ABSTRACT
In recent years, many experiments have been conducted for the production and evaluation of anticancer glycoconjugated vaccines in developed countries and many achievements have been accomplished with Globo H derivatives. In the current experiment, a new chemically designed triplicate version of (Globo H)3-diethylenetriamine pentaacetic acid (DTPA)-KLH antigen was synthesized and characterized. Immunization with (Globo H)3-DTPA-KLH, a hexasaccharide that is a member of a family of antigenic carbohydrates that are highly expressed in various types of cancers conjugated with DTPA and KLH protein, induced a high level of antibody titer along with an elevated level of IL-4 in mice. Treatment of tumors with the collected sera from immunized mice decreased the tumor size in nude mice as well. None of the immunized mice illustrated any sign of tumor growth after injection of MCF-7 cells compared to the control animals. These findings, based on the newly presented structure of the Globo H antigen, lend exciting and promising evidence for clinical advancement in the development of a therapeutic vaccine in the future.
Subject(s)
Antigens, Tumor-Associated, Carbohydrate/chemistry , Antigens, Tumor-Associated, Carbohydrate/immunology , Cancer Vaccines/chemistry , Hemocyanins/chemistry , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/therapy , Pentetic Acid/chemistry , Adjuvants, Immunologic , Animals , Cancer Vaccines/immunology , Drug Design , Female , Hemocyanins/immunology , Humans , MCF-7 Cells , Mice , Mice, Nude , Molecular Structure , Pentetic Acid/immunologyABSTRACT
D2B is a new monoclonal antibody directed against an extracellular domain of prostate-specific membrane antigen (PSMA), which is overexpressed in prostate cancer. The potential of D2B IgG, and F(ab')2 and Fab fragments of this antibody for targeting prostate cancer was determined in mice bearing subcutaneous prostate cancer xenografts. The optimal time point for imaging was determined in biodistribution and microSPECT imaging studies with (111)In-D2B IgG, (111)In-capromab pendetide, (111)In-D2B F(ab')2 and (111)In-D2B Fab fragments in mice with PSMA-expressing LNCaP and PSMA-negative PC3 tumors at several time points after injection. All (111)In-labeled antibody formats specifically accumulated in the LNCaP tumors, with highest uptake of (111)In-D2B IgG and (111)In-capromab pendetide at 168 h p.i. (94.8 ± 19.2% injected dose per gram (ID/g) and 16.7 ± 2.2% ID/g, respectively), whereas uptake of (111)In-D2B F(ab')2 and (111)In-D2B Fab fragments peaked at 24 h p.i. (12.1 ± 3.0% ID/g and 15.1 ± 2.9% ID/g, respectively). Maximum LNCaP tumor-to-blood ratios were 13.0 ± 2.3 (168 h p.i.), 6.2 ± 0.7 (24 h p.i.), 23.0 ± 4.0 (24 h p.i.) and 4.5 ± 0.6 (168 h p.i.) for (111)In-D2B IgG, (111)In-F(ab')2, (111)In-Fab and (111)In-capromab pendetide, respectively. LNCaP tumors were clearly visualized with microSPECT with all antibody formats. This study demonstrates the feasibility of D2B IgG, F(ab')2 and Fab fragments for targeting PSMA-expressing prostate cancer xenografts.
Subject(s)
Antibodies, Monoclonal/immunology , Antigens, Surface/immunology , Glutamate Carboxypeptidase II/immunology , Immunoglobulin G/administration & dosage , Pentetic Acid/analogs & derivatives , Prostatic Neoplasms/therapy , Animals , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/chemistry , Antigens, Surface/biosynthesis , Cell Line, Tumor , Glutamate Carboxypeptidase II/biosynthesis , Humans , Immunoglobulin Fab Fragments/administration & dosage , Immunoglobulin Fab Fragments/immunology , Immunoglobulin G/chemistry , Immunoglobulin G/immunology , Male , Mice , Pentetic Acid/administration & dosage , Pentetic Acid/chemistry , Pentetic Acid/immunology , Prostatic Neoplasms/immunology , Prostatic Neoplasms/pathology , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: Bombesin has been used to target Bombesin receptor, a growth receptor, which is over-expressed in many cancers, including prostate cancer. Polymer-anti-neoplastic-drug-conjugates (PDC) were also developed to reduce non-specific toxicity and increase tumor toxicity utilizing the enhanced permeability and retention effect, benefitting treatment of large tumors with well-established vasculature. PURPOSE: If PDCs were delivered by targeted delivery to cancer cells, tumor toxicity would be enhanced and non-specific toxicity decreased. METHODS: Cardiocyte toxicity was assessed in H9c2 cardiocytes with doxorubicin (Dox) or N-terminal DTPA-modified-Doxorubicin-loaded-polyglutamic acid polymers (D-Dox-PGA). Therapeutic efficacy of targeted D-Dox-PGA after pretargeting with Bombesin-conjugated anti-DTPA-antibody Bispecific Complexes (Bom-BiSpCx) was compared to that of Dox in PC3 cells. Bom-BiSpCx was generated by thioether bond between Bombesin to Anti-DTPA antibody. RESULTS: D-Dox-PGA was demonstrated to have less cardiocyte toxicity (IC50 = 20 µg/ml) than free Dox (1.55 µg/ml, p < 0.001). However, after pre-targeting of human prostate cancer PC3 cells with Bom-BiSpCx and targeting with D-Dox-PGA, IC50 (13.2 µg/ml) was about two times less than that of Dox (28.5 µg/ml, p < 0.0001). DISCUSSION: Targeted delivery of PDCs having lower cardiocyte toxicity enabled higher efficiency cancer cell therapy. CONCLUSION: This study may allow development of very efficient targeted prostate cancer pro-drug therapy.
Subject(s)
Bombesin/pharmacology , Doxorubicin/pharmacology , Drug Delivery Systems , Prostatic Neoplasms/drug therapy , Animals , Antibiotics, Antineoplastic/administration & dosage , Antibiotics, Antineoplastic/pharmacology , Antibiotics, Antineoplastic/toxicity , Antibodies, Bispecific/immunology , Bombesin/administration & dosage , Cell Line , Cell Line, Tumor , Doxorubicin/administration & dosage , Doxorubicin/toxicity , Drug Carriers/chemistry , Humans , Inhibitory Concentration 50 , Male , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/metabolism , Pentetic Acid/immunology , Polyglutamic Acid/chemistry , Prostatic Neoplasms/pathology , Rats , Receptors, Bombesin/metabolismABSTRACT
CONTEXT: The advent of monoclonal antibodies such as Rituximab, in recent years, has brought about decisive progress in the treatment of aggressive and indolent non-Hodgkin's lymphoma. AIMS: A further tried and tested improvement to the unmodified antibody has been its coupling to the beta-emitters Y-90. The optimization of 90 Y-antiCD20 radioimmunoconjugate production and quality control methods for future clinical studies in the country was targeted in this work. MATERIALS AND METHODS: The antibody was labeled with 90 Y-yttrium chloride (185 MBq) after conjugation with freshly prepared ccDTPA. Y-90 chloride was obtained by thermal neutron flux (4 × 10 13 n/cm 2 /s) of a natural Y 2 O 3 sample, dissolved in acidic media. Radiolabeling was completed in 24 h by the addition of DTPA-Rituximab conjugate at room temperature. STATISTICAL ANALYSIS USED: All values were expressed as mean ± standard deviation (mean ± SD), and the data were compared using Student's t-test. Statistical significance was defined as P < 0.05. RESULTS: Radiochemical purity of 96% was obtained by using ITLC method for the final radioimmunoconjugate (specific activity = 440-480 MBq/mg). The final isotonic 90 Y-Rituximab complex was checked by gel electrophoresis for protein integrity retention. Biodistribution studies in normal rats were carried out to determine the radioimmunoconjugate distribution up to 72 h. CONCLUSION: The results showed that 90 Y-DTPA-Rituximab could be considered for further evaluation in animals and possibly in humans as a radiopharmaceutical for use in radioimmunotherapy against non-Hodgkin's lymphomas. Because of the importance of developing anti-lymphoma B agents in nuclear medicine for country use, an optimized radiolabeling method has been introduced.
Subject(s)
Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/immunology , Antigens, CD20/immunology , Immunoconjugates/administration & dosage , Radioimmunotherapy/methods , Yttrium Radioisotopes/administration & dosage , Animals , Antibodies, Monoclonal, Murine-Derived/administration & dosage , Antibodies, Monoclonal, Murine-Derived/immunology , Antigens, CD20/administration & dosage , Humans , Immunoconjugates/immunology , Lymphoma, Non-Hodgkin/immunology , Lymphoma, Non-Hodgkin/radiotherapy , Male , Pentetic Acid/administration & dosage , Pentetic Acid/immunology , Radiopharmaceuticals/administration & dosage , Radiopharmaceuticals/immunology , Rats , RituximabABSTRACT
UNLABELLED: The prognosis of medullary thyroid carcinoma (MTC) varies from long- to short-term survival based on such prognostic factors as serum calcitonin and carcinoembryonic antigen (CEA) doubling times (DTs). This prospective phase II multicenter trial evaluated the efficacy and safety of anti-CEA pretargeted radioimmunotherapy (pRAIT) in rapidly progressing metastatic MTC patients and also how serum biomarker DTs correlate with clinical outcome. METHODS: From June 2004 to January 2008, 42 patients were treated with anti-CEA × anti-diethylenetriaminepentaacetic acid (DTPA) bispecific antibody (hMN-14 × m734) (40 mg/m(2)), followed by (131)I-di-DTPA-indium bivalent hapten (1.8 GBq/m(2)) 4-6 d later. RESULTS: The disease control rate (durable stabilization plus objective response) was 76.2%. Grade 3-4 hematologic toxicity was observed in 54.7% of patients and myelodysplastic syndrome in 2, including 1 heavily treated previously. After pRAIT, 21 of 37 assessed patients (56.7%) showed a significant impact on DT (≥100% increase of pre-pRAIT calcitonin or CEA DT or prolonged decrease of the biomarker concentration after pRAIT). Pre-pRAIT DT and post-pRAIT DT were significant independent predictors for overall survival (OS) from pRAIT (pre-pRAIT: hazard ratio [HR], 0.46; 95% confidence interval [CI], 0.24-0.86; P = 0.016; and post-pRAIT: HR, 5.32; 95% CI, 1.63-17.36; P = 0.006) and OS from diagnosis (pre-pRAIT: HR, 0.21; 95% CI, 0.08-0.51; P = 0.001; and post-pRAIT: HR, 6.16; 95% CI, 1.81-20.98; P = 0.004). CONCLUSION: pRAIT showed antitumor activity, with manageable hematologic toxicity in progressive MTC. Increased biomarker DT after treatment correlated with increased OS.
Subject(s)
Antibodies, Bispecific/therapeutic use , Biomarkers, Tumor/metabolism , Carcinoembryonic Antigen/immunology , Disease Progression , Radioimmunotherapy/methods , Thyroid Neoplasms/pathology , Thyroid Neoplasms/radiotherapy , Adult , Aged , Aged, 80 and over , Animals , Antibodies, Bispecific/adverse effects , Antibodies, Monoclonal, Humanized/adverse effects , Antibodies, Monoclonal, Humanized/therapeutic use , Carcinoma, Neuroendocrine , Humans , Male , Mice , Middle Aged , Neoplasm Metastasis , Pentetic Acid/immunology , Radioimmunotherapy/adverse effects , Survival Analysis , Thyroid Neoplasms/metabolism , Treatment Outcome , Young AdultABSTRACT
PURPOSE: Pretargeting with bispecific monoclonal antibodies (bsMAb) for tumor imaging was developed to enhance target to background activity ratios. Visualization of tumors was achieved by the delivery of mono- and divalent radiolabeled haptens. To improve the ability to image tumors with bsMAb, we have combined the pretargeting approach with targeting of high specific activity radiotracer labeled negatively charged polymers. The tumor antigen-specific antibody was replaced with bombesin (Bom), a ligand that binds specifically to the growth receptors that are overexpressed by many tumors including prostate cancer. Bomanti- diethylenetriaminepentaacetic acid (DTPA) bispecific antibody complexes were used to demonstrate pretargeting and imaging of very small human prostate cancer xenografts targeted with high specific activity ¹¹¹In- or 99mTc-labeled negatively charged polymers. METHODS: Bispecific antibody complexes consisting of intact anti-DTPA antibody or Fab' linked to Bom via thioether bonds (Bom-bsCx or Bom-bsFCx, respectively) were used to pretarget PC-3 human prostate cancer xenografts in SCID mice. Negative control mice were pretargeted with Bom or anti-DTPA Ab. 111In-Labeled DTPA-succinyl polylysine (DSPL) was injected intravenously at 24 h (7.03 ± 1.74 or 6.88 ± 1.89 MBq ¹¹¹In-DSPL) after Bom-bsCx or 50 ± 5.34 MBq of 99mTc-DSPL after Bom-bsFCx pretargeting, respectively. Planar or single photon emission computed tomography (SPECT)/CT gamma images were obtained for up to 3 h and only planar images at 24 h. After imaging, all mice were killed and biodistribution of 111In or 99mTc activities were determined by scintillation counting. RESULTS: Both planar and SPECT/CT imaging enabled detection of PC-3 prostate cancer lesions less than 1-2 mm in diameter in 1-3 h post 111In-DSPL injection. No lesions were visualized in Bom or anti-DTPA Ab pretargeted controls. 111In-DSPL activity in Bom-bsCx pretargeted tumors (1.21 ± 0.36 %ID/g) was 5.4 times that in tumors pretargeted with Bom or anti-DTPA alone (0.22 ± 0.08, p = 0.001). PC-3 xenografts pretargeted with Bom-bsFCx and targeted with 99mTc-DSPL were visualizable by 1-3 h. Exquisite tumor uptake at 24 h (6.54 ± 1.58 %ID/g) was about 15 times greater than that of Bom pretargeted controls (0.44 ± 0.17, p = 0.002). CONCLUSION: Pretargeting prostate cancer with Bom-bsCx or Bom-bsFCx enabled fast delivery of high specific radioactivity ¹¹¹In- or 99mTc-labeled polymer-drug conjugates resulting in visualization of lesions smaller than 1- 2 mm in diameter within 3 h.
Subject(s)
Antibodies, Bispecific/metabolism , Bombesin/immunology , Cell Transformation, Neoplastic , Molecular Imaging/methods , Polylysine/chemistry , Prostatic Neoplasms/pathology , Tumor Burden , Animals , Antibodies, Bispecific/chemistry , Antibodies, Bispecific/immunology , Antibodies, Monoclonal/immunology , Antineoplastic Agents/chemistry , Cell Line, Tumor , Humans , Immunoglobulin Fab Fragments/immunology , Indium Radioisotopes , Isotope Labeling , Male , Mice , Mice, Inbred C57BL , Organotechnetium Compounds , Pentetic Acid/immunology , Sulfides/chemistryABSTRACT
BACKGROUND: Antibody-based imaging agents are available commercially, but their success has been limited, mainly because of low contrast and the emergence of 2-fluoro-2-deoxy-D-glucose-positron emission tomography (FDG-PET) scanning. In pretargeting, administration of the radionuclide is separated from the antibody, thereby enhancing image contrast and allowing detection at earlier time points after injection. METHODS: The authors conducted an open-label, single-arm trial that assessed a pretargeting procedure in which an anticarcinoembryonic antigen x (anti-CEA x) anti-diethylenetriaminepentaacetic acid (anti-DTPA)-indum (In) antibody was used in combination with a (111)In-labeled di-DTPA peptide for the diagnostic imaging of CEA-expressing colorectal cancer. Three patients received the (111)In peptide alone to investigate tumor targeting, organ distribution, and clearance of the peptide. Thereafter, 11 patients received the bispecific antibody (bsAb) (5 mg) to pretarget the tumor. After 3 to 5 days, patients were injected with 185 megabecquerels of (111)In-labeled peptide to assess the optimal interval for best image quality. RESULTS: Fourteen patients with primary colorectal cancer were enrolled. One of 3 patients who received (111)In peptide alone had low-level tumor uptake. In 9 of 11 other patients, tumors were observed. In 1 patient, FDG-PET-positive lymph nodes were observed clearly with pretargeted immunoscintigraphy. Peptide pharmacokinetics revealed enhanced circulating levels of (111)In-labeled peptide in patients in the 3-day interval cohort compared with the other cohorts. Tumor-to-background ratios ranged from 3.5 to 6.4 in the 3-day interval group, from 5.1 to 14.2 in the 4-day interval group, and from 3.5 to 3.9 in the 5-day interval group. The best images were acquired with a 4-day interval at 24 hours after injection of the radiolabeled peptide. Grade 1 adverse events were observed in 2 patients. CONCLUSIONS: Imaging of colorectal cancer using a 2-step, pretargeting system produced the best imaging results 24 hours after peptide administration using a 4-day interval between injection of the bsAb and the peptide.
Subject(s)
Antibodies, Bispecific , Carcinoembryonic Antigen/immunology , Colorectal Neoplasms/diagnostic imaging , Immunoglobulin Fab Fragments , Pentetic Acid/immunology , Radioimmunodetection/methods , Receptors, Immunologic , Aged , Aged, 80 and over , Feasibility Studies , Female , Haptens , Humans , Indium Radioisotopes , Male , Middle Aged , Radioimmunodetection/adverse effectsABSTRACT
OBJECTIVE: The purpose of this study was to explore the development of a pre-clinical nuclear imaging model as a tool for testing novel radiopharmaceutical agents for imaging and/or delivery systems to human tissues. Here we report for the first time the imaging of human synovial tissue transplanted into SCID mice using a radiolabelled anti-E-selectin antibody and NanoSPECT/CT technology. METHODS: Human synovium was transplanted into SCID mice. Two to three weeks post-transplantation tissue vasculature was stimulated with TNF-alpha by intra-graft injection 5 h prior to intravenous injection of (111)In-labelled anti-E-selectin or isotype control antibody. At 1, 4, 24 and 48 h animals were imaged and transplant activity quantified. RESULTS: Activity was detectable in the grafts at all time points, with clear delineation of the transplants in the reconstructed images. A significant difference in graft radioactivity was observed at 4 and 24 h with a significantly higher uptake (P < 0.05) of (111)In-anti-E-selectin compared with isotype control antibody. CONCLUSIONS: This article highlights NanoSPECT/CT imaging in the SCID mouse chimeric model as a powerful tool for the pre-clinical development of radiopharmaceutical and delivery agents targeting human synovial tissue in vivo.
Subject(s)
Osteoarthritis/immunology , Synovial Membrane/blood supply , Animals , Autoantibodies/immunology , E-Selectin/immunology , Enzyme-Linked Immunosorbent Assay/methods , Humans , Immunohistochemistry , Indium Radioisotopes , Mice , Mice, SCID , Models, Animal , Pentetic Acid/immunology , Radiopharmaceuticals , Synovial Membrane/transplantation , Tomography, Emission-Computed, Single-Photon/methods , Transplantation, HeterologousABSTRACT
Gene expression analysis showed that a human mindin homologue, mindin/RG-1, is expressed selectively in prostate tissues and that its expression level is elevated in some prostate tumors. Mindin/RG-1 protein expression is maintained in >80% of prostate cancers metastatic to bone or lymph nodes as well as in locally recurrent tumors in androgen-unresponsive patients. In contrast, mindin/RG-1 expression in other normal tissues is significantly lower than that seen in the prostate. A fully human antibody, 19G9, was generated against mindin/RG-1 protein and was shown to accumulate at high abundance in LNCaP tumor xenografts. Conjugates of this antibody with the chelator CHX-A''-DTPA were generated and radiolabeled with either 111In, 90Y, or 86Y. Small animal positron emission tomography imaging with the 86Y-radiolabeled conjugate showed very specific accumulation of the antibody in LNCaP tumor xenografts with clear tumor delineation apparent at 4 hours. The therapeutic efficacy of [90Y]-CHX-A''-DTPA-19G9 was evaluated in mice bearing LNCaP xenografts. A dose-finding study identified a nontoxic therapeutic dose to be approximately 75 microCi. Significant antitumor effects were seen with a single administration of radiolabeled antibody to animals bearing 200 to 400 mm3 tumors. Inhibition of tumor growth was observed in all treated animals over a 49-day period. At 49 days posttreatment, slow tumor growth recurred but this could be prevented for an additional 40-day period by a second administration of a 75 microCi dose at day 49. We conclude that [90Y]-CHX-A''-DTPA-19G9 is a novel antibody conjugate that has considerable promise for therapy of metastatic prostate cancer in androgen-unresponsive patients.
Subject(s)
Antibodies, Monoclonal/immunology , Extracellular Matrix Proteins/immunology , Immunotoxins/immunology , Prostatic Neoplasms/radiotherapy , Radioimmunotherapy/methods , Amino Acid Sequence , Animals , Antibodies, Monoclonal/pharmacokinetics , Antibodies, Monoclonal/pharmacology , Antibody Specificity , Bone Neoplasms/metabolism , Bone Neoplasms/secondary , CHO Cells , Cricetinae , Dose-Response Relationship, Immunologic , Extracellular Matrix Proteins/biosynthesis , Extracellular Matrix Proteins/genetics , Humans , Immunotoxins/pharmacokinetics , Immunotoxins/pharmacology , Isothiocyanates/immunology , Isothiocyanates/pharmacokinetics , Isothiocyanates/pharmacology , Male , Molecular Sequence Data , Pentetic Acid/analogs & derivatives , Pentetic Acid/immunology , Pentetic Acid/pharmacokinetics , Pentetic Acid/pharmacology , Positron-Emission Tomography , Prostatic Neoplasms/diagnostic imaging , Prostatic Neoplasms/immunology , Prostatic Neoplasms/metabolism , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Tissue Distribution , Xenograft Model Antitumor Assays , Yttrium Radioisotopes/administration & dosage , Yttrium Radioisotopes/pharmacokinetics , Yttrium Radioisotopes/pharmacologyABSTRACT
The molecular characterization of antibodies directed against metal-chelate complexes will provide important insights into the design and development of radiotherapeutic and radioimaging reagents. In this study, two monoclonal antibodies directed against different metal-chelate complexes were expressed as recombinant Fab fragments. Covalent modification and site-directed mutagenesis were employed to ascertain those residues important in antigen recognition. Antibody 5B2 was raised to a Pb(II)-loaded isothiocyanatobenzyl-diethylenetriamine pentaacetic acid (DTPA)-protein conjugate. The native antibody bound to complexes of Pb(II)-p-aminobenzyl-DTPA with an affinity of 4.6 x 10(-9) M. A monovalent Fab fragment prepared from the native protein and a bivalent recombinant fragment exhibited comparable affinities for the same Pb(II)-chelate complex, approximately 6-fold lower than that of the intact antibody. Covalent modification and molecular modeling predicted that Lys(58) in the heavy chain contacted the Pb(II)-chelate ligand. Mutational analysis supported a role for Lys(58) in ion pair or hydrogen bond formation with the carboxylate groups on the chelate. Antibody E5 was directed toward an isothiocyanatobenzyl-ethylenediamine tetraacetic acid (EDTA)-protein conjugate loaded with ionic Cd(II). The native immunoglobulin recognized Cd(II)-p-aminobenzyl-EDTA with an affinity of 8.2 x 10(-12) M. A proteolytically derived fragment and a bivalent recombinant fragment bound to the same Cd(II)-chelate complex with affinities that were comparable to that of the native antibody. Homology modeling and mutagenesis identified three residues (Trp(52) and His(96) in the heavy chain and Arg(96) in the light chain) that were important for Cd(II)-chelate recognition. His(96) likely mediates a direct ligation to the Cd(II) ion and Trp(52) appears to be involved in hydrophobic stacking with the benzyl moiety of the chelator. Arg(96) appeared to mediate an electrostatic or hydrogen bond to the chelate portion of the complex. These studies demonstrate that antibody recognition of metal-chelate haptens occurs through a limited number of molecular contacts and that these molecular interactions involve both direct ligation between the antibody and the metal ion and interactions between the antibody and the chelator.
Subject(s)
Amino Acids/chemistry , Antibodies, Monoclonal/chemistry , Binding Sites, Antibody , Chelating Agents/chemistry , Metals, Heavy/chemistry , Amino Acid Substitution/genetics , Amino Acids/genetics , Animals , Antibodies, Monoclonal/genetics , Antigens/chemistry , Antigens/immunology , Binding Sites, Antibody/genetics , Edetic Acid/chemistry , Edetic Acid/immunology , Immunoassay , Immunoglobulin G/chemistry , Lead/chemistry , Lead/immunology , Lysine/chemistry , Metals, Heavy/immunology , Mice , Mice, Inbred BALB C , Models, Molecular , Pentetic Acid/chemistry , Pentetic Acid/immunology , Protein Structure, Tertiary/genetics , Recombinant Proteins/chemistryABSTRACT
La tos ferina todavía es una enfermedad no bien controlada desde el punto de vista epidemiológico. En España su incidencia ha disminuido notablemente, en paralelo con el aumento de cobertura vacunal de la población infantil, pero en el año 2000 aún se declararon 906 casos. Aunque la cobertura vacunal de la primovacunación es muy buena, la administración de las dosis de refuerzo de los 18 meses y los 4-6 años ha sido poco frecuente en la mayoría de comunidades, por lo que el nivel de inmunidad de la población es deficiente. Esta falta de control de la tos ferina se debe a una subóptima cobertura vacunal, consecuente a la elevada frecuencia de efectos adversos de la vacuna de tos ferina de célula completa (Pw) y a la pérdida de inmunidad protectora en adolescentes y adultos, que constituyen la principal fuente de contagio de los niños pequeños. La solución a estos problemas pasa por la utilización de las nuevas vacunas acelulares de tos ferina (Pa), tanto en la primovacunación como en las dosis de refuerzo, para conseguir mantener una inmunidad prolongada. Las vacunas DTPa con Pa de tres o más componentes protegen mejor que las de uno o dos componentes, y con una eficacia similar a la de las vacunas DTPw. Los efectos adversos de las vacunas DTPa son menores que los de las vacunas DTPw. En los países desarrollados, como España, las ventajas de las vacunas combinadas con Pa justifican su uso en sustitución de las vacunas con Pw (AU)
No disponible
Subject(s)
Child , Child, Preschool , Female , Humans , Male , Diphtheria-Tetanus-Pertussis Vaccine/immunology , Diphtheria-Tetanus-Pertussis Vaccine/therapeutic use , Pentetic Acid/immunology , Whooping Cough/immunology , Pertussis Vaccine/adverse effects , Pertussis Vaccine/immunology , Efficacy/trends , Mass Vaccination/organization & administration , Vaccination/methods , Immunization Programs/standardsABSTRACT
Background The introduction of polysaccharide-protein conjugate vaccines against Haemophilus influenzae type b (Hib) in the 1990's resulted in a dramatic reduction in the number of cases of invasive Hib disease. In Germany, following the launch of DTPa/Hib combination vaccines in late 1996 and DTPa-IPV/Hib in 1998, the majority of Hib immunisations are made with such combinations. It is well established that such combinations elicit lower anti-Hib antibody concentrations than the equivalent Hib conjugate administered as a separate injection. While there are good reasons to assume that this is of no clinical relevance the clinical impact of this phenomenon has not yet been fully evaluated. Methods and Findings To assess the impact of DTPa/Hib combination vaccines on the incidence of invasive Hib disease in Germany two independent, one hospital- and one laboratory-based, surveillance systems were used during 1998-99 for detection of cases. Tetra- and penta- valent DTPa-Hib vaccines accounted for 92.1 % of all Hib vaccines used in 1999. During the 2 year study period the annual number of invasive Haemophilus influenzae b disease decreased further from 28 in 1998 to 13 in 99. National vaccination coverage rates for 1998/99 revealed that only 70 % of children given DTPa/Hib or DTPa-IPV/Hib received the recommended 3 doses in their first year of life, the overall effectiveness of the most used combination vaccine was high at 97.4 % (95 % CI: 96.0-98.3) for those who had received at least one dose of such a vaccine. In subjects who received the full 3-dose schedule effectiveness was 98.8 % (95 % CI: 98.1-99.2). Conclusion These data show that the lower anti-Hib antibody concentrations observed with DTPa/Hib combination vaccines are of no clinical relevance (AU)
No disponible
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Pentetic Acid/immunology , Polysaccharides, Bacterial/immunology , Vaccination/methods , Immunization Programs/standards , Immunization Programs , Haemophilus influenzae/immunology , Haemophilus Vaccines/immunology , Haemophilus Vaccines/standards , Haemophilus Infections/epidemiology , Haemophilus Infections/immunology , Mass Vaccination/standards , Immunization/statistics & numerical data , Vaccination/standards , Immunization/standards , Germany/epidemiologyABSTRACT
Uno de los progresos alcanzados por la vacunología en el último decenio ha sido la comercialización de preparados pertenecientes a nuevos grupos de vacunas, entre los que destacan las conjugadas y las combinadas. El mayor beneficio de las vacunas combinadas para los niños es la disminución del número de inyecciones para la protección frente a más enfermedades. La vacuna combinada hexavalente contiene una vacuna combinada trivalente, la DTPa, de 2 o 3 componentes antigénicos de Bordetella pertussis (Pa), en las dos especialidades farmacéuticas comercializadas (Hexavac ® e Infanrix Hexa ®, respectivamente); una vacuna combinada trivalente, pero de un solo microorganismo (poliovirus 1, 2 y 3); una vacuna recombinante (hepatitis B) y una vacuna conjugada de Haemophilus influenzae tipo b (PRP o antígeno polisacarídico capsular conjugado con toxoide tetánico). La inclusión de la vacuna hexavalente en el calendario de inmunizaciones sistemáticas permitirá alcanzar seis importantes objetivos: introducción de la vacuna acelular de la tos ferina en la primovacunación y la revacunación, sustitución de la vacuna antipoliomielítica oral de virus atenuados por la parenteral inactivada, unificación de la vacunación frente a la hepatitis B en el lactante, mantenimiento de la pauta de 4 dosis en la inmunización anti-H. influenzae tipo b, disminución del número de pinchazos y armonización de los 19 calendarios de vacunaciones sistemáticas existentes en España. Con los datos disponibles en la ficha técnica de las dos especialidades farmacéuticas de vacuna hexavalente y de acuerdo con los calendarios de vacunaciones sistemáticas de las comunidades autónomas y el propuesto por el Comité Asesor de Vacunas de la Asociación Española de Pediatría (AEP) (2001-2002), la introducción de la vacuna combinada hexavalente en el calendario vacunal se puede realizar según las pautas siguientes: 1. Lactantes vacunados de hepatitis B en el período neonatal incluidos los hijos de madre HBsAg-positivas o de estatus desconocido. La única vacuna hexavalente que se puede utilizar en esta pauta es Infanrix Hexa ®. 2. Lactantes no vacunados de hepatitis B en el período neonatal (AU)
No disponible
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Vaccines, Combined/immunology , Vaccines, Combined/therapeutic use , Vaccination/methods , Immunization Programs/methods , Hepatitis B/immunology , Haemophilus influenzae type b/immunology , Poliomyelitis/epidemiology , Poliomyelitis/immunology , Vaccines, Combined/administration & dosage , Vaccines, Combined/standards , Pentetic Acid/immunology , Bordetella/immunology , Bordetella Infections/immunology , Immunization Programs/standards , Immunization Programs , Spain/epidemiologyABSTRACT
El número de vacunas recomendadas para los niños es cada vez mayor, por lo que resulta necesario simplificar su administración. Se precisan vacunas combinadas, que contengan múltiples antígenos, para garantizar un adecuado cumplimiento del calendario de vacunaciones, tanto por parte de los padres como de los profesionales de la salud. Recientemente se han comercializado en España dos vacunas combinadas hexavalentes que permiten la vacunación frente a seis antígenos con una sola inyección. Este artículo revisa la experiencia clínica en España con la vacuna hexavalente de los laboratorios GlaxoSmithKline (Infanrix Hexa ®) (AU)
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Subject(s)
Female , Humans , Infant , Male , Vaccines, Combined/administration & dosage , Vaccines, Combined/immunology , Pentetic Acid/immunology , Vaccination/methods , Immunization Programs/methods , Immunization Programs , Diphtheria Toxoid/immunology , Diphtheria-Tetanus-Pertussis Vaccine/immunology , Diphtheria-Tetanus Vaccine/immunology , Vaccines, Combined/standards , Immunization Programs/organization & administration , Spain/epidemiology , Follow-Up StudiesABSTRACT
Detailed equilibrium binding studies were conducted on a monoclonal antibody directed against Pb(II) complexed with a protein conjugate of diethylenetriaminepentaacetic acid (DTPA). Binding curves obtained with DTPA and a cyclohexyl derivative of DTPA in the presence and absence of metal ions were consistent with the anticipated one-site homogeneous binding model. Binding curves obtained with aminobenzyl-DTPA or its complexes with Ca(II), Sr(II), and Ba(II) were highly sigmoidal, characterized by Hill coefficients of 2.3-6.5. Binding curves obtained with the Pb(II) and In(III) complexes of aminobenzyl-DTPA were hyperbolic, but in each case the apparent affinity of the antibody for the chelator-metal complex was higher in the presence of excess chelator than it was in the presence of excess metal ion. In the presence of excess chelator, the equilibrium dissociation constant for the binding of aminobenzyl-DTPA-Pb(II) to the antibody was 9.5 x 10(-)(10) M. Binding curves obtained with the Hg(II) and Cd(II) complexes of aminobenzyl-DTPA were biphasic, indicative of negative cooperativity. Further binding studies demonstrated that aminobenzyl-DTPA-Hg(II) opposed the binding of additional chelator-metal complexes to the antibody more strongly than did aminobenzyl-DTPA-Cd(II). The Fab fragment differed from the intact antibody only in that the apparent affinity of the Fab was generally lower for a given chelator-metal complex. These data are interpreted in terms of a model in which (i) aminobenzyl-DTPA and its complexes bind both to the antigen binding site and to multiple charged sites on the surface of the compact immunoglobulin; and (ii) the bound, highly charged ligands interact in a complicated fashion through the apolar core of the folded antibody.
Subject(s)
Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/metabolism , Binding Sites, Antibody , Immunoglobulin Fab Fragments/chemistry , Immunoglobulin Fab Fragments/metabolism , Pentetic Acid/analogs & derivatives , Allosteric Regulation , Amino Acid Sequence , Animals , Binding, Competitive , Cadmium/immunology , Enzyme-Linked Immunosorbent Assay , Female , Haptens/administration & dosage , Haptens/immunology , Hemocyanins/administration & dosage , Hemocyanins/immunology , Hybridomas , Injections, Intravenous , Lead/administration & dosage , Lead/immunology , Mercury Compounds/immunology , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Pentetic Acid/administration & dosage , Pentetic Acid/immunologyABSTRACT
A monoclonal antibody (2C12) that recognizes a Pb(II)-cyclohexyldiethylenetriamine pentaacetic acid complex was produced by the injection of BALB/c mice with a Pb(II)-chelate complex covalently coupled to a carrier protein. The ability of purified antibody to interact with a variety of metal-free chelators and metal-chelate complexes was assessed by measuring equilibrium dissociation constants. The antibody bound to metal-free trans-cyclohexyldiethylenetriamine pentaacetic acid (CHXDTPA) with an equilibrium dissociation constant of 2.3 x 10(-)(7) M. Addition of Pb(II) increased the affinity of the antibody for the complex by 25-fold; Pb(II) was the only metal cation (of 15 different di-, tri-, and hexavalent metals tested) that increased the affinity of the antibody for CHXDTPA. The increased affinity was due primarily to an increase in the association rate constant. The antibody also had the ability to interact with ethylenediamine tetraacetic acid (EDTA), diethylenetriamine pentaacetic acid (DTPA), and structurally related derivatives, but with affinities from 50- to 10000-fold less than that determined for CHXDTPA. Addition of metals to EDTA-based chelators reduced the affinity of the antibody for these ligands. However, when DTPA was used as the chelator, addition of Pb(II) increased the affinity of the antibody for the complex by 200-fold. The sensitivity of prototype immunoassays for Pb(II) could be modulated by changing the structure of the immobilized metal-chelate complex and/or the soluble chelator used to complex Pb(II) in the test solution.
Subject(s)
Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/chemistry , Chelating Agents , Lead , Organometallic Compounds/immunology , Amino Acid Sequence , Animals , Antibodies, Monoclonal/isolation & purification , Antibody Specificity , Base Sequence , Chelating Agents/chemistry , Female , Hybridomas , Immunoassay/methods , Kinetics , Lead/chemistry , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Organometallic Compounds/chemistry , Pentetic Acid/analogs & derivatives , Pentetic Acid/immunology , Sequence AnalysisABSTRACT
As small cell lung carcinoma (SCLC) is frequently a widespread disease at diagnosis, highly radiosensitive and often only partially responsive to chemotherapy, radioimmunotherapy (RIT) would appear to be a promising technique for treatment. We report the preliminary results of a Phase I/II trial of RIT in SCLC using a two-step method and a myeloablative protocol with circulating stem cells transplantation. Fourteen patients with proved SCLC relapse after chemotherapy were treated with RIT. They were first injected i.v. with a bispecific (anti-carcinoembryonic antigen/anti-diethylenetriaminepentaacetic acid) monoclonal antibody (20-80 mg in 100 ml of saline solution) and then 4 days later with di-(In-diethylenetriaminepentaacetic acid)-tyrosyl-lysine hapten labeled with 1.48-6.66 GBq (40-180 mCi) of I-131 and diluted in 100 ml of saline solution. In patients receiving 150 mCi or more, circulating stem cells were harvested before treatment and reinfused 10-15 days later. Treatment response was evaluated by CT and biochemical data during the month before and 1, 3, 6, and 12 months after treatment. All patients received the scheduled dose without immediate adverse reactions to bispecific antibody or 1-131 hapten. Toxicity was mainly hematological, with two cases of grade 2 leukopenia and three cases of grade 3 or 4 thrombopenia. Body scanning 8 days after injection of the radiolabeled hapten generally showed good uptake at the tumor sites. Estimated tumor dose was 2.6-32.2 cGy/mCi. Among the 12 patients evaluated to date, we have observed 9 progressions, 2 partial responses (one almost complete for 3 months), and 1 stabilization of more than 24 months. Efficiency and toxicity were dose-related. The maximal tolerable dose without hematological rescue was 150 mCi. These preliminary results are encouraging, and dose escalation is currently continuing to reach 300 mCi. RIT should prove to be an interesting therapeutic method for SCLC, although repeated injections and hematological rescue will probably be required, as well as combination with other treatment modalities.
Subject(s)
Antibodies, Bispecific/therapeutic use , Carcinoembryonic Antigen/immunology , Carcinoma, Small Cell/radiotherapy , Haptens/therapeutic use , Iodine Radioisotopes/therapeutic use , Lung Neoplasms/radiotherapy , Pentetic Acid/immunology , Radioimmunotherapy , Aged , Animals , Antibodies, Anti-Idiotypic/blood , Female , Humans , Male , Mice , Middle Aged , Radiotherapy DosageABSTRACT
UNLABELLED: The alpha-particle-emitting radionuclides have several physical characteristics that make them attractive candidates for radioimmunotherapy: (a) high linear energy transfer; (b) short path lengths (50-80 microm); and (c) limited ability of cells to repair damage to DNA. This article describes the pharmacokinetic, bioactivity, toxicity and chemical characteristics of alpha-particle-emitting, 213Bi and 212Bi radiometal conjugated HuM195 (anti-CD33) constructs. Conjugation of HuM195 to SCN-CHX-A-DTPA resulted in the attachment of up to 10 chelating ligand molecules per antibody. RESULTS: Radiolabeling efficiency of the CHX-A-DTPA-HuM195 construct with 213Bi was 78%+/-10% (n = 46) after 10 min at specific activities of up to 1110 MBq/mg. The immunoreactivity of the 213Bi-labeled CHX-A-DTPA-HuM195 construct was 84%+/-10% (n = 28) and was independent of the specific activity. The bismuth-labeled CHX-A-DTPA-HuM195 construct was rapidly internalized into the cell in a time-dependent manner ranging from 50% at 1 h to 65% at 24 h. 205Bi/206Bi-labeled constructs were stable for at least 2 d in vitro in the presence of human serum at 37 degrees C. After injection into mice, there was no uptake or loss of bismuth to mouse tissues, which do not express CD33, or to the kidney, which has avidity for free bismuth. Mice injected intraperitoneally with doses of (213Bi)CHX-A-DTPA-HuM1 95 ranging from 18.5 to 740 MBq/kg showed no toxicity, but at 2590 MBq/kg, two of the three mice died within 2 wk and a third mouse showed significant reductions in white blood cell counts. Mice injected intravenously with doses of (213Bi)CHX-A-DTPA-HuM195 up to 370 MBq/kg exhibited little toxicity, but 666 MBq/kg was above the MTD for mice. Leukemia cell killing in vitro with bismuth-labeled HuM1 95 showed dose- and specific activity-dependent killing of CD33+ HL60 cells; approximately 50% killing was observed when two bismuth atoms (50 fM radiolabeled antibody) were initially bound onto the target cell surface. CONCLUSION: Alpha-emitting antibodies are among the most potent cytotoxic agents known, yet are specific and appear safe in vivo. The physical and biochemical characteristics of the 213Bi isotope and its generation, as well as the biochemistry of the 213Bi-labeled CHX-A-DTPA-HuM195 construct, make it possible to use the constructs safely and feasibly in humans at therapeutic levels.
Subject(s)
Antibodies, Monoclonal , Antigens, CD/immunology , Antigens, Differentiation, Myelomonocytic/immunology , Pentetic Acid/analogs & derivatives , Alpha Particles , Animals , HL-60 Cells/pathology , Humans , Mice , Mice, Inbred BALB C , Pentetic Acid/chemistry , Pentetic Acid/immunology , Pentetic Acid/pharmacokinetics , Pentetic Acid/toxicity , Radioimmunotherapy , Recombinant Proteins , Sialic Acid Binding Ig-like Lectin 3ABSTRACT
UNLABELLED: The purpose of this study was to compare the toxicity and efficacy of two-step radioimmunotherapy using a bispecific anticarcinoembryonic antigen (CEA)/anti-diethylenetriamine pentaacetic acid (DTPA) antibody (F6-734 bispecific monoclonal antibodies (BsMAbs) and an 131I-di-DTPA-TL bivalent hapten with F(ab')2 fragments of the same directly labeled anti-CEA 131-F6. METHODS: Eight groups of nude mice subcutaneously grafted with the human TT medullary thyroid cancer cell line were injected once tumor volume reached about 200 mm3. Two groups received 37 or 92.5 MBq (1 or 2 nmol) 131I-di-DTPA-TL 48 h after injection of 2 or 4 nmol F6-734 BsMAb and two groups received 37 or 92.5 MBq (250 microg) 131I-F6. Four control groups were treated respectively with (a) 92.5 MBq nonspecific 131I-734 fragments, (b) 92.5 MBq 131I-di-DTPA-TL 48 h after injection of a mixture of irrelevant F6-679 (anti-CEA/anti-histamine) and G7A5-734 (anti-melanoma/anti-DTPA) BsMAb, (c) 250 microg nonradiolabeled F6, and 250 microg F6-734 BsMAb and then 48 h later 1.25 nmol of nonradiolabeled hapten. A control group received no injections. Toxicity was evaluated by determining animal weight and the number of leukocytes and platelets, and efficacy by variation in tumor volume and thyrocalcitonin during a 90-d period. Histological analysis of tumors and statistical studies were performed. RESULTS: The time required for the tumor to double in size was respectively 57 and 86 d with 37 and 92.5 MBq F6-734/131I-di-DTPA-TL and 44 and 65 d with 37 and 92.5 MBq 131I-F6. Changes in thyrocalcitonin levels were parallel to those in tumor volume. Weight loss was 5%, leukocyte nadirs respectively 1640+/-838 and 1560+/-1160/mm3 and platelet nadirs 1.46+/-0.52 10(6)/mm3 and 0.73+/-0.38 10(6)/mm3 after injections of 37 and 92.5 MBq F6-734/1311-di-DTPA-TL. Weight loss was respectively 8% and 16%, leukocyte nadirs 50+/-100/mm3 and 175+/-50/mm3 and platelet nadirs 0.71+/-0.18 10(6)/mm3 and 0.48+/-0.11 10(6)/mm3 after injections of 37 and 92.5 MBq 131I-F6. CONCLUSION: Two-step radioimmunotherapy was as efficient as the one-step system and markedly less toxic.
Subject(s)
Antibodies, Bispecific/therapeutic use , Antibodies, Monoclonal/therapeutic use , Carcinoembryonic Antigen/biosynthesis , Carcinoma, Medullary/radiotherapy , Haptens/therapeutic use , Radioimmunotherapy , Thyroid Neoplasms/radiotherapy , Animals , Carcinoembryonic Antigen/immunology , Carcinoma, Medullary/metabolism , Carcinoma, Medullary/pathology , Humans , Immunoglobulin Fab Fragments/immunology , Iodine Radioisotopes/therapeutic use , Mice , Mice, Nude , Neoplasm Transplantation , Pentetic Acid/immunology , Thyroid Neoplasms/metabolism , Thyroid Neoplasms/pathology , Transplantation, Heterologous , Tumor Cells, CulturedABSTRACT
UNLABELLED: Two-step pretargeting strategies have been designed to deliver radioisotopes to tumors more selectively than directly labeled antibodies or fragments. In this article, we compare quantitatively the potential of these strategies for the radioimmunotherapy of solid tumors. METHODS: Direct targeting was performed using iodine-labeled IgG and F(ab')2. As two-step strategies, we used the sequential injection of anti-CEA x anti-DTPA-In bispecific F(ab')2 (BsF(ab')2) and monovalent and bivalent DTPA derivatives labeled with iodine. The biodistribution of iodine in nude mice grafted with the LS174T human colorectal carcinoma was monitored in time and used for calculating radiation doses. RESULTS: In agreement with earlier studies, the IgG was more effective for delivering a radiation dose to the tumor than the F(ab')2 (7.8 versus 0.76 Gy/MBq, respectively) and both were moderately selective with respect to normal tissues (tumor:blood of 2.9 and 1.7, respectively). At their MTD, they should deliver 86 and 34 Gy, respectively, to the tumor. Using a nM-affinity DTPA-In bivalent hapten, the two-step protocol was optimized by varying the dosage of the BsF(ab')2, the stoichiometry of the reagents and the pretargeting time. The saturation of the tumor was obtained by injecting 5 nmol (500 microg) of BsF(ab')2. The pretargeted BsF(ab')2 was saturated by the injection of 0.5 mol of bivalent hapten per mole of antibody. With a 48-hr pretargeting time, the selectivity of the irradiation of the tumor was optimized (tumor:blood of 7.8) but only at the price of a lower efficiency (0.35 versus 0.86 Gy/MBq, 48-hr and 20-hr pretargeting time, respectively). Attempts to increase selectivity by using a microM-affinity DTPA-Y bivalent hapten or by chasing excess circulating radiolabeled hapten with an excess of unlabeled hapten also reduced tumor exposure. The use of a monovalent hapten resulted in both lower efficiency and selectivity. However, the two-step pretargeting of high-affinity bivalent hapten (Affinity Enhancement System, AES) should deliver 30-60 Gy to the tumor with less than 9 Gy to the blood in tumor-bearing mice. CONCLUSION: Radioimmunotherapy with AES is predicted to be as efficient and with lower hematological toxicity than direct targeting.