos , Linfócitos T/imunologia , Relação Dose-Resposta Imunológica , Caranguejos Ferradura/imunologia , Humanos , Cinética , Ativação Linfocitária , Moluscos/imunologia , Linfócitos T/metabolismo , Timidina/metabolismo
13.
Hum Immunol ; 3(3): 209-30, 1981 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-6975768

RESUMO

Activation of human T cells requires presentation of antigen by Ia (HLA-DR in man) bearing cells of the mononuclear phagocytic series (macrophages, MO, and more recently Langerhans cells, dendritic cells, and vascular endothelial cells. Since T cells must cross endothelial barriers to enter extravascular tissues during immune reactions, we investigated the role of endothelial cells in antigen presentation. Endothelial cells were cultured from human umbilical veins and identified by classic morphology and specific markers (factor VIII related antigen, and so on). Antigen-pulsed endothelial cells were used to present antigen to MO-depleted human T cells; activation was assessed by 3H-thymidine uptake. The HLA-DR compatible endothelial cells were as effective as MO in reconstituting MO-depleted T-cell responses. The endothelial cell reconstituted responses were antigen specific, HLA-DR restricted, and blocked by monoclonal antibodies to HLA-DR framework structures. Moreover, the T-cell responses were clonal with respect to HLA-DR. A monoclonal antibody completely eliminated MO reconstitution of the MO-depleted response without diminution of endothelial cell reconstitution of the same response. Fibroblasts and smooth muscle cells cultured from the same umbilical veins could not reconstitute the MO-depleted T-cell response. These data indicate that endothelial cells play an important and distinctive role in lymphocyte triggering.


Assuntos
Antígenos , Ativação Linfocitária , Linfócitos T/imunologia , Veias Umbilicais/imunologia , Antígenos/genética , Separação Celular , Células Cultivadas , Células Clonais/imunologia , Endotélio/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Soros Imunes/farmacologia , Macrófagos/imunologia
14.
Autoimmunity ; 9(2): 91-7, 1991.
Artigo em Inglês | MEDLINE | ID: mdl-1718460

RESUMO

This paper describes the idiotypic specificities of eight murine monoclonal antibodies directed to three independent epitopes on retinal S-antigen. The antigenic sites recognised by these monoclonal antibodies have previously been localised to a small region near the C-terminal of bovine S-antigen. Xenogeneic, site-related anti-idiotypes prepared against each of the monoclonal antibodies recognised common idiotypes only amongst those monoclonal antibodies which reacted with the same epitope on S-antigen. Two of the three idiotypes were detected in the sera of BALB/c mice but not in two strains of rat immunised with xenogeneic S-antigen and none could be detected in the sera of patients with anti-photoreceptor autoantibodies. Our results demonstrate that the idiotypes of murine monoclonal antibodies to retinal S-antigen exhibit restricted epitope specificity but are species-restricted and imply that the S-antigen lacks a dominant antigenic epitope.


Assuntos
Anticorpos Monoclonais/imunologia , Antígenos/imunologia , Autoantígenos/imunologia , Epitopos/imunologia , Proteínas do Olho/imunologia , Idiótipos de Imunoglobulinas/análise , Especificidade de Anticorpos , Arrestina , Ligação Competitiva , Humanos , Imunoensaio , Técnicas In Vitro , Doenças Retinianas/imunologia , Vasculite/imunologia
15.
Arch Ophthalmol ; 106(1): 111-4, 1988 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-2892482

RESUMO

The effect of an additional adjuvant, Bordetella pertussis, on the clinical and histopathologic features of experimental autoimmune uveitis in black-hooded Lister rats was investigated. Disease was induced by a single footpad injection of purified retinal S-antigen in Freund's complete adjuvant. In those animals that did not receive B Pertussis the clinical features were those of a retinal vasculitis with disc edema, periphlebitis, and deep retinal infiltrates. In contrast, animals that received B pertussis developed lesions in the pigment epithelium and choroid. Histopathologic studies disclosed focal photoreceptor necrosis associated with mononuclear cell infiltration in both groups of animals. However, in the group that did not receive B pertussis the disease was predominantly a retinitis associated with perivascular infiltration of retinal vessels, whereas in the group that did receive B pertussis the main feature was a focal choroiditis, with superficial retinal lesions being rarely observed. Retinal photoreceptors were the target tissue in both groups of rats, but the route by which they were damaged was altered from predominantly retinal to choroidal by the addition of Bordetella pertussis as an adjuvant. This change may be ascribed to the ability of B pertussis toxin to sensitize vascular endothelium to local mast cell products, these cells being plentiful around choroidal vessels but absent in the retinal circulation.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Bordetella pertussis/imunologia , Uveíte/patologia , Animais , Antígenos/administração & dosagem , Arrestina , Autoantígenos/administração & dosagem , Doenças Autoimunes/patologia , Corioide/patologia , Proteínas do Olho/administração & dosagem , Angiofluoresceinografia , Adjuvante de Freund/administração & dosagem , Masculino , Mycobacterium tuberculosis/imunologia , Disco Óptico/patologia , Epitélio Pigmentado Ocular/patologia , Ratos , Retina/patologia , Vasos Retinianos/patologia , Uveíte/etiologia
16.
J Clin Pathol ; 24(6): 533-6, 1971 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-5094689

RESUMO

Inhibition of buffy layer peripheral leucocyte migration by tuberculin purified protein derivative (PPD) from micro-capillaries mounted in small tissue culture chambers correlates in all cases with a positive Mantoux test. This quick and reproducible test of cellular hypersensitivity in man requires only 5-15 ml of blood and is applicable for study in children and in clinical situations where repeated monitoring of cellular immunity may be required.


Assuntos
Inibição de Migração Celular , Leucócitos/imunologia , Teste Tuberculínico , Adolescente , Adulto , Idoso , Antígenos , Criança , Pré-Escolar , Técnicas de Cultura , Humanos , Imunidade Celular , Lactente , Métodos , Microquímica , Pessoa de Meia-Idade
17.
Am J Ophthalmol ; 113(6): 697-701, 1992 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-1598962

RESUMO

Using a modified enzyme-linked immunosorbent antibody method that included dissociation of antigen antibody complexes with sodium thiocyanate, we examined the functional affinity of antibody to retinal S-antigen in 48 patients with retinal vasculitis and 46 age-matched healthy control subjects. Antibody affinity was markedly lower in patients with retinal vasculitis than in healthy subjects. Low-affinity antibody was more prevalent in acute retinal vasculitis and in patients with normal levels of circulating immune complexes. We found distinct differences between the antiretinal antibodies found in patients with retinal vasculitis and those in control subjects. The association of low-affinity antibody with normal levels of circulating immune complexes may suggest defective regulation of antiretinal autoimmunity and have important pathogenic implications.


Assuntos
Afinidade de Anticorpos/imunologia , Antígenos/imunologia , Autoantígenos/imunologia , Proteínas do Olho/imunologia , Vasos Retinianos/imunologia , Vasculite/imunologia , Complexo Antígeno-Anticorpo/imunologia , Arrestina , Autoanticorpos/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Doenças Retinianas/imunologia
18.
Br J Ophthalmol ; 76(9): 553-9, 1992 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-1420062

RESUMO

S-antigen induced experimental autoimmune uveoretinitis (EAU) was produced in the Royal College of Surgeons (RCS) strain of rat which develops a photoreceptor dystrophy within 2 weeks of birth. Animals were sensitised at 60, 90, and 105 days of age: all animals developed disease, but onset was significantly delayed in older (105 day) animals compared with those aged 60 days at sensitisation (p 0.003). Disease was characterised by the early development of complete serous retinal detachment which resolved in a few days: the prevalence of retinal detachment was increased to 80% in dystrophic animals compared with 10% in the congenic, non-dystrophic controls (p < 0.001). Anterior uveitis was seen in 17/30 control strain eyes, but in none of 30 dystrophic eyes (p < 0.001). Genetically determined photoreceptor and retinal pigment epithelium dysfunction in the RCS rat, which may involve the local accumulation of altered S-antigen, predisposes the dystrophic strain to display an acute retinal detachment in the early stages of EAU. This phenomenon illustrates how biochemical dysfunction of a target organ may influence susceptibility, form, and severity of an experimental autoimmune disease.


Assuntos
Doenças Autoimunes/etiologia , Degeneração Retiniana/complicações , Retinite/etiologia , Uveíte/etiologia , Fatores Etários , Animais , Antígenos/imunologia , Arrestina , Autoantígenos/imunologia , Doenças Autoimunes/patologia , Proteínas do Olho/imunologia , Angiofluoresceinografia , Ratos , Ratos Endogâmicos , Retina/patologia , Descolamento Retiniano/etiologia , Retinite/patologia , Uveíte/patologia
19.
Br J Ophthalmol ; 72(6): 442-7, 1988 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-3390420

RESUMO

Fifty-two patients with retinal vasculitis--26 with idiopathic disease and 26 with associated systemic inflammatory disease--were followed up for periods ranging from six months to 12 years. The aim of the study was to determine the relationship between relapse of uveitis, visual outcome, and the occurrence of circulating immune complexes (CIC) and antiretinal antibodies. In a total of 69 relapses, CIC were increased in one-third of patients and antiretinal antibodies in one-half. In those 34 patients who expressed antiretinal antibodies 27 (79%) of the relapses were characterised by antiretinal antibodies in the absence of raised CIC levels (p less than 0.01). These findings support our previous hypothesis that CIC may have a protective role in autoimmune retinal vasculitis and that antiretinal autoimmunity is of pathogenetic importance in relapse. In individual patients the visual outcome was not related to the number of relapses or to the CIC-autoantibody pattern, suggesting the operation of additional features which merit identification.


Assuntos
Complexo Antígeno-Anticorpo/análise , Autoanticorpos/análise , Doenças Retinianas/imunologia , Vasos Retinianos/imunologia , Vasculite/imunologia , Adolescente , Adulto , Idoso , Seguimentos , Humanos , Pessoa de Meia-Idade , Recidiva
20.
Br J Ophthalmol ; 82(9): 1017-21, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9893591

RESUMO

BACKGROUND/AIMS: Toxoplasma retinochoroiditis (TR) is an important cause of blindness and visual morbidity, affecting young adults. It has been postulated that some of the retinal damage observed in TR is due to antiretinal autoimmune mechanisms. METHODS: Humoral antiretinal autoimmunity in TR was investigated by indirect immunofluorescence (IIF) on normal human cadaveric retina and by a human retinal S-antigen ELISA. 36 patients with TR were separated on clinical grounds into those with first recurrence of disease (n = 18) or those with multiple recurrences (n = 18). Patients were also segregated into those with active (n = 28) or quiescent disease (n = 8). Serum from 16 normal controls (six with positive toxoplasma serology and 10 without) with no evidence of eye disease and 12 patients with idiopathic retinal vasculitis (IRV) were also tested. RESULTS: Sera from 34 of the 36 patients (94%) with TR demonstrated photoreceptor layer reactivity by IIF contrasting with six of 16 normal controls (p = < 0.001) and three of 12 IRV patients (p = < 0.001). Titres of antiphotoreceptor antibody were also higher among TR patients than controls. Sera from 27 of the 36 TR patients, 10 of 16 normals, and nine of 12 retinal vasculitis patients possessed anti-human retinal S-antigen antibodies at a titre of 1:400 or more as assessed by ELISA (p = > 0.05). Antiretinal autoantibody as detected by IIF did not run in parallel with S-antigen reactivity. CONCLUSIONS: The data indicate that the extent of antiretinal reactivity within TR is not accounted for by anti-S-antigen antibodies alone. This remarkably high prevalence of antiphotoreceptor antibody in TR as opposed to that found in either healthy or disease controls suggest that these antibodies may be co-pathogenic in toxoplasma retinochoroiditis.


Assuntos
Autoanticorpos/análise , Pan-Uveíte/imunologia , Retina/imunologia , Retinite/imunologia , Toxoplasmose Ocular/imunologia , Adulto , Arrestina/imunologia , Autoanticorpos/sangue , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Células Fotorreceptoras/imunologia , Recidiva , Vasos Retinianos/imunologia , Vasculite/imunologia
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