Fully human IgG and IgM antibodies directed against the carcinoembryonic antigen (CEA) Gold 4 epitope and designed for radioimmunotherapy (RIT) of colorectal cancers

BACKGROUND: Human monoclonal antibodies (MAbs) are needed for colon cancer radioimmunotherapy (RIT) to allow for repeated injections. Carcinoembryonic antigen (CEA) being the reference antigen for immunotargeting of these tumors, we developed human anti-CEA MAbs. METHODS: XenoMouse(®)-G2 animals were immunized with CEA. Among all the antibodies produced, two of them, VG-IgG2κ and VG-IgM, were selected for characterization in vitro in comparison with the human-mouse chimeric anti-CEA MAb X4 using flow cytometry, surface plasmon resonance, and binding to radiolabeled soluble CEA and in vivo in human colon carcinoma LS174T bearing nude mice. RESULTS: Flow cytometry analysis demonstrated binding of MAbs on CEA-expressing cells without any binding on NCA-expressing human granulocytes. In a competitive binding assay using five reference MAbs, directed against the five Gold CEA epitopes, VG-IgG2κ and VG-IgM were shown to be directed against the Gold 4 epitope. The affinities of purified VG-IgG2κ and VG-IgM were determined to be 0.19 ± 0.06 × 10(8 )M(-1 )and 1.30 ± 0.06 × 10(8 )M(-1), respectively, as compared with 0.61 ± 0.05 × 10(8 )M(-1 )for the reference MAb X4. In a soluble phase assay, the binding capacities of VG-IgG2κ and VG-IgM to soluble CEA were clearly lower than that of the control chimeric MAb X4. A human MAb concentration of about 10(-7 )M was needed to precipitate approximatively 1 ng (125)I-rhCEA as compared with 10(-9 )M for MAb X4, suggesting a preferential binding of the human MAbs to solid phase CEA. In vivo, 24 h post-injection, (125)I-VG-IgG2κ demonstrated a high tumor uptake (25.4 ± 7.3%ID/g), close to that of (131)I-X4 (21.7 ± 7.2%ID/g). At 72 h post-injection, (125)I-VG-IgG2κ was still concentrated in the tumor (28.4 ± 11.0%ID/g) whereas the tumor concentration of (131)I-X4 was significantly reduced (12.5 ± 4.8%ID/g). At no time after injection was there any accumulation of the radiolabeled MAbs in normal tissues. A pertinent analysis of VG-IgM biodistribution was not possible in this mouse model in which IgM displays a very short half-life due to poly-Ig receptor expression in the liver. CONCLUSION: Our human anti-CEA IgG2κ is a promising candidate for radioimmunotherapy in intact form, as F(ab')(2 )fragments, or as a bispecific antibody

Radioprotection associée aux nouvelles évolutions, diagnostiques et thérapeutiques, en médecine nucléaire

Radiation protection for innovative diagnostic and therapeutic approaches in nuclear medicine A real technological revolution has deeply modified the field of application and perspectives of nuclear medicine, and nuclear oncology in particular, during the past 5 years. Diagnostic applications such as positron emission tomography (PET) with 18F-fluorodeoxyglucose (FDG) have had a significant impact on the diagnostic strategy adopted by medical oncologists, with the addition of invaluable functional data to already available anatomical data provided by conventional imaging modalities. Numerous other 18F-labeled tracers currently under clinical evaluation have been developed to study various tumor functions (tumor proliferation, hypoxia, chemotherapy-induced apoptosis, etc.). These tracers may have a considerable impact on therapeutic strategies. Other positron-emitting radionuclides, such as copper-64, iodine-124, and yttrium-86 (whose respective half-lives are 12.7 hours, 4.2 days, and 14.7 hours) will soon be available for certain clinical indications, such as immuno-PET (with monoclonal antibodies or antibody fragments used as carriers) or pretreatment dosimetry, which cannot be performed with fluorine-18 due its short half-life. As far as therapeutic applications are concerned, the use of internal radiotherapy, which has been restricted to thyroid cancer for a long time, was recently extended to other cancers as new carriers, such as monoclonal antibodies (radioimmunotherapy) or peptides (radiopeptide therapy), new targeting methods (pretargeting), and new radionuclides, especially alpha particle emitters (alpha therapy), became available. These technological advances require that specific radiation safety regulations be implemented to protect nuclear medicine personnel, patients' close relatives, and the environment. Most current regulations concern diagnostic applications with technetium-99m and therapeutic applications with iodine-131. Regulations pertaining to the clinical use of 18F-FDG were recently enacted (2001). Regarding exposure nuclear medicine personnel, the amount of radioactivity used for therapeutic purposes is currently limited to 740 MBq, which requires that the patient be kept in a shielded room. Radiation exposure can be roughly estimated by measuring the exposure rate (in μSv/h) delivered at a distance of 1 meter from a source of one MBq. When considering both the exposure rate and half-life, it appears that similar doses of iron-52, yttrium-86, and iodine-124 deliver a high dose rate immediately after injection and during the following few hours. Current regulations on environmental exposure limit radioactivity levels in hospital sewage outfalls to 1000 Bq/L, for technetium 99m, and 100 Bq/L, for iodine-131. Forthcoming regulations for other radionuclides should not be established solely based on general rules such as the 100 Bq/L limit. Impact studies similar to those required in the nuclear industry, should be conducted. © 2006 EDP Sciences.Une véritable révolution technologique a, au cours des 5 dernières années, profondément modifié le périmètre d’application et les perspectives de la médecine nucléaire, en particulier de la cancérologie nucléaire. Dans le cadre des applications diagnostiques, la tomographie par émission de positons (TEP) avec le 18F-fluoro déoxyglucose (FDG) a eu un impact déterminant sur la stratégie diagnostique des oncologues en ajoutant des informations fonctionnelles très précieuses aux informations morphologiques de l’imagerie conventionnelle. De nombreux autres traceurs fluorés, en cours d’évaluation clinique, sont destinés à étudier des fonctions tumorales diverses (prolifération tumorale, hypoxie, apoptose chimioinduite, etc.) pouvant avoir des répercussions thérapeutiques importantes. D’autres émetteurs de positons seront disponibles à terme, comme le 64Cu (période de 12 heures), le 124I (période de 4,1 jours) et le 86Y (période de 15 heures) dans des indications comme l’immuno-TEP ou la dosimétrie pré-thérapeutique qui ne peuvent être développées avec le 18F du fait de sa période trop courte. Dans le cadre des applications thérapeutiques, la radiothérapie interne, longtemps limitée au cancer thyroïdien différencié, s’est étendue à beaucoup d’autres types de cancers du fait de la disponibilité de nouveaux vecteurs de radionucléides comme les anticorps monoclonaux (radioimmunothérapie) ou des peptides (radiopeptide-thérapie), de nouvelles méthodes de ciblage tumoral et de nouveaux radionucléides, en particulier des émetteurs de particules alpha (alpha-thérapie). Toutes ces évolutions technologiques ont bien sûr une implication en radioprotection en ce qui concerne l’exposition du personnel de médecine nucléaire, de l’entourage familial proche et plus généralement de l’environnement. Actuellement l’essentiel des dispositions règlementaires s’applique au 99mTc pour les applications diagnostiques et au 131I en thérapie, elles ont été étendues en 2001 au 18F dans le cadre des contraintes de radioprotection liées à l’utilisation de FDG. Concernant l’exposition de l’entourage proche du malade, la règlementation est actuellement limitée au seuil d’activité de 740 MBq de 131I qui impose un confinement hospitalier du malade. Une indication indirecte de cette exposition peut être donnée par le débit d’exposition (en µSv/h) à 1 mètre pour une source de 1 MBq. Si on prend en compte la valeur du débit et la période physique, il apparaît, qu’à activité identique, le 52Fe, le 86Y et le 124I sont des radionucléides émetteurs de positons qui présentent un débit élevé à la sortie du service et pendant les heures qui suivent l’injection. Concernant l’exposition de l’environnement, la règlementation actuelle impose une concentration radioactive, à l’émissaire de l’établissement, de 1000 Bq/L pour le 99mTc et de 100 Bq/L pour le 131I. Pour les nouveaux radionucléides, d’autres hypothèses que celle d’une concentration permanente de 100 Bq/L doivent être retenues et des études d’impact, similaires à celles pratiquées dans le domaine du nucléaire devraient être effectuées

Ovarian cancer management. Practice guidelines for nuclear physicians

Ovarian cancer is a frequent and severe malignancy. Over 75% of cases are diagnosed at an advanced stage with disease spread beyond the ovaries. Despite the high response rates of initial treatments (i.e.,70-80%), the median progression-free survival of advanced ovarian cancer is 16-22 months, and the 5-year overall survival, 20-30%. The majority of these patients relapse, with metastatic peritoneal spread, unresectable, or drug resistant disease. Our goal was to outline current knowledge about diagnosis, prognostic factors, and treatments, and to dwell on non-nuclear medicine and nuclear-medicine diagnostic procedures

Mechanisms of Cell Sensitization to Alpha-Radioimmunotherapy by Doxorubicin or Paclitaxel in MultipleMyeloma Cell Lines

Purpose:The purpose of this studywas to analyze differentmechanisms (cell cycle synchronization, DNA damage, and apoptosis) that might underlie potential synergy between chemotherapy (paclitaxel or doxorubicin) and radioimmunotherapy with a radionuclides. Experimental Design:Threemultiplemyeloma cell lines (LP1, RMI 8226, and U266)were treatedwith 213Bi-radiolabeledB-B4, amonoclonal antibody that recognizes syndecan-1 (CD138) 24 hours after paclitaxel (1nmol/L) or doxorubicin (10 nmol/L) treatment.Cell survivalwas assessed using a clonogenic survival assay. Cell cycle modifications were assessed by propidium iodide staining and DNA strand breaks by the comet assay. Level of apoptosis was determined by Apo 2.7 staining. Results: Radiation enhancement ratio showed that paclitaxel and doxorubicin were synergistic with a radioimmunotherapy.After a 24-hour incubation, paclitaxel and doxorubicinarrested all cell lines in the G2-M phase of the cell cycle. Doxorubicin combined with a radioimmunotherapy increased tail DNA in the RPMI 8226 cell line but not the LP1or U266 cell lines compared with doxorubicin alone or a radioimmunotherapy alone. Neither doxorubicin nor paclitaxel combined with a radioimmunotherapy increased the level of apoptosis induced by either drug alone or a radioimmunotherapy alone. Conclusion: Both cell cycle arrest in the G2-M phase and an increase in DNA double-strand breaks could lead to radiosensitization of cells by doxorubicin or paclitaxel, but apoptosis would not be involved in radiosensitizationmechanisms.JRC.E.5-Nuclear chemistr