RESUMO
BACKGROUND: Although ocular adverse events are frequent in AD patients treated with dupilumab, their characterization remains limited due to a lack of prospective studies with a systematic ophthalmological examination. OBJECTIVE: To examine the incidence, characteristics and risk factors of dupilumab-induced ocular adverse events. METHODS: A prospective, multicenter, and real-life study in adult AD patients treated with dupilumab. RESULTS: At baseline, 27 out of 181 patients (14.9%) had conjunctivitis. At week 16 (W16), 25 out of 27 had improved their conjunctivitis and 2 remained stable and 34 out of 181 patients (18.7%) had dupilumab-induced blepharoconjunctivitis: either de novo (n = 32) or worsening of underlying blepharoconjunctivitis (n = 2). Most events (27/34; 79.4%) were moderate. A multivariate analysis showed that head and neck AD (OR = 7.254; 95%CI [1.938-30.07]; p = 0.004), erythroderma (OR = 5.635; 95%CI [1.635-21.50]; p = 0.007) and the presence of dry eye syndrome at baseline (OR = 3.51; 95%CI [3.158-13.90]; p = 0.031) were independent factors associated with dupilumab-induced blepharoconjunctivitis. LIMITATIONS: Our follow-up period was 16 weeks and some late-onset time effects may still occur. CONCLUSION: This study showed that most dupilumab-induced blepharoconjunctivitis cases are de novo. AD severity and conjunctivitis at baseline were not found to be associated risk factors in this study.
Assuntos
Conjuntivite , Dermatite Atópica , Adulto , Humanos , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/diagnóstico , Estudos Prospectivos , Anticorpos Monoclonais Humanizados/efeitos adversos , Conjuntivite/induzido quimicamente , Conjuntivite/epidemiologia , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
CONTEXT: Injecting intracameral cefuroxime has been found beneficial in reducing the risk of postoperative endophthalmitis but its use has been limited through a lack of approved marketing and of ready-to-use single-units as well as the problem of aseptic compounding. OBJECTIVE: Our aim was to assess a new automated primary packaging system which should ensure a higher level of sterility, thanks to its closed, sterile, ready-to-use polymer vial called "Crystal® vial". The chemical stability of a 10 mg/mL cefuroxime solution was compared in 1 mL Crystal® vials and 1 mL Luer-lock polypropylene syringes (actual reference) to eliminate any potential and specific interactions with its cyclic olefin copolymer (COC) body and elastomer stopper. METHODS: Cefuroxime solution was introduced into vials and syringes and stored at -20 °C, +5 °C and +25°C/60% Relative Humidity. Cefuroxime concentration and the relative amount of the main degradation product (descarbamoyl-cefuroxime) were both determined by an HPLC/UV method indicating stability. Solutions were considered steady if the concentration remained at over 90% of the initial value. In the adapted storage conditions, the evolution of osmolality, pH and sterility was assessed. RESULTS: Stability profiles were identical between vials and syringes in all storage and temperature conditions. The solution was stable (cefuroxime concentration, pH and osmolality) and still sterile for 365 days at -20°C. The concentration fell below 90% after 21 days at +5 °C and after 16 h at +25°C/60%s relative humidity. CONCLUSIONS: The COC and thermoplastic elastomer of the vials had no impact on the degradation process confirming its possible use for a ready-to-use cefuroxime solution single-unit dose.
Assuntos
Cefuroxima/química , Cicloparafinas/química , Embalagem de Medicamentos/métodos , Polipropilenos/química , Antibacterianos/administração & dosagem , Antibacterianos/química , Antibacterianos/uso terapêutico , Cefuroxima/administração & dosagem , Cefuroxima/uso terapêutico , Composição de Medicamentos/normas , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Endoftalmite/prevenção & controle , Estudos de Viabilidade , Vidro , Injeções Intraoculares , Concentração Osmolar , Soluções Farmacêuticas/administração & dosagem , Soluções Farmacêuticas/química , Soluções Farmacêuticas/uso terapêutico , Complicações Pós-Operatórias/prevenção & controle , Seringas , TemperaturaRESUMO
Constitutive COL2A1 mutations are associated with a wide variety of clinical manifestations known as type II collagenopathies. Among them is Kniest dysplasia, which is phenotypically variable and includes both skeletal (short trunk and limbs, kyphoscoliosis, prominent joints, and osteoarthritis) and craniofacial characteristics. Kniest dysplasia mutations primarily arise in the triple-helicoidal region of the alpha 1 (II) chain in COL2A1 between exons 12 and 24. Somatic COL2A1 mutations have been identified in chondrosarcoma, a rare cartilage forming neoplasm, with a hypermutability of the gene reported in 37% of cases. However, to the best of our knowledge, there is no reported increase in predisposition to chondrosarcoma in human collagenopathies, and no reported clinical association between these congenital diseases and cartilaginous tumors. In the case study presented here, we report the first description of an association between these two rare diseases involving COL2A1, in a child presenting with Kniest dysplasia and a grade I sphenoethmoidal chondrosarcoma. We also describe a new constitutive mutation in COL2A1.
Assuntos
Neoplasias Ósseas/genética , Condrossarcoma/genética , Fissura Palatina/genética , Doenças do Colágeno/genética , Colágeno Tipo II/genética , Nanismo/genética , Face/anormalidades , Doença da Membrana Hialina/genética , Mutação/genética , Osteocondrodisplasias/genética , Neoplasias Ósseas/patologia , Condrossarcoma/patologia , Fissura Palatina/patologia , Doenças do Colágeno/patologia , Nanismo/patologia , Face/patologia , Humanos , Doença da Membrana Hialina/patologia , Recém-Nascido , Masculino , Osteocondrodisplasias/patologia , PrognósticoRESUMO
Sjögren's disease (SD), also known as Sjögren's syndrome (SS) or Gougerot-Sjögren's syndrome in France, is a rare systemic autoimmune disease in its primary form and is characterised by tropism for the exocrine glandular epithelia, particularly the salivary and lacrimal glands. The lymphocytic infiltration of these epithelia will clinically translate into a dry syndrome which, associated with fatigue and pain, constitutes the symptom triad of the disease. In about one third of patients, SD is associated with systemic complications that can affect the joints, skin, lungs, kidneys, central or peripheral nervous system, and lymphoid organs with an increased risk of B-cell lymphoma. SD affects women more frequently than men (9/1). The peak frequency is around the age of 50. However, the disease can occur at any age, with paediatric forms occurring even though they remain rare. SD can occur alone or in association with other systemic autoimmune diseases. In its isolated or primary form, the prevalence of SD is estimated to be between 1 per 1000 and 1 per 10,000 inhabitants. The most recent classification criteria were developed in 2016 by EULAR and ACR. The course and prognosis of the disease are highly variable and depend on the presence of systemic involvement and the severity of the dryness of the eyes and mouth. The current approach is therefore to identify at an early stage those patients most at risk of systemic complications or lymphoma, who require close follow-up. On the other hand, regular monitoring of the ophthalmological damage and of the dental status should be ensured to reduce the consequences.
Assuntos
Síndrome de Sjogren , Humanos , Feminino , Criança , Síndrome de Sjogren/complicações , Síndrome de Sjogren/diagnóstico , Síndrome de Sjogren/epidemiologia , Olho , Pele , França/epidemiologiaRESUMO
PURPOSE: To assess the physico-chemical stability of Voriconazole Eye-Drops (VED), when stored frozen and refrigerated once thawed, in 3 containers: Amber glass with a Low-Density PolyEthylene (LDPE) eyedropper, and two types of LDPE bottles: one classical and one with an innovative insert that maintains sterility after opening (Novelia® from Nemera). METHODS: Three batches of 1% VED (10â¯mL) were aseptically compounded from marketed injectable voriconazole (Vfend®) diluted in sterile water for injection. VEDs were stored for three months at -20⯰C in amber glass (nâ¯=â¯32), classical LDPE (nâ¯=â¯32) or innovative LDPE (nâ¯=â¯31) bottles. Stability-indicating (HPLC-UV-DAD) and chiral chromatography methods were developed. The stability study was conducted according to GERPAC-SFPC guidelines. At each study time, the following parameters were controlled: visual aspect, voriconazole concentration, pH and osmolality. In addition, non-visible particle count, sterility and absence of racemisation (impurity D - (2S,3R)-voriconazole) were assessed at the beginning and end of the study. Results are expressed as mean ± standard deviation. Statistical analyses were performed using non-parametric tests (α < 5%) to compare containers. RESULTS: When stored frozen, concentration was between 95.2⯱â¯1.4% and 103.6⯱â¯1.3% of the initial concentration (C0) with no difference between the three containers (pâ¯=â¯0.564; non-significant). Fifteen days after thawing, concentration was between 97.1⯱â¯1.6% and 98.6⯱â¯0.8% of C0 with no difference between containers (pâ¯=â¯0.278 and 0.368 for VED thawed at room temperature and at 2-8⯰C, respectively). pH remained stable between each time. Osmolality was slightly higher in glass (533.17⯱â¯8.93 mOsm/Kg) than in plastic containers (522.17±3.31mOsm/Kg, classical LDPE; 517.5⯱â¯12.42 mOsm/Kg, innovative LDPE) (pâ¯=â¯0.022). Sterility was preserved. Degradation product areas increased slightly but remained below the limit of quantification. Impurity D was never detected. CONCLUSION: We have demonstrated that the ability of the innovative container Novelia® to maintain VED physicochemical and microbiological stability does not differ from that of amber glass and classical LDPE containers. Real life studies are required to find out if there is a potential difference between Novelia® and other containers in terms of sterility preservation.
Assuntos
Antifúngicos/química , Soluções Oftálmicas/química , Voriconazol/química , Embalagem de Medicamentos , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Congelamento , Vidro/química , Polietileno/químicaRESUMO
The present paper describes a case of bilateral panuveitis due to Borrelia burgdorferi sensu lato diagnosed by a PCR approach using cerebral spinal fluid. Since the culture of B. burgdorferi takes a long time to grow and the accuracy of serological tests is doubtful in patients, the PCR method of amplifying a B. burgdorferi flagellin could be suitable to make a positive diagnosis in a case of atypical clinical history of Lyme disease.
Assuntos
Grupo Borrelia Burgdorferi/classificação , Grupo Borrelia Burgdorferi/isolamento & purificação , Líquido Cefalorraquidiano/microbiologia , Doença de Lyme , Reação em Cadeia da Polimerase/métodos , Uveíte , Grupo Borrelia Burgdorferi/genética , Feminino , Flagelina/genética , Humanos , Doença de Lyme/diagnóstico , Doença de Lyme/microbiologia , Doença de Lyme/fisiopatologia , Pessoa de Meia-Idade , Uveíte/diagnóstico , Uveíte/microbiologia , Uveíte/fisiopatologiaRESUMO
Microsporidia are protists close to the kingdom of fungi that may cause eye infections. Most cases are reported in Asia and affect both immunocompromised and immunocompetent patients. Here, we report a rare case of microsporidial keratoconjunctivitis in an immunocompetent French patient 3 weeks after returning from India. In our patient, Weber trichrome staining of conjunctival scrapings revealed rounded elements approximately 1-3 µm in size. Conventional polymerase chain reaction analysis by ribosomal RNA subunit sequencing showed 100% identity with Vittaforma corneae. Treatment by corneal debridement combined with fluoroquinolone eye drops allowed complete resolution of the lesions. Although rare, ocular microsporidiosis should be investigated in a patient who is native to Asia or has returned from an endemic area and presents with keratoconjunctivitis of undetermined etiology.
Assuntos
Antifúngicos/uso terapêutico , Infecções Oculares Fúngicas/diagnóstico , Fluoroquinolonas/uso terapêutico , Ceratoconjuntivite/diagnóstico , Microsporidiose/diagnóstico , Córnea/efeitos dos fármacos , Córnea/microbiologia , Córnea/patologia , Córnea/cirurgia , Desbridamento/métodos , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Fúngicas/cirurgia , França , Humanos , Índia , Ceratoconjuntivite/tratamento farmacológico , Ceratoconjuntivite/microbiologia , Ceratoconjuntivite/cirurgia , Masculino , Microsporidiose/tratamento farmacológico , Microsporidiose/microbiologia , Microsporidiose/cirurgia , Pessoa de Meia-Idade , Viagem , Vittaforma/efeitos dos fármacos , Vittaforma/crescimento & desenvolvimento , Vittaforma/patogenicidadeRESUMO
PURPOSE: Macular edema is the leading cause of vision loss in bilateral chronic noninfectious posterior uveitis, and is currently being treated using corticosteroids, immunosuppressive agents, and biotherapies. The aim of this trial was to assess and compare the efficacy and safety of corticosteroids and interferon-α (IFN-α) in adults with such conditions. DESIGN: Randomized controlled trial. METHODS: Subjects: Adult patients with bilateral posterior autoimmune noninfectious and nontumoral uveitis complicated by macular edema in at least 1 eye. INTERVENTION: Patients received either subcutaneous IFN-α2a, systemic corticosteroids, or no treatment for 4 months. The efficacy and safety were assessed for up to 4 months. MAIN OUTCOME MEASURES: The main endpoint was the change of the central foveal thickness (CFT) obtained by optical coherence tomography. RESULTS: Forty-eight patients were included. In intention-to-treat analysis, the median CFT change showed no significant difference. However, the per-protocol analysis showed a significant difference between groups for both eyes (OD and OS), and for the worse and better eyes. Statistically significant difference was found between the control and corticosteroid groups for the OD (P = .0285), and between the control and IFN-α groups for the OD (P = .0424) and worse eye (P = .0354). Serious adverse events occurred in 2 patients in the IFN group, in 1 patient in the corticosteroid group, and in 2 patients in the control group and were completely resolved after switch. CONCLUSIONS: IFN-α and systemic corticosteroids, compared with no treatment, were associated with significant anatomic and visual improvement shown in the per-protocol study.
Assuntos
Glucocorticoides/administração & dosagem , Interferon-alfa/administração & dosagem , Edema Macular/tratamento farmacológico , Uveíte Posterior/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Crônica , Relação Dose-Resposta a Droga , Feminino , Seguimentos , Fóvea Central/patologia , Humanos , Fatores Imunológicos/administração & dosagem , Injeções Subcutâneas , Interferon alfa-2 , Edema Macular/diagnóstico , Edema Macular/etiologia , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes/administração & dosagem , Fatores de Tempo , Tomografia de Coerência Óptica , Resultado do Tratamento , Uveíte Posterior/complicações , Uveíte Posterior/diagnóstico , Acuidade Visual , Adulto JovemRESUMO
PURPOSE: To describe the clinical features in patients presenting with ocular toxoplasmosis after the fifth decade and to analyze laboratory findings in comparison to uveitis history and clinical data. DESIGN: Prospective consecutive observational case series. METHODS: A prospective clinical analysis of 27 consecutive patients older than 50 years of age with primary or recurrent ocular toxoplasmosis was performed during a period of 8 years. These cases account for 12% of all ocular toxoplasmosis cases irrespective of age indexed in our institution during the same period. Paired serum and aqueous humor samples were tested for anti-Toxoplasma gondii IgG, IgM, and IgA antibodies. The presence of T. gondii DNA in aqueous humor was determined by polymerase chain reaction followed by DNA hydridization method. RESULTS: Although similar in age, two groups were distinguished clinically: 12 patients (44%) presented with usual forms of retinochoroiditis (mean +/- SD, 1.6 +/- 0.5 disk areas [DA] in size); 15 patients (56%) presented with atypical lesions, greater than 3 DA in size (mean +/- SD, 5.0 +/- 2.0 DA). The second group showed a higher rate of complications (P =.028) and a poorer visual outcome (P =.015). Twenty-four patients (89%) had intraocular IgG production, 17 (63%) had intraocular IgA production, 3 (11%) had intraocular IgM production, and 12 (44%) had a positive T. gondii DNA detection. CONCLUSIONS: After the fifth decade, ocular toxoplasmosis remains an important cause of posterior uveitis. The combination of antibody detection by immunocapture tests with T. gondii DNA detection, both in aqueous humor, allowed the diagnosis of toxoplasmic infection in the atypical cases with large ocular lesions.
Assuntos
Toxoplasmose Ocular/complicações , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos Antiprotozoários/sangue , Antiprotozoários/uso terapêutico , Humor Aquoso/imunologia , Humor Aquoso/parasitologia , DNA de Protozoário/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Estudos Prospectivos , Recidiva , Toxoplasma/genética , Toxoplasma/imunologia , Toxoplasma/isolamento & purificação , Toxoplasmose Ocular/diagnóstico , Toxoplasmose Ocular/tratamento farmacológico , Uveíte/diagnóstico , Uveíte/tratamento farmacológico , Uveíte/etiologiaRESUMO
The KIT gene encodes c-kit, a transmembrane receptor that has tyrosine kinase activity and plays a role in haematopoiesis, gametogenesis and melanogenesis. The c-kit protein is found in normal cutaneous and choroidal melanocytes, and there is evidence that expression is lost in melanoma. Expression of c-kit was analysed in 57 paraffin-embedded sections of choroidal melanoma specimens and three choroidal melanoma cell lines using immunochemistry and Western blotting. Of the tumour specimens, 75% stained positively for c-kit with a membrane pattern of reactivity. Of the six patients who underwent proton beam therapy before enucleation, five tumours exhibited no c-kit immunoreactivity and the other tumour demonstrated weak staining. Of the three melanoma cell lines used, c-kit expression was observed in only one. No correlations between c-kit positivity and parameters such as cell type, largest macroscopic tumour dimension, scleral invasion or pigmentation were observed. In contrast, a significant positive association was found between c-kit staining and mitotic activity (P = 0.02). However, c-kit expression did not significantly influence survival when evaluated by univariate analysis. In conclusion, c-kit is expressed in most choroidal melanoma tumours. Further analysis should provide new insights into the mechanisms underlying the molecular and cellular changes in choroidal melanomas.
Assuntos
Neoplasias da Coroide/metabolismo , Melanoma/metabolismo , Proteínas Proto-Oncogênicas c-kit/biossíntese , Western Blotting , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Humanos , Immunoblotting , Imuno-Histoquímica , Mitose , Fatores de TempoRESUMO
BACKGROUND: Optical coherence tomography has emerged as a new tool for quantifying axonal loss in multiple sclerosis (MS). A reduction in retinal nerve fiber layer (RNFL) thickness is correlated with Expanded Disability Status Scale score and brain atrophy. OBJECTIVE: To investigate RNFL and macular volume measurements using optical coherence tomography in the clinically isolated syndrome population. DESIGN: Prospective case series. Settings Neurologic clinics at the university hospitals of Lille and Strasbourg (France). PARTICIPANTS: Fifty-six consecutive patients with clinically isolated syndrome (18 with optic neuritis and 38 without optic neuritis) and 32 control subjects. MAIN OUTCOME MEASURES: Macular volume and RNFL thickness. RESULTS: Mean (SD) overall RNFL thickness (98.98 [10.26] microm) and macular volume (6.86 [0.32] microm(3)) in the clinically isolated syndrome population were not significantly different compared with the controls (98.71 [9.08] mum and 6.92 [0.38] microm(3), respectively). No link was noted between atrophy of the RNFL or macula and conversion to MS at 6 months. CONCLUSIONS: Optical coherence tomography does not reveal retinal axonal loss at the earliest clinical stage of MS and does not predict conversion to MS at 6 months.
Assuntos
Axônios/patologia , Doenças Desmielinizantes/patologia , Esclerose Múltipla/patologia , Retina/patologia , Adulto , Potenciais Evocados Visuais , Feminino , Humanos , Masculino , Tomografia de Coerência ÓpticaRESUMO
PURPOSE: The treatment of primary central nervous system lymphoma (PCNSL) and its subset, primary intraocular lymphoma (PIOL), remains of limited efficiency, and salvage therapies are often used without prior testing in adequate animal models. Most PNCSL/PIOL are aggressive B-cell malignancies. Two animal models that closely mimic the human situation were established to evaluate the efficiency of intravitreal and intracerebral anti-CD20 monoclonal antibody (rituximab) injections. METHODS: Human CD20-transfected murine B-lymphoma cells (38C13 CD20(+)) were inoculated in the vitreous through the pars plana or in the caudate nucleus with the use of a stereotaxic frame in immunocompetent syngeneic mice. Animals were monitored clinically and by funduscopic and histologic examination. Rituximab was injected intravitreally or intracerebrally. Occurrences of exophthalmia, neurologic disturbance, and weight loss were monitored over 2 months. RESULTS: Inoculation of 38C13 CD20(+) cells in the eye or the brain resulted in tumor occurrence after a median of 15 days or 22 days, respectively, with histologic characteristics closely resembling those of PIOL and PCNSL. Local rituximab injections eradicated tumor colonization in more than half the graft recipients and inhibited tumor progression significantly in the others compared with progression in mice that underwent grafting with the control 38C13 cell line (no human CD20 expression) and in mice that underwent grafting with 38C13 CD20(+) cells that received local injections of an irrelevant antibody (trastuzumab). CONCLUSIONS: Inoculation of native or human CD20-transfected murine 38C13 cells in the vitreous or the brain of immunocompetent mice provides useful novel models for evaluating the biology and treatment of PIOL and PCNSL. Intravitreal and intracerebral rituximab injections reduced tumor occurrence and growth in each model.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Antígenos CD20/imunologia , Antineoplásicos/administração & dosagem , Neoplasias Encefálicas/tratamento farmacológico , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Animais , Anticorpos Monoclonais Murinos , Antígenos CD20/biossíntese , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Cérebro , Citometria de Fluxo , Imuno-Histoquímica , Injeções , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/patologia , Camundongos , Camundongos Endogâmicos C3H , Neoplasias Experimentais , Rituximab , Resultado do Tratamento , Corpo VítreoRESUMO
BACKGROUND: Juvenile xanthogranuloma is a rare non-Langerhans cell histiocytosis that could occasionally produce diffuse systemic disease in young children, but associated posterior uveal lesions are very uncommon. METHODS: Case report of an 8-month-old boy with macrocephaly who presented with chronic subdural haematoma and intracranial hypertension. A combination of nodular skin lesions and bilateral yellowish choroidal infiltration with limited retinal involvement suggested juvenile xanthogranuloma. RESULTS: The diagnosis was confirmed by a skin biopsy, and oral corticosteroid therapy was introduced. Progression of the disease to involve an anterior uveitis with hypopyon and numerous other systemic lesions, including the central nervous system, lung, liver, spleen, kidney and testis, was also suggestive of Letterer-Siwe disease. Histiocytes were negative for Langerhans cell markers (S-100 and CD1a) and positive for macrophage marker (CD68). Electron microscopy failed to show Birbeck granules. Ocular lesions regressed under prolonged corticosteroid treatment, but resurgence of the other lesions required chemotherapy with vinblastine. CONCLUSION: In this atypical systemic variant of juvenile xanthogranuloma with bilateral uveal involvement, the immunohistochemical and ultrastructural findings were crucial in distinguishing juvenile xanthogranuloma from Langerhans cell histiocytosis.
Assuntos
Doenças da Úvea/complicações , Xantogranuloma Juvenil/complicações , Biópsia , Hematoma Subdural Crônico/complicações , Hematoma Subdural Crônico/diagnóstico , Humanos , Lactente , Hipertensão Intracraniana/complicações , Hipertensão Intracraniana/diagnóstico , Masculino , Prednisona/uso terapêutico , Pele/patologia , Dermatopatias/complicações , Dermatopatias/diagnóstico , Doenças da Úvea/diagnóstico , Xantogranuloma Juvenil/diagnósticoRESUMO
Fuchs heterochromic cyclitis (FHC) is characterized by a predominant CD8(+) T cell subset infiltrating the anterior chamber, but the clonal composition of these T cells is unknown. In the present study, T cell repertoire diversity of the accumulating T cells was analyzed to investigate if a high degree of restriction could indicate antigen-driven immune response. Aqueous humor (AH) and peripheral blood cells were collected in two patients with FHC. T cell repertoire diversity was screened by T cell receptor (TCR) BV family expression. In one patient, several BV gene segments were used by lymphocytes from the AH but with over-representation of BV16 that accounted for around half of the expressed intraocular repertoire. In the other patient, a more restricted TCRBV usage was found in AH, as only BV15 and BV18 were expressed in the ocular sample. In this patient, virtually all AH CD8(-) T-cells were CD28- and CD57-negative by three-color flow cytometry, an immunophenotype suggestive of past antigenic stimulation. High resolution immunoscope analysis of TCRBV CDR3 profiles and sequencing of subcloned TCRBV-BJ PCR products evidenced a highly restricted TCRBV-BJ usage, since virtually all the intraocular cells comprise only five clonotypes in this patient. Unique peaks of CDR3 length were found in BV15 joined to BJ2S1, BJ2S3 and especially BJ2S5, in AH but did not predominate in blood. Conversely, identical clonotypes using rearranged BV18 and BJ2S7 gene segments were detected in both AH and peripheral blood. Maintenance of the TCRBV18-BJ2S7 clonotypes in aqueous humor was demonstrated over 6 months in this patient, with a switch in the predominance of two clonotypes. Our results show the presence of a finite number of CD8(+)CD28(neg) T cell clonotypes, which suggests an antigen-driven process and the involvement of these T cells in the pathogenesis of FHC.
Assuntos
Humor Aquoso/imunologia , Linfócitos T CD8-Positivos/imunologia , Iridociclite/imunologia , Subpopulações de Linfócitos T/imunologia , Antígenos CD28/análise , Regiões Determinantes de Complementaridade/genética , Seguimentos , Rearranjo Gênico da Cadeia beta dos Receptores de Antígenos dos Linfócitos T , Humanos , Imunofenotipagem , Masculino , Receptores de Antígenos de Linfócitos T/genética , Células-Tronco/imunologiaRESUMO
PURPOSE: The protein encoded by the Microphthalmia gene (MITF) is a transcription factor essential for the development and survival of melanocytes. It serves as a master regulator in modulating extracellular signals. Because of its central role in melanocytes survival and to assess its potential use as a histopathological marker for melanoma, MITF expression was examined in human choroidal melanomas. METHODS: Fifty-seven paraffin-embedded sections of choroidal melanoma specimens and 1 choroidal melanoma cell line were analyzed using immunochemistry and RT-PCR. Normal choroids and normal choroidal melanocyte cells were used as control. RESULTS: Sixty-five percent of the tumoral specimens stained positively for MITF with a predominant nuclear pattern of reactivity. MITF-M and MITF-A isoforms were detected by RT-PCR in all specimens examined. Using a chimeric protein resulting from the fusion of each Mitf protein with the GFP, Mitf-M exhibited an exclusive nuclear staining whereas Mitf-A exhibited a mixed nuclear and cytoplasmic staining. No correlation between MITF-positivity and parameters such as cell type, largest tumor diameter, sclera invasion, mitotic figures was observed. In contrast, a significant negative association was found between MITF staining and the pigmentation (p=0.02) and a positive correlation between MITF staining and the proliferative marker Ki67 was found (p=0.02). CONCLUSION: MITF may be implicated in choroidal melanoma pigmentation and proliferation. Further analysis should provide new insights into the mechanisms underlying the molecular and cellular changes of choroidal melanomas.
Assuntos
Neoplasias da Coroide/metabolismo , Proteínas de Ligação a DNA/análise , Melanoma/metabolismo , Fatores de Transcrição/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/análise , Neoplasias da Coroide/mortalidade , Neoplasias da Coroide/patologia , Proteínas de Ligação a DNA/genética , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica/métodos , Masculino , Melanoma/mortalidade , Melanoma/patologia , Fator de Transcrição Associado à Microftalmia , Pessoa de Meia-Idade , Pigmentação , Isoformas de Proteínas/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Taxa de Sobrevida , Fatores de Transcrição/genética , Células Tumorais CultivadasRESUMO
Toxoplasma gondii infection is an important cause of chorioretinitis in Europe and the United States. Ophthalmological examination and a good clinical response to adequate therapy mainly support ocular toxoplasmosis diagnosis. However, clinical diagnostic may be difficult in some atypical cases. In these cases, laboratory confirmation, based on detection of local specific antibodies and parasite DNA by conventional PCR, is therefore important to confirm the disease etiology. More recently, real-time PCR has been developed to improve prenatal congenital toxoplasmosis diagnosis. We therefore examined the diagnostic value of quantitative real-time PCR for the detection of T. gondii in aqueous humor samples, associated with quantification of human beta-globin to control sample quantitative quality, by using a double fluorescence resonance energy transfer hybridization probes system with a double fluorescence reading. Of the 23 the clinically toxoplasmosis suspect patients, 22 showed serological evidence of exposure to Toxoplasma; one had a serological profile indicative of active infection. The analysis of paired aqueous humor and serum samples revealed an intraocular antibody production in 9 of 23 cases (39.1%). The quantitative real-time PCR revealed positive and high parasite numbers and high Toxoplasma/human genome ratios in three cases. Furthermore, PCR was the only positive confirmatory test in two cases (11.1%). None of the patients included in the control group (n = 7) had evidence of either local specific antibody production or T. gondii DNA detection, suggesting a good relative assay specificity. On the whole, quantitative real-time PCR appears to be useful for diagnosing atypical ocular toxoplasmosis presentations.