RESUMEN
33 patients with advanced malignant melanoma were studied after intravenous administration of 131I-labeled Fab fragments specific for p97, an oncofetal glycoprotein of human melanoma. In all, 47 gamma camera imaging studies were performed for the purpose of localization of metastatic deposits. In addition to tumor, 131I-Fab uptake was also seen in liver and kidney. 20 of these studies included simultaneous administration of both an 131I-labeled Fab specific for p97, and an 125I-labeled Fab not specific for p97. Blood clearance of p97-specific Fab was significantly more rapid than for nonspecific Fab. Eight of these patients had biopsies of subcutaneous nodules at 48 and 72 h postinjection in order to assess whether localization of radioactivity was antigen specific. Antigen-specific localization was observed with average ratios of specific/nonspecific uptake of 3.7 (48 h) and 3.4 (72 h); uptake was strongly correlated with tumor p97 concentration (r = 0.81, P less than 0.01). Also, imaging studies of the bio-distribution of 131I-labeled anti-p97 Fab in patients selected for high p97 tumor concentration showed avid tumor uptake and more prolonged retention of labeled Fab in tumor than in normal tissues. Based on these studies, we estimated that total 131I doses of 500 mCi could be safely given to patients before dose-limiting toxicity would be observed. Accordingly, in seven selected patients, phase I radiotherapeutic trials were begun. For improved radiation safety, we developed automated methods to label Fab fragments with up to 200 mCi of 131I. So far, a total of 12 individual therapeutic doses, ranging from 34 to 197 mCi of 131I-labeled to 5 to 10 mg of Fab, have been administered with excellent tumor localization and without major target organ toxicity. Cumulative doses ranged from 132 to 529 mCi 131I. Side effects attributable to the radiation were mild, with a transient drop slightly greater than 50% in platelet and absolute neutrophil counts being observed in the two patients who received cumulative doses greater than 500 mCi. In the combined series of 47 diagnostic and 12 therapeutic studies, four acute reactions were observed: one episode each of transient chills and fever; flushing and hypotension; and two skin rashes. All of these reactions responded promptly to symptomatic therapy. After multiple administrations of 131I-(anti-p97) Fab (IgG1), isotype-specific immunity was observed in three patients. In two of these patients it was possible to successfully reinfuse after immunity had developed with 131I-(anti-p97) Fab of a different isotype (IgG2a). Dosimetry estimates were performed based on the biodistribution of (131)I-Fab in these patients,and for every 100 mCi of (131)I-Fab given, tumor receives 1,040 rads; liver. 325 rads; and bone marrow, 30 rads. Marrow would be expected to be the critical organ, if doses >500 mCi (131)I-Fab are given. These studies demonstrated that, with proper precautions, large doses (of an (131)I-labeled murine Fab fragments immunologically specific for a human melanoma-associated antigen) could be safely given to humans by using repetitive intravenous injections.
Asunto(s)
Fragmentos Fab de Inmunoglobulinas/análisis , Radioisótopos de Yodo , Melanoma/diagnóstico por imagen , Proteínas de Neoplasias/análisis , Animales , Anticuerpos Antiidiotipos/análisis , Antígenos de Neoplasias , Epítopos , Humanos , Riñón/inmunología , Melanoma/inmunología , Melanoma/radioterapia , Antígenos Específicos del Melanoma , Ratones , Metástasis de la Neoplasia , Radiometría , Cintigrafía , Distribución Tisular , Vejiga Urinaria/inmunologíaRESUMEN
A 186Re-labeled monoclonal antibody (MAb), NR-LU-10, was used for the radioimmunotherapy of a subcutaneous human small cell lung carcinoma xenograft, SHT-1, in nude mice. Biodistribution with specific and irrelevant labeled MAb demonstrated peak tumor uptake of 8% and 3% of the injected dose/g at 2 days, respectively. Dosimetry analysis predicted tumor:whole-body radiation-absorbed dose ratios of 2.43:1 for NR-LU-10 and 0.62:1 for irrelevant MAb. Single-dose toxicity screening estimated a 50% lethal dose within 30 days of 600 microCi (880 cGy of whole-body radiation). As anticipated, a multiple-dose regimen of 490 microCi in four doses over 10 days (720 cGy of whole-body radiation, eight of eight surviving greater than 30 days) was less toxic than a single bolus dose of 430 microCi (644 cGy of whole-body radiation), six of eight surviving greater than 30 days). A multidose radioimmunotherapy regimen was initiated in nude mice bearing 66-mm3 tumors (total dose, 500 to 600 microCi). Complete remissions (greater than 140 days) were achieved in three of 16 mice, and the remainder showed a mean tumor growth delay of 53 days. Matched doses with irrelevant MAb produced one remission, one treatment-related death, and a mean growth delay of only 20 days in six of eight mice. Thus, in this nonoptimal radioimmunotherapy model, significant antitumor responses were observed using a mildly toxic multiple dosing regimen.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Carcinoma de Células Pequeñas/terapia , Neoplasias Pulmonares/terapia , Radioisótopos/uso terapéutico , Renio/uso terapéutico , Animales , Autorradiografía , Carcinoma de Células Pequeñas/patología , Carcinoma de Células Pequeñas/radioterapia , Línea Celular , Femenino , Humanos , Inmunoterapia , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/radioterapia , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Renio/farmacocinética , Distribución Tisular , Trasplante HeterólogoRESUMEN
The relative rates of degradation of the outer lipid bilayer of large multilamellar and small unilamellar bovine brain sphingomyelin/cholesterol (2:1; mol/mol) liposomes in the livers of Balb/c mice were compared. The rate of the release of entrapped In-111 ions from the aqueous reservoir of small unilamellar liposomes or from the outermost aqueous compartment of multilamellar liposomes was used to monitor the rate of degradation of the exterior lipid bilayer surface of these liposomes. The technique of gamma-ray perturbed angular correlation and a method for loading In-111 ions into the outermost aqueous compartment of liposomes were used in this investigation. It was found that in the liver the exterior lipid bilayer of large multilamellar liposomes was degraded more rapidly than the bilayer of small unilamellar liposomes in vivo. In contrast to the situation for small unilamellar liposomes, the degradative process for large multilamellar liposomes in the liver was not maintained under ischemic conditions. Our results suggest that multiple pathways operate in the degradation of liposomes in the liver. The rate of degradation of liposomes in the liver may depend on accessibility of liposomes to degradative sites.
Asunto(s)
Encéfalo/metabolismo , Colesterol/metabolismo , Liposomas/metabolismo , Hígado/metabolismo , Esfingomielinas/metabolismo , Animales , Bovinos , Membrana Dobles de Lípidos/metabolismo , Ratones , Ratones Endogámicos BALB C , Factores de TiempoRESUMEN
The encapsulation of radioactive metalic cations, such as 111In3+ or 67Ga3+, in the internal aqueous compartment of liposomes can be achieved with an efficiency of about 90%. The efficient loading of a high specific activity of cations into liposomes involves the transport of 111In3+ or 67Ga3+ through the lipid bilayer to an encapsulated strong chelate, such as nitrilotriacetic acid, by 8-hydroxyquinoline, in conjunction with an efficient anion-exchange resin technique for the removal of the external cations. The efficiency of loading cations to liposomes is affected markedly by the concentration of 8-hydroxyquinoline-metal, and the presence of the chelating agents in the loading incubation mixture. However, the loading efficiency is not affected by the pH of the internal aqueous compartment of liposomes over a range of pH 5-9, the concentration of the liposomes, the method of liposomal preparation, the lamellar structure of the liposomes, and the composition of liposomes. Furthermore, the loading procedures do not appear to affect the size and the permeability of liposomes. There is a good agreement in the tissue distributions of the liposomes prepared by the present loading methods and those by the conventional method of encapsulation by sonication. Liposomes entrapping high specific activity of 67Ga3+ or 111In3+ will be useful for future studies of the in vivo kinetics of liposomes by the combined techniques of scintigraphic imaging and the gamma-ray perturbed angular correlation.
Asunto(s)
Radioisótopos de Galio , Indio , Liposomas , Radioisótopos , Animales , Cationes , Citratos , Ácido Cítrico , Radioisótopos de Galio/metabolismo , Concentración de Iones de Hidrógeno , Indio/metabolismo , Liposomas/metabolismo , Métodos , Ratones , Ratones Endogámicos BALB C , Ácido Nitrilotriacético , Oxiquinolina , Distribución TisularRESUMEN
The distribution of liposomes within the intravascular space and the extent to which they escape into extravascular space strongly impact on the application of lipid vesicles as a carrier for pharmacologically active agents. The present study investigates how intact small unilamellar vesicles (SUV) may be taken up by different tissues after intravenous injection into mice, using various types of SUV with different entrapped markers, lipid composition, size, doses of liposomal lipids and stability in the blood. Our focus was specifically on sphingomyelin (or distearoyl phosphatidylcholine)/cholesterol (2:1, mol/mol) SUV, which are known to be stable in the blood circulation. Our results indicated that, in addition to the reticuloendothelial tissues, intact SUV were taken up in several other parts of the body, including intestine, skin, carcass and legs. It appears that the accumulation of SUV in the intestine and the skin increases with time post-injection. Furthermore, from the kinetic data, the process of uptake of SUV by the skin and intestine is compatible with a non-saturable pathway, which follows first-order kinetics. This suggests that the cells involved in the uptake of SUV in the intestine and skin are not phagocytic cells, which are normally saturable.
Asunto(s)
Liposomas/metabolismo , Animales , Colesterol , Mucosa Intestinal/metabolismo , Cinética , Lípidos de la Membrana , Ratones , Ratones Endogámicos BALB C , Sistema Mononuclear Fagocítico/metabolismo , Fosfatidilcolinas , Piel/metabolismo , Esfingomielinas , Distribución TisularRESUMEN
The efficient loading of high levels of In-111 into the inner aqueous compartment of liposomes can be achieved using a water-soluble lipophilic chelate, acetylacetone. The loading method involves the incubation of the acetylacetone-In-111 complex with liposomes entrapping 1mM nitrilotriacetic acid. The loading process is carried out in isotonic saline, 10 mM Tris-HCI, pH 7.6, at room temperature. Results indicate that In-111 ions are concentrated in the internal aqueous compartment of liposomes by acetylacetone, which mediates the transport of In-111 ions across the outermost lipid bilayer of liposomes, permitting subsequent transfer to the encapsulated nitrilotriacetic acid. As much as 90% of the acetylacetone-chelated In-111 becomes internalized in the aqueous compartment of liposomes and bound by entrapped nitrilotriacetic acid. Using liposomes made from bovine brain shingomyelin/cholesterol, our results indicate that the loaded liposomes retain the entrapped internally bound In-111 even after incubation with serum at 37 degrees C for 24 hr.
Asunto(s)
Indio , Cetonas , Liposomas , Pentanonas , Radioisótopos , MétodosRESUMEN
The Oyen et al. study is valuable in that it systematically evaluates several of the factors involved in radiolabeled protein uptake and retention in infectious foci. The role of particular proteins and their receptor specific interactions seems to be inconsequential in agreement with the findings of other. However, the role of the radiolabel was shown to be important and significant differences were delineated from comparisons of the radionuclides and their associated chemistries. The conclusion implicating radionuclide chemistry and associated linkages underscores the need to optimize the attachment and labeling chemical modifications of protein carriers. Evaluation criteria should include serum stability, determination and assessment of the effect of molar substitution ratio, and potential for improving blood clearance without reducing the target-to-non-target ratio. Important areas for future study include characterization of radioactive metabolites and the design and synthesis of new ligands which direct the disposition of metabolites reducing retention in normal organs or accelerating renal excretion. Additionally, intracellular processing of radiolabel, compartmental distribution and strategies for augmenting internalization and retention within the target cell merit detailed exploration. For each radionuclide of interest, 111In, radioiodines, 99mTc and others, improved chemical moieties exist for controlling radiolabel fate. When carrying out mechanistic and evaluative studies, clear-cut conclusions will only be reached when defined and controlled chemistry is used. Having established a "gold standard," simplifications in radiolabeling and other chemical refinements can then be pursued with a quantitative understanding of the trade-offs in targeting agent performance versus other considerations such as cost reduction, simplicity, and convenience.
Asunto(s)
Infecciones Bacterianas/metabolismo , Inmunoglobulina A/metabolismo , Inmunoglobulina G/metabolismo , Radioisótopos de Indio/farmacocinética , Radioisótopos de Yodo/farmacocinética , Albúmina Sérica/farmacocinética , Tecnecio/farmacocinética , Infecciones Bacterianas/diagnóstico por imagen , Cintigrafía , Distribución TisularRESUMEN
Two different murine monoclonal antibody Fab fragments specific for p97, a melanoma-associated antigen, were labeled with I-131 at high activity levels without excessive chemical damage. Up to 20 mg of Fab were labeled with up to 300 mCi of I-131 using the chloramine-T method and large working volumes at room temperature. As much as 90% of the initial activity was recovered as labeled product. The labeled Fabs varied in their sensitivity to radioiodination damage, as measured by an in vitro cell-binding assay. Radioiodination was performed safely using a remote iodination apparatus. The final product was of radiopharmaceutical quality suitable for clinical diagnosis and experimental radiotherapy in humans.
Asunto(s)
Anticuerpos Monoclonales/análisis , Fragmentos Fab de Inmunoglobulinas/análisis , Radioisótopos de Yodo/uso terapéutico , Melanoma/radioterapia , Proteínas de Neoplasias/análisis , Compuestos de Tosilo , Animales , Anticuerpos Monoclonales/administración & dosificación , Antígenos de Neoplasias , Cloraminas , Exposición a Riesgos Ambientales , Humanos , Marcaje Isotópico/métodos , Melanoma/inmunología , Antígenos Específicos del Melanoma , Ratones , Dosis de RadiaciónRESUMEN
The tumor targeting capacity of monoclonal antibody Fab fragments was explored in nude mice bearing human melanoma xenografts. Radioiodinated Fab 8.2 and 96.5, specific for melanoma-associated antigen p97, were tested in vitro for immunoreactivity and in vivo for tumor localization relative to a co-administered control, Fab 1.4. Fab was cleared rapidly from the blood with a T1/2 of 3-3.5 hr and greater than 90% of the injected radioactivity was excreted by 16 hr. The mean specific Fab in tumor reached a maximum of 3.5% injected dose/g at 4 hr and decreased to 1.5% at 16 hr. Over the same period, the ratio of specific/control Fab in tumor normalized to blood, the localization index, rose from 3 to 25 compared with ratios near unity for all other tissues. The concentration of specific Fab in tumor could be correlated to the amount of Fab protein administered as well as its immunoreactivity.
Asunto(s)
Anticuerpos Antineoplásicos , Fragmentos Fab de Inmunoglobulinas , Radioisótopos de Yodo , Melanoma/diagnóstico por imagen , Animales , Anticuerpos Monoclonales , Anticuerpos Antineoplásicos/metabolismo , Antígenos de Neoplasias/inmunología , Antígenos de Superficie/inmunología , Autorradiografía , Línea Celular , Femenino , Semivida , Humanos , Fragmentos Fab de Inmunoglobulinas/metabolismo , Marcaje Isotópico , Cinética , Melanoma/inmunología , Antígenos Específicos del Melanoma , Ratones , Ratones Desnudos , Proteínas de Neoplasias/inmunología , Cintigrafía , Distribución TisularRESUMEN
A kit has been developed for 99mTc antibody radiolabeling via defined chemistry using an N2S2 diamide dimercaptide bifunctional chelating agent and the performed chelate method. The process involved efficient transchelation of 99mTc from gluconate to 2,3,5,6-tetrafluorophenyl 4,5-bis-S-(1-ethoxyethyl) mercaptoacetamidopentanoate as an active ester ligand and subsequent conjugation to antibody lysine amine functional groups. The use of the ethoxyethyl group for sulfur protection allowed optimum yields of 99mTc N2S2 chelate formation with complete retention of the active ester. Subsequent addition of antibody Fab fragment gave 99mTc chelate conjugates indistinguishable from the stepwise in situ esterification and purification of the 99mTc N2S2 complex followed by conjugation as previously shown to give stable 99mTc antibody fragments with retained immunoreactivity and tumor-targeting properties.
Asunto(s)
Aminoácidos Diaminos , Aminoácidos Sulfúricos , Fragmentos Fab de Inmunoglobulinas , Juego de Reactivos para Diagnóstico , Tecnecio , Aminoácidos Diaminos/síntesis química , Aminoácidos Diaminos/farmacocinética , Aminoácidos Sulfúricos/síntesis química , Aminoácidos Sulfúricos/farmacocinética , Animales , Estudios de Evaluación como Asunto , Fragmentos de Inmunoglobulinas , Marcaje Isotópico , Ratones , Ratones Desnudos , Distribución TisularRESUMEN
Rhenium-186 is a beta-emitting radionuclide that has been studied for applications in radioimmunotherapy. Its 137 keV gamma photon is ideal for imaging the biodistribution of the immunoconjugates and for obtaining gamma camera data for estimation of dosimetry. Methods used for determining radiation absorbed dose are described. We have estimated absorbed dose to normal organs and tumors following administration of two different 186Re-labeled immunoconjugates, intact NR-LU-10 antibody and the F(ab')2 fragment of NR-CO-02. Tumor dose estimates in 46 patients varied over a wide range, 0.4-18.6 rads/mCi, but were similar in both studies. Accuracy of activity estimates in superficial tumors was confirmed by biopsy. Prediction of 186Re dosimetry from a prior 99mTc imaging study using a tracer dose of antibody was attempted in the NR-CO-02 (Fab')2 study. Although 99mTc was an accurate predictor of tumor localization and the mean predicted and observed radiation absorbed doses to normal organs compared favorably, 186Re dosimetry could not be reliably predicted in individual patients. The methods described nevertheless provide adequate estimates of 186Re dosimetry to tumor and normal organs.
Asunto(s)
Radioinmunodetección , Radioinmunoterapia , Radioisótopos/uso terapéutico , Renio/uso terapéutico , Humanos , Radioisótopos de Yodo/uso terapéutico , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/radioterapia , Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/radioterapia , Radiometría , Tecnecio , Distribución TisularRESUMEN
Lymphoscintigraphy was performed at 3 and 20 hr following subcutaneous injection of 131I anti-melanoma antibody (Fab) in 11 patients who had surgical resection of lymph nodes (neck, axilla, groin) at 24 hr for suspected metastatic melanoma. Comparable amounts of 125I nonspecific control antibody (Fab) were co-administered. Six patients had nodal metastases and three showed positive images at both time periods. Five patients had no metastases though one was image positive. Four other nondiseased inguinal node groups were image negative. A total of 28 tumored nodes and 110 normal nodes were removed, counted and histologically examined. All metastatic tumors expressed antigen against which the specific Fab was directed. The concentrations of both specific and nonspecific Fab were similar in tumored nodes and both were significantly greater than in normal nodes showed essentially identical intranodal spatial distribution of the specific and control Fab in areas containing tumor. These preliminary results suggest the increased concentration of murine immunoglobulin (Fab) retained in diseased nodes was a nonspecific phenomenon.
Asunto(s)
Anticuerpos Monoclonales , Radioisótopos de Yodo , Ganglios Linfáticos/diagnóstico por imagen , Melanoma/diagnóstico por imagen , Adulto , Anticuerpos Monoclonales/análisis , Autorradiografía/métodos , Axila , Ingle , Humanos , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/cirugía , Metástasis Linfática , Melanoma/secundario , Melanoma/cirugía , Persona de Mediana Edad , Cuello , CintigrafíaRESUMEN
UNLABELLED: Pretargeted radioimmunotherapy (PRIT) was evaluated using an antibody-streptavidin conjugate, followed by a biotin-galactose-human serum albumin clearing agent and 90Y-dodecane tetraacetic acid (DOTA)-biotin as the final step for therapy. The objective was to develop a clinical protocol that could show an improved tumor-to-red marrow therapeutic ratio compared with conventional radioimmunotherapy (RIT) and at the same time preserve the efficiency of tumor targeting. METHOD: Forty-three patients with adenocarcinomas reactive to NR-LU-10 murine monoclonal antibody received the 3 components. Doses and timing parameters were varied to develop an optimized schema. In some patients, the conjugate was radiolabeled with 186Re as an imaging tracer to assess biodistribution of the conjugate and effectiveness of the clearing agent. 111In-DOTA-biotin was coinjected with 90Y-DOTA-biotin for quantitative imaging. Safety, biodistribution, pharmacokinetics, dosimetry, and antiglobulin formation were evaluated. RESULTS: The optimal schema was defined as a conjugate dose of 125 microg/mL plasma volume followed at 48 h by a clearing agent in a 10:1 molar ratio of clearing agent to serum conjugate. The therapeutic third step was 0.5 mg radiobiotin administered 24 h later. No significant adverse events were observed after administration of any of the components. The mean tumor-to-marrow absorbed dose ratio when using the optimized PRIT schema was 63:1, compared with a 6:1 ratio reported previously for conventional RIT. Antiglobulin to murine antibody and to streptavidin developed in most patients. CONCLUSION: This initial study confirmed that the PRIT approach is safe and feasible and achieved a higher therapeutic ratio than that achieved with conventional RIT using the same antibody.
Asunto(s)
Adenocarcinoma/radioterapia , Radioinmunoterapia/métodos , Radioisótopos de Itrio/uso terapéutico , Estudios de Factibilidad , Femenino , Humanos , Masculino , Factores de TiempoRESUMEN
A convenient, rapid, and reproducible assay was developed to evaluate the immunoreactivity of radiolabeled monoclonal antibodies against three different human melanoma-associated antigens, p97, a proteoglycan and a GD3 ganglioside. A cloned melanoma cell line (M 2669 CL 13) was selected as the target and, when fixed with paraformaldehyde, showed binding as good as or better than that obtained with live cells for the three antigens. Fixed cells retained good binding properties stored at 4 degrees C for over 6 mo. This assay has general applicability to other antigen-antibody systems for testing chemically modified monoclonal antibodies or fragments during the development of a radiopharmaceutical or as a routine quality control measure for clinical agents.
Asunto(s)
Anticuerpos Monoclonales/análisis , Inmunoensayo/métodos , Melanoma/inmunología , Antígenos de Neoplasias , Recuento de Células , Línea Celular , Estudios de Evaluación como Asunto , Gangliósido G(M3) , Humanos , Fragmentos Fab de Inmunoglobulinas/análisis , Antígenos Específicos del Melanoma , Proteínas de Neoplasias , ProteoglicanosRESUMEN
A mouse-human chimeric monoclonal antibody (NR-LU-13), with the same pancarcinoma antigen recognition site as a previously studied murine monoclonal antibody (NR-LU-10), was radiolabeled with 186Re using a bifunctional chelate. Nine patients (ages 31-81 yr) with metastatic adenocarcinoma received 186Re NR-LU-13. A single intravenous dose of 42 mg NR-LU-13 labeled with 25 mCi/m2 (two patients) or 60 mCi/m2 (seven patients) was administered. Mean serum disappearance half-time values for the chimeric 186Re NR-LU-10). Fifty percent of the radiolabel was excreted in the urine by 6 days. Tumor localization was demonstrated by gamma camera imaging in seven of nine patients. The percent injected dose per gram in a single tumor biopsy specimen was 0.003% at 72 hr postinjection. Absorbed dose to bone marrow was 1.5 +/- 0.7 rads/mCi and resulted in reversible myelosuppression in five of six evaluable patients who received 60 mCi/m2: median WBC nadir = 2500/microliters; median platelet nadir = 85,500/microliters. Low grade fever, nausea, slight elevations of liver function tests and mild allergic reactions were seen in some patients. The chimeric antibody elicited low levels of anti-NR-LU-13 antibody in six of eight evaluable patients (75%), in contrast to NR-LU-10 which elicited higher levels of human anti-mouse antibody in all patients. This pilot study demonstrates the ability of the chimeric antibody to target tumors with reduced (but not absent) immunogenicity and delayed clearance relative to the murine antibody.
Asunto(s)
Anticuerpos Monoclonales/metabolismo , Radioinmunoterapia , Radioisótopos/farmacocinética , Proteínas Recombinantes de Fusión/farmacocinética , Renio/farmacocinética , Adenocarcinoma/diagnóstico por imagen , Adenocarcinoma/inmunología , Adenocarcinoma/metabolismo , Adenocarcinoma/radioterapia , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales/inmunología , Anticuerpos Antineoplásicos/inmunología , Formación de Anticuerpos , Semivida , Humanos , Persona de Mediana Edad , Radioisótopos/efectos adversos , Cintigrafía , Proteínas Recombinantes de Fusión/inmunología , Renio/efectos adversos , Renio/inmunología , Distribución TisularRESUMEN
L6 is a murine monoclonal antibody (MAb) binding to cells of most human carcinomas, mediating antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity, and inhibiting tumor growth in nude mice [10]. Fab and F(ab')2 fragments of L6, as well as intact MAb, have been evaluated for immunospecific localization in nude mice xenografted with a human lung carcinoma. They were compared with preparations of an isotype-matched control immunoglobulin, P1.17, after labelling with 125I or 131I. L6 Fab fragments prepared from MAb L6 and labelled with 67Ga via desferrioxamine were also tested. The data suggest that MAb L6 may be useful for in vivo detection of human carcinomas.
Asunto(s)
Anticuerpos Monoclonales , Neoplasias Experimentales/metabolismo , Animales , Femenino , Radioisótopos de Galio , Fragmentos Fab de Inmunoglobulinas , Radioisótopos de Yodo , Tasa de Depuración Metabólica , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias Experimentales/inmunología , Distribución TisularRESUMEN
This study investigated the biodistribution of bovine brain sphingomyelin (SM)/cholesterol (CH) (2/1; M/M) small unilamellar vesicles (SUV) in mice, addressing specifically the volume of distribution and transcapillary passage of the SUV. The complex of nitrilotriacetic acid with In-111 or Ga-67 ions was encapsulated in the SUV as the radioactive marker for various studies. The structural integrity of liposomes in vitro and in vivo was monitored by the technique of gamma ray perturbed angular correlation. Our data suggested that initially the SM/CH SUV remained within the vascular system and occupied a volume of distribution approximately 1.28 times larger than that of erythrocytes in the vascular system of mice. However, our data also indicated that with time the SM/CH SUV could get out of the vascular system of mice and were taken up by surrounding tissues over a period of 24 hours.
Asunto(s)
Liposomas/administración & dosificación , Esfingomielinas/sangre , Animales , Inyecciones Intravenosas , Cinética , Ratones , Ratones Endogámicos BALB C , Distribución TisularRESUMEN
Mouse monoclonal antibody L6 (IgG2a subtype) recognizes a ganglioside antigen expressed at the surface of cells from human non-small-cell lung carcinomas, breast carcinomas, and colon carcinomas. We now show that this antibody can lyse L6 antigen-positive human tumor cells in the presence of Leu-11b-positive human lymphocytes (i.e., mediate antibody-dependent cellular cytotoxicity) or human serum (mediate complement-dependent cytotoxicity) and that it can inhibit the outgrowth of an L6 antigen-positive human tumor transplanted onto nude mice.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Anticuerpos Antineoplásicos/inmunología , Inmunoglobulina G/inmunología , Neoplasias/inmunología , Animales , Citotoxicidad Celular Dependiente de Anticuerpos , Antígenos de Neoplasias/análisis , Proteínas del Sistema Complemento/inmunología , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones DesnudosRESUMEN
The effect of the liposome dose of bovine brain sphingomyelin/cholesterol (2/1; mol/mol) small unilamellar vesicles (mean diameter 187 42A) on the rate of elimination of the vesicles from the circulation of mice was investigated. The results of the study indicated that the relative rate of elimination of the vesicles from the blood depended on the amount of intravenously administered liposomal lipid. The distribution of the liposomes in vivo was followed by monitoring entrapped In-111. In all tissues examined, the uptake of the liposomes was a dose-dependent process. An examination of the dose dependency of the distribution of the administered liposomes in the blood and liver at 23 hours post-injection, and of the kinetics of the elimination of these vesicles from the blood, suggests a hepatic uptake process involving two parallel pathways. One pathway is apparently a capacity-limited Michaelis-Menten process; the other pathway is a linear, non-saturable process. These pathways operate in parallel but respectively dominate at the low end and the high end of the dose range examined.
Asunto(s)
Colesterol/sangre , Liposomas/metabolismo , Esfingomielinas/sangre , Animales , Bovinos , Relación Dosis-Respuesta a Droga , Inyecciones Intravenosas , Cinética , Liposomas/administración & dosificación , Hígado/metabolismo , Ratones , Ratones Endogámicos BALB C , Distribución TisularRESUMEN
The kinetics of hepatic uptake and degradation of sphingomyelin/cholesterol (2:1, M/M) small unilamellar liposomes were investigated in a BALB/c mouse. The tissue distribution of liposomes was determined by scintillation spectrometry. The percentage of intact liposomes in tissues was estimated by the technique of gamma-ray perturbed angular correlation. A kinetic model was developed to analyze the above data. A remarkable agreement was noted between the experimental data and the corresponding theoretical values. Our results indicate that the sphingomyelin/cholesterol unilamellar liposomes had an unusually long half-life of 16.5 hr in the circulation after intravenous administration to mice. The hepatic degradation of the liposomes in vitro at 37 degrees C followed first-order kinetics, with a half-life of 3.5 +/- 0.2 (SEM) hr. Furthermore, the rate of the in vivo degradation of liposomes in the liver was found to be quite similar to that in vitro, with a half-life of 3.6 +/- 0.4 hr. The rate of release of the liposome-encapsulated agent, indium-111, in the liver was not constant, and reached a maximum at about 8 hr after the administration of liposomes. The approach developed in the present study is general and can be applied to the investigation of factors that may control the release of pharmacologically active agents in any tissue.