RESUMO
In our previous works, we demonstrated that human neural stem cells (NSCs) transduced with the cytosine deaminase (CD) gene showed remarkable 'bystander killer effect' on glioma and medulloblastoma cells after administration of the prodrug 5-fluorocytosine (5-FC). In addition, herpes simplex virus thymidine kinase (TK) is a widely studied enzyme used for suicide gene strategies, for which the prodrug is ganciclovir (GCV). To apply this strategy to brain metastasis treatment, we established here a human NSC line (F3.CD-TK) expressing the dual suicide genes CD and TK. We examined whether F3.CD-TK cells intensified the antitumor effect on lung cancer brain metastases. In vitro studies showed that F3.CD-TK cells exerted a marked bystander effect on human lung cancer cells after treatment with 5-FC and GCV. In a novel experimental brain metastases model, intravenously administered F3 cells migrated near lung cancer metastatic lesions, which were induced by the injection of lung cancer cells via the intracarotid artery. More importantly, F3.CD-TK cells in the presence of prodrugs 5-FC and GCV decreased tumor size and considerably prolonged animal survival. The results of the present study indicate that the dual suicide gene-engineered, NSC-based treatment strategy might offer a new promising therapeutic modality for brain metastases.
Assuntos
Neoplasias Encefálicas/secundário , Técnicas de Transferência de Genes , Genes Transgênicos Suicidas , Neoplasias Pulmonares/patologia , Células-Tronco Neurais/metabolismo , Administração Intravenosa , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Neoplasias Encefálicas/terapia , Efeito Espectador , Artérias Carótidas/metabolismo , Movimento Celular , Sobrevivência Celular , Citosina Desaminase/genética , Citosina Desaminase/metabolismo , Flucitosina/administração & dosagem , Flucitosina/farmacologia , Ganciclovir/administração & dosagem , Ganciclovir/farmacologia , Engenharia Genética/métodos , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Estimativa de Kaplan-Meier , Camundongos , Camundongos Nus , Neoplasias Experimentais/terapia , Pró-Fármacos/administração & dosagem , Pró-Fármacos/farmacologia , Simplexvirus/enzimologia , Simplexvirus/genética , Timidina Quinase/genética , Timidina Quinase/metabolismoRESUMO
Primitive neuroectodermal tumors (PNETs) are a family of primary malignant brain tumors that include medulloblastomas. Although genetic models of a subset of medulloblastomas are documented over the past decade, the molecular basis of other subclasses of PNET remains unclear. As elevated c-Myc expression, activation of Wnt/beta-catenin signaling and dysfunction of p53 are seen in human PNETs, we investigated what role these abnormalities have in the formation of PNETs. Incorporating these abnormalities, we generated supratentorial PNET (sPNET) in mice using somatic cell gene transfer. We show that sPNETs arise from GFAP-expressing cells by forced c-Myc expression combined with p53 inactivation. beta-catenin activation promotes tumor progression and induces divergent differentiation. These c-Myc+beta-catenin-induced PNETs are histologically similar to large cell/anaplastic medulloblastomas and can occur in both cerebrum and cerebellum. Furthermore, we have obtained one PNET with marked epithelial differentiation having histological resemblance to choroid plexus carcinoma in this series. Our results in mice suggest that sPNET with varied differentiation and large cell/anaplastic medulloblastomas may be two tumor groups with similar genetic foundations. These data provide insights into the biology and classification of human PNETs and suggest that multiple tumor types or variants can be generated from a fixed set of genetic abnormalities.
Assuntos
Tumores Neuroectodérmicos Primitivos/etiologia , Proteínas Proto-Oncogênicas c-myc/fisiologia , Neoplasias Supratentoriais/etiologia , Proteína Supressora de Tumor p53/fisiologia , beta Catenina/fisiologia , Animais , Diferenciação Celular , Modelos Animais de Doenças , Genes myc , Meduloblastoma/etiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Tumores Neuroectodérmicos Primitivos/classificação , Tumores Neuroectodérmicos Primitivos/patologia , Transdução de Sinais , Proteína Supressora de Tumor p53/genéticaRESUMO
AIM: Dmo1 (Diabetes Mellitus OLETF type I) is a major quantitative trait locus for dyslipidaemia, obesity and diabetes phenotypes in the Otsuka Long Evans Tokushima Fatty (OLETF) rat strain. To evaluate possible metabolic and pathological improvements generated by correction of the Dmo1 genetic pathway, we produced congenic lines, in which both OLETF Dmo1 alleles are replaced by the F344-derived genome. METHODS: Congenic animals were produced by introgressing F344-derived Dmo1 alleles into the OLETF rat. Congenic animals of the fourth generation (BC4) were intercrossed to obtain F1 animals (BC4:F1). Animals of the next generation, BC4:F2, were used for this study. We used 23 BC4:F2 males harbouring homozygous replacement of the OLETF Dmo1 region with the F344-derived genome. Seven animals with OLETF-derived Dmo1 alleles were used as controls. RESULTS: Dmo1-F344/F344 congenic rats showed significant decreases in body weight, abdominal fat weight, serum triacylglycerols, total cholesterol, food consumption and blood glucose after glucose loading (13%, 39%, 45%, 27%, 18% and 27% respectively; p < 0.05) compared with Dmo1-OLETF/OLETF animals. Furthermore, histopathological analysis of the kidney showed that mesangial sclerosis, hyalin deposits and deposition of PAS-positive substance were significantly lower in Dmo1-F344/F344 animals (p < 0.05). CONCLUSION: Improvements in metabolic parameters and histopathological scores show that correction of the Dmo1 genetic pathway in the diabetic and mildly obese OLETF rat strain produces wide-ranging therapeutic effects. Thus, this pathway might represent a new drug target also applicable to humans.
Assuntos
Diabetes Mellitus/genética , Obesidade/genética , Animais , Animais Congênicos , Diabetes Mellitus/metabolismo , Nefropatias Diabéticas/genética , Hiperglicemia/metabolismo , Hiperlipidemias/metabolismo , Rim/ultraestrutura , Fígado/ultraestrutura , Masculino , Obesidade/metabolismo , Fenótipo , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos OLETFRESUMO
Nine new sesquiterpene pyridine alkaloids [wilfornines A (1), B (2), C (3), D (4), E (5), F (8), and G (9); wilfordinines I (6) and J (7)] and six known compounds (10-15) were isolated from a clinically used extract (T(II)) of Tripterygium wilfordii. The structures of 1-9 were elucidated by spectroscopic and chemical methods. The inhibitory effects on cytokine production of 1-3 and several related compounds were evaluated. Compounds 10 and 14 showed significant inhibitory effects on cytokine production.
Assuntos
Alcaloides/farmacologia , Imunossupressores/farmacologia , Plantas Medicinais/química , Sesquiterpenos/farmacologia , Alcaloides/isolamento & purificação , Anti-Inflamatórios/farmacologia , China , Citocinas/biossíntese , Humanos , Imunossupressores/isolamento & purificação , Espectroscopia de Ressonância Magnética , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Prednisolona/farmacologia , Sesquiterpenos/isolamento & purificação , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Espectrofotometria InfravermelhoRESUMO
Whole-genome scans have identified Dmo1 as a major quantitative trait locus (QTL) for obesity and dyslipidaemia in the Otsuka Long Evans Tokushima Fatty (OLETF) rat. We have produced congenic rats for the Dmo1 locus, using marker-assisted speed congenic protocols, enforced by selective removal of other QTL regions (QTL-marker-assisted counterselection), to efficiently transfer chromosomal segments from non-diabetic Fischer 344 (F344) rats into the OLETF background. In the third generation of congenic animals, we observed a substantial therapeutic effect of the Dmo1 locus on lipid metabolism, obesity control and plasma glucose homeostasis. We conclude that single-allele correction of an impaired genetic pathway can generate a substantial therapeutic effect, despite the complex polygenic nature of type II diabetic syndromes.
Assuntos
Hiperglicemia/genética , Hiperlipidemias/genética , Obesidade/genética , Alelos , Animais , Glicemia/metabolismo , Mapeamento Cromossômico , Cruzamentos Genéticos , Diabetes Mellitus Tipo 2/genética , Feminino , Marcadores Genéticos , Genótipo , Masculino , Fenótipo , Característica Quantitativa Herdável , Ratos , Ratos Endogâmicos F344 , Ratos Long-Evans , Ratos MutantesRESUMO
Aerial parts of Daphne oleoides Schreber ssp. oleoides (Thymelaeaceae) are used to treat rheumatoid arthritis and lumbago in Turkish folk medicine. In order to evaluate folkloric utilization, in vitro inhibitory effects of the ethyl acetate extract and fractions obtained from this extract on interleukin 1 (IL-1alpha, IL-1beta) and tumour necrosis factor (TNF-alpha) biosynthesis were studied. Through chemical isolation techniques and activity-guided fractionation process, seventeen compounds were isolated and their structures were elucidated (numbered 1-17). Diterpenoids genkwadaphnin (3) and 1,2-dehydrodaphnetoxin (6) and a coumarin derivative daphnetin (9) showed potent inhibitory activity and were found to be the main active ingredients. Furthermore, gnidilatin (4), gnidilatin-20 palmitate (5), genkwadaphnin-20-palmitate (7) and gnidicin-20-palmitate (8), having diterpenoid structure, and eudesmine (12), wikstromol (13) and matairesinol (14), having lignan structure, were determined to possess moderate inhibitory activity and may have a contributory role in the effect of the remedy.
Assuntos
Citocinas/metabolismo , Extratos Vegetais/farmacologia , Acetatos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Diterpenos/farmacologia , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Sequestradores de Radicais Livres/farmacologia , Furanos/farmacologia , Humanos , Interleucina-1/biossíntese , Interleucina-1/sangue , Lignanas/farmacologia , Modelos Químicos , Plantas Medicinais/química , Fator de Necrose Tumoral alfa/biossíntese , Umbeliferonas/farmacologiaRESUMO
1. Whole-genome scans have identified Dmo1 as a major quantitative trait locus for dyslipidaemia and obesity in the Otsuka Long Evans Tokushima Fatty (OLETF) rat. 2. We have produced congenic rats for the Dmo1 locus through successive back-cross breeding with diabetic OLETF rats. Marker-assisted speed congenic protocols were applied to efficiently transfer chromosomal segments from non-diabetic Brown Norway (BN) rats into the OLETF background. 3. In the fourth generation of congenic animals, we observed a substantial therapeutic effect of the Dmo1 locus on lipid metabolism, obesity control and plasma glucose homeostasis. 4. We have concluded that Dmo1 primarily affects lipid homeostasis, obesity control and/or glucose homeostasis at fasting and is secondarily involved in glucose homeostasis after loading. 5. The results of the present study show that single-allele correction of a genetic defect of the Dmo1 locus can generate a substantial therapeutic effect, despite the complex polygenic nature of type II diabetic syndromes.
Assuntos
Glicemia/genética , Peso Corporal/genética , Mapeamento Cromossômico/métodos , Diabetes Mellitus Tipo 2/genética , Hiperlipidemias/genética , Obesidade/genética , Alelos , Animais , Animais Congênicos , Insulina/sangue , Masculino , Fenótipo , Ratos , Ratos Long-EvansRESUMO
1. The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is a model of type II diabetes with accompanying dyslipidaemia and obesity. 2. To define chromosomal intervals associated with obesity (abdominal fat weight and plasma leptin levels), dyslipidaemia (plasma triglyceride, cholesterol and free fatty acids) and hyperglycaemia (plasma glucose levels), we have performed genome-wide quantitative traits loci (QTL) analyses of 115 male OLETF x (OLETF x Fischer 344) backcross animals at 16 weeks of age. 3. The Diabetes Mellitus OLETF type I (Dmo1) locus on rat chromosome 1 showed statistically significant involvement in elevations of plasma levels of triglycerides (P = 4.87 x 10(-6) at D1Rat90) and total cholesterol (P = 1.16 x 10(-5) at D1Rat306). 4. No other loci produced significant linkage to these observed phenotypes. 5. These analyses have confirmed the importance of Dmo1 in lipid homeostasis at younger ages as well as during overt diabetes, which appears later. Thus, alterations at the Dmo1 locus are a major risk factor for pathogenesis in the strain, a finding that agrees with physiological studies that indicate a role for dyslipidaemia in the type II diabetic syndrome of OLETF rats.
Assuntos
Mapeamento Cromossômico , Hiperlipidemias/genética , Metabolismo dos Lipídeos , Obesidade/genética , Característica Quantitativa Herdável , Animais , Colesterol/sangue , Feminino , Marcadores Genéticos , Genótipo , Hiperlipidemias/sangue , Lipídeos/sangue , Masculino , Obesidade/sangue , Fenótipo , Reação em Cadeia da Polimerase , Ratos , Ratos Endogâmicos F344 , Ratos Long-Evans , Triglicerídeos/sangueRESUMO
In the early stage of infection, Sendai virus delivers its genome into the cytoplasm by fusing the viral envelope with the cell membrane. Although the adsorption of virus particles to cell surface receptors has been characterized in detail, the ensuing complex process that leads to the fusion between the lipid bilayers remains mostly obscure. In the present study, we identified and characterized cell lines with a defect in the Sendai virus-mediated membrane fusion, using fusion-mediated delivery of fragment A of diphtheria toxin as an index. These cells, persistently infected with the temperature-sensitive variant Sendai virus, had primary viral receptors indistinguishable in number and affinity from those of parental susceptible cells. However, they proved to be thoroughly defective in the Sendai virus-mediated membrane fusion. We also found that viral HN protein expressed in the defective cells was responsible for the interference with membrane fusion. These results suggested the presence of a previously uncharacterized, HN-dependent intermediate stage in the Sendai virus-mediated membrane fusion.
Assuntos
Fusão de Membrana/fisiologia , Respirovirus/fisiologia , Animais , Antígenos Virais/análise , Células COS , Linhagem Celular , Vírus Defeituosos/genética , Vírus Defeituosos/fisiologia , Toxina Diftérica/análise , Toxina Diftérica/farmacocinética , Técnica Indireta de Fluorescência para Anticorpo , Genoma Viral , Células HeLa , Humanos , Cinética , Lipossomos , Fragmentos de Peptídeos/análise , Fragmentos de Peptídeos/farmacocinética , Respirovirus/genética , Transfecção , Replicação ViralRESUMO
The extract (T(II)) of Tripterygium wilfordii Hook f. afforded four triterpenoids: wilforic acid D (3beta,24-epoxy-2alpha-hydroxy-24R*-ethoxy-29-friedelanoic acid); (E) 3beta,24-epoxy-2-oxo-3alpha-hydroxy-29-friedelanoic acid; (F) 2beta-hydroxy-3-oxo-friedelan-29-oic acid; 29-hydroxy-3-oxo-olean-12-en-28-oic acid and 17 known triterpenoids. Their structures were established on the basis of spectroscopic studies. In a bioactivity analysis, only the known dulcioic acid compound showed a significant inhibitory effect on cytokine production.
Assuntos
Plantas Medicinais/química , Triterpenos/química , Imunossupressores/química , Estrutura Molecular , Análise EspectralRESUMO
A clinically used extract of Tripterygium wilfordii afforded three new diterpenoids-3beta,19-dihydroxyabieta-8,11,13-triene (triptobenzene L) (1); 12,19-dihydroxy-3-oxoabieta-8,11,13-triene (triptobenzene M) (2); and 19-hydroxy-3,7-dioxo-abieta-8,11, 13-triene (triptobenzene N) (3)-along with 14 known diterpenoids. The structures of 1-3 were established on the basis of spectroscopic studies. Of the known compounds, the stereochemistry at C-4 of triptonediol (4) was reassigned. Tripterifordin (8) and 13-epi-manoyl oxide-18-oic acid (9) showed significant inhibitory effects on cytokine production.
Assuntos
Abietanos , Imunossupressores/isolamento & purificação , Medicina Tradicional Chinesa , Fenantrenos/isolamento & purificação , Plantas Medicinais/química , Anti-Inflamatórios/farmacologia , Citocinas/biossíntese , Citocinas/sangue , Depressão Química , Humanos , Imunossupressores/farmacologia , Técnicas In Vitro , Espectroscopia de Ressonância Magnética , Fenantrenos/farmacologia , Prednisolona/farmacologia , Espectrofotometria Infravermelho , Espectrofotometria UltravioletaRESUMO
The pathogenesis and clinical course of traumatic cerebral vasospasm (TCV) are not yet fully understood. We report a case of delayed symptomatic TCV exaggerated by hyponatremia in spite of minor subarachnoid hemorrhage. A 77-year-old woman was admitted to our hospital with multiple injury caused by a traffic accident. Glasgow coma scale on admission was 8, and CT scan revealed right temporal lobe contusion, thin sub-arachnoid hemorrhage in the right sylvian cistern and right temporal bone fracture. The patient's consciousness level and CT findings improved gradually, but on the 11th day, she suddenly fell into a comatose state. No apparent change was observed on the CT scan, but her serum sodium level was markedly low (113 mEq/L). Under the suspicion of hyponatremia induced consciousness disturbance, sodium replacement therapy was commenced. She showed transient neurological recovery, but on day 13, worsening of left hemiparesis and a new low density area on CT were observed. Vasospasms of the right M2 proximal portion were confirmed by cerebral angiogram, so we performed intraarterial papaverine infusion leading to good extension of spastic vessels, but regrettably, there was no neurological recovery. In general, subarachnoid blood plays an important role in the delayed development of cerebral vasospasm, following not only aneurysmal rupture but also head injury. TCV tends to pass subclinically, but secondary water and electrolyte imbalance may unexpectedly cause TCV to manifest itself clinically. We can confirm that with this patient management of electrolyte balance following head injury was the most important strategy to avoid symptomatic TCV.
Assuntos
Traumatismos Craniocerebrais/complicações , Hiponatremia/complicações , Vasoespasmo Intracraniano/etiologia , Idoso , Progressão da Doença , Feminino , Humanos , Papaverina/uso terapêutico , Sódio/uso terapêutico , Hemorragia Subaracnóidea/complicações , Vasoespasmo Intracraniano/tratamento farmacológicoRESUMO
A method for scalp closure to prevent alopecia along a suture line is described. Only the galea is sutured. By tacking a sufficient width of the galea on both sides with an absorbable suture material, the sutured wound forms a ridge. The outer layer is then closed with skin staples to keep the blood circulation undisturbed. This procedure contrasts with the traditional method in which the galea is sutured with the overlying subcutaneous tissue and consequently the hair follicles are strangled. By adopting the method of suturing the galea, development of alopecia along a suture line has been effectively prevented and scarring has become less conspicuous.
Assuntos
Couro Cabeludo/cirurgia , Técnicas de Sutura , Alopecia/prevenção & controle , Alopecia/cirurgia , Humanos , Procedimentos Cirúrgicos Operatórios/métodos , Grampeadores CirúrgicosRESUMO
HIV-infected cells aberrantly express a high level of antigenic glycosidic structures such as GM2 and Gg4. Some normal sera containing natural IgM Abs to GM2 and/or Gg4 cause C-mediated cytolysis of HIV-infected cells. In the present study we demonstrated that a human IgM anti-GM2 mAb (L55 Ab) can induce cytolysis of HIV-infected cells. Increased GM2 expression by HIV-1 infection of a human T cell line (MOLT4), a human monocyte cell line (U937), and human lymphoblastoid cells was confirmed by immunofluorescence staining with L55 Ab. These infected cells were readily lysed by L55 Ab in the presence of fresh human serum as a C source that alone did not cause cytolysis. L55 Ab also had the ability to destroy HIV-1 particles via C-mediated lysis. By adding L55 Ab together with human C to mixed culture of HIV-infected cells and naive cells, HIV-1 replication was significantly suppressed, and this effect was synergistic when L55 Ab was combined with a reverse transcriptase inhibitor and a proteinase inhibitor. Therefore, a human IgM anti-GM2 mAb may be effective in treating HIV-infected patients, especially when used together with chemotherapeutic agents.
Assuntos
Fármacos Anti-HIV/imunologia , Anticorpos Monoclonais/farmacologia , Proteínas do Sistema Complemento/fisiologia , Gangliosídeo G(M2)/imunologia , HIV-1/imunologia , Imunoglobulina M/farmacologia , Anticorpos Monoclonais/toxicidade , Membrana Celular/metabolismo , Membrana Celular/virologia , Citotoxicidade Imunológica , Gangliosídeo G(M2)/biossíntese , HIV-1/crescimento & desenvolvimento , Humanos , Imunoglobulina M/toxicidade , Leucemia-Linfoma de Células T do Adulto , Leucócitos Mononucleares/virologia , Ativação Linfocitária , Linfócitos/imunologia , Linfócitos/virologia , Células Tumorais Cultivadas , Células U937 , Vírion/imunologia , Replicação ViralRESUMO
Neurinoma arising from the abducens nerve independent of neurofibromatosis has been rarely reported in the existing literature. We present a case of abducens neurinoma which was confirmed during a surgical excision. A 31-year-old female had experienced a hearing disturbance for the past 8 months. Abduction of the right eye ball was almost full. Magnetic resonance images showed a tumor having a size of 44 x 33 X 33 mm at the right cerebellopontine angle. Neuro-otological examination revealed the findings specific to acoustic neurinoma. Surgical excision confirmed that the tumor originated from the abducens nerve. The tumor located at the cavernous sinus or cerebellopontine angle might originate from the abducens nerve, though the abduction of eye ball is not in any way impaired.
Assuntos
Nervo Abducente/patologia , Neoplasias dos Nervos Cranianos/diagnóstico , Neurilemoma/diagnóstico , Neuroma Acústico/diagnóstico , Adulto , Neoplasias Cerebelares/diagnóstico , Ângulo Cerebelopontino/patologia , Neoplasias dos Nervos Cranianos/cirurgia , Diagnóstico Diferencial , Movimentos Oculares , Feminino , Transtornos da Audição/diagnóstico , Humanos , Imageamento por Ressonância Magnética , Neurilemoma/cirurgiaRESUMO
The expression of staphylococcal epidermal cell differentiation inhibitor (EDIN), an ADP-ribosyltransferase targeting the small GTP-binding protein rho p21, was examined using Bacillus subtilis. A recombinant plasmid, containing B. licheniformis alpha-amylase promoter flanking either a beta-glucanase or a B. cereus sphingomyelinase signal sequence, and a DNA fragment corresponding to mature EDIN were constructed and used to transform B. subtilis KN2. Transformants were designated ED7 and ED8, respectively. ED7 extracellularly produced recombinant protein, which was purified to homogeneity through column chromatography using SP-Toyopearl 650 cation-exchange gel and the HA1000 hydroxyapatite HPLC column. ED8 did not grow in broth culture. Biochemical and biological studies of purified protein revealed that ED7 produced a correctly processed recombinant EDIN, indistinguishable from natural EDIN.
Assuntos
Difosfato de Adenosina/metabolismo , Bacillus subtilis/química , Proteínas de Bactérias/isolamento & purificação , Células 3T3 , Aminoácidos/análise , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Western Blotting , Chlorocebus aethiops , Cromatografia Líquida de Alta Pressão , Primers do DNA/química , Relação Dose-Resposta a Droga , Eletroforese em Gel de Poliacrilamida , Cinética , Camundongos , Plasmídeos , Reação em Cadeia da Polimerase , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Staphylococcus aureus/química , Fatores de Tempo , Células VeroRESUMO
With the aim of developing an effective immunotherapy for malignant glioma, glioma cells were incubated with tumor necrosis factor-alpha (TNF-alpha) to increase their susceptibility to lysis by lymphokine-activated killer (LAK) cells. Treatment with exogenous TNF-alpha induced the expression of intercellular adhesion molecule-1 (ICAM-1) on the surface of glioma cells. In addition, the cytolytic activity of LAK cells toward exogenous TNF-alpha treated glioma cells was significantly greater than LAK cell activity toward untreated glioma cells. This increase in cytolytic activity was blocked by anti-ICAM-1 monoclonal antibodies (MAb). Furthermore, co-treatment with a bifunctional antibody (BFA) composed of anti-CD3 (UCHT1) and antiglioma (G-22) antibodies synergistically increased the cytolytic activity of LAK cells towards TNF-alpha-treated glioma cells. These results indicate that a combination of exogenous TNF-alpha and anti-CD3/antiglioma BFA may provide an effective modified adoptive immunotherapy for patients with malignant glioma.
Assuntos
Anticorpos Biespecíficos/imunologia , Anticorpos Antineoplásicos/imunologia , Complexo CD3/imunologia , Citotoxicidade Imunológica , Glioma/imunologia , Células Matadoras Ativadas por Linfocina/imunologia , Fator de Necrose Tumoral alfa/farmacologia , Anticorpos Biespecíficos/uso terapêutico , Anticorpos Monoclonais/farmacologia , Anticorpos Antineoplásicos/uso terapêutico , Complexo CD3/uso terapêutico , Citotoxicidade Imunológica/efeitos dos fármacos , Citometria de Fluxo , Glioma/terapia , Humanos , Imunoterapia Adotiva/métodos , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/imunologia , Células Matadoras Ativadas por Linfocina/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêutico , Células Tumorais Cultivadas , Fator de Necrose Tumoral alfa/uso terapêuticoRESUMO
The T-cell receptor (TCR) can acquire a new antigen binding site by treatment with a bifunctional antibody (BFA) prepared with mAb against a specified antigen and an epitope of the TCR. Lymphocytes armed with BFA directed to CD3 and an HIV antigen were able to eliminate all HIV antigen-positive cells during incubation with a mixture of HIV-infected and uninfected cells. HIV antigen-positive cells even from persistently infected cells were undetectable with immunofluorescence staining although HIV genes were detectable by polymerase chain reaction (PCR) amplification indicating that only dormantly infected or low producer cells, if any, survived. This suggests that HIV antigen-positive cells could be eliminated by administration of BFA-armed lymphocytes leaving HIV patients with only dormantly infected or low producer cells.
Assuntos
Complexo CD3/imunologia , Proteína gp120 do Envelope de HIV/imunologia , HIV-1/imunologia , Linfócitos T/imunologia , Anticorpos Biespecíficos/imunologia , Linhagem Celular , Células Cultivadas , Infecções por HIV/terapia , HumanosRESUMO
With the aim of developing an effective cancer immunotherapy for common epithelial cancer, a new class of bifunctional antibody (BFA) was developed; one arm of this BFA recognized c-erbB-2 gene product, and the other arm recognized CD3 epsilon, a T-cell specific surface antigen. Application of this BFA with human peripheral blood lymphocytes exhibited specific anti-tumor activity in vitro on a breast tumor cell line, ZR-75-1, which expressed abundant c-erbB-2 gene product on its cell surface. These results indicate that BFA recognizing an oncogene product on cell surface is a potential new agent for cancer immunotherapy.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Anticorpos Antineoplásicos/uso terapêutico , Antígenos de Diferenciação de Linfócitos T/imunologia , Citotoxicidade Imunológica , Proteínas Proto-Oncogênicas/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Anticorpos Monoclonais/química , Anticorpos Antineoplásicos/química , Complexo CD3 , Reagentes de Ligações Cruzadas , Humanos , Imunidade Celular , Fragmentos Fab das Imunoglobulinas/química , Fragmentos Fab das Imunoglobulinas/uso terapêutico , Técnicas In Vitro , Receptor ErbB-2 , Linfócitos T/imunologia , Células Tumorais CultivadasRESUMO
The Fab' fragment of a monoclonal antibody (mAb) directed to CD3 (a portion of the T cell receptor) and the Fab' or F(ab')2 fragment of an mAb to HIV were combined to generate bifunctional antibody (BFA) consisting of antiCD3-Fab' conjugated with anti-HIV-Fab' (Fab'/Fab') or antiCD3-Fab' conjugated with anti-HIV-F(ab')2 (Fab'/F(ab')2), respectively. In the presence of these BFA, HIV-infected target cells were cytolysed by peripheral blood lymphocytes. Treatment of lymphocytes with Fab'/F(ab')2 type BFA rendered the lymphocytes significantly cytotoxic to HIV-infected target cells. Since BFA-armed lymphocytes can react on HIV-infected cells regardless of histocompatibility, lymphocytes from healthy donors could be armed with BFA for treatment of HIV-infected patients including those who do not exhibit histocompatibility with the lymphocyte donor.