Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 19 de 19
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Cancer Immunol Immunother ; 69(11): 2247-2257, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32500232

RESUMO

Cancer vaccines induce cancer-specific T-cells capable of eradicating cancer cells. The impact of cancer peptide vaccines (CPV) on the tumor microenvironment (TME) remains unclear. S-588410 is a CPV comprising five human leukocyte antigen (HLA)-A*24:02-restricted peptides derived from five cancer testis antigens, DEPDC1, MPHOSPH1, URLC10, CDCA1 and KOC1, which are overexpressed in esophageal cancer. This exploratory study investigated the immunologic mechanism of action of subcutaneous S-588410 emulsified with MONTANIDE ISA51VG adjuvant (median: 5 doses) by analyzing the expression of immune-related molecules, cytotoxic T-lymphocyte (CTL) response and T-lymphocytes bearing peptide-specific T-cell receptor (TCR) sequencing in tumor tissue or blood samples from 15 participants with HLA-A*24:02-positive esophageal cancer. Densities of CD8+, CD8+ Granzyme B+, CD8+ programmed death-1-positive (PD-1+) and programmed death-ligand 1-positive (PD-L1+) cells were higher in post- versus pre-vaccination tumor tissue. CTL response was induced in all patients for at least one of five peptides. The same sequences of peptide-specific TCRs were identified in post-vaccination T-lymphocytes derived from both tumor tissue and blood, suggesting that functional peptide-specific CTLs infiltrate tumor tissue after vaccination. Twelve (80%) participants had treatment-related adverse events (AEs). Injection site reaction was the most frequently reported AE (grade 1, n = 1; grade 2, n = 11). In conclusion, S-588410 induces a tumor immune response in esophageal cancer. Induction of CD8+ PD-1+ tumor-infiltrating lymphocytes and PD-L1 expression in the TME by vaccination suggests S-588410 in combination with anti-PD-(L)1 antibodies may offer a clinically useful therapy.Trial registration UMIN-CTR registration identifier: UMIN000023324.


Assuntos
Vacinas Anticâncer/uso terapêutico , Neoplasias Esofágicas/tratamento farmacológico , Neoplasias Esofágicas/imunologia , Linfócitos do Interstício Tumoral/imunologia , Linfócitos T Citotóxicos/imunologia , Idoso , Antígenos de Neoplasias/imunologia , Feminino , Antígeno HLA-A24/imunologia , Humanos , Linfócitos do Interstício Tumoral/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Linfócitos T Citotóxicos/efeitos dos fármacos , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/imunologia , Vacinas de Subunidades Antigênicas/uso terapêutico
2.
Epidemiol Infect ; 143(13): 2721-32, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25600435

RESUMO

A large outbreak of enterohaemorrhagic Escherichia coli (EHEC) O111 and O157 occurred in Japan in April 2011. We conducted an unmatched case-control study and trace-back investigation to determine the source of EHEC O111 infection and risk factors for severe complications. Pulsed-field gel electrophoresis was performed to help define cases. A total of 86 individuals met the case definition. Of these, 40% experienced haemolytic uraemic syndrome (HUS), 24% acute encephalopathy, and 6% died. Illness was significantly associated with eating the raw beef dish yukhoe (odds ratio 19·64, 95% confidence interval 7·03-54·83), the likely food vehicle. EHEC O111 and its closely related stx-negative variants were found in the beef. HUS occurred most frequently in individuals aged 5-9 years, and this age group was significantly associated with acute encephalopathy. The prevalence of HUS and acute encephalopathy was higher than in previous non-O157-related outbreaks, indicating a high risk of severe complications.


Assuntos
Encefalopatias/epidemiologia , Encefalopatias/microbiologia , Surtos de Doenças , Escherichia coli Êntero-Hemorrágica/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/microbiologia , Síndrome Hemolítico-Urêmica/epidemiologia , Síndrome Hemolítico-Urêmica/microbiologia , Carne/microbiologia , Doença Aguda , Adolescente , Adulto , Estudos de Casos e Controles , Criança , Pré-Escolar , Eletroforese em Gel de Campo Pulsado , Escherichia coli O157/isolamento & purificação , Feminino , Microbiologia de Alimentos , Humanos , Lactente , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco
3.
Clin Exp Dermatol ; 38(8): 897-903, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24252082

RESUMO

BACKGROUND: The Kv1.3 voltage-gated potassium channel is selectively upregulated upon activation in effector memory T (TEM ) cells in inflamed tissue, and plays an important role in maintenance of T-cell activation. Although Kv1.3 blockers have been shown to ameliorate allergic contact dermatitis (ACD) in a rat model, it remains unknown whether the effect of Kv1.3 blockers on ACD is mediated by suppressing TEM cell function and/or whether naive T-cells or central memory T (TCM ) cells are influenced. AIM: To analyse the detailed mechanism of Kv1.3 blockers in a rat model of ACD. METHODS: We examined the effects of a Kv1.3 blocker on inflammation and production of the effector cytokine interferon (IFN)-γ in inflamed tissue in rat ACD. Single-cell suspensions were isolated from inflamed rat ears (TEM cells), and regional lymph nodes (naive T/TCM cells), and the effect of Kv1.3 blockers on anti-CD3-stimulated IFN-γ production in vitro was measured. RESULTS: The Kv1.3 blocker significantly suppressed ear inflammation and IFN-γ production at the protein level in vivo. It also suppressed in vitro IFN-γ production from TEM cells from inflamed tissues, but did not suppress the function of naive T/TCM cells from lymph nodes. CONCLUSIONS: We found that the Kv1.3 blocker ameliorated ACD by inhibiting TEM cell functions only, thus Kv1.3 blockers could be a potentially selective therapeutic agent for TEM cell-mediated inflammatory skin diseases without producing harmful side-effects.


Assuntos
Dermatite Alérgica de Contato/tratamento farmacológico , Ficusina/farmacologia , Memória Imunológica/efeitos dos fármacos , Canal de Potássio Kv1.3/antagonistas & inibidores , Bloqueadores dos Canais de Potássio/farmacologia , Linfócitos T/efeitos dos fármacos , Animais , Células Cultivadas , Dermatite Alérgica de Contato/imunologia , Dermatite Alérgica de Contato/metabolismo , Modelos Animais de Doenças , Orelha , Feminino , Interferon gama/metabolismo , Canal de Potássio Kv1.3/fisiologia , Linfonodos/citologia , Ratos
4.
Phys Rev Lett ; 93(25): 257207, 2004 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-15697937

RESUMO

We report measurements of linear dichroism in x-ray absorption at Ti L(2,3) edges of a Mott-insulating ferromagnet YTiO3, where orbital ordering occurs in the triply degenerate Ti 3d t(2g) states. Dichroic spectra and their integrated intensities are obtained for the incident electric field with polarizations parallel to a, b, and c axes. The comparison of the spectra with atomic multiplet calculations removes the ambiguity about the orbital polarization, i.e., the relative weights of |xy>, |yz>, and |zx> orbits, which are crucial for the origin of ferromagnetism. The result is consistent with the previous analysis of nuclear magnetic resonance in the Mizokawa-Fujimori scheme.

5.
Virology ; 264(2): 422-6, 1999 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-10562503

RESUMO

Persistent infection of human immunodeficiency virus (HIV) takes place in the secondary lymphoid tissues even during clinically latent stages. The CC chemokines secondary lymphoid tissue chemokine (SLC) and EBI1-ligand chemokine (ELC) are constitutively expressed in the secondary lymphoid tissues. They share CCR7 expressed on lymphocytes and mature dendritic cells and play key roles in the trafficking of these types of cells into the secondary lymphoid tissues. Here we report that growth of both X4 and R5 strains of HIV-1 in activated peripheral blood T cells was enhanced by SLC. The enhancing effect of SLC was abrogated by pretreatment of cells with pertussis toxin, indicating the involvement of signaling via a receptor coupled with a Galphai class of G-protein. Furthermore, SLC was found to enhance the promoter activity of HIV-1 LTR. These results suggest that signaling via CCR7 has a strong positive effect on HIV growth. Thus, SLC and ELC may contribute to persistent infection of HIV in the secondary lymphoid tissues by promoting viral replication in activated T cells.


Assuntos
Quimiocinas CC/metabolismo , HIV-1/fisiologia , Replicação Viral , Adulto , Linhagem Celular , Células Cultivadas , Quimiocina CCL21 , Quimiocina CCL4 , Quimiocina CCL5/farmacologia , Quimiocinas CC/farmacologia , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP , HIV-1/efeitos dos fármacos , HIV-1/genética , HIV-1/crescimento & desenvolvimento , Humanos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/virologia , Tecido Linfoide , Proteínas Inflamatórias de Macrófagos/farmacologia , Toxina Pertussis , Receptores CCR7 , Receptores CXCR3 , Receptores de Quimiocinas/metabolismo , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/virologia , Fatores de Virulência de Bordetella/farmacologia
6.
Int Immunol ; 11(1): 81-8, 1999 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10050676

RESUMO

Helper T cells are classified into Th1 and Th2 subsets based on their profiles of cytokine production. Th1 cells are involved in cell-mediated immunity, whereas Th2 cells induce humoral responses. Selective recruitment of these two subsets depends on specific adhesion molecules and specific chemoattractants. Here, we demonstrate that the T cell-directed CC chemokine thymus and activation-regulated chemokine (TARC) was abundantly produced by monocytes treated with granulocyte macrophage colony stimulating factor (GM-CSF) or IL-3, especially in the presence of IL-4 and by dendritic cells derived from monocytes cultured with GM-CSF + IL-4. The receptor for TARC and another macrophage/dendritic cell-derived CC chemokine macrophage-derived chemokine (MDC) is CCR4, a G protein-coupled receptor. CCR4 was found to be expressed on approximately 20% of adult peripheral blood effector/memory CD4+ T cells. T cells attracted by TARC and MDC generated cell lines predominantly producing Th2-type cytokines, IL-4 and IL-5. Fractionated CCR4+ cells but not CCR4- cells also selectively gave rise to Th2-type cell lines. When naive CD4+ T cells from adult peripheral blood were polarized in vitro, Th2-type cells selectively expressed CCR4 and vigorously migrated toward TARC and MDC. Taken together, CCR4 is selectively expressed on Th2-type T cells and antigen-presenting cells may recruit Th2 cells expressing CCR4 by producing TARC and MDC in Th2-dominant conditions.


Assuntos
Células Apresentadoras de Antígenos/imunologia , Quimiocinas/imunologia , Quimiotaxia de Leucócito , Receptores de Quimiocinas/imunologia , Células Th2/imunologia , Timo/imunologia , Antígenos de Diferenciação de Linfócitos T , Linfócitos T CD4-Positivos , Quimiocina CCL17 , Quimiocina CCL22 , Quimiocinas CC/imunologia , Células Dendríticas/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Memória Imunológica , Interleucina-3/farmacologia , Interleucina-4/farmacologia , Antígenos Comuns de Leucócito , Macrófagos/imunologia , Monócitos/imunologia , Receptores CCR4 , Subpopulações de Linfócitos T
7.
Int Immunol ; 10(7): 901-10, 1998 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9701028

RESUMO

EBI1-ligand chemokine (ELC) is a CC chemokine constitutively expressed in various lymphoid tissues and a high-affinity functional ligand for EBI1/CCR7, a seven transmembrane G-protein-coupled receptor originally identified as an Epstein-Barr virus (EBV)-inducible gene. Here we examined chemotactic activity of ELC on peripheral blood leukocytes. ELC attracted both CD4+ and CD8+ T cells, particularly efficiently after activation with IL-2 or with phytohemagglutinin (PHA) plus IL-2, as well as CD19+ B cells, but not CD16+ NK cells, CD14+ monocytes or neutrophils. Among CD3+ T cells, ELC attracted both CD45RO- naive and CD45RO+ memory subsets. ELC also induced vigorous calcium mobilization in T cells stimulated with IL-2 with an ED50 of 3 nM. ELC fused with the secreted form of alkaline phosphatase (ELC-SEAP) specifically bound to lymphocytes and this binding was blocked only by ELC among 10 CC chemokines so far tested. Notably, lymphocytes stimulated with IL-2 or T cells expanded by PHA plus IL-2 showed much higher levels of binding than fresh lymphocytes. Consistently, CCR7 mRNA was detected in CD4+ and CD8+ T cells as well as B cells, but not in NK cells, monocytes or neutrophils, and was dramatically increased in T cells upon treatment with IL-2 or with PHA plus IL-2. Like ELC mRNA, CCR7 mRNA was expressed in various lymphoid tissues. By in situ hybridization, ELC and CCR7 mRNA were detected in the parafollicular and inner cortical regions of a lymph node, and in the parafollicular regions of an appendix. Collectively, ELC and CCR7 may be involved in the trafficking of a broad spectrum of lymphocytes, especially activated T cells, into and within various lymphoid tissues.


Assuntos
Quimiocinas CC/fisiologia , Quimiotaxia de Leucócito/fisiologia , Ativação Linfocitária/fisiologia , Receptores de Quimiocinas/fisiologia , Linfócitos T/fisiologia , Adulto , Movimento Celular/fisiologia , Quimiocina CCL19 , Fatores Quimiotáticos/biossíntese , Fatores Quimiotáticos/metabolismo , Fatores Quimiotáticos/fisiologia , Humanos , Hibridização In Situ , RNA Mensageiro/metabolismo , Receptores CCR7 , Receptores de Quimiocinas/biossíntese , Receptores de Quimiocinas/metabolismo , Linfócitos T/imunologia , Regulação para Cima/fisiologia
8.
Eur J Immunol ; 28(5): 1516-23, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9603456

RESUMO

Secondary lymphoid tissue chemokine (SLC) is a CC chemokine expressed mainly in lymph nodes, appendix and spleen, and specifically chemotactic for lymphocytes (Nagira et al., J. Biol. Chem. 1997. 272: 19518-19524). Here, we carried out transendothelial migration assays to determine the classes and subsets of lymphocytes migrating toward SLC. SLC attracted freshly isolated B cells with high efficiency and T cells modestly. Thus, SLC is the first CC chemokine with a strong chemotactic activity on fresh B cells. Among T cell types and subsets, SLC broadly attracted CD4+ and CD8+ cells, CD45RO- (naive) and CD45RO+ (memory) cells, and CD26high (activated) and CD26low- (resting) cells. SLC also attracted both L-selectin+ and L-selectin- subpopulations of various T cell subsets and B cells. Furthermore, mitogenic stimulation strongly enhanced migratory responses of T cells and B cells toward SLC. By in situ hybridization, SLC mRNA was detected in the cortical parafollicular regions (the T cell areas) of a lymph node and an appendix. Collectively, SLC may be a basic chemokine supporting homeostatic migration of a broad spectrum of lymphocytes into the secondary lymphoid tissues. SLC may also be involved in immune responses by inducing highly efficient migration of T and B cells following antigenic stimulation.


Assuntos
Subpopulações de Linfócitos B/imunologia , Quimiocinas CC/fisiologia , Quimiotaxia de Leucócito/imunologia , Ativação Linfocitária , Subpopulações de Linfócitos T/imunologia , Adulto , Quimiocina CCL21 , Quimiocinas CC/sangue , Quimiocinas CC/genética , Humanos , Ativação Linfocitária/efeitos dos fármacos , Tecido Linfoide/citologia , Tecido Linfoide/metabolismo , Fito-Hemaglutininas/farmacologia , RNA Mensageiro/biossíntese , RNA Mensageiro/metabolismo , Subpopulações de Linfócitos T/metabolismo
9.
J Biol Chem ; 273(12): 7118-22, 1998 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-9507024

RESUMO

Secondary Lymphoid-tissue Chemokine (SLC) is a recently identified CC chemokine that is constitutively expressed in various lymphoid tissues and is a potent and specific chemoattractant for lymphocytes. The SLC gene and the gene encoding another lymphocyte-specific CC chemokine, EBI1-ligand chemokine (ELC), form a mini-cluster at human chromosome 9p13. Here, we show that SLC is a high affinity functional ligand for chemokine receptor 7 (CCR7) that is expressed on T and B lymphocytes and a known receptor for ELC. SLC induced a vigorous calcium mobilization in murine L1.2 cells stably expressing human CCR7. SLC tagged with the secreted form of alkaline phosphatase (SLC-SEAP) showed specific binding to CCR7 that was fully competed by SLC with an IC50 of 0.5 nM. SLC also induced a vigorous chemotactic response in CCR7-expressing L1.2 cells with a typical bell-shaped dose-response curve and a maximal migration at 10 nM. When assessed using CCR7-transfected L1.2 cells, SLC and ELC were essentially equivalent in terms of cross desensitization in calcium mobilization via CCR7, cross-competition in binding to CCR7, and induction of chemotaxis via CCR7. SLC and ELC were also shown to fully share receptors expressed on cultured normal T cells known to express CCR7. Notably, however, SLC was somehow less efficient in cross-desensitization against ELC in calcium mobilization and in cross-competition with ELC for binding when assessed using cultured normal T cells. Thus, SLC and ELC, even though sharing only 32% amino acid identity, constitute a genetically and functionally highly related subgroup of CC chemokines.


Assuntos
Quimiocinas CC/metabolismo , Tecido Linfoide/metabolismo , Receptores de Quimiocinas/metabolismo , Sequência de Aminoácidos , Cálcio/metabolismo , Células Cultivadas , Quimiocina CCL19 , Quimiocina CCL21 , Quimiotaxia de Leucócito , Humanos , Ligantes , Dados de Sequência Molecular , Receptores CCR7 , Receptores de Quimiocinas/química , Receptores de Quimiocinas/genética , Alinhamento de Sequência , Linfócitos T/metabolismo , Transfecção
10.
Stud Health Technol Inform ; 52 Pt 1: 193-6, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-10384445

RESUMO

An integrated multimedia medical information network system at Shimane Medical university has been developed to organize medical information generated from each section and provide information services useful for education, research and clinical practice. The report describes the outline of our system. It is designed to serve as a distributed database for electronic medical records and images. We are developing the MML engine that is to be linked to the world wide web (WWW) network system. To the users, this system will present an integrated multimedia representation of the patient records, providing access to both the image and text-based data required for an effective clinical decision making and medical education.


Assuntos
Redes de Comunicação de Computadores , Sistemas Computadorizados de Registros Médicos/organização & administração , Multimídia , Centros Médicos Acadêmicos , Segurança Computacional , Sistemas de Informação Hospitalar/organização & administração , Humanos , Internet , Japão , Redes Locais , Integração de Sistemas
11.
Cell ; 91(4): 521-30, 1997 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-9390561

RESUMO

Leukocyte trafficking at the endothelium requires both cellular adhesion molecules and chemotactic factors. Fractalkine, a novel transmembrane molecule with a CX3C-motif chemokine domain atop a mucin stalk, induces both adhesion and migration of leukocytes. Here we identify a seven-transmembrane high-affinity receptor for fractalkine and show that it mediates both the adhesive and migratory functions of fractalkine. The receptor, now termed CX3CR1, requires pertussis toxin-sensitive G protein signaling to induce migration but not to support adhesion, which also occurs without other adhesion molecules but requires the architecture of a chemokine domain atop the mucin stalk. Natural killer cells predominantly express CX3CR1 and respond to fractalkine in both migration and adhesion. Thus, fractalkine and CX3CR1 represent new types of leukocyte trafficking regulators, performing both adhesive and chemotactic functions.


Assuntos
Adesão Celular/imunologia , Movimento Celular/imunologia , Quimiocinas CX3C , Quimiocinas/metabolismo , Leucócitos/citologia , Proteínas de Membrana/metabolismo , Receptores de Citocinas/metabolismo , Receptores de HIV/metabolismo , Antígenos CD/análise , Receptor 1 de Quimiocina CX3C , Membrana Celular/imunologia , Células Cultivadas , Quimiocina CX3CL1 , Endotélio Vascular/imunologia , Proteínas de Ligação ao GTP/metabolismo , Humanos , Leucemia Eritroblástica Aguda , Subpopulações de Linfócitos , RNA Mensageiro/análise , Receptores de Quimiocinas/metabolismo , Receptores de Citocinas/genética , Receptores de HIV/genética , Transdução de Sinais/imunologia , Células Tumorais Cultivadas , Veias Umbilicais
12.
J Biol Chem ; 272(31): 19518-24, 1997 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-9235955

RESUMO

By searching the Expressed Sequence Tag (EST) data base, we identified partial cDNA sequences potentially encoding a novel human CC chemokine. We determined the entire cDNA sequence which encodes a highly basic polypeptide of 134 amino acids total with a putative signal peptide of 23 amino acids. The predicted mature protein of 111 amino acids has the four canonical cysteine residues and shows 21-33% identity to other human CC chemokines, but has a unique carboxyl-terminal extension of about 30 amino acids which contains two extra cysteine residues. The mRNA was expressed strongly in tissues such as the lymph nodes, Appendix, and spleen. The recombinant protein, which was produced by the baculovirus system and purified to homogeneity, was a highly efficient chemoattractant for certain human T cell lines and a highly potent one for freshly isolated peripheral blood lymphocytes and cultured normal T cells expanded by phytohemagglutinin and interleukin 2. Unlike most other CC chemokines, however, this novel chemokine was not chemotactic for monocytes or neutrophils, suggesting that it is specific for lymphocytes. From these results, we designated this novel CC chemokine as SLC from secondary lymphoid-tissue chemokine. SLC fused with the secreted form of alkaline phosphatase (SLC-SEAP) was used to characterize the SLC receptor. Binding of SLC-SEAP to freshly isolated lymphocytes was blocked by SLC (IC50, 0.12 nM) but not by any other CC chemokine so far tested, suggesting that resting lymphocytes express a class of receptors highly specific for SLC. By using somatic cell hybrids, radiation hybrids, and selected yeast and bacterial artificial chromosome clones, we mapped the SLC gene (SCYA21) at chromosome 9p13 and between chromosomal markers, D9S1978(WI-8765) and AFM326vd1, where the gene for another novel CC chemokine termed ELC from EBI1-ligand chemokine (SCYA19) also exists. Collectively, SLC is a novel CC chemokine specific for lymphocytes and, together with ELC, constitutes a new group of chemokines localized at chromosome 9p13.


Assuntos
Quimiocinas CC/genética , Mapeamento Cromossômico , Cromossomos Humanos Par 9 , Linfócitos/imunologia , Sequência de Aminoácidos , Sequência de Bases , Cálcio/metabolismo , Linhagem Celular , Quimiocina CCL21 , Clonagem Molecular , Humanos , Dados de Sequência Molecular , Proteínas Recombinantes/biossíntese
13.
Cell Immunol ; 157(1): 144-57, 1994 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8039242

RESUMO

C33 Ag (CD82) is a member of the transmembrane 4 superfamily (TM4SF) and an activation Ag of T-cells. Recent studies have shown that CD82 associates with CD4 or CD8 and delivers costimulatory signals for the TCR/CD3 pathway. We have isolated cDNA and genomic clones of mouse CD82. Mouse CD82 has 266 amino acid residues with 76% identity to human CD82. The mouse CD82 gene consists of nine exons and spans more than 20 kb of genomic DNA. The genomic organization of CD82 is quite similar to that of three other TM4SF members whose genomic structures were described, i.e., Tapa-1 (CD81), CD53, and CD63. By mapping the 5' end of CD82 transcripts, we found a single major transcription initiation site 144 bp upstream of the ATG initiation codon. We also determined the sequence of the 5' flanking region of CD82 gene for about 2 kb. The 5' flanking sequence has a housekeeping promoter with potential binding motifs for various transcriptional factors. Northern blot analysis showed quite variable expression of the CD82 gene among different organs. The highest expression was seen in the spleen and the kidney. The expression was low in skeletal muscle and hardly detectable in the heart. Northern blot analysis was also carried out for CD81, CD53, and CD63. The expression of the CD81 gene was ubiquitous and similar among different organs, while that of CD53 was seen only in the spleen. The expression of the CD63 gene was ubiquitous, with the highest expression in the kidney. These results together with the comparison of the structures of 5' flanking sequences of these genes indicate distinct regulations of gene expression for these four members of TM4SF.


Assuntos
Antígenos CD , Glicoproteínas de Membrana/genética , Família Multigênica , Proteínas Proto-Oncogênicas , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Southern Blotting , Cloranfenicol O-Acetiltransferase/genética , DNA Complementar/química , Éxons , Expressão Gênica , Biblioteca Genômica , Células HeLa , Humanos , Íntrons , Proteína Kangai-1 , Células L , Glicoproteínas de Membrana/biossíntese , Camundongos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Homologia de Sequência de Aminoácidos , Transcrição Gênica
14.
J Immunol ; 149(9): 2879-86, 1992 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-1401919

RESUMO

C33 Ag was originally identified by mAb inhibitory to syncytium formation induced by human T cell leukemia virus type 1. The Ag was shown to be a highly heterogeneous glycoprotein consisting of a 28-kDa protein and N-linked oligosaccharides ranging from 10 to 50 kDa. In the present study, cDNA clones were isolated from a human T cell cDNA expression library in Escherichia coli by using mAb C33. The identity of cDNA was verified by immunostaining and immunoprecipitation of transfected NIH3T3 cells with mAb. The cDNA contained an open reading frame of a 267-amino acid sequence which was a type III integral membrane protein of 29.6 kDa with four putative transmembrane domains and three putative N-glycosylation sites. The C33 gene was found to belong to a newly defined family of genes for membrane proteins, such as CD9, CD37, CD53, CD63, and TAPA-1, and was identical to R2, a cDNA recently isolated because of its strong up-regulation after T cell activation. Availability of mAb for C33 Ag enabled us to define its distribution in human leukocytes. C33 Ag was expressed in CD4+ T cells, CD19+ B cells, CD14+ monocytes, and CD16+ granulocytes. Its expression was low in CD8+ T cells and mostly negative in CD16+ NK cells. PHA stimulation enhanced the expression of C33 Ag in CD4+ T cells by about 5-fold and in CD8+ T cells by about 20-fold. PHA stimulation also induced the dramatic size changes in the N-linked sugars previously shown to accompany human T cell leukemia virus type 1-induced transformation of CD4+ T cells.


Assuntos
Antígenos de Diferenciação/imunologia , Antígenos de Superfície/imunologia , Células Gigantes/imunologia , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Glicoproteínas de Membrana , Proteínas Proto-Oncogênicas , Sequência de Aminoácidos , Anticorpos Monoclonais , Antígenos CD/imunologia , Antígenos de Diferenciação/genética , Antígenos de Superfície/biossíntese , Sequência de Bases , Northern Blotting , Linhagem Celular , Clonagem Molecular , Citometria de Fluxo , Biblioteca Gênica , Glicosilação , Humanos , Proteína Kangai-1 , Ativação Linfocitária/imunologia , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , RNA Mensageiro/análise , Linfócitos T/imunologia , Acetato de Tetradecanoilforbol/farmacologia , Transfecção , Regulação para Cima
15.
Int J Cancer ; 52(3): 418-27, 1992 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-1356935

RESUMO

Sixteen human T-cell lines were studied for the expression of a cell-adhesion molecule ICAM-1 and its counter-receptor LFA-1. The cell lines included 3 human T-cell-leukemia-virus-type-I (HTLV-1)-negative cell lines derived from acute lymphoblastic leukemia (ALL) and 13 HTLV-1-positive cell lines, 7 of them established from cord- or peripheral-blood T cells by in vitro transformation with HTLV-1, 2 derived from HTLV-1 carriers, and 4 derived from patients with adult T-cell leukemia (ATL). In sharp contrast to a basal level of ICAM-1 in 3 HTLV-1-negative ALL cell lines, strong induction of ICAM-1 was seen in all HTLV-1-positive T-cell lines except for MT-1, one of the 4 ATL cell lines used in the present study. On the other hand, the expression of LFA-1 (CD11a and CD18) was more or less similar among the cell lines with and without HTLV-1. Interestingly, however, 3 out of 4 ATL cell lines (TL-Om1, H582, HUT102) revealed striking depression of LFA-1 expression. Several lines of evidence strongly argued against direct involvement of the viral transactivator p40tax or some autocrine cytokines in the induction of ICAM-1 in HTLV-1-positive T-cell lines. It was also found that ICAM-1 and LFA-1 were involved in syncytium formation induced in the co-culture of HTLV-1-positive and HTLV-1-negative human T-cell lines. Implications of constitutive expression of ICAM-1 for certain clinical manifestations of ATL and of depression of either ICAM-1 or LFA-1 during progression of ATL are discussed.


Assuntos
Moléculas de Adesão Celular/biossíntese , Vírus Linfotrópico T Tipo 1 Humano/patogenicidade , Leucemia de Células T/metabolismo , Antígeno-1 Associado à Função Linfocitária/análise , Linfócitos T/metabolismo , Animais , Anticorpos Monoclonais/imunologia , Sequência de Bases , Northern Blotting , Moléculas de Adesão Celular/genética , Transformação Celular Viral , Citocinas/fisiologia , Citometria de Fluxo , Produtos do Gene tax/fisiologia , Humanos , Molécula 1 de Adesão Intercelular , Leucemia de Células T/patologia , Camundongos , Dados de Sequência Molecular , RNA Mensageiro/análise , Células Tumorais Cultivadas
16.
J Pharmacobiodyn ; 15(8): 455-9, 1992 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-1479546

RESUMO

MH61 is an acrosome-reacted sperm-specific monoclonal antibody. The antibody-coated beads effectively analyze the acrosomal status of human sperm. However, the nature of the antigen was not known. The antigen was purified by immunoprecipitation and SDS-PAGE followed by blotting on PVDF paper. The N-terminal sequence of the antigen was analyzed by an automated protein sequencer. An exactly matching sequence was found in CD46, that is also known as a membrane cofactor protein.


Assuntos
Antígenos CD/química , Antígenos de Superfície/química , Glicoproteínas de Membrana/química , Espermatozoides/imunologia , Acrossomo/fisiologia , Sequência de Aminoácidos , Anticorpos Monoclonais , Antígenos de Superfície/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Humanos , Masculino , Proteína Cofatora de Membrana , Dados de Sequência Molecular , Testes de Precipitina , Sêmen , Homologia de Sequência de Aminoácidos
17.
Mol Reprod Dev ; 32(4): 389-93, 1992 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-1497887

RESUMO

A monoclonal antibody (MAb) against human acrosome-reacted sperm was attached to paramagnetic polystyrene beads. Human sperm prepared by the swim-up method were 1) incubated in m-BWW, 2) incubated and ionophore treated, or 3) incubated with 5% seminal fluid. After treatment, sperm were mixed with the beads and incubated for 1 hr. Variously treated sperm showed different binding abilities to the beads. Sperm bound to the beads were collected by a magnet and subjected to triple staining. Most of the collected sperm were acrosome reacted. The results suggested that the beads can be used to estimate the acrosomal status of sperm, and that the use of antibody-coated paramagnetic beads provides a convenient way of collecting acrosome-reacted sperm. The acrosomal status detected by the beads was also compared with the ability of sperm to fuse with zona-free hamster eggs. It was found that greater bead-binding ability correlated with more sperm fusing with zona-free hamster eggs.


Assuntos
Acrossomo/metabolismo , Separação Celular/métodos , Espermatozoides/metabolismo , Acrossomo/imunologia , Acrossomo/ultraestrutura , Animais , Anticorpos Monoclonais/imunologia , Calcimicina/farmacologia , Cricetinae , Feminino , Humanos , Magnetismo , Masculino , Microesferas , Poliestirenos , Capacitação Espermática , Interações Espermatozoide-Óvulo , Espermatozoides/efeitos dos fármacos , Espermatozoides/ultraestrutura
18.
Fertil Steril ; 54(6): 1121-6, 1990 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-1700957

RESUMO

A monoclonal antibody to human sperm (MH61) was established. The antibody did not attach to ejaculated sperm but to a head region of some sperm that was incubated in medium for 4 to 12 hours. A23187 treatment significantly increased the number of sperm that were reactive to the antibody. When sperm that bound to the zona-free hamster egg were subjected to the indirect immunofluorescence staining, all the sperm reacted to MH61 with their entire head region. The addition of the antibody to the medium before sperm addition reduced the fusion rate of human sperm to zona-free hamster egg.


Assuntos
Antígenos/imunologia , Óvulo/imunologia , Interações Espermatozoide-Óvulo , Espermatozoides/imunologia , Acrossomo/fisiologia , Animais , Anticorpos Monoclonais , Calcimicina/farmacologia , Cricetinae , Eletroforese em Gel de Poliacrilamida , Feminino , Imunofluorescência , Humanos , Masculino , Cabeça do Espermatozoide/imunologia , Espermatozoides/efeitos dos fármacos , Coloração e Rotulagem , Zona Pelúcida
19.
Experientia ; 45(2): 193-4, 1989 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-2920807

RESUMO

In order to study the sperm-egg recognition mechanism on the surface of the plasma membrane, zonae were removed from mouse eggs by exposure to acidic conditions. Sperm binding to denuded eggs was then observed in the presence of various sugars. Among several carbohydrates tested, only glucosamine (GlcN) was found to increase the number of sperm bound to eggs while inhibiting sperm-egg fusion. The inhibition was reversible; when denuded eggs were transferred to a GlcN free medium, a high rate of polyspermy was observed.


Assuntos
Glucosamina/farmacologia , Interações Espermatozoide-Óvulo/efeitos dos fármacos , Animais , Carboidratos/farmacologia , Membrana Celular/fisiologia , Feminino , Masculino , Camundongos , Óvulo/fisiologia , Zona Pelúcida/fisiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...