RESUMO
BACKGROUND: Skin health declines with age and this is partially attributed to immunosenescence. Mast cells (MCs) are innate immune cells that coordinate tissue immune responses integral to skin homeostasis and disease. OBJECTIVES: To understand how MCs contribute to human skin ageing, we investigated how intrinsic ageing impacts MC phenotype and MC relationships with other immune cells and skin structures. METHODS: In photoprotected skin biopsies from young (≤ 30 years) and aged (≥ 75 years) individuals, immunostaining and spatial morphometry were performed to identify changes in MC phenotype, number, distribution and interaction with the vasculature and nerve fibres. Quantitative polymerase chain reaction was used to measure changes in gene expression related to immune cell activity and neuropeptide signalling. RESULTS: Skin MCs, macrophages and CD8+ T cells increased in number in intrinsically aged vs. young skin by 40%, 44% and 90%, respectively (P < 0·05), while CD4+ T cells and neutrophils were unchanged. In aged skin, MCs were more numerous in the papillary dermis and showed a reduced incidence of degranulation (50% lower than in young, P < 0·01), a conserved tryptase-chymase phenotype and coexpression of granzyme B. In aged skin, MCs increased their association with macrophages (~ 48% vs. ~27%, P < 0·05) and nerve fibres (~29% vs. 16%, P < 0·001), while reducing their interactions with blood vessels (~34% vs. 45%, P < 0·001). Additionally, we observed modulation of gene expression of vasoactive intestinal peptide (VIP; increased) and substance P (decreased) with age; this was associated with an increased frequency of VIP+ nerve fibres (around three times higher in aged skin, P < 0·05), which were strongly associated with MCs (~19% in aged vs. 8% in young, P < 0·05). CONCLUSIONS: In photoprotected skin we observed an accumulation of MCs with increasing age. These MCs have both altered functionality and distribution within the skin, which supports a role for these cells in altered tissue homeostasis during ageing.
Assuntos
Comunicação Celular/imunologia , Macrófagos/imunologia , Mastócitos/imunologia , Envelhecimento da Pele/imunologia , Pele/citologia , Adulto , Idoso , Biópsia , Linfócitos T CD8-Positivos , Contagem de Células , Perfilação da Expressão Gênica , Humanos , Fibras Nervosas/imunologia , Fibras Nervosas/metabolismo , Pele/imunologia , Pele/patologia , Peptídeo Intestinal Vasoativo/metabolismoRESUMO
BACKGROUND: Sexual dysfunction is a frequent, yet underrated, symptom of neurological disease. While knowledge of non-motor comorbidity in focal dystonia is growing rapidly, there is no information on the prevalence of sexual dysfunction in cervical dystonia (CD) or blepharospasm (BL). METHODS: In this controlled study, we examined sexual dysfunction in 65 patients with CD and 54 patients with BL by the Arizona Sexual Experience Scale, a validated self-rating scale. RESULTS: Sexual dysfunction was significantly higher in CD patients (45%) than in controls (24%), and frequent in BL (39%). Interestingly, variables of dystonia such as disease duration or severity did not influence sexuality; yet, 23% of CD patients ascribed worsening of their sexual life to dystonia. Symptoms of depression were identified as the most important predictors for sexual dysfunction, followed by age, and personal status (single). CONCLUSION: Our observations establish sexual dysfunction as a frequent non-motor symptom in CD and BL that is perceived as a burden. It should be considered when investigating patients with adult-onset focal dystonia.
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Causes of cardiovascular autonomic dysfunction in cervical dystonia (CD) are poorly understood. Studies examining effects of botulinum neurotoxin (BoNT) therapy on heart rate variability (HRV) yielded contradictory results. There is compelling evidence that depression shifts autonomic balance towards sympathetic predominance. As depression is the most frequent non-motor symptom in CD, we sought to determine if it is associated to dysfunction of cardiovascular autonomic regulation. Standardized interviews, clinical examinations, self-rating forms, autonomic symptom questionnaire, and automated autonomic testing in outpatients with idiopathic CD were used. Cardiovascular autonomic screening encompassed five different analyses of HRV, and testing of orthostasis. 85 CD patients participated in the study. 21% of them had HRV impairment, 14% orthostatic hypotension. 30% of CD patients had symptoms of depression. In those, decreased HRV was more frequent than in CD patients without mood disturbance (40 vs. 13%; p = 0.008). CD patients with and without depression had no other significant differences, including demographics, dystonia severity, comorbidity, medication, or BoNT therapy. Cardiovascular autonomic imbalance with sympathetic predominance is a non-motor manifestation of CD, associated to depression. Impaired HRV is a cardiovascular risk factor, moreover, emphasizing the need to identify and treat depression in dystonia.
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Depressão/fisiopatologia , Frequência Cardíaca , Torcicolo/fisiopatologia , Torcicolo/psicologia , Adulto , Idoso , Doenças do Sistema Nervoso Autônomo/complicações , Doenças do Sistema Nervoso Autônomo/fisiopatologia , Doenças do Sistema Nervoso Autônomo/psicologia , Toxinas Botulínicas/uso terapêutico , Doenças Cardiovasculares/complicações , Doenças Cardiovasculares/fisiopatologia , Doenças Cardiovasculares/psicologia , Comorbidade , Depressão/complicações , Feminino , Frequência Cardíaca/fisiologia , Humanos , Entrevistas como Assunto , Masculino , Pessoa de Meia-Idade , Fármacos Neuromusculares/uso terapêutico , Autorrelato , Índice de Gravidade de Doença , Torcicolo/complicações , Torcicolo/tratamento farmacológicoAssuntos
Transtornos Relacionados ao Uso de Anfetaminas/complicações , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Dopamina/metabolismo , Músculos Faciais/fisiopatologia , Transtornos dos Movimentos/etiologia , Adulto , Transtornos Relacionados ao Uso de Anfetaminas/diagnóstico por imagem , Humanos , Masculino , Transtornos dos Movimentos/patologia , Tomografia Computadorizada de Emissão de Fóton Único , Tropanos/farmacocinéticaRESUMO
INTRODUCTION: Limbic encephalitis (LE) associated with voltage-gated potassium channel-complex antibodies (VGKC-LE) is frequently non-paraneoplastic and associated with marked improvement following corticosteroid therapy. Mesial temporal lobe abnormalities are present in around 80 % of patients. If associated or preceded by faciobrachial dystonic seizures, basal ganglia signal changes may occur. In some patients, blurring of the supratentorial white matter on T2-weighted images (SWMB) may be seen. The purpose of this study was to evaluate the incidence of SWMB and whether it is specific for VGKC-LE. METHODS: Two experienced neuroradiologists independently evaluated signal abnormalities on FLAIR MRI in 79 patients with LE while unaware on the antibody type. RESULTS: SWMB was independently assessed as present in 10 of 36 (28 %) compared to 2 (5 %) of 43 non-VGKC patients (p = 0.009). It was not related to the presence of LGI1 or CASPR2 proteins of VGKC antibodies. MRI showed increased temporomesial FLAIR signal in 22 (61 %) VGKC compared to 14 (33 %) non-VGKC patients (p = 0.013), and extratemporomesial structures were affected in one VGKC (3 %) compared to 11 (26 %) non-VGKC patients (p = 0.005). CONCLUSION: SWMB is a newly described MRI sign rather specific for VGKC-LE.
Assuntos
Cérebro/patologia , Imagem de Tensor de Difusão/métodos , Encefalite Límbica/imunologia , Encefalite Límbica/patologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana/imunologia , Substância Branca/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Autoanticorpos/imunologia , Cérebro/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Substância Branca/imunologia , Adulto JovemRESUMO
Focal lesions of brainstem, thalamus, and subcortical white matter may cause movement disorders that are clinically indistinguishable from cerebellar symptoms. It is suspected that ataxia in these cases is due to damage of efferent or afferent pathways of the cerebellum. However, the precise anatomical correlate often remains undefined. We used deterministic diffusion tensor magnetic resonance imaging (DTI) tractography to study the anatomical relationship between lesions causing ataxia and efferent cerebellar pathways. Study subjects were six male patients with focal lesions of different etiology (demyelination, hemorrhage, ischemia, neoplasm) outside the cerebellum. Five patients had cerebellar-like ataxia with prominent contralateral upper limb involvement. One patient with an almost midline mesencephalic lesion had a symmetrical ataxic syndrome. We used 3T MRI (Intera, Philips Medical Systems, Best, Netherlands) and DTI tractography (32 directions, StealthViz DTI, Medtronic Navigation, Louisville, USA) to delineate the dentato-rubro-thalamo-cortical tract (DRT). In all patients, tractography demonstrated focal lesions affecting the DRT in different locations. We conclude that in vivo mapping of cerebral pathways using DTI tractography in patients with focal extracerebellar brain lesions may provide direct evidence of circumscribed damage to the DRT, causing unilateral cerebellar-like ataxia. Also, a unilateral mesencephalic lesion at the level of the crossing of the DRT may cause bilateral ataxia.
Assuntos
Ataxia/patologia , Núcleos Cerebelares/patologia , Córtex Cerebral/patologia , Imagem de Tensor de Difusão/métodos , Tálamo/patologia , Tremor/patologia , Adolescente , Idoso , Idoso de 80 Anos ou mais , Ataxia Cerebelar/patologia , Vias Eferentes/patologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Mast cells (MCs) play a central role in allergic and inflammatory disorders by rapid degranulation and release of inflammatory mediators upon antigen-driven engagement of the FcεRI. Receptor-mediated MC responses are controlled by the activation of different isoforms of phosphoinositide-3-kinase (PI3K) and the downstream signaling processes. Recent evidence suggests that miRNAs are important molecular players regulating the PI3K/Akt pathway. METHODS: The role of miR-155 in the regulation of MC functions in vivo was studied in the passive cutaneous anaphylaxis (PCA) MC-dependent model. WT and miR-155(-/-) mice were injected intradermally with anti-DNP-IgE and intravenously with the antigen DNP-HSA. Ear swelling was assessed to evaluate the anaphylactic response. All investigations, to characterize miR-155 specific activities in MCs, were conducted comparing WT and miR-155(-/-) bone marrow-derived MCs (BMMCs). RESULTS: We report that miR-155(-/-) mice display enhanced anaphylaxis reactions. Although miR-155(-/-) BMMCs show normal development, proliferation, and survival, miR-155 deficiency enhances FcεRI-mediated degranulation and release of TNF-α, IL-13, and IL-6. Interestingly, the level of Akt phosphorylation on both of its regulatory residues Thr308 and Ser473 was increased in miR-155(-/-) compared to WT BMMCs. Gene expression profiling showed that miR-155(-/-) BMMCs exhibited significantly increased expression of the adapter PI3Kγ subunits Pik3r5 (p101) and Pik3r6 (p84, p87(PIKAP) ). Furthermore, selective blockade of the PI3Kγ pathway inhibited degranulation in miR-155(-/-) BMMCs. CONCLUSIONS: Thus, we suggest that miR-155 plays a critical role in FcεRI-mediated MC responses by modulating components of the PI3Kγ pathway. This newly identified mechanism of miRNA-controlled MC activation may affect the initiation and maintenance of allergic disorders.
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Anafilaxia/etiologia , Classe Ib de Fosfatidilinositol 3-Quinase/metabolismo , Mastócitos/imunologia , Mastócitos/metabolismo , MicroRNAs/genética , Transdução de Sinais , Animais , Degranulação Celular/genética , Degranulação Celular/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica , Camundongos , Camundongos Knockout , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismoRESUMO
BACKGROUND AND PURPOSE: Conventional scales measure the effect of botulinum toxin (BT) therapy only at specific points in time. The Dystonia Discomfort Scale (DDS), a novel, easy-to-use, self-assessment scale to record temporal profiles of the effect of BT therapy in cervical dystonia (CD), is introduced and evaluated against the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS). METHODS: Seventy-six patients with CD (age 54.4 ± 10.9 years, 34% male) receiving ≤5 cycles of incobotulinumtoxinA (Xeomin); Merz Pharmaceuticals, Frankfurt am Main, Germany) injections at intervals ≥10 weeks used DDS to record the severity of their symptoms daily. DDS data were compared with TWSTRS-Total scores and patients' subjective estimation (SE) of the onset (TO) and waning (TW) of the treatment effect. RESULTS: The Toronto Western Spasmodic Torticollis Rating Scale - Total scores correlated significantly with DDS (P ≤ 0.028 at all visits evaluated). TO-DDS and TO-SE were 7.9 ± 8.6 and 7.1 ± 4.1 days, respectively; TW-DDS and TW-SE were 41.8 ± 19.2 and 45.1 ± 21.5 days, respectively. CONCLUSION: The Dystonia Discomfort Scale is a novel, easy-to-use, self-assessment scale for valid and sensitive monitoring of the temporal profile of the effect of BT therapy in patients with CD. DDS provides important additional information about onset, duration, waning, stability and reproducibility of the effects of BT therapy.
Assuntos
Toxinas Botulínicas Tipo A/uso terapêutico , Monitoramento de Medicamentos , Fármacos Neuromusculares/uso terapêutico , Índice de Gravidade de Doença , Torcicolo/tratamento farmacológico , Adulto , Idoso , Feminino , Alemanha , Humanos , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Estudos Prospectivos , Reprodutibilidade dos Testes , Autoavaliação (Psicologia) , Resultado do TratamentoRESUMO
A common subset of genetic risk factors for Parkinson's disease (PD) and essential tremor (ET) has been postulated. Recently, an association between the dopamine D(3) receptor (DRD3) Ser9Gly polymorphism and ET has been reported. We studied whether PD tremor is influenced by Ser9Gly in a genetic association study based on the gene bank of the German Competence Network on Parkinson's disease. The study included analyses of motor predominance (mixed, hypokinetic, and tremor), and tremor type (resting, postural, and action). We did not identify any effect of DRD3 Ser9Gly on tremor in PD, even when regarding various symptom combinations to avoid missing a weak effect on the phenotype. Additional studies incorporating symptoms at disease onset, and grading of tremor response to dopaminergic therapy, are warranted.
Assuntos
Predisposição Genética para Doença/genética , Doença de Parkinson/genética , Polimorfismo de Nucleotídeo Único , Receptores de Dopamina D3/genética , Tremor/genética , Idoso , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Doença de Parkinson/complicações , FenótipoRESUMO
Although therapy of CD30-positive lymphomas such as classical Hodgkin lymphoma and anaplastic large cell lymphoma has been improved considerably during the last decades, patients suffer from high toxicity of current therapeutic regimens. Since CD30 expression is very restricted, CD30-positive tumors are well suited for immunotherapeutic approaches. Several distinct immunotherapeutic approaches with chimeric, humanized, and bispecific antibodies as well as immunotoxins are already described. In this report, we give a short overview of CD30-targeting approaches in humans. Furthermore, we introduce two novel anti-CD30 fusion proteins consisting of the single chain variable fragment of the CD30 monoclonal antibody Ber-H2 and human interleukin-2, evaluate their biological activity in a human CD30-positive syngeneic murine model, and demonstrate the immunological mechanisms leading to tumor rejection by these reagents. The data indicate that there are several promising approaches in CD30-targeted immunotherapy. The findings of the anti-CD30 IL-2 constructs suggest that these fusion proteins are particularly useful to remove small, residual tumors.
Assuntos
Anticorpos Monoclonais/imunologia , Sobrevivência Celular/efeitos dos fármacos , Interleucina-2/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Animais , Sistemas de Liberação de Medicamentos , Humanos , Interleucina-2/imunologia , Antígeno Ki-1/imunologia , Proteínas Recombinantes de Fusão/imunologiaRESUMO
Interleukin (IL)-21 is a T cell-derived cytokine which uses a heterodimeric receptor, composed of the common gamma-chain (CD132) and an IL-21Ralpha-chain. IL-21 activates lymphoid T and B cells, modulates antibody production but also suppresses maturation of myeloid dendritic cells; however, its role in the differentiation and function of other myeloid cells remains less clear. In this study we analysed IL-21/IL-21Ralpha effects on macrophage (MPhi) differentiation and function. MPhi could be generated readily from bone marrow with MPhi-colony-stimulating factor in the presence of IL-21 (designated IL-21MPhi) or from IL-21Ralpha-/- mice. IL-21Ralpha-/- mice had normal MPhi numbers, suggesting a non-essential role of both IL-21 and the IL-21Ralpha for MPhi generation. We could demonstrate that mature MPhi express the IL-21Ralpha and the common gamma-chain. However, short-term IL-21 stimulation did not enhance MPhi proliferation but induced anti-apoptotic cell-cycle regulators p21(waf1)/p27(Kip1) and expression of suppressors of cytokine signalling (SOCS)2/SOCS3. Moreover, IL-21 enhanced phagocytosis by MPhi via IL-21Ralpha signalling and supports protease activity and matrix metalloproteinase 12 expression. Stimulating MPhi with IL-21 enhanced their capacity to induce antigen-specific CD4+ T cell proliferation in dependence from the IL-21Ralpha, which was not the case for CD8+ T cells. Taken together, IL-21 plays a previously unrecognized role in modulating innate and acquired effector mechanisms of murine MPhi by linking these different functions to support CD4+ T cell-mediated immune responses.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Interleucinas/imunologia , Macrófagos/imunologia , Animais , Proteínas de Ciclo Celular/metabolismo , Diferenciação Celular/imunologia , Proliferação de Células , Epitopos de Linfócito T/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Subunidade p40 da Interleucina-12/imunologia , Subunidade alfa de Receptor de Interleucina-21/imunologia , Ativação Linfocitária/imunologia , Macrófagos/enzimologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Peptídeo Hidrolases/metabolismo , Fagocitose/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
BACKGROUND: It has been recently demonstrated that apoptosis is involved in beta-cell destruction in the NOD mouse model of diabetes. The aim of the present study was to investigate whether IL-15, a cytokine involved in the modulation of the apoptotic process, is capable of modifying the natural history of diabetes and/or insulitis in pre-diabetic NOD mice. The rationale for the use of IL-15-IgG2b recombinant cytokine is related to its long half-life (28 +/- 4 h). METHODS: At 10 weeks of age, 2 groups of 24 female mice were treated with single or multiple i.p. doses of IL-15-IgG2b respectively. As control, 2 groups of 24 age- and litter-matched female mice were injected intra-peritoneally with single or multiple doses of IgG2b immunoglobulin. RESULTS: Diabetes incidence at 33 weeks of age was lower in the group of mice treated with multiple doses than in the control group (p = 0.03). The cumulative incidence of diabetes at 33 weeks of age between single-dose treated mice and the control group was similar. No significant differences in the calculated index of insulitis were observed in all treated and control mice. CONCLUSIONS: We conclude that IL-15-IgG2b reduces the cumulative incidence of diabetes, without affecting the extent and severity of the insulitis process. Considering this and the well-defined anti-apoptotic effects of IL-15, we suggest that the reduction of diabetes incidence could be due to a down-regulation of beta-cell apoptosis.
Assuntos
Diabetes Mellitus Tipo 1/prevenção & controle , Interleucina-15/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Animais , Diabetes Mellitus Tipo 1/epidemiologia , Glicosúria , Humanos , Imunoglobulina G/farmacologia , Incidência , Camundongos , Camundongos Endogâmicos NOD , Fatores de TempoRESUMO
We report a 73-year-old patient with acid maltase deficiency who initially had been suspected of having motor neuron disease. We discuss clinical and electrophysiological features of muscular lysosomal glycogenosis, with special emphasis on the histopathological differential diagnosis of the disease.
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Doença de Depósito de Glicogênio Tipo II/diagnóstico , Doenças Neuromusculares/diagnóstico , Idoso , Biópsia , Diagnóstico Diferencial , Eletromiografia , Feminino , Glucana 1,4-alfa-Glucosidase/deficiência , Doença de Depósito de Glicogênio Tipo II/patologia , Humanos , Debilidade Muscular/diagnóstico , Debilidade Muscular/patologia , Músculo Esquelético/patologia , Atrofia Muscular/diagnóstico , Atrofia Muscular/patologia , Exame Neurológico , Doenças Neuromusculares/patologia , Paralisia/diagnóstico , Paralisia/patologia , alfa-GlucosidasesAssuntos
Doenças Desmielinizantes/terapia , Mielite/terapia , Troca Plasmática , Esteroides/uso terapêutico , Adulto , Terapia Combinada , Doenças Desmielinizantes/diagnóstico , Progressão da Doença , Feminino , Gadolínio , Humanos , Imageamento por Ressonância Magnética , Debilidade Muscular/etiologia , Mielite/diagnóstico , Pescoço , Parestesia/etiologia , Prednisolona/uso terapêutico , Recuperação de Função Fisiológica , Medula Espinal/efeitos dos fármacos , Medula Espinal/patologiaAssuntos
Antineoplásicos/farmacologia , Hidroxiureia/farmacologia , Neoplasias Meníngeas/tratamento farmacológico , Meningioma/tratamento farmacológico , Neoplasias do Nervo Óptico/tratamento farmacológico , Administração Oral , Antineoplásicos/administração & dosagem , Feminino , Humanos , Hidroxiureia/administração & dosagem , Neoplasias Meníngeas/patologia , Meningioma/patologia , Pessoa de Meia-Idade , Neoplasias do Nervo Óptico/patologia , Resultado do TratamentoRESUMO
A functional hybrid receptor associating the common gamma chain (gammac) with the granulocyte/macrophage colony-stimulating factor receptor beta (GM-CSFRbeta) chain is found in mobilized human peripheral blood (MPB) CD34+ hematopoietic progenitors, SCF/Flt3-L primed cord blood (CB) precursors (CBPr CD34+/CD56-), and CD34+ myeloid cell lines, but not in normal natural killer (NK) cells, the cytolytic NK-L cell line or nonhematopoietic cells. We demonstrated, using CD34+ TF1beta cells, which express an interleukin (IL)-15Ralpha/beta/gammac receptor, that within the hybrid receptor, the GM-CSFRbeta chain inhibits the IL-15-triggered gammac/JAK3-specific signaling controlling TF1beta cell proliferation. However, the gammac chain is part of a functional GM-CSFR, activating GM-CSF-dependent STAT5 nuclear translocation and the proliferation of TF1beta cells. The hybrid receptor is functional in normal hematopoietic progenitors in which both subunits control STAT5 activation. Finally, the parental TF1 cell line, which lacks the IL-15Rbeta chain, nevertheless expresses both a functional hybrid receptor that controls JAK3 phosphorylation and a novel IL-15alpha/gammac/TRAF2 complex that triggers nuclear factor kappaB activation. The lineage-dependent distribution and function of these receptors suggest that they are involved in hematopoiesis because they modify transduction pathways that play a major role in the differentiation of hematopoietic progenitors.
Assuntos
Antígenos CD34/metabolismo , Células-Tronco Hematopoéticas/imunologia , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/metabolismo , Receptores de Interleucina-2/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Anticorpos Monoclonais/metabolismo , Divisão Celular/fisiologia , Linhagem Celular , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Células Matadoras Naturais/metabolismo , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/genética , Receptores de Interleucina-15 , Receptores de Interleucina-2/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Transdução de Sinais/fisiologiaRESUMO
Cytokine-immunoglobulin (Ig)-fusion proteins have attracted increasing interest as antitumour agents. Here, we have investigated the antimetastatic and antitumour responses elicited in vivo by mammary adenocarcinoma cells (TS/A) engineered to secrete interleukin (IL)-2-IgG fusion proteins. TS/A cells were transfected with DNA coding for IL-2-IgG2b, IgG2b or IL-2, and injected subcutaneously into syngeneic mice. Animals injected with TS/A-IL-2 or TS/A-IL-2-IgG2b both efficiently rejected tumours, whereas treatment with parental cells or TS/A-IgG2b was lethal. Interestingly, only mice vaccinated with IL-2-IgG2b fusion protein-secreting cells showed a long-lasting protective immunity against a later challenge with parental tumour cells. Moreover, the metastatic potential of TS/A-IL-2-IgG2b-transfected cells was dramatically decreased compared with TS/A-IL-2-cells, with a virtual absence of lung metastases after intravenous injection. Adenocarcinomas secreting IL-2-IgG2b exhibited a more prominent, early and persistent infiltration of CD4+, CD8+ and natural killer (NK) cells than TS/A-IL-2 cells. Therefore, upon transfection into adenocarcinoma cells, the IgG2b part of IL-2 fusion protein exerts intriguing added antitumour properties over IL-2 alone, thus contributing to a long-lasting tumour immunity, probably by the recruitment of specific immune effector cells. These findings suggest a promising new oncotherapeutic strategy for poorly immunogenic tumours: vaccination with tumour cells engineered to secrete IL-2-IgG2b fusion protein.
Assuntos
Adenocarcinoma/imunologia , Imunoglobulina G/imunologia , Interleucina-2/imunologia , Neoplasias Mamárias Experimentais/imunologia , Proteínas Recombinantes de Fusão/imunologia , Adenocarcinoma/patologia , Adenocarcinoma/terapia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Feminino , Inibidores do Crescimento/genética , Inibidores do Crescimento/imunologia , Imunoglobulina G/genética , Imuno-Histoquímica , Imunoterapia , Interleucina-2/genética , Células Matadoras Naturais/imunologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/prevenção & controle , Neoplasias Pulmonares/secundário , Neoplasias Mamárias Experimentais/patologia , Neoplasias Mamárias Experimentais/terapia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes de Fusão/genética , Transfecção , Células Tumorais CultivadasRESUMO
BACKGROUND: The immunoreaction after corneal transplantation is caused by the T cell receptor interacting with the major histocompatibility complex (MHC) receptor of the antigen-presenting cell. The signal is amplified by the CD4 receptor and the costimulatory signal interactions of CD28-B7 and CD40-CD154. We investigated the influence of costimulatory signal blocking on corneal transplant survival in mice. METHODS: Seven groups of 6 BALB/c mice received an orthotopic corneal transplant from C3H mice differing in minor and major MHC and were postoperatively treated as follows: (1) 80 micrograms of CTLA4 fusion protein intraperitoneally (i.p.) for 6 days; (2) 50 microliters of PBS i.p. for 6 days; (3) 1 mg of Solu-Decortin H i.p. for 5 days + dexamethasone AT 0.1% for 35 days; (4) therapy (3) + 50 micrograms of CTLA4 fusion protein i.p. for 6 days; (5) CTLA4-Ig as in (1) + 15 micrograms of anti-CD154 subconjunctivally (s.c.) on days 0, 2, 4, 6, and 8; (6) CTLA4-Ig as in (1) + 25 micrograms of anti-CD154 s.c. for 9 days; and (7) 25 microliters of PBS s.c. for 9 days. RESULTS: All animals had an immunoreaction on the following days: (1) day 18 +/- 3.1; (2) day 13.6 +/- 1.6; (3) day 48 +/- 6.6; (4) day 65 +/- 41; (5) day 23.5 +/- 8.5; (6) day 16.2 +/- 3.6; (7) day 13.8 +/- 2.7. CONCLUSION: The significant prolongation of transplant survival achieved by corticosteroids alone (P < 0.001) is again significantly increased by combining them with CTLA4-Ig (P < 0.001). Specific immunotherapy combined with nonspecific steroid therapy may also improve clinical corneal transplantation results. Compared to the two control groups, CTLA4-Ig and anti-CD154 only influenced transplant survival at a low dosage (P < 0.001).
Assuntos
Adjuvantes Imunológicos , Anti-Inflamatórios/uso terapêutico , Antígenos de Diferenciação/imunologia , Ligante de CD40/imunologia , Transplante de Córnea , Desoxicorticosterona/uso terapêutico , Dexametasona/uso terapêutico , Imunoconjugados , Imunossupressores/uso terapêutico , Abatacepte , Animais , Anti-Inflamatórios/administração & dosagem , Antígenos CD , Antígeno CTLA-4 , Desoxicorticosterona/administração & dosagem , Dexametasona/administração & dosagem , Feminino , Sobrevivência de Enxerto , Imunossupressores/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Fatores de TempoRESUMO
The alpha-chain of the IL-15R (IL-15Ralpha) serves as the specific, high-affinity receptor for IL-15. It is expressed by lymphoid and nonlymphoid cells, including B cell lymphoma lines. In this study, we have further explored IL-15Ralpha-mediated signaling in activated primary B cells and in Raji cells, a human B-lymphoblastoid cell line which expresses the IL-15Ralpha and IL-2Rgamma chains, but lacks the IL-2Rbeta chain. Stimulation of Raji cells with IL-15 induces their proliferation and rescues them from C2-ceramide-induced apoptosis. By immunoprecipitation and Western blotting, we show that treatment of Raji cells and activated primary B cells with IL-15 induces coprecipitation of Syk kinase with the IL-15Ralpha chain. Upon association, the activated Syk kinase phosphorylates the IL-15Ralpha chain as well as phospholipase Cgamma, which coprecipitates with Syk. Furthermore, transfection of Raji cells with stem-loop Syk antisense oligonucleotides prevents IL-15Ralpha and phospholipase Cgamma phosphorylation as well as the inhibition of apoptosis by IL-15. Mutation of a defined region of the intracellular signaling portion of IL-15Ralpha (Tyr227) abrogates both the IL-15Ralpha/Syk association and IL-15Ralpha phosphorylation. Taken together, this suggests that Syk kinase physically and functionally associates with the IL-15Ralpha chain in B cells and that Syk plays a key role in mediating IL-15-induced signal transduction, thus accounting for the distinct functional consequences of IL-15 vs IL-2 binding to B cells.
Assuntos
Subpopulações de Linfócitos B/imunologia , Subpopulações de Linfócitos B/metabolismo , Precursores Enzimáticos/metabolismo , Interleucina-15/metabolismo , Proteínas Tirosina Quinases/metabolismo , Receptores de Interleucina-2/fisiologia , Transdução de Sinais/imunologia , Esfingosina/análogos & derivados , Apoptose/efeitos dos fármacos , Apoptose/genética , Apoptose/imunologia , Subpopulações de Linfócitos B/efeitos dos fármacos , Subpopulações de Linfócitos B/enzimologia , Cálcio/antagonistas & inibidores , Sinalização do Cálcio/genética , Sinalização do Cálcio/imunologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/imunologia , Ativação Enzimática/imunologia , Precursores Enzimáticos/antagonistas & inibidores , Precursores Enzimáticos/genética , Precursores Enzimáticos/fisiologia , Humanos , Interleucina-15/antagonistas & inibidores , Interleucina-15/fisiologia , Peptídeos e Proteínas de Sinalização Intracelular , Isoenzimas/antagonistas & inibidores , Isoenzimas/metabolismo , Células Jurkat , Células K562 , Ativação Linfocitária , Mutagênese Sítio-Dirigida , Oligonucleotídeos Antissenso/farmacologia , Fosfolipase C gama , Fosforilação , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/fisiologia , Receptores de Interleucina-15 , Receptores de Interleucina-2/antagonistas & inibidores , Receptores de Interleucina-2/biossíntese , Receptores de Interleucina-2/metabolismo , Transdução de Sinais/genética , Esfingosina/farmacologia , Quinase Syk , Subpopulações de Linfócitos T/enzimologia , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Células Tumorais Cultivadas , Fosfolipases Tipo C/antagonistas & inibidores , Fosfolipases Tipo C/metabolismo , Domínios de Homologia de src/imunologiaRESUMO
PURPOSE: To define the role of MRI in the diagnosis of Creutzfeldt-Jakob disease (CJD). METHODS: 14 patients with suspected CJD were studied within 3 years. MRI findings were correlated with WHO established diagnostic criteria (clinical findings, EEG, CSF with 14-3-3 protein assay). RESULTS: 12 patients had CJD. One patient each suffered from Hashimoto's encephalitis and ALS dementia complex, respectively. Nine of 12 CJD patients had increased signal intensity of the striatum (n = 8), pulvinar thalami (n = 5) and/or cerebellar and cerebral cortex (n = 3), respectively. Signal intensity was most pronounced on FLAIR sequences; six patients were studied with diffusion-weighted MRI and showed impaired diffusion in these areas. Both patients without CJD did not show the abovementioned signal changes (sensitivity 75%, specificity and positive predictive value 100%, respectively). CONCLUSION: If patients with suspected CJD are studied with FLAIR and diffusion-weighted sequences, this disorder can reliably be proven or ruled out. Typical MRI findings narrow down the differential diagnosis and should be included in the WHO diagnostic criteria.