ABSTRACT
Annexin A1 (AnxA1) is a protein that displays potent anti-inflammatory properties, but its expression in eye tissue and its role in ocular inflammatory diseases have not been well studied. We investigated the mechanism of action and potential uses of AnxA1 and its mimetic peptide (Ac2-26) in the endotoxin-induced uveitis (EIU) rodent model and in human ARPE-19 cells activated by LPS. In rats, analysis of untreated EIU after 24 and 48 h or EIU treated with topical applications or with a single s.c. injection of Ac2-26 revealed the anti-inflammatory actions of Ac2-26 on leukocyte infiltration and on the release of inflammatory mediators; the systemic administration of Boc2, a formylated peptide receptor (fpr) antagonist, abrogated the peptide's protective effects. Moreover, AnxA1(-/-) mice exhibited exacerbated EIU compared with wild-type animals. Immunohistochemical studies of ocular tissue showed a specific AnxA1 posttranslational modification in EIU and indicated that the fpr2 receptor mediated the anti-inflammatory actions of AnxA1. In vitro studies confirmed the roles of AnxA1 and fpr2 and the protective effects of Ac2-26 on the release of chemical mediators in ARPE-19 cells. Molecular analysis of NF-κB translocation and IL-6, IL-8, and cyclooxygenase-2 gene expression indicated that the protective effects of AnxA1 occur independently of the NF-κB signaling pathway and possibly in a posttranscriptional manner. Together, our data highlight the role of AnxA1 in ocular inflammation, especially uveitis, and suggest the use of AnxA1 or its mimetic peptide Ac2-26 as a therapeutic approach.
Subject(s)
Annexin A1/genetics , Anti-Inflammatory Agents/pharmacology , Peptides/pharmacology , Uveitis/genetics , Animals , Annexin A1/administration & dosage , Annexin A1/chemistry , Annexin A1/metabolism , Annexin A1/pharmacology , Anti-Inflammatory Agents/administration & dosage , Aqueous Humor/cytology , Aqueous Humor/immunology , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolism , Cytokines/biosynthesis , Cytokines/genetics , Cytokines/immunology , Disease Models, Animal , Endotoxins/adverse effects , Gene Expression Regulation/drug effects , Lipopolysaccharides/immunology , Male , Mice , Mice, Knockout , Models, Biological , NF-kappa B/metabolism , Neutrophil Infiltration/immunology , Neutrophils/drug effects , Neutrophils/immunology , Oligopeptides/pharmacology , Peptides/administration & dosage , Phosphorylation , Protein Transport/drug effects , Rats , Receptors, Formyl Peptide/genetics , Receptors, Formyl Peptide/metabolism , Uveitis/chemically induced , Uveitis/immunologyABSTRACT
AIM: To evaluate the antioxidant capacity and concentrations of vascular endothelial growth factor (VEGF) and interleukin 6 (IL-6) in aqueous humor from patients with type 2 diabetes mellitus (T2DM) with or without retinopathy. METHODS: Aqueous humor was obtained during elective cataract surgery from T2DM patients with or without retinopathy and from healthy subjects. Reducing response was evaluated by MTT dye reduction and the generation of reactive oxygen species (ROS) was determined by chemiluminescence assay. Granulocytes were treated with phorbol dibutyrate (PDB)-stimulated. Cytokines were quantified by ELISA. RESULTS: Antioxidant capacity of aqueous humor from patients with retinopathy was greater (P<0.05) than that of healthy controls or persons with diabetes without retinopathy. ROS production in PDB (protein kinase C activator)-stimulated granulocytes from T2DM patients with or without retinopathy was inhibited by autologous aqueous humor. Concentrations of VEGF and IL-6 were similar in aqueous humor from healthy controls and from patients without retinopathy, but lower (P<0.05) than those from T2DM patients with retinopathy. Plasma levels of VEGF and IL-6 were similar (P>0.05) in healthy controls and in T2DM patients with and without retinopathy. CONCLUSION: Aqueous humor from T2DM patients with retinopathy exhibits elevated antioxidant activity with significant suppressive effect on ROS production and enhanced levels of locally secreted VEGF and IL-6 in comparison with T2DM patients without retinopathy. These results suggest an inflammatory profile in the absence of typical oxidative stress for T2DM patients with retinopathy, possibly resulting from the compensatory antioxidant response detected in the aqueous humor improving the ocular redox state.
Subject(s)
Antioxidants/metabolism , Aqueous Humor/metabolism , Diabetic Retinopathy/metabolism , Interleukin-6/metabolism , Reactive Oxygen Species/metabolism , Vascular Endothelial Growth Factor A/metabolism , Adult , Aged , Aged, 80 and over , Aqueous Humor/immunology , Brazil , Cataract/metabolism , Diabetic Retinopathy/immunology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Luminescent Measurements , Male , Middle Aged , Phorbol 12,13-Dibutyrate , Reactive Oxygen Species/immunologyABSTRACT
A Leishmaniose Visceral Canina (LVC) é uma zoonose sistêmica grave, causada pelo protozoário intracelular obrigatório Leishmania infantum. Os cães são considerados os principais reservatórios da doença e podem desenvolver desde a infecção assintomática, a uma variedade de sinais clínicos, tais como linfadenopatia periférica, perda de peso, letargia, lesões ulcerativas de pele e febre. Alterações oculares, ortopédicas e neurológicas também podem ser observadas associadas à LVC, tais como ceratoconjuntivite, uveíte, convulsões, alterações comportamentais, artralgia e claudicação. Tal fato inclui a doença no diagnóstico diferencial das afecções oftálmicas, osteoarticulares e neurológicas, principalmente em áreas consideradas endêmicas para a infecção. O objetivo deste trabalho foi realizar o diagnóstico da leishmaniose visceral canina utilizando amostras de conjuntiva, humor aquoso, líquido sinovial, membrana sinovial e líquido cefalorraquidiano (LCR). O teste ELISA foi aplicado em amostras de LCR, líquido sinovial e humor aquoso; e avaliações histológica e imunohistoquímica foram realizadas na conjuntiva e na membrana sinovial. As amostras foram coletadas de 20 cães sorologicamente positivos para leishmaniose visceral. Através do Ensaio Imunoenzimático foi possível detectar anticorpos IgG anti-Leishmania em uma das amostras de LCR e de humor aquoso, e em seis amostras de líquido sinovial. A imunohistoquímica apresentou marcação positiva de formas amastigotas de L. infantum em cinco membranas sinoviais e cinco conjuntivas estudadas. Diante dos resultados, conclui-se que formas amastigotas de Leishmania sp. são encontradas parasitando a membrana sinovial e a conjuntiva de cães sorologicamente positivos para a doença, bem como anticorpos IgG específicos no humor aquoso, no LCR e no líquido sinovial desses animais.
Canine Visceral Leishmaniasis (CVL) is a severe systemic zoonosos caused by the obligate intracellular protozoan Leishmania infantum. The dogs are the major reservoir of the disease and may develop since asymptomatic infection to a variety of clinical signs, such as peripheral lymphadenopathy, weight loss, lethargy, ulcerative lesions of the skin and fever. Ocular, orthopedic and neurological disorders can also be seen associated with CVL, such as keratitis, seizures, behavioral changes, joint pain and lameness. This fact includes the disease in the differential diagnosis of ophthalmic disorders, osteoarticular and nervous system, especially in endemic areas to infection. The aim of this study was the diagnosis of canine visceral leishmaniasis using samples of conjunctiva, aqueous humor, synovial fluid, synovial membrane and cerebrospinal fluid (CSF). The ELISA test was applied to samples of CSF, synovial fluid and aqueous humor, and histological and immunohistochemical evaluations were performed in the conjunctiva and in the synovial membrane of these animals. The samples were collected from 20 dogs serologically positive for visceral leishmaniasis. Through the immunoenzymatic assay could detect anti-Leishmania IgG antibodies in a sample of CSF and aqueous humor, and six samples of synovial fluid. Immunohistochemistry showed positive staining of amastigotes of L. infantum five synovial membranes and connective five studied. From the results, we conclude that amastigotes of Leishmania sp. are found parasitizing the synovial membrane and the conjunctiva of dogs serologically positive for the disease, as well as specific IgG antibodies in aqueous humor, CSF and synovial fluid of these animals.
Subject(s)
Animals , Dogs , Aqueous Humor/immunology , Leishmaniasis, Visceral/diagnosis , Cerebrospinal Fluid/immunology , Synovial Fluid/immunology , Synovial Membrane/immunology , Conjunctiva/immunology , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay/veterinaryABSTRACT
The genetic and immunophenotypic characteristics of a 3-year-old patient with Blau syndrome (BS), an early onset sarcoidosis caused by mutations in NOD2, were investigated. Molecular analysis of NOD2 gene was achieved by PCR and direct nucleotide sequencing. Immunophenotyping included cytometric analysis of memory-effector markers on T-cells, and cytokine in serum, aqueous humour and vitreous. A novel M513R mutation in NOD2 was demonstrated. Immunophenotyping revealed higher frequency of CCR4+ cells and CCR9+ cells on CD4+ cells; most CD8+ cells were CCR7- and CCR9+. IL6 and IL-8 were detected in a gradient manner: vitreous humour>aqueous humour>serum. The immunophenotype in this patient was characterized by a differential expression of chemokine receptors on T cells and by a particular ocular microenvironment enriched in IL-6 and IL-8. To our knowledge, this is the first study analysing the immunological features of BS at aqueous humour, vitreous and blood levels. Our results expand the knowledge of the genetic and immunopathological basis of BS.
Subject(s)
Aqueous Humor/immunology , Cranial Nerve Diseases/genetics , Cranial Nerve Diseases/immunology , Immunophenotyping , Leukocytes, Mononuclear/immunology , Mutation/genetics , Nod2 Signaling Adaptor Protein/genetics , Synovitis/genetics , Synovitis/immunology , Uveitis/genetics , Uveitis/immunology , Arthritis , Base Sequence , Child, Preschool , Cranial Nerve Diseases/pathology , Cytokines/biosynthesis , Cytokines/immunology , Female , Heterozygote , Humans , Leukocytes, Mononuclear/metabolism , Phenotype , Receptors, Chemokine/genetics , Receptors, Chemokine/immunology , Sarcoidosis , Synovitis/pathology , Toll-Like Receptors/immunology , Toll-Like Receptors/metabolism , Uveitis/pathology , Vitreous Body/immunologyABSTRACT
BACKGROUND: Ocular brucellosis is usually diagnosed by clinical criteria and serological tests. Little is known with regard to the ocular immunology of brucellosis and the use of intraocular diagnostic tests. We report retrospectively the laboratory findings of patients with ocular involvement associated with brucellosis. MATERIALS AND METHODS: Patients with uveitis with no evident etiologic diagnosis were evaluated at the Instituto de Medicina Tropical "Alexander von Humboldt" of the Universidad Peruana Cayetano Heredia and the Hospital Nacional Cayetano Heredia. Patients were tested for brucellosis, tuberculosis, syphilis, toxoplasmosis, toxocariasis, and human T-cell lymphotropic virus-1. Blood and intraocular fluid samples were examined. Patients with a diagnosis of brucellar uveitis were selected as cases and patients with a diagnosis of uveitis of other etiology were included as controls. The Goldmann-Witmer coefficient was determined. RESULTS: Twelve patients with clinical and laboratory findings suggestive of brucellar uveitis were considered as cases. Seven patients with uveitis of other etiology were selected as controls. Four (33.3%) patients with ocular brucellosis had negative ocular agglutinations and eight (66.7%) had positive agglutinations. No control cases had positive agglutinations for Brucella melitensis. The sensitivity of the test was 66.7% and the specificity 100%. Only one patient had a positive culture for B. melitensis in subretinal fluid. The Goldmann-Witmer coefficient was calculated in six cases of brucellosis uveitis and five uveitis controls. It was highly positive in three patients with ocular brucellosis. Tissue samples showed lymphoplasmacytic infiltrates. CONCLUSIONS: Intraocular serological tests could be used to support the diagnosis of ocular brucellosis.
Subject(s)
Antibodies, Bacterial/blood , Aqueous Humor , Brucella melitensis , Brucellosis , Uveitis , Adolescent , Adult , Agglutination Tests , Aqueous Humor/immunology , Aqueous Humor/microbiology , Biopsy , Brucella melitensis/immunology , Brucella melitensis/isolation & purification , Brucellosis/diagnosis , Brucellosis/immunology , Brucellosis/microbiology , Female , Humans , Immunoglobulin G/blood , Male , Middle Aged , Peru , Sensitivity and Specificity , Uveitis/diagnosis , Uveitis/immunology , Uveitis/microbiology , Young AdultABSTRACT
Toxoplasma gondii, an obligate intracellular protozoan parasite, infects most species of warm-blooded animals, and in humans it causes toxoplasmosis. Healthy people that become infected rarely present clinical symptoms because the immune system prevents the parasite from causing illness. Congenital toxoplasmosis may result in abortion, hydrocephalus, as well as neurological and ocular disease (most frequently retinochoroiditis) of the newborn. In immunocompromised patients, reactivation of latent disease can cause encephalitis. Cell-mediated immunity to T. gondii antigens involves innate acute inflammatory responses and antigen-specific adaptive immunity. Considering the complexity of the immunological events triggered during toxoplasmosis, systemic and local responses were evaluated by cytokine measurements. Aqueous humour and serum were obtained from non-infected and T. gondii Me-49 strain infected C57BL/6 mice for cytokine quantification. Histopathological analyses were made with eyes enucleated from mice after 30 days of infection. ELISA assays showed an increase of IFN-gamma levels both in serum and aqueous humour of infected mice in opposition to a decrease in IL-10 levels. On the other hand, TGF-beta was high, whereas IL-12 and TNF-alpha were present in small levels in both groups. We also detected higher levels of IL-4 and IL-6 in aqueous humour than in serum of infected mice when compared to the control group. MIP-2 presented no significant differences between the two groups. Fas and Fas-L were also present in similar levels in serum of non-infected and infected mice, but both chemokines were increased in the aqueous humour of infected mice. Histopathological analysis of infected mice showed inflammatory infiltrates around blood vessels and alteration of the outer photoreceptor segments, on the external and inner nuclear layer. Parasites were observed in 82% of eyes, inside the blood vessels associated with inflammatory infiltrate. Edema, characterized by the increase of interstitial spaces between the FTR, forming lacunae was also noted. These alterations take the form of projections (retino-vitreal), characteristic of retinochoroiditis. In conclusion, T. gondii infection of C57BL/6 mice revealed that cytokine patterns alone do not assure susceptibility or resistance against infection, thus reinforcing the notion that it is necessary more than cytokine dosage to determine Th1 or Th2 profile in this model.
Subject(s)
Aqueous Humor/immunology , Cytokines/biosynthesis , Toxoplasmosis, Ocular/immunology , Toxoplasmosis, Ocular/pathology , Animals , Cytokines/blood , Female , Fluorescent Antibody Technique, Indirect , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Interleukin-4/biosynthesis , Mice , Mice, Inbred C57BL , Transforming Growth Factor beta1/biosynthesis , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
PURPOSE: To study the applicability (sensitivity, specificity) of polymerase chain reaction (PCR) tests in the detection of cytomegalovirus (CMV), herpes virus (HSV) and varicella zoster (VZV), Epstein-Barr virus (EBV), Mycobacterium sp and Toxoplasma gondii in the diagnosis of patients with or without AIDS, with presumably infectious uveitis, using serum, aqueous humor and vitreous humor samples. METHODS: Twenty individuals with uveitis of presumed infectious origin were evaluated. Sixteen of them had AIDS, four were immunocompetent individuals. We also evaluated 4 normal controls who underwent vitrectomy surgery. Clinical evaluation of the patients was performed together by three clinicians. PCR evaluations of the serum, aqueous, and vitreous humor were performed in a masked fashion by the laboratory staff. RESULTS: Twelve patients had a clinical diagnosis of CMV retinitis. Of these 6 (50 percent) had a positive PCR for CMV in the vitreous, three (25 percent) had a positive PCR for CMV in the serum, and none were positive in the aqueous. Five patients had a clinical diagnosis of acute retinal necrosis (ARN). Three (60 percent) of these had positive PCR for HSV/VZV in the vitreous. One of these patients had a positive PCR reaction for both EBV and HSV/VZV in the vitreous samples. One patient with cutaneous herpes zoster had a positive PCR reaction for HSV/VZV in the serum. Four patients had a presumed diagnosis of ocular toxoplasmosis, one patient (25 percent) had a positive PCR for Toxoplasma gondii in the serum, 3 (75 percent) had positive results in the aqueous, and 2 (50 percent) had positive results in the vitreous. One patient with presumed ocular tuberculosis had a positive PCR reaction both in the serum and in the vitreous samples. Finally, none of the four control individuals revealed any positive PCR reaction. CONCLUSION: PCR is an auxiliary diagnostic procedure that should be evaluated together with ophthalmological aspects...
OBJETIVOS: Avaliar a aplicabilidade (especificidade, sensibilidade) do teste da reação da cadeia de polimerase (PCR) na detecção de citomegalovírus (CMV), herpes vírus e varicela zoster (HSV, VZV), Epstein-Barr vírus (EBV), Mycobacterium sp e Toxoplasma gondii no diagnóstico de pacientes com ou sem AIDS, com uveíte infecciosa presumível, utilizando amostras de humor aquoso, humor vítreo e soro. MÉTODOS: Vinte pacientes com uveíte infecciosa presumível foram estudados. Dezesseis destes apresentavam AIDS e quatro eram imunocompetentes. Foram utilizados quatro pacientes como grupo controle que se submeteram a vitrectomia. A avaliação clínica foi feita conjuntamente com três oftalmologistas. O exame do PCR do soro, aquoso e vítreo foi feito sem o conhecimento da hipótese diagnóstica pela equipe do laboratório. RESULTADOS: Doze pacientes tinham o diagnóstico clínico de retinite por CMV. Deste subgrupo 6 (50 por cento) eram PCR positivo para CMV no vítreo, 3 (25 por cento) eram PCR positivos para CMV no soro e nenhum destes foi positivo no aquoso. Cinco pacientes tinham o diagnóstico clínico de necrose aguda de retina (ARN). Três (60 por cento) destes eram PCR positivos para HSV/VZV no vítreo. Um destes pacientes era PCR positivo tanto para EBV e HSV/VZV na amostra do vítreo. Um destes pacientes com herpes zoster cutâneo era PCR positivo para HSV/VZV no soro. Quatro pacientes tinham o diagnóstico de toxoplasmose ocular presumida, um paciente (25 por cento) era PCR positivo para Toxoplasma gondii no soro, 3 (75 por cento) eram positivos no aquoso e 2 (50 por cento) eram positivo no vítreo. Um paciente com tuberculose ocular presumível era PCR positivo tanto no soro quanto no vítreo. Nenhum dos pacientes do grupo controle era PCR positivo em qualquer amostra. CONCLUSÃO: O exame do PCR é procedimento diagnóstico auxiliar que deve ser utilizado conjuntamente com os aspectos clínicos.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , AIDS-Related Opportunistic Infections/diagnosis , Aqueous Humor , Polymerase Chain Reaction/methods , Uveitis/diagnosis , Vitreous Body , AIDS-Related Opportunistic Infections/blood , AIDS-Related Opportunistic Infections/microbiology , AIDS-Related Opportunistic Infections/virology , Aqueous Humor/immunology , Aqueous Humor/microbiology , Case-Control Studies , Sensitivity and Specificity , Uveitis/blood , Uveitis/microbiology , Uveitis/virology , Vitreous Body/microbiology , Vitreous Body/virologyABSTRACT
The present study examined the outcomes of systemic or topical treatment with thalidomide, a compound that possesses anti-inflammatory, immunomodulatory and anti-angiogenic properties, in rats subjected to endotoxin-induced uveitis (EIU). The effects of thalidomide were evaluated on endotoxin-induced leucocyte and protein infiltration and also on the production of interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha in rat aqueous humour (AqH). Moreover, the actions of thalidomide were assessed on the cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) protein expression in retinal tissue. EIU was produced by a hindpaw injection of lipopolysaccharide (LPS), in male Wistar rats. Thalidomide (5, 25 and 50 mg/kg) was administered orally 1 h before LPS injection. In another set of experiments, to evaluate the therapeutic efficacy, 5% thalidomide was applied topically to both eyes at 6, 12 and 18 h after LPS administration. The oral pre-treatment with thalidomide decreased, in a dose-dependent manner, the number of inflammatory cells, the protein concentration, and the levels of IL-1beta and TNF-alpha in the AqH. Similar results were found in the AqH of rats that received a topical application of thalidomide. Furthermore, oral (50 mg/kg) and local (5%) thalidomide treatment also reduced expression of the pro-inflammatory proteins COX-2 and iNOS in the posterior segment of the eye. Thalidomide exhibited marked preventive and curative ocular effects in EIU in rats, a property that might be associated with its ability to inhibit the production of inflammatory cytokines and the expression of COX-2 and iNOS. This assembly of data provides additional molecular and functional insights into beneficial effects of thalidomide as an agent for the management of ocular inflammation.
Subject(s)
Anti-Inflammatory Agents/therapeutic use , Aqueous Humor/immunology , Thalidomide/therapeutic use , Uveitis/drug therapy , Administration, Oral , Administration, Topical , Animals , Aqueous Humor/metabolism , Blotting, Western/methods , Cyclooxygenase 2 , Dexamethasone/therapeutic use , Dose-Response Relationship, Drug , Eye Proteins/metabolism , Interleukin-1/immunology , Leukocytes/immunology , Lipopolysaccharides , Male , Membrane Proteins , Models, Animal , Nitric Oxide Synthase Type II/analysis , Nitric Oxide Synthase Type II/metabolism , Rats , Rats, Wistar , Tumor Necrosis Factor-alpha/analysis , Tumor Necrosis Factor-alpha/immunology , Uveitis/immunology , Uveitis/metabolismABSTRACT
Tests were performed for antibodies IgG, IgM and IgA anti-Toxoplasma gondii antibodies in serum and intraocular fluids (Aqueous and vitreous humor) from patients with ocular toxoplasmosis. By the results obtained, it was verified that intraocular IgG and IgA anti-Toxoplasma gondii antibodies can be important markers for the diagnosis of ocular toxoplasmosis.
Subject(s)
Antibodies, Protozoan/analysis , Aqueous Humor/immunology , Toxoplasma/immunology , Toxoplasmosis, Ocular/immunology , Vitreous Body/immunology , Adult , Animals , Aqueous Humor/parasitology , Biomarkers/analysis , Enzyme-Linked Immunosorbent Assay , Female , Fluorescent Antibody Technique, Indirect , Humans , Vitreous Body/parasitologyABSTRACT
Foi realizada pesquisa de anticorpos IgG, IgM e IgA anti-Toxoplasma gondii no soro e fluidos intra-oculares (humor aquoso e vítreo) de pacientes com toxoplasmose ocular. A partir dos resultados obtidos verificou-se que anticorpos IgG e IgA intraocular anti-Toxoplasma gondii podem vir a ser importantes marcadores no diagnóstico de toxoplasmose ocular.
Subject(s)
Adult , Animals , Female , Humans , Antibodies, Protozoan/analysis , Aqueous Humor/immunology , Toxoplasma/immunology , Toxoplasmosis, Ocular/immunology , Vitreous Body/immunology , Aqueous Humor/parasitology , Biomarkers/analysis , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique, Indirect , Vitreous Body/parasitologyABSTRACT
Propósito: Evaluar el rol del estudio de los fluidos intraoculares, por medio del test ELISA, en casos de uveítis difusa o posterior severa, con evolución desfavorable y amenaza visual. Métodos: Pacientes portadores de uveítis difusa o posterior, con una evolución maligna fueron seleccionados. Luego de haber firmado un consentimiento informado, se procedió a tomar muestra de humor acuosos y vítreo, así como muestras de líquido cefalorraquídeo en los casos que lo ameritaran. Cada caso fue ingresado a una planilla computarizada, donde se registraron los datos clínicos y de laboratorio. Resultados: Se estudio una cohorte formada por 9 pacientes, de sexo femenino en 2 casos y masculino en 7 casos. 2 de 9 casos eran portadores de virus de la inmunodeficiencia humana (HIV). El rango de edad fue de 7 a 61 años, con una media de 36 años. En cuatro casos, el estudio de la muestra intraocular fue decisiva en la confirmación diagnóstica. Estos casos correspondieron a sospechosos de toxoplasmosis, que demostraron altos niveles de anticuerpos en los fluidos intraoculares, especialmente en humor vítreo, probablemente debido a la cercanía con el foco coriorretinal. Estos niveles fueron significativamente más altos que los niveles sanguíneos, lo que confirma el diagnósticos de coriorretinitis toxoplásmica. Otros diagnósticos fueron: sifilis en tres casos, en los caules el estudio intraocular contribuyó a un diagnóstico más preciso. La etiología viral fue estudiada mediante cultivo en monocapa celular y fue negativa en todos los casos, al igual que el test de ELISA. Conclusión: El estudio de los fluidos intraoculares en casos de uveítis severa de etiología desconocida, demostró ser una herramienta segura y efectiva, entregando evidencia diagnóstica en 6 de 9 casos. Aunque no se pudo establecer la etiología final en tres casos, a lo menos se pudieron descartar las etiologías infecciosas más frecuentes en nuestro medio, lo que permitió un mejor enfoque del tratamiento.
Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Vitreous Body/immunology , Aqueous Humor/immunology , Panuveitis/diagnosis , Uveitis, Posterior/diagnosis , Uveitis, Posterior/immunology , Case-Control Studies , Chile , HIV , Immunologic Tests/methods , Toxoplasma/pathogenicity , Treponema pallidum/pathogenicityABSTRACT
El ojo funciona como un compartimiento aislado del resto del organismo desde el punto de vista inmunológico, lo que explica que. ante la presencia de enfermedades inflamatorias y/o infecciosas dentro del globo ocular, muchas veces no se encuentra evidencia de estas a nivel plamático. Es necesario conocer entonces las características inmunológicas normales en los fluidos intraoculares para un mejor entendimiento y manejo de las enfermedades oculares inflamatorias y/o infecciosas. En este trabajo se logró evaluar y comprobar la idemnidad de la barrera hematoocular a través de la determinación de la relación IgG/albúmina tanto en humor acuoso como en plasma cuyo coeficiente resultó ser 0,59. Además, ante la presencia de títulos positivos en sangre para toxocariasis y toxoplasmosis, no se encontró presencia de anticuerpos contra estas en los fluidos intraoculares. Finalmente los títulos de anticuerpos para lúes fueron negativos en ambos compartimentos
Subject(s)
Humans , Male , Female , Middle Aged , Aqueous Humor/immunology , Eye/immunology , Uveitis/diagnosis , Serum Albumin , Cataract/immunology , Glaucoma/immunology , Immunoglobulin G , Immunoglobulin G/immunology , Nephelometry and Turbidimetry/methods , Toxocara/isolation & purification , Toxoplasma/isolation & purificationABSTRACT
En las uveítis en un porcentaje importante de casos no logra determinarse su etiología a pesar de ser sometidos a un exhaustivo estudio sistémico. Las uveítis posteriores están asociadas en forma considerable a agentes etiológicos de tipo infeccioso. No existe en nuestro medio experiencia en el estudio de fluidos intraoculares como ayuda diagnóstica en estos casos. Esta serie que consta de 13 pacientes pone de manifiesto que el conocimiento del resultado de exámenes de laboratorio de fluidos intraoculares puede ser de gran utilidad en aproximarse al diagnóstico etiológico en más del 60 por ciento de ellos, y con elllo llevar a cabo una terapia más racional. Es importante conocer las dificultades técnicas económicas, y otras que se presentan frente a estas nuevas alternativas de evaluación diagnóstica. Sin embargo, el beneficio que puede significar para los pacientes nos obliga al desarrollo de estas técnicas basadas en la utilización de micrométodos que permiten aprovechar de forma racional las muestras de fluidos intraoculares
Subject(s)
Humans , Child , Adolescent , Adult , Middle Aged , Panuveitis/diagnosis , Uveitis, Posterior/diagnosis , Aqueous Humor/immunology , Vitreous Body/immunology , Fundus Oculi , Immunologic Tests/methodsABSTRACT
Using a modified enzyme-linked immunosorbent assay that included dissociation of antigen antibody complexes with 6M urea solution, we analyzed the avidity of Toxoplasma-specific IgG in aqueous humor and serum samples from 24 patients with toxoplasmic chorioretinitis. As a control, we studied aqueous humor and serum samples from 14 cataract patients without history of uveitis and serum samples from 10 patients with recent primary systemic toxoplasmic infection without ocular lesions. IgG avidity was markedly lower in aqueous humor samples from patients with toxoplasmic chorioretinitis than in serum samples, despite those samples presenting higher levels of Toxoplasma-specific IgG than in serum samples. The detection of the low-avidity Toxoplasma-specific antibodies can offer a valuable aid to make a specific etiologic diagnosis and perhaps contribute to understand the pathogenic mechanisms of ocular toxoplasmosis.
Subject(s)
Antibody Affinity , Aqueous Humor/immunology , Immunoglobulin G/analysis , Toxoplasma/immunology , Toxoplasmosis, Ocular/immunology , Adolescent , Adult , Aged , Animals , Antibodies, Protozoan/analysis , Antibodies, Protozoan/blood , Antibodies, Protozoan/immunology , Cataract/immunology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin G/blood , Immunoglobulin G/immunology , Male , Middle Aged , Toxoplasmosis/immunology , Toxoplasmosis, Ocular/diagnosisABSTRACT
Com o objetivo de se verificar as respostas imune sérica e intra-ocular relativa aos anticorpos IgA específicos na uveíte de provável etiologia toxoplasmática, assim como a associaçäo desse anticorpo com os anticorpos IgG e IgM específicos, foram comparadas amostras de soro e humor aquoso de 24 pacientes portadores de uveíte de provável etiologia toxoplasmática e de 12 pacientes com catarata, através da reaçäo imunoenzimática (ELISA). No soro de pacientes com uveíte de provável etiologia toxoplasmática, foram detectadas 3,2% de amostras positivas para o anticorpo IgG. No humor aquoso de pacientes com uveíte, foram detectados anticorpos IgA, IgG e IgM específicos em 60% e 0% das amostras respectivamente. Nos pacientes com catarata näo foram detectadas amostras de humor aquoso positivas para o anticorpo IgA específico, no grupo de pacientes com uveíte, e apenas 4,2% de positividade nas respectivas amostras de soro, sugerem que houve produçäo intra-ocular deste anticorpo, e reforça a hipótese desta uveíte ser uma infecçäo crônica, provavelmente congênita
Subject(s)
Humans , Antibodies/biosynthesis , Aqueous Humor/immunology , Cataract/etiology , Immunoglobulin A/immunology , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Drug Interactions/classification , Toxoplasmosis, Ocular/diagnosis , Uveitis/etiology , Brazil , Uveitis/congenitalABSTRACT
No humor aquoso de pacientes com uveite posterior ativa (25 pacientes) foi encontrada uma concentraçäo de imunoglobulinas significativamente maior do que em pacientes com catarata senil (10 pacientes). No humor aquoso de mais de 90% de pacientes portadores de uveite, foram detectados anticorpos anti-T. gondii, enquanto que nos pacientes com catarata isto näo ocorreu em nenhum. Näo houve diferença significativa entre os títulos de anticorpos anti-T. gondii, detectados pela reaçäo de imunofluorescencia indireta, no soro dos pacientes com uveite e dos com catarata. A reaçäo de imunofluorescência indireta para toxoplasmose, realizada com conjugado anti-IgM, foi quase sempre negativa, näo tendo colaborado para o esclarecimento diagnóstico dos casos
Subject(s)
Humans , Adolescent , Adult , Middle Aged , Aqueous Humor/analysis , Cataract/immunology , Immunoglobulins/analysis , Toxoplasmosis, Ocular/immunology , Uveitis/immunology , Aqueous Humor/immunology , Fluorescent Antibody Technique , Immunoglobulin A/analysis , Immunoglobulin G/analysis , Immunoglobulin M/analysisABSTRACT
O autor calculou o coeficiente de anticorpos do humor aquoso (C) em 25 pacientes portadores de uveíte posterior ativa, com a finalidade de confirmar a suspeita clínica de toxoplasmose ocular. Para isso, foram dosados imunoglobulinas A,G e M, através da imunodifusäo radial, e anticorpos anti-T. gondii, através da reaçäo de imunofluorescência indireta, no soro e no humor aquoso desses pacientes. As mesmas dosagens foram também realizadas em um grupo de 10 pacientes portadores de catarata senil, a fim de se comparar os resultados obtidos. O coeficiente de anticorpos do humor aquoso (C) foi calculado utilizando-se a seguinte fórmula: título de anticorpos anti-T. gondii no humor aquoso título de anticorpos anti-T. gondii no soro C= concentraçäo de imunoglobulinas no humor aquoso concentraçäo de imunoglobulinas no soro. Coeficientes de 2 a 7 foram considerados sugestivos e aqueles iguais ou acima de 8 indicativos de etiologia toxoplásmica. No grupo de paciente portadores de uveíte posterior, 56 por cento dos coeficientes obtidos foram superiores a 2, sendo que na metade das vezes acima de 7. Enquanto isso, no grupo de pacientes portadores de catarata senil, näo foram detectados anticorpos anti-T. gondii no humor aquoso, impedindo assim o cálculo do coeficiente. Quando se fez uma comparaçäo entre os resultados obtidos nos pacientes com menos de 30 dias de evoluçäo da uveíte e naqueles com mais de 30 dias, verificou-se existir uma diferença significativa no sentido de coeficientes mais altos nos pacientes com doença mais crônica. O estudo permitiu concluir que o exame do humor aquoso é útil no diagnóstico das uveítes posteriores. Sugeriu ou confirmou a etiologia toxoplásmica na maior parte de um grupo de pacientes com uveíte onde havia clinicamente esta suspeita. Além disso, demonstrou que é naqueles casos com maior tempo de duraçäo que se obtém os coeficientes mais significativos.