RESUMO
BACKGROUND: The purpose of this study was to investigate the usefulness of a hydrocolloid dressing containing ceramide for hand-foot skin reaction (HFSR) on the soles of the feet in metastatic renal cell carcinoma (RCC) patients treated with sorafenib. PATIENTS AND METHODS: Patients with grade 1 HFSR on the soles of the feet were randomly assigned in to two groups. One group received a hydrocolloid dressing containing ceramide (arm A) and the other received 10% urea cream (arm B). Patients in both groups applied treatment to the affected sites on the soles of the feet, but not to the hands. The primary end point was the incidence of grade 2 or 3 HFSR on the soles of the feet in the first 4 weeks. RESULTS: Thirty-three patients were assessed (17 in arm A and 16 in arm B), and there were no significant differences in baseline characteristics between the two groups. During the observation period of this study, grade 2 or 3 HFSR on the soles of the feet was found in 29% of patients in arm A and was significantly less than the 69% in arm B (P=0.03). The incidence of HFSR on the hands, however, was similar in both arms. The median time to grade 2 or 3 HFSR on the soles of the feet was also significantly longer in arm A than in arm B (P=0.03). CONCLUSIONS: These results indicate that a hydrocolloid dressing containing ceramide prevented the worsening of HFSR caused by sorafenib in metastatic RCC patients. CLINICAL TRIAL REGISTRATION NUMBER: UMIN000002016.
Assuntos
Antineoplásicos/efeitos adversos , Carcinoma de Células Renais/tratamento farmacológico , Ceramidas/administração & dosagem , Síndrome Mão-Pé/terapia , Neoplasias Renais/tratamento farmacológico , Niacinamida/análogos & derivados , Compostos de Fenilureia/efeitos adversos , Idoso , Antineoplásicos/uso terapêutico , Curativos Hidrocoloides , Carcinoma de Células Renais/secundário , Feminino , Humanos , Estimativa de Kaplan-Meier , Neoplasias Renais/patologia , Masculino , Pessoa de Meia-Idade , Niacinamida/efeitos adversos , Niacinamida/uso terapêutico , Compostos de Fenilureia/uso terapêutico , Sorafenibe , Propriedades de Superfície , Resultado do TratamentoRESUMO
At 4 wk after intraperitoneal inoculation of murine cytomegalovirus (MCMV) in adult BALB/c mice, MCMV remained detectable only in the salivary glands. When T cells of these mice were activated by a single injection of anti-CD3 epsilon monoclonal antibody, mice died of interstitial pneumonitis at 24-48 h after injection, accompanied by elevation of serum levels of TNF-alpha and IFN-gamma. However, MCMV remained undetectable in the lungs during the period. Simultaneous injection of cyclosporin A reduced such effects of anti-CD3. In conclusion, although the presence of MCMV in the host may be required, MCMV-associated pneumonitis is not mediated by virus in the lung but probably by the cytokines released from T cells, of which responsiveness to stimulation via CD3 molecule has been presumably modified by MCMV infection.
Assuntos
Infecções por Citomegalovirus/patologia , Citomegalovirus/isolamento & purificação , DNA Viral/análise , Pulmão/patologia , Pneumonia Viral/microbiologia , Reação em Cadeia da Polimerase/métodos , Animais , Anticorpos Monoclonais/farmacologia , Sequência de Bases , Complexo CD3/imunologia , Ciclosporina/farmacologia , Citomegalovirus/genética , Citomegalovirus/patogenicidade , Infecções por Citomegalovirus/sangue , Infecções por Citomegalovirus/microbiologia , Primers do DNA , Interferon gama/sangue , Pulmão/microbiologia , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Pneumonia Viral/sangue , Pneumonia Viral/patologia , Glândulas Salivares/microbiologia , Glândulas Salivares/patologia , Baço/patologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Timo/patologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Patients with infective endocarditis (IE) often have renal complications which may include infarcts, abscesses and glomerulonephritis (GN). Furthermore, it is generally accepted that there is an association between IE and anti-neutrophil cytoplasmic antibody (ANCA). Here, we report the case of a 24-year-old man who developed rapidly progressive GN in the course of IE due to infection with alpha-streptococcus. The initial clinical manifestation of the condition was severe sacroiliitis without fever. Sandwich ELISA showed that the patient was positive for PR3-ANCA at low titer, and the classical complement pathway was also activated. Renal biopsy demonstrated several lesions: focal embolic GN, GN with immune deposits and focal and segmental crescentic necrotizing GN. Treatment with antibiotics and steroids led to eradication of the infection, and resolution of the renal disease was accompanied by immediate disappearance of PR3-ANCA and hypocomplementemia. During a 4-year follow-up period, no recurrence was observed. There have only been 7 case reports of GN associated with IE and PR3-ANCA in which the renal pathology has been described, and the current report is the first to document renal pathology in a patient with isolated pulmonic valve IE and PR3-ANCA. Moreover, this report is the first to show a change in renal biopsy findings in response to treatment. A review of the 7 literature cases and that of our patient showed that none involved pauci-immune GN. Hence, further studies are needed to clarify the prevalence of pauci-immune GN in ANCA-positive IE patients.
Assuntos
Anticorpos Anticitoplasma de Neutrófilos/metabolismo , Endocardite Bacteriana/complicações , Glomerulonefrite/complicações , Rim/patologia , Mieloblastina/metabolismo , Valva Pulmonar/microbiologia , Adulto , Biópsia , Progressão da Doença , Ecocardiografia Transesofagiana , Endocardite Bacteriana/diagnóstico por imagem , Humanos , Rim/ultraestrutura , Masculino , Metanálise como AssuntoRESUMO
Compared to an untransformed rat cell line, 3Y1, adenovirus 12-transformed cell lines of 3Y1 were highly sensitive to the cytotoxic action of an imidazole antibiotic, clotrimazole. In contrast, SV40-transformed rat cell lines derived from the 3Y1 cell line showed no appreciable difference in the response to clotrimazole when compared to 3Y1 cells. Clotrimazole-resistant clones, WCT-1 and WCT-2, which were spontaneously isolated from the adenovirus 12-transformed cell line W5, showed 5- to 10-fold higher resistance than did W5; the dose-response curves of clotrimazole-resistant clones were similar to those of the untransformed 3Y1 cells. The growth of 3Y1 cells was blocked in the presence of 1% serum, whereas those of W5, WCT-1, and WCT-2 cells were only slightly affected by the same dose of serum in the medium. The membrane fractions of 3Y1, W5, and WCT-1 cells were found to contain similar cholesterol:phospholipid ratios. The phospholipid composition in the membrane fraction of line WCT-1 was similar to that of line W5 but not to that of line 3Y1. By contrast, the fatty acid composition was specifically altered in clotrimazole-resistant clones; cellular contents and some species of fatty acid such as 16:1, 18:2, and 20:4 in WCT-1 and WCT-2 cells were similar to those of 3Y1 but not to those of W5 cells. Differential sensitivities of various cell lines to clotrimazole are discussed in relation to the lipid composition.
Assuntos
Transformação Celular Viral , Clotrimazol/toxicidade , Imidazóis/toxicidade , Animais , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Membrana Celular/análise , Células Cultivadas , Cromatografia Gasosa , Resistência a Medicamentos , Ácidos Graxos/análise , Cinética , Lipídeos de Membrana/análise , Camundongos , Camundongos Endogâmicos BALB C , Fenótipo , Fosfolipídeos/análise , RatosRESUMO
The imidazole antimycotics clotrimazole and miconazole were tested on untransformed rat cell line 3Y1-B clone 1-6 (3Y1) and six transformed cell lines, which were independently isolated from 3Y1 or 3Y1-B clone 1 after infection with adenovirus 12 (AD-12) or with SV40, to determine their sensitivities to these drugs. The relative plating efficiency of three cell lines (T3, W4, and W5) transformed with AD-12 was reduced to 10(-1( of the initial value by clotrimazole (2 to 4 microgram/ml), whereas that of the parental cell line 3Y1 was reduced to 10(-1) of the initial value by clotrimazole (20 to 25 microgram/ml). By contrast, the differential effect of miconazole on 3Y1 and AD-12-transformed cell lines was found to be a factor of 2. The sensitivity of the SV40-transformed 3Y1 and the AD-12-transformed cell lines. The cellular sensitivity of untransformed 3Y1 cells to clotrimazole was significantly enhanced when exposed to various doses of the unsaturated fatty acid, linoleic acid. The untransformed and transformed cell lines showed sensitivities similar to the cytocidal activity of sterol-binding antimycotics, amphotericin B and filipin.
Assuntos
Transformação Celular Neoplásica/efeitos dos fármacos , Clotrimazol/farmacologia , Imidazóis/farmacologia , Lipídeos de Membrana/metabolismo , Anfotericina B/farmacologia , Animais , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Transformação Celular Neoplásica/patologia , Filipina/farmacologia , Miconazol/farmacologia , RatosRESUMO
The colony-forming ability of rat 3Y1 fibroblasts transformed by adenovirus type 12 (Ad12) was drastically reduced when the cells were cultivated for 18 h in medium augmented with 300 micrograms/ml of liposomes composed of either phosphatidylcholine (PC) or phosphatidylinositol. In contrast, those of untransformed 3Y1 cells and simian virus 40-transformed and polyomavirus-transformed 3Y1 cells were not. The cytotoxicity of PC liposomes was also observed in 3Y1 cells transformed by plasmid DNA containing Ad12-E1A gene but not in those transformed by adenovirus type 2, Rous avian sarcoma virus, or plasmid DNA carrying v-Ha-ras oncogene. The extensive killing of Ad12-transformed and E1A-transformed 3Y1 cells occurred in liposomes of dioleoyl-PC and of dilinoleoyl PC but not those of dipalmitoyl PC, distearoyl-PC, or diarachidonyl PC, suggesting that the acyl groups of phospholipids play an important role in cytotoxicity. Dilinoleoylglycerol, 60 micrograms/ml, was also cytotoxic selectively to Ad12-transformed and E1A-transformed 3Y1 cells, although the toxicity of lysophosphatidylcholine or linoleic acid was not specific to these transformants. These results suggest that cell transformation by Ad12 is characterized by a high sensitivity to exogenously administered phospholipids and diacylglycerol that contain oleoyl or linoleoyl acyl groups and that the sensitivity is attributable to the expression of E1A gene of Ad12.
Assuntos
Divisão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Transformação Celular Viral , Diglicerídeos/toxicidade , Glicerídeos/toxicidade , Fosfolipídeos/toxicidade , Proteínas Precoces de Adenovirus , Adenovírus Humanos , Ácidos Graxos não Esterificados/toxicidade , Lipossomos , Lisofosfatidilcolinas/toxicidade , Proteínas Oncogênicas Virais , Fosfatidilcolinas/toxicidade , Relação Estrutura-AtividadeRESUMO
Previous studies demonstrated that injecting mice with the cytokine interleukin 12 (IL-12) could significantly suppress the growth of a number of tumors, including murine B16 melanoma. In this report, the persistence of the antitumor effects of IL-12 is investigated. The i.p. injection of IL-12 (0.1 microg) on days 14, 16, 18, 20, and 22 was found to significantly suppress the growth of s.c. inoculated B16 melanoma for up to 2 weeks after the last injection of IL-12. Interestingly, the IL-12 serum level 4 days after the last injection of IL-12 was significantly elevated in tumor-bearing mice compared with that of IL-12-treated normal mice. The in vivo depletion of either CD4+ or CD8+ T cells abrogated the antitumor activity of IL-12 and diminished the apparent autocrine stimulation of IL-12 release seen after IL-12 treatment. Resection of the tumor-draining lymph nodes (LNs) but not of the spleen abrogated the antitumor effect of IL-12 treatment as well as the elevation of serum IL-12. Expression of mRNA encoding IL-12 as well as CD40 ligand (CD40L) was detected in the tumor-draining LNs but not in the spleen of tumor-bearing mice after IL-12 treatment. Furthermore, the antitumor activity observed after IL-12 treatment was diminished by the in vivo administration of either anti-IL-12 or anti-CD40L monoclonal antibodies. Collectively, these results suggest that the endogenous production of IL-12 resulting from the CD40-CD40L interaction between antigen-presenting cells and CD4+ T cells in the tumor-draining LNs may play a role in the persistence of the antitumor effects seen after IL-12 treatment.
Assuntos
Imunoterapia , Interleucina-12/administração & dosagem , Melanoma Experimental/terapia , Animais , Linfócitos T CD4-Positivos/imunologia , Antígenos CD40/imunologia , Antígenos CD40/metabolismo , Linfócitos T CD8-Positivos/imunologia , Feminino , Interleucina-12/biossíntese , Interleucina-12/sangue , Interleucina-12/imunologia , Melanoma Experimental/sangue , Melanoma Experimental/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Células Tumorais CultivadasRESUMO
Rat 3Y1 fibroblasts transformed by the E1A gene of adenovirus type 12 (E1A-3Y1 cells) are highly sensitive to the cell-killing effect of 1,3-dilinoleoylglycerol (DLG) administered in a culture medium, whereas the parental 3Y1 cells are less sensitive (H. Shimura et al., Cancer Res., 48: 578-583, 1988). The selective cytotoxicity of DLG to E1A-3Y1 cells was markedly reduced by the simultaneous administration of nonspecific antioxidants such as vitamin E, butylated hydroxytoluene, and ascorbic acid. Specific scavengers for oxygen radicals had no effect. Lipoxygenase inhibitors (nordihydroguaiaretic acid, esculetin, and baicalein) reduced the DLG-mediated selective cytotoxicity, whereas cyclooxygenase inhibitors (acetylsalicylic acid and indomethacin) showed no effect. The intracellular and extracellular contents of the products from lipid peroxidation as measured by the thiobarbituric acid test were significantly greater in E1A-3Y1 cells than in the parental 3Y1 cells. In comparison with DLG, linoleic acid and monolinoleoylglycerol were equally toxic to E1A-3Y1 and parental 3Y1, and trilinoleoylglycerol was weakly toxic to both types of cells. Scanning electron microscopy revealed that numerous holes about 0.2 micron in diameter were scattered all over the surface of the E1A-3Y1 cells after treating the cultures with DLG. These results suggest that; (a) the DLG-mediated cytotoxicity to the E1A-transformed cells is attributable to lipid peroxidation; (b) the structural property of DLG is essential to the E1A specificity of cytotoxicity; and finally (c) the destruction of the cell membrane is the basis of cytotoxicity of DLG.
Assuntos
Antineoplásicos , Transformação Celular Neoplásica/patologia , Diglicerídeos/toxicidade , Glicerídeos/toxicidade , Proteínas Precoces de Adenovirus , Animais , Antioxidantes/farmacologia , Divisão Celular/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Diglicerídeos/farmacologia , Ácido Linoleico , Ácidos Linoleicos/farmacologia , Peróxidos Lipídicos/metabolismo , Inibidores de Lipoxigenase , Lipídeos de Membrana/metabolismo , Microscopia Eletrônica de Varredura , Proteínas Oncogênicas Virais/genética , Ratos , Células Tumorais CultivadasRESUMO
The purpose of this study was to elucidate a possible immune response to tumor cells mediated by tumor-infiltrating lymphocytes (TIL) in lung cancer. In flow cytometry, the majority of T-cells of TIL were CD45RA-, CD45RO+, and CDw29high, and expressed HLA-DR. The expression of interleukin 2 receptor beta chain increased in both CD4+ and CD8+ TIL compared with both types of T-cells in peripheral blood. These results indicate that the major population of TIL is activated memory T-cells. The TIL preparation, which was usually contaminated with 5 to 10% tumor cells, did not exhibit any response in autologous mixed lymphocyte-tumor culture even in the presence of interleukin 2 (IL-2) in all five cases tested. Although purified T-cells from TIL showed the positive response in only 1 of 10 cases tested without addition of IL-2, it occurred in 7 of 10 cases in the addition of a low concentration of IL-2. The IL-2-dependent response to irradiated autologous tumor cells was suppressed when nonirradiated autologous tumor cells were added to the culture. Culture supernatants of four lung cancer cell lines and freshly prepared lung cancer cells obtained from 6 cases exhibited suppressive activity against anti-CD3 antibody-induced mitogenesis of peripheral blood mononuclear cells from healthy donors. We suggest that, taken together, (a) the major population of TIL in lung cancer are activated memory T-cells, and they include tumor-reactive ones, and that (b) the function of the TIL is impaired by unavailability of IL-2 and/or by suppression due to lung cancer cell-derived factor(s).
Assuntos
Neoplasias Pulmonares/imunologia , Linfócitos do Interstício Tumoral/imunologia , Linfócitos T/imunologia , Antígenos CD/análise , Antígenos CD4/análise , Antígenos CD8/análise , Carcinoma/imunologia , Carcinoma/patologia , Carcinoma/cirurgia , Linhagem Celular , Separação Celular , Citometria de Fluxo , Fluoresceína-5-Isotiocianato , Antígenos HLA-DR/análise , Humanos , Interleucina-2/farmacologia , Pulmão/imunologia , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/cirurgia , Ativação Linfocitária , Linfócitos do Interstício Tumoral/efeitos dos fármacos , Fenótipo , Receptores de Interleucina-2/análise , Proteínas Recombinantes/farmacologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/patologia , Células Tumorais CultivadasRESUMO
In this study, we investigated the cytolytic activity of peripheral blood T-cells (PBT) obtained from nine patients with primary lung cancer treated by surgical adjuvant adoptive immunotherapy (AIT) with lymphokine-activated killer cells and low-dose recombinant interleukin 2 at the time of rebound lymphocytosis (24-48 h after AIT). In eight of nine patients, nonspecific cytotoxicity of peripheral blood lymphocytes significantly increased as compared with that of pre-AIT peripheral blood lymphocytes. However, purified PBT showed much less activity to kill tumor cells although they increased in number and were activated well in terms of increases in the expression of HLA-DR and interleukin 2 receptor. The cytolytic activity of post-AIT PBT was significantly enhanced when they were targeted to Fc receptor-bearing tumor cells (K562 or Daudi) with anti-CD3 (NU-T3) or anti-T-cell receptor (TCR)alpha beta (WT31) monoclonal antibody in all five patients examined. Phenotypically, the targeted cytotoxicity was predominantly mediated by CD8+ cells. The results indicated that in vivo-activated PBT by AIT could not exhibit direct cytotoxicity, but they acquired cytolytic potential, the effect of which was expressed by targeting to tumor cells.
Assuntos
Citotoxicidade Imunológica , Imunoterapia Adotiva , Interleucina-2/uso terapêutico , Células Matadoras Ativadas por Linfocina/imunologia , Glicoproteínas de Membrana , Neoplasias/terapia , Linfócitos T/imunologia , Idoso , Feminino , Humanos , Ativação Linfocitária , Masculino , Proteínas de Membrana/genética , Pessoa de Meia-Idade , Neoplasias/imunologia , Perforina , Fenótipo , Proteínas Citotóxicas Formadoras de PorosRESUMO
We studied the antitumor immune response in gut-associated lymphoid tissue (GALT), which is the tolerance-inducing site for numerous dietary antigens. The mice inoculated with colon 26 carcinoma (C-26) into the subserosal space of the cecum (i.c.) showed a more rapid tumor growth than did the mice inoculated s.c. with C-26 into the flank. In addition, the serum of the i.c. C-26-inoculated mice showed a more potent suppressive activity, and their plasma contained a higher level of transforming growth factor than the s.c. C-26-inoculated mice. We also evaluated the tumor-specific T-cell response in the GALT by utilizing B7-transfected P815 mastocytoma (B7/P815). The rejection of i.c. inoculated B7/P815 was delayed compared to that of the s.c. inoculated B7/P815. The draining axillary lymph node (LN) cells of the s.c. B7/P815-inoculated mice exhibited a CD4+ T-cell-dependent proliferative response to in vitro restimulation, whereas the draining mesenteric LN cells of the i.c. B7/P815-inoculated mice exhibited no apparent response even with the addition of interleukin 2. However, such draining mesenteric LN cells did produce higher levels of interleukin 2 and transforming growth factor beta than the draining axillary LN without any stimulation, and their production of such cytokines depend on the CD4+ and CD8+ cells, respectively. Collectively, our results suggest the possibility that the impaired antitumor T-cell response in the GALT may be attributed to "bystander suppression" by TGF-beta-producing CD8+ T cells.
Assuntos
Carcinoma/imunologia , Neoplasias do Colo/imunologia , Intestinos/imunologia , Linfócitos T/imunologia , Fator de Crescimento Transformador beta/fisiologia , Animais , Carcinoma/patologia , Endotoxinas/sangue , Feminino , Tolerância Imunológica , Imunidade Celular , Imunidade nas Mucosas , Linfonodos/patologia , Ativação Linfocitária , Mesentério/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Albumina Sérica/metabolismo , Baço/patologia , TransfecçãoRESUMO
p56lck, a member of the src family of non-receptor protein tyrosine kinases (PTKs), is expressed predominantly in T-lymphocytes. Association of p56lck with CD4 and CD8 T-cell receptor (TcR) accessory molecules suggests that p56lck may play a specialized role in antigen-induced T-cell activation. CD4 and CD8 molecules are known to stabilize the interaction between TcR and the major histocompatibility complex during T-cell activation. To examine the role of p56lck in the dynamics of the CD4 molecule, p56lck-expressing transfectant cell clones were prepared by the transfection of an lck-gene plasmid containing an inducible promoter into a CD4+lck- human monocytoid cell line. When these transfectant cells were stimulated with phorbol ester, CD4 internalization on these p56lck-expressing cell lines was selectively and markedly retarded, as compared to p56lck-negative control cell lines. When cell-surface CD4 and intracellular CD4 were selectively precipitated after stimulation, the intracellular CD4 molecules were dissociated from p56lck whereas the surface-retained CD4 molecules were still associated with p56lck. Moreover, the dissociation of p56lck from CD4 appeared to occur prior to the PMA-induced internalization of CD4. These data indicate that p56lck regulates the PMA-induced internalization of CD4 possibly via its association with CD4. Treatment with genistein, a PTK inhibitor, revealed that the PTK activity of p56lck might not be involved in this regulatory effect of p56lck on CD4 internalization.
Assuntos
Antígenos CD4/metabolismo , Linfócitos/metabolismo , Proteínas Tirosina Quinases/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Western Blotting , Linhagem Celular , Eletroforese em Gel de Poliacrilamida , Expressão Gênica , Genisteína , Humanos , Isoflavonas/farmacologia , Proteína Tirosina Quinase p56(lck) Linfócito-Específica , Linfócitos/efeitos dos fármacos , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/genética , RNA/metabolismo , TransfecçãoRESUMO
Osmotic water movement across the toad urinary bladder in response to both vasopressin and cyclic AMP was inhibited by 10(-5) to 10(-4) M colchicine on the serosal but not on the mucosal side. This inhibitory effect was found to be time- and dose-dependent. Colchicine alone did not change basal osmotic flow and a baseline of the short-circuit current (Isc) and also did not affect a vasopressin-induced rise of the Isc. The inhibitory effect was not prevented by the addition of pyruvate. The osmotic water movement produced by 360 mM Urea (mucosal), 360 mM mannitol (serosal) or 2 mug/ml amphotericin B (mucosal), was not affected by 10(-4) M colchicine. These results suggest that colchicine inhibits some biological process subsequent to the formation of cyclic AMP except a directional cytoplasmic streaming process where microtubules may be involved.
Assuntos
Água Corporal/metabolismo , Colchicina/farmacologia , Bexiga Urinária/metabolismo , Anfotericina B/farmacologia , Animais , Bufo bufo , AMP Cíclico/farmacologia , Humanos , Manitol/farmacologia , Concentração Osmolar , Pressão Osmótica , Ureia/farmacologia , Bexiga Urinária/efeitos dos fármacos , Vasopressinas/farmacologiaRESUMO
Engagement of T-cell receptor (TcR)/CD3 complexes on T-cells rapidly provokes tyrosine phosphorylation of cellular proteins, which is thought to be an essential step to the following events of T-cell activation. p56lck, a member of src-related, non-receptor type protein tyrosine kinases, is expressed predominantly in lymphocytes. Accumulating data suggest that p56lck is one of the kinases responsible for TcR-mediated protein tyrosine phosphorylation. To investigate the role of p56lck in TcR-signaling in detail, we injected anti-Lck monoclonal antibody (mAb), MOL171 or MOL294, both specifically suppress Lck kinase activity in vitro, into Jurkat T-cells by the erythrocyte-ghost procedure in order to block the activity of p56lck. In Jurkat cells injected with anti-Lck mAb, intracellular Ca2+ mobilization induced by TcR-stimulation was markedly reduced in comparison with control mouse IgG-injected samples. This block of Ca2+ influx seems to be specific for TcR-signaling because anti-Lck mAb-injection did not cause significant suppression of phytohaemagglutinin-induced Ca2+ increase. Furthermore, injection of anti-Lck mAb inhibited TcR-mediated protein tyrosine phosphorylation of 100 kDa protein and phospholipase C gamma 1. These results confirm that p56lck is an indispensable element of TcR-signaling and p100 and phospholipase C gamma 1 are strongly presumed to be candidates for substrates for p56lck.
Assuntos
Anticorpos Monoclonais/imunologia , Cálcio/metabolismo , Proteínas Tirosina Quinases/imunologia , Receptores de Antígenos de Linfócitos T/fisiologia , Transdução de Sinais , Linfócitos T/imunologia , Linhagem Celular , Membrana Eritrocítica , Humanos , Proteína Tirosina Quinase p56(lck) Linfócito-Específica , Microinjeções , Fosforilação , Proteínas Tirosina Quinases/antagonistas & inibidores , Tirosina/metabolismoRESUMO
BACKGROUND: Recently, we found that sodium restriction shifted the circadian rhythm of blood pressure from nondipper to dipper in patients with the sodium-sensitive essential hypertension. This study examined whether diuretics can transform the circadian rhythm of blood pressure from nondipper to dipper. METHODS AND RESULTS: We studied 21 patients with essential hypertension during both a baseline period and a period of treatment with hydrochlorothiazide (25 mg daily). The periods lasted 4 weeks each. Twenty-four hour ambulatory blood pressures were measured on the same day of the week at the end of the each period. In nondippers (n=11), but not in dippers (n=10), a significant interaction existed between diuretic therapy and nocturnal fall in systolic and diastolic blood pressure, which indicated that the degree of nocturnal blood pressure fall was affected by diuretic therapy. Nocturnal fall, which was diminished in nondippers, was restored by diuretic therapy with hydrochlorothiazide, indicating that the circadian rhythm of blood pressure shifted from nondipper to dipper patterns. CONCLUSIONS: The present study demonstrated that diuretics can restore nocturnal blood pressure decline in a manner similar to sodium restriction, which suggests that the kidneys and sodium metabolism may play important roles in the genesis of the circadian rhythm of blood pressure. Diuretic-based treatment may have an additional therapeutic advantage of reducing the risk for cardiovascular complications by transforming the circadian rhythm of blood pressure.
Assuntos
Pressão Sanguínea/efeitos dos fármacos , Ritmo Circadiano/efeitos dos fármacos , Hidroclorotiazida/farmacologia , Hipertensão/fisiopatologia , Natriurese/efeitos dos fármacos , Inibidores de Simportadores de Cloreto de Sódio/farmacologia , Idoso , Anti-Hipertensivos/farmacologia , Anti-Hipertensivos/uso terapêutico , Pressão Sanguínea/fisiologia , Diuréticos , Feminino , Frequência Cardíaca/efeitos dos fármacos , Humanos , Hidroclorotiazida/uso terapêutico , Hipertensão/classificação , Hipertensão/tratamento farmacológico , Rim/efeitos dos fármacos , Rim/fisiopatologia , Masculino , Pessoa de Meia-Idade , Sódio/metabolismo , Inibidores de Simportadores de Cloreto de Sódio/uso terapêuticoRESUMO
BACKGROUND-Variations in the ventricular response interval (VRI) during atrial fibrillation (AF) may be reduced in patients with adverse clinical outcomes. The properties of VRI dynamics associated with prognosis remain undetermined. METHODS AND RESULTS-In 107 patients with chronic AF (age, 64+/-9 years), we analyzed a 24-hour ambulatory ECG for VRI variability (SD, SD of successive differences, and SD of 5-minute averages) and VRI irregularity (Shannon entropy of histogram, symbolic dynamics, and approximate entropy of beat-to-beat and minute-to-minute fluctuations [ApEn(b-b) and ApEn(m-m)]). During a follow-up period of 33+/-16 months, 18 patients died (17%), 9 from cardiac causes, 7 from fatal strokes, and 2 from malignancies. Reductions in all VRI variability and irregularity measures were associated with an increased risk for cardiac death but not for fatal stroke. A significant association with cardiac death was also found for ejection fraction (relative risk, 1.10; 95% confidence interval [CI], 1.04 to 1.17, per 1% decrement) and ischemic AF (relative risk, 6.52; 95% CI, 1.62 to 26. 3). After adjustment for these clinical variables, all irregularity measures except symbolic dynamics had predictive value (relative risks [95% CIs] per 1SD decrement: Shannon entropy of histogram, 2. 03 [1.14 to 3.61]; ApEn(b-b), 1.72 [1.14 to 2.60]; and ApEn(m-m), 1. 90 [1.03 to 3.52]); however, the predictive power of variability measures was no longer significant. When the patients were stratified with the 33rd and 67th percentile values of ApEn(b-b) (1. 83 and 1.94, respectively), the 5-year cardiac mortality rates for the upper, middle, and lower tertiles were 0%, 13%, and 43%, respectively (log-rank test, P=0.04). CONCLUSIONS-Reduced VRI irregularity in a 24-hour ambulatory ECG has an independent prognostic value for cardiac mortality during long-term follow-up in patients with chronic AF.
Assuntos
Fibrilação Atrial/complicações , Fibrilação Atrial/mortalidade , Disfunção Ventricular/etiologia , Adulto , Idoso , Doença Crônica , Eletrocardiografia Ambulatorial , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Prognóstico , Fatores de Risco , Volume Sistólico , Análise de Sobrevida , Disfunção Ventricular/fisiopatologiaRESUMO
A 20-year-old Japanese man was referred because of severe pancytopenia with 14% of abnormal blasts in hypocellular bone marrow. After treatment by granulocyte colony-stimulating factor (G-CSF) and transfusions of red blood cells, spontaneous remission was subsequently achieved. After 3 months' remission, however, the patient developed AML characterized by the abnormal karyotype: 46XY,+8,t(9;11)(p22;q23). FISH study revealed the presence of trisomy 8 clone also in the hypoplastic state. While MLL-AF9 chimeric mRNA was observed in leukemic cells, it was not detectable in bone marrow cells from the hypoplastic state by RT-PCR. This is the first report of a trisomy 8 clone which evolved into one with a MLL gene rearrangement.
Assuntos
Cromossomos Humanos Par 21 , Proteínas de Ligação a DNA/genética , Leucemia Mieloide Aguda/patologia , Síndromes Mielodisplásicas/genética , Proto-Oncogenes , Fatores de Transcrição , Trissomia , Histona-Lisina N-Metiltransferase , Humanos , Hibridização in Situ Fluorescente , Leucemia Mieloide Aguda/genética , Masculino , Proteína de Leucina Linfoide-Mieloide , Proteínas Nucleares/genética , Translocação GenéticaRESUMO
We describe two new human leukemia cell lines, MOLM-13 and MOLM-14, established from the peripheral blood of a patient at relapse of acute monocytic leukemia, FAB M5a, which had evolved from myelodysplastic syndrome (MDS). Both cell lines express monocyte-specific esterase (MSE) and MLL-AF9 fusion mRNA. Gene fusion is associated with a minute chromosomal insertion, ins(11;9)(q23;p22p23). MOLM-13 and MOLM-14 are the first cell lines with, and represent the third reported case of, MLL gene rearrangement arising via chromosomal insertion. Both cell lines carry trisomy 8 which was also present during the MDS phase, as well as the most frequent trisomies associated with t(9;11), ie, +6, +13, +19 variously present in different subclones. Despite having these features in common, differences in antigen expression were noted between the two cell lines: that of MOLM-13 being CD34+, CD13-, CD14-, CD15+, CD33+; whereas MOLM-14 was CD4+, CD13+, CD14+, CD15+, CD33+. Differentiation to macrophage-like morphology could be induced in both cell lines after stimulation with INF-gamma alone, or in combination with TNF-alpha, which treatment also induced or upregulated, expression of certain myelomonocyte-associated antigens, including CD13, CD14, CD15, CD64, CD65 and CD87. Together, these data confirm that both cell lines are likely to be novel in vitro models for studying monocytic differentiation and leukemogenesis.
Assuntos
Aberrações Cromossômicas/genética , DNA de Neoplasias/genética , Proteínas de Ligação a DNA/genética , Leucemia Monocítica Aguda/genética , Proteínas Nucleares/genética , Proto-Oncogenes , Fatores de Transcrição , Células Tumorais Cultivadas , Adulto , Antígenos CD/metabolismo , Bandeamento Cromossômico , Transtornos Cromossômicos , Cromossomos Humanos Par 11 , Cromossomos Humanos Par 9 , Citocinas/farmacologia , Regulação Neoplásica da Expressão Gênica , Histona-Lisina N-Metiltransferase , Humanos , Imunofenotipagem , Hibridização in Situ Fluorescente , Cariotipagem , Masculino , Proteína de Leucina Linfoide-Mieloide , Fenótipo , RNA Mensageiro/genéticaRESUMO
We present the case of a 69-year-old man with nephrotic syndrome and renal insufficiency, who developed lobular glomerulonephritis. An electron microscopy examination of a renal biopsy showed microtubular structures of 24 nm in diameter in the subendothelial space and the paramesangial area. These deposits were PAS-positive and Congo red-negative, and revealed predominantly positive staining for kappa light chain. There was no evidence of diseases with highly organized glomerular deposits, such as amyloidosis, cryoglobulinemia, systemic lupus erythematosus or paraproteinemia. Therefore, the patient was diagnosed to have immunotactoid glomerulopathy (ITG). During a seven-year course he has not developed any disease known to be associated with organized glomerular immune deposits. Hence, we believe ITG occurred as a primary glomerular disease in this case. We also highlight cases of ITG with microtubular deposits that have been reported in Japan, compare these cases to previous reports, and show that the characteristics of the Japanese cases are male predominance; a high incidence of membranoproliferative glomerulonephritis (MPGN); a low incidence of monoclonal gammopathy and hematological malignancies and a higher incidence of hypocomplementemia.
Assuntos
Glomerulonefrite Membranoproliferativa/imunologia , Glomerulonefrite Membranoproliferativa/patologia , Imunoglobulina G/análise , Microtúbulos/ultraestrutura , Idoso , Alprostadil/uso terapêutico , Biópsia por Agulha , Análise Química do Sangue , Transfusão de Sangue , Terapia Combinada , Dilazep/uso terapêutico , Seguimentos , Glomerulonefrite Membranoproliferativa/terapia , Humanos , Imuno-Histoquímica , Japão , Testes de Função Renal , Masculino , Proteinúria/diagnóstico , Proteinúria/terapia , Medição de Risco , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
The orientation (chemotaxis) and locomotion (chemokinesis) of human polymorphonuclear leukocytes (PMNs) are generated by an internal movement mechanism that involves active cytoplasmic movement; they are influenced by external environmental and ionic conditions. We have studied the degree to which the orientation and movement mechanisms of PMNs are self-contained within the cell and the degree to which they are under membrane control. PMNs were partially and selectively demembranated by treatment with the non-ionic detergent, octyl-phenoxyl-polyethoxyethanol (commercially known as Triton X-100) under controlled conditions. The tritonated PMNs (referred to in the literature as models) were non-motile and non-locomotory. Addition of ATP/Mg++ with a trace amount of Ca++ to the medium was followed by reactivation of the tritonated PMN models to move again as motile cells. Although these reactivated PMN models actively locomoted, they could no longer orient to chemoattractants. Thus, the reactivation process restored the physical self-contained movement parameters but could not reestablish the orientation capacity (chemotactic responsiveness) that was characteristic of live PMNs. The demembranation process apparently destroyed the chemotactic receptors and/or eradicated the coordination function of the membrane. Videotapes of normal (control) as well as reactivated PMN movement were analyzed for movement characteristics. These characteristics were objectively analyzed with a newly designed computer-assisted micro-image-processing technique whereby the videotapes were digitized and quantified and the actual PMN movement printed out in computer-graphics and tracings (Freeman codes) for confirmation of orientation and movement arising as a result of reactivation.