RESUMO
OBJECTIVE: To evaluate the influence of continent external urinary diversion type Mitrofanoff on male sexuality. MATERIAL AND METHODS: Between 1992 and 2011, 140 patients underwent continent urinary diversion type Mitrofanoff at an academic hospital. Among 76 men, 46 were interviewed about their sexuality after this operation. This study was performed using a set of validated questionnaires (IIEF, DAN PSS and Urolife), grouped by the model of the CTMH. Patients were divided according to their marital status: group 1: patients married before surgery (15 cases), group 2: patients married after surgery (7 cases) and group 3: singles (24 cases). RESULTS: In the first group, the functional dimension of sexuality was positive with an overall score of 81%, the sexual discomfort score was assessed at 26 % and the sexual satisfaction score was 77%. In the second group, sexual function was considered conserved in all cases with a satisfaction score estimated at 98%. These patients reported a feeling of well-being following the disappearance of urinary incontinence with integrity of their body images. In contrast, in the last group, relatively impaired sexual function was noted (65%) with a satisfaction score estimated at 59%. These disorders were multifactorial, mainly related to neurological causal pathology. CONCLUSION: To our knowledge, this is the first study about male sexuality in patients with a continent urinary diversion type Mitrofanoff. Marital status has a major role in the sexuality of these patients. A prospective study with pre- and postoperative evaluation will better clarify the factors affecting sexuality in these patients.
Assuntos
Sexualidade , Derivação Urinária , Coletores de Urina , Adulto , Idoso , Imagem Corporal , Humanos , Masculino , Estado Civil , Pessoa de Meia-Idade , Sexualidade/psicologia , Inquéritos e Questionários , Adulto JovemRESUMO
We have previously characterized a large panel of provirus insertion Notch1 mutant alleles and their products arising in thymomas of MMTV(D)/myc transgenic mice. Here, we show that these Notch1 mutations represent two clearly distinct classes. In the first class (type I), proviral integrations were clustered just upstream of sequences encoding the transmembrane domain. Type I Notch1 alleles produced two types of mutant Notch1 RNA, one of which encoded the entire Notch1 cytoplasmic domain [N(IC)] and the other of which encoded a soluble ectodomain [N(EC)(Mut)] which, in contrast to the processed wild-type ectodomain [N(EC)(WT)], did not reside at the cell surface and became secreted in a temperature-dependent manner. A second, novel class of mutant Notch1 allele (type II) encoded a Notch1 receptor with the C-terminal PEST motif deleted (DeltaCT). The type II Notch1(DeltaCT) protein was expressed as a normally processed receptor [N(EC)(WT) and N(IC)(DeltaCT)] at the cell surface, and its ectodomain was found to be shed into the extracellular medium in a temperature- and calcium-dependent manner. These data suggest that both type I and type II mutations generate two structurally distinct Notch1 N(EC) and N(IC) proteins that may participate in tumor formation, in collaboration with the c-myc oncogene, through distinct mechanisms. Constitutive type I N(IC) and type II N(IC)(DeltaCT) expression may enhance Notch1 intracellular signaling, while secreted or shed type I N(EC)(Mut) and type II N(EC) proteins may differentially interact in an autocrine or paracrine fashion with ligands of Notch1 and affect their signaling.
Assuntos
Alelos , Leucemia de Células T/etiologia , Proteínas de Membrana/genética , Proteínas Proto-Oncogênicas c-myc/fisiologia , Receptores de Superfície Celular , Timoma/etiologia , Neoplasias do Timo/etiologia , Fatores de Transcrição , Sequência de Aminoácidos , Animais , Linhagem Celular Transformada , Membrana Celular/metabolismo , Camundongos , Camundongos Transgênicos , Dados de Sequência Molecular , Mutagênese Insercional , Proteínas Proto-Oncogênicas c-myc/genética , Provírus/genética , RNA , Processamento Pós-Transcricional do RNA , Receptor Notch1 , Linfócitos TRESUMO
To identify which segments of the beta 1 domain of the E beta k polypeptide control T cell recognition of antigen, E beta genes were constructed with in-phase insertion mutations. Five independent mutants, with insertions mapping to positions 24, 50 and 93 of the E beta k polypeptide, were obtained. Cell lines expressing these mutated genes were analysed by microfluorometry using a panel of 20 anti-Ek monoclonal antibodies. None of the tested in-phase insertions has resulted in the loss of antibody binding sites. In striking contrast, mutations at position 93 and at a lesser level 50 were indicative of a crucial role of the corresponding regions in T-cell recognition, because they led to significant or complete loss of antigen-presenting function with all but one of the T hybridomas tested. These data are discussed with regard to a model of the foreign antigen binding site of Ia molecules.
Assuntos
Genes MHC da Classe II , Antígenos de Histocompatibilidade Classe II/genética , Mutação , Sequência de Aminoácidos , Animais , Sequência de Bases , Elementos de DNA Transponíveis , Substâncias Macromoleculares , Camundongos , Dados de Sequência Molecular , Plasmídeos , Linfócitos T/imunologia , TransfecçãoRESUMO
A solid-phase radioimmunoanalysis of plasma membrane molecules solubilized in the presence of detergent was developed. From a crude cell lysate, membrane molecules were specifically immobilized by solid-phase adsorbed monoclonal antibodies. Analysis of these molecules was easily performed with radiolabeled monoclonal antibodies of different specificities. This technique was used to analyze the HLA-DR and HLA-A,B,C molecules expressed by RAJI cells. It was possible (a) to determine which epitopes are expressed on the same molecules; (b) to define subsets of HLA molecules according to the epitopes which they express, and (c) to perform quantitative analysis of HLA molecules on a crude cell lysate.
Assuntos
Epitopos/análise , Antígenos HLA/análise , Polietilenoglicóis/farmacologia , Radioimunoensaio/métodos , Animais , Anticorpos Monoclonais/imunologia , Linhagem Celular , Epitopos/imunologia , Antígenos HLA/imunologia , Antígenos HLA-B , Antígenos HLA-C , Antígenos HLA-DR , Antígenos de Histocompatibilidade Classe II/análise , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , SolubilidadeRESUMO
Four independent PLT clones displaying an apparently identical class II specificity (i.e., Dw/DR3) were found to give a heterogeneous pattern of inhibition in relation to 15 anti-class II mAb and an anti-beta 2m mAb used as a control. Some clones were inhibited by all anti-class II mAbs, irrespective of the cluster of molecular subsets with which they reacted. Such clones were also inhibited by the control anti-beta 2m mAb. Other clones were inhibited by only a few of the mAb tested. Within this group of T clones following the addition of a limiting amount of conditioned medium the inhibitory data of all independent clones with an identical specificity were inhibited by the same mAb; under these conditions, it was possible to relate the mAb inhibition patterns with the specificity of the T cell clones and these T cell specificities with the epitopic cluster/molecular subsets defined by these mAbs. A new level of T cell subset heterogeneity within T cell clones with apparent identical proliferative specificities is demonstrated, in relation to the T cell clone "susceptibility" or "resistance" to the effects of anti-class II mAbs directed towards their own Ia-like antigens.
Assuntos
Antígenos de Histocompatibilidade Classe II/imunologia , Complexo Principal de Histocompatibilidade , Linfócitos T/imunologia , Anticorpos Monoclonais , Especificidade de Anticorpos , Células Clonais , Epitopos , Antígenos HLA-DR , Humanos , Ativação LinfocitáriaRESUMO
A T cell growth factor-dependent alloreactive human T cell line has been used to generate a monoclonal antibody B1.49.9 that reacts with an antigen present on most if not all mitogen or alloantigen activated T cells but not on resting T cells. The T lymphoblastoid cell line HUT-102 is also strongly reactive with B1.49.9 but all other T and non-T leukemia-lymphoma cell lines tested were negative. The B1.49.9 antigen is a glycoprotein of 55,000 Mr on mitogen or alloantigen activated T cells and 50,000 Mr on the cell line HUT-102. Pulse labeling experiments showed that a 40,000 Mr precursor (at approximately 0.7 h) which does not bind to ricin lectin precedes the appearance of the ricin-binding 55,000 Mr form. Comparisons of the monoclonal antibody anti-Tac, which recognizes the IL-2 receptor, to B1.49.9 suggest that B1.49.9 also recognizes a structure similar or identical to the IL-2 receptor.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Superfície/isolamento & purificação , Ativação Linfocitária , Linfócitos T/imunologia , Animais , Anticorpos Monoclonais/análise , Reações Antígeno-Anticorpo , Antígenos de Diferenciação de Linfócitos T , Antígenos de Superfície/imunologia , Fenômenos Químicos , Química , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Fito-Hemaglutininas/farmacologiaRESUMO
The reovirus type 3 S1 gene product (type 3 hemagglutinin; HA3) is the viral protein responsible for binding to a mammalian cell-surface receptor. It has been shown that HA3 binding to its receptor inhibits cell growth, even in the continuous presence of serum mitogens. Here, receptor-mediated signal transduction leading to growth arrest was studied after binding with synthetic or recombinant ligands in the absence of viral infection. Receptor ligation caused rapid inactivation of p21(ras), a decrease in Raf phosphorylation and in mitogen-activated protein kinase (MAPK) enzymatic activity, and G1 cell cycle arrest. Transfection and expression of constitutively active v-Has-ras prevented the G1 arrest, indicating that inactivation of p21(ras) is causative. Interestingly, v-Has-ras expression also decreased the efficiency of reoviridae replication, suggesting that inactivation of p21(ras) signals is required at some step of the viral cycle. This study may define new mechanisms regulating cell growth and support the approach of using viral proteins to identify and study cellular receptors. Synthetic receptor ligands with antiproliferative properties may be useful in drug development with the aim of blocking mitosis.
Assuntos
Anticorpos Monoclonais/farmacologia , Proteínas do Capsídeo , Proteínas de Ciclo Celular , Fase G1/efeitos dos fármacos , Inibidores do Crescimento/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Orthoreovirus Mamífero 3/fisiologia , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteína Oncogênica p21(ras)/fisiologia , Peptídeos Cíclicos/farmacologia , Proteínas Proto-Oncogênicas p21(ras)/fisiologia , Receptores Virais/fisiologia , Proteínas Virais/fisiologia , Animais , Anticorpos Monoclonais/imunologia , Divisão Celular/efeitos dos fármacos , Efeito Citopatogênico Viral , Genes ras , Inibidores do Crescimento/química , Guanosina Difosfato/metabolismo , Guanosina Trifosfato/metabolismo , Humanos , Ligantes , Camundongos , Proteína Oncogênica p21(ras)/antagonistas & inibidores , Fosforilação/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Estrutura Terciária de Proteína , Proteínas Proto-Oncogênicas c-raf/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/antagonistas & inibidores , Receptores Virais/agonistas , Proteínas Recombinantes de Fusão/fisiologia , Transfecção , Células Tumorais Cultivadas , Proteínas Virais/química , Proteínas Virais/imunologia , Replicação Viral/efeitos dos fármacosAssuntos
Divisão Celular , Orthoreovirus Mamífero 3/metabolismo , Receptores Virais/metabolismo , Infecções por Reoviridae/virologia , Transdução de Sinais , Animais , Sítios de Ligação , Hemaglutininas Virais/metabolismo , Humanos , Ligantes , Orthoreovirus Mamífero 3/genética , Receptores Virais/genética , Infecções por Reoviridae/patologiaAssuntos
Modelos Animais de Doenças , Infecções por HIV , HIV-1 , Camundongos Transgênicos , Animais , Antígenos CD4/genética , Infecções por HIV/patologia , HIV-1/genética , Hibridização In Situ , Rim/patologia , Pulmão/patologia , Tecido Linfoide/patologia , Camundongos , Camundongos Transgênicos/genética , Especificidade de Órgãos , Especificidade da Espécie , TransgenesRESUMO
The monoclonal antibodies B1.23.2 and B9.12 were developed to characterize human class I major histocompatibility gene products. They detect two different epitopes and define on a given lymphoblastoid B cell line two subsets of beta 2 microglobulin-complexed HLA class I molecules. One subset reacts with B9.12 and B1.23.2 and the other one expresses only the B9.12 epitope. Binding assays were performed on C3H mouse L cells which had been transformed with various single HLA-class I genes and the two detected molecular subsets were shown to be encoded by different genetic loci. Unlike B1.23.2, B9.12 detects all the beta 2m-complexed molecules expressed in human B cell lines and recognizes an epitope different from the one defined by W6/32. In contrast to the vast majority of the other reported anti-class I monoclonal antibodies (including B9.12), B1.23.2 recognizes an epitope expressed on both beta 2m-associated and -free HLA class I heavy chains.
Assuntos
Antígenos HLA/classificação , Animais , Anticorpos Monoclonais/imunologia , Epitopos , Antígenos HLA/genética , Antígenos HLA/imunologia , Humanos , Substâncias Macromoleculares , Camundongos , Especificidade da Espécie , Relação Estrutura-Atividade , Microglobulina beta-2/imunologiaRESUMO
To identify the regulatory elements controlling expression of the human CD4 (hCD4) gene in different cell types of the immune system, deletion and chimeric (human/murine) reporter genes were constructed and tested in transgenic (Tg) mice. Regulatory elements required for the proper hCD4 expression in the immature double-positive thymic T cells were identified in the enhancer and in the 3' end of intron 1. Expression of hCD4 in macrophages is controlled by at least 2 sets of regulatory elements: one present in front of exon 1 and the second at the 5' end of intron 1. The hCD4 elements required for expression on both myeloid and lymphoid CD8alpha(+) dendritic cells (DCs) from lymph node and thymus were found to be different from those required for macrophage expression. The results indicate that expression of hCD4 in T cells, macrophages, and DCs is controlled by distinct regulatory elements.
Assuntos
Antígenos CD4/genética , Células Dendríticas/metabolismo , Genes Reguladores/genética , Macrófagos/metabolismo , Linfócitos T/metabolismo , Animais , Northern Blotting , Antígenos CD4/metabolismo , DNA/metabolismo , Citometria de Fluxo , Regulação da Expressão Gênica/genética , Humanos , Camundongos , Camundongos Transgênicos , Distribuição TecidualRESUMO
Monoclonal antibodies (mAb) derived from BALB/c mice immunized with human anti-HLA-A2 cloned cytotoxic T lymphocytes (CTL) were screened for their ability to block, in the absence of complement, the cytolytic activity of the immunizing CTL clone. Eight cytolysis-inhibiting mAb have been derived. One of these was directed against a monomorphic determinant expressed on HLA-class I molecules and thus probably inhibited cytolysis via a target cell antigen-masking effect. The 7 other mAb recognized "CTL function-associated structures" and did not have to interfere with target cell antigens in order to inhibit cytolysis. F(ab')2 and Fab fragments of these 7 mAb were also inhibitory. Competitive inhibition of binding and preliminary biochemical analysis suggested that these 7 mAb defined on cloned CTL various epitopes of a structure of 30 kDa disulfide-bonded into several multimeric forms when analyzed without reduction. The inhibitory effect of these 7 mAb has been investigated on a series of short-term (1 month) and long-term (greater than 10 months) expanded cloned CTL lines derived in vitro from an in vivo allosensitized individual exhibiting various specificities. Unexpectedly, only 10% of these CTL clones were inhibited. Flow cytofluorimetric analysis further revealed that noninhibited and inhibited CTL clones expressed similar amounts of the 30-kDa structure. Consequently, the inhibition of CTL was heterogeneous when analyzed at the clonal level and not simply related to the presence or absence of this structure. Furthermore, the ability of CTL clones to be inhibited appeared to be unrelated to their HLA-A, B or C specificity or to their lytic activity. The connections between this mAb-defined structure and CTL function are discussed.
Assuntos
Anticorpos Monoclonais , Citotoxicidade Imunológica , Linfócitos T/imunologia , Animais , Células Clonais/imunologia , Antígenos HLA/imunologia , Antígeno HLA-A2 , Humanos , Fragmentos Fab das Imunoglobulinas , Camundongos , Camundongos Endogâmicos BALB CRESUMO
A surface receptor complex of Mr approximately 65 000 (p65) and approximately 95 000 (p95) is expressed in cells of the central nervous system of mice. This receptor is recognized by monoclonal antibody 87.92.6 or by reovirus type 3 haemagglutinin as unnatural ligands. The p65/p95 receptor is expressed mostly in neural embryonic precursors undergoing proliferation, especially those in the S-G2 phase of the cell cycle. Receptor expression decreases progressively throughout embryogenesis to low but detectable levels in the adult brain. Biochemical characterization revealed that the neural p65/p95 receptor complex is indistinguishable from the p65/p95 receptor expressed in T cells, where receptor ligation leads to a mitogenic block. In neural and lymphoid tissues the p65/p95 receptor (or an associated protein) possesses a tyrosine kinase enzymatic activity. Receptor ligation in neural cells resulted in the rapid tyrosine phosphorylation of cellular proteins which are different from substrates phosphorylated in T cells. Differential substrate coupling to the receptor may account for differences in signal transduction and biology between neural cells and T cells. Further study of this receptor complex may help define important features of neural proliferation, differentiation and survival.
Assuntos
Proteínas de Ligação ao Cálcio , Fase G2 , Glicoproteínas de Membrana/biossíntese , Proteínas do Tecido Nervoso/biossíntese , Neurônios/citologia , Fase S , Animais , Anticorpos Monoclonais/imunologia , Encéfalo/citologia , Encéfalo/embriologia , Encéfalo/crescimento & desenvolvimento , Células Cultivadas , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Regulação da Expressão Gênica no Desenvolvimento , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Camundongos , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/imunologia , Neurônios/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo , Receptores Virais/classificação , Transdução de Sinais , Sinaptotagmina I , SinaptotagminasRESUMO
The Pr60gag protein of the murine AIDS (MAIDS) defective virus promotes the proliferation of the infected target B cells and is responsible for inducing a severe immunodeficiency disease. Using the yeast two-hybrid system, we identified the SH3 domain of c-Abl as interacting with the proline-rich p12 domain of Pr60gag. The two proteins were shown to associate in vitro and in vivo in MAIDS virus-infected B cells. Overexpression of Pr60(gag) in these cells led to a detectable increase of the levels of c-Abl protein and to its translocation at the membrane. These results suggest that this viral protein serves as a docking site for signaling molecules and that c-Abl may be involved in the proliferation of infected B cells.
Assuntos
Produtos do Gene gag/metabolismo , Vírus da Leucemia Murina/metabolismo , Precursores de Proteínas/metabolismo , Proteínas Proto-Oncogênicas c-abl/metabolismo , Domínios de Homologia de src , Sequência de Aminoácidos , Animais , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Produtos do Gene gag/genética , Vírus da Leucemia Murina/genética , Camundongos , Dados de Sequência Molecular , Ligação Proteica , Precursores de Proteínas/genética , Proteínas Proto-Oncogênicas c-abl/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismoRESUMO
We used stepwise photochemical cross-linking for specifically assembling soluble and covalent complexes made of a T-cell antigen receptor (TCR) and a class I molecule of the major histocompatibility complex (MHC) bound to an antigenic peptide. For that purpose, we have produced in myeloma cells a single-chain Fv construct of a TCR specific for a photoreactive H-2Kd-peptide complex. Photochemical cross-linking of this TCR single-chain Fv with a soluble form of the photoreactive H-2Kd-peptide ligand resulted in the formation of a ternary covalent complex. We have characterized the soluble ternary complex and showed that it reacted with antibodies specific for epitopes located either on the native TCR or on the Kd molecules. By preventing the fast dissociation kinetics observed with most T cell receptors, this approach provides a means of preparing soluble TCR-peptide-MHC complexes on large-scale levels.