DTPA Groups Research Articles

A green and pollution-free heterogeneous ruthenium catalyst anchoring on magnetic nanoparticles with FPBA and DTPA groups for oxidation of aromatic alcohols

Using ruthenium chloride as catalyst precursor, magnetic silicon spheres as support, 4-formylphenylboronic acid and ethylenediaminepentaacetic acid as functional ligands, a magnetic heterogeneous nano-ruthenium catalyst (Fe3O4@SiO2-FPBA-DTPA-Ru) was prepared for the first time. It was nano level with 20.17 emu/g saturation magnetic strength and thermal stability under 249.63 °C. Fe3O4@SiO2-FPBA-DTPA-Ru contained 0.38 mmol/g Ru and was successfully used to catalyze the oxidation reactions of 15 aromatic secondary alcohols. Results proved that Fe3O4@SiO2-FPBA-DTPA-Ru as catalyst had good catalytic activity for the oxidation of secondary aromatic alcohols with yields exceeding 90 % for 12 products and 95 % for 10 products. High TON and TOF values (6386 and 9158 h−1) in 1-phenylethanol oxidation were obtained. All the reactions of aromatic alcohols were carried out in a solvent-free system, so the prepared catalyst had the characteristics of green and pollution-free. It was’nt suitable for oxidation of aromatic primary alcohols. If there was steric hindrance around the hydroxyl group in substrate, the catalysis also may be limited. The Fe3O4@SiO2-FPBA-DTPA-Ru catalyst also exhibited the advantages of energy saving, satisfactory reusability and stability in 9 consecutive runs. It showed great potential for future large-scale industrial production.

Adsorption removal of cationic dyes from wastewater using the corn straw modified with diethylenetriaminepentacetic acid ligand

Taking the thiazide cationic dye methylene blue (MB), triphenylmethane cationic dye crystal violet (CV), monoazo cationic dye cationic red 46 (R-46), and polycarboxycyanine cationic dye cationic rosé FG (P-FG) as the research objects, the adsorption behaviors of a self-made corn straw modified adsorbent HQ-DTPA-I for the dyes were investigated in depth. Under optimized conditions, HQ-DTPA-I can quickly adsorb most dyes within 3 min and reach equilibrium adsorption in 15–20 min. The removal rates of HQ-DTPA-I to MB, CV, R-46 and AP-FG can reach 95.28 %, 99.78 %, 99.28 % and 98.53 %, respectively. It also has good anti-interference ability for common ions present in most actual dye wastewater. For six consecutive adsorption-desorption cycles, the adsorption performance of HQ-DTPA-I can still reach 80.17 %, 81.61 %, 90.77 % and 83.48 % of the initial adsorption capacity, indicating good recovery performance. Based on Gaussian density functional theory to calculate its surface potential energy, it is found that the adsorption mechanism of HQ-DTPA-I for the cationic dyes is mainly due to the electrostatic interaction between the carboxyl groups in ligand DTPA and amino groups in dye molecules.

The Enhancement of Hepatic Parenchyma and Portal Vein in different CLD Groups: A Comparative Study of Gadoxetic Acid and Gadopentetate Dimeglumine.

Chronic liver disease (CLD) will affect the enhancement of hepatic parenchyma and portal vein on abdominal-enhanced MRI. To investigate the difference in liver parenchyma and portal vein enhancement in patients with CLD of different liver function grades between Gd- EOB-DTPA and Gd-DPTA in the portal venous phase (PVP). This retrospective study included 218 patients with CLD who had undergone abdominal enhanced MRI from January 2019 to June 2020. Patients with various degrees of liver dysfunction were identified with Child-Turcotte-Pugh and albumin-bilirubin grade. Two readers measured the precontrast and PVP signal intensities of liver parenchyma, portal vein, spleen, and psoas muscle. Relative liver enhancement, liver-to-spleen contrast index, portal vein image contrast, and portal vein-to-liver contrast were calculated. The relative enhancement of liver parenchyma was significantly lower for the Gd-EOB-DTPA group in any degree of liver function than the Gd- DTPA group in the PVP. The Gd-EOB-DTPA group showed significantly lower portal vein-to-liver contrast in the overall study population, CTP class B, and ALBI grade 2 patients compared to the group of Gd-DTPA at PVP. No significant difference was noted in the portal vein image contrast between the two contrast agents, regardless of CTP and ALBI grading. In CLD patients, Gd-EOB-DTPA yielded lower liver parenchymal enhancement and similar portal vein image contrast compared to Gd-DTPA in the PVP. Portal vein-to-liver contrast in the Gd-EOB-DTPA group was lower in the CTP class B and ALBI grade 2 subgroups compared to the Gd- DTPA group.

Granulocyte Colony-Stimulating Factor Nanocarriers for Stimulation of the Immune System (Part I): Synthesis and Biodistribution Studies.

In the field of cancer immunotherapy, an original approach consists of using granulocyte colony-stimulating factor (G-CSF) to target and activate neutrophils, cells of the innate immune system. G-CSF is a leukocyte stimulating molecule which is commonly used in cancer patients to prevent or reduce neutropenia. We focused herein on developing a G-CSF nanocarrier which could increase the in vivo circulation time of this cytokine, keeping it active for targeting the spleen, an important reservoir of neutrophils. G-CSF-functionalized silica and gold nanoparticles were developed. Silica nanoparticles of 50 nm diameter were functionalized by a solid phase synthesis approach. The technology enabled us to incorporate multiple functionalities on the surface such as a PEG as hydrophilic polymer, DTPA as 111In chelating agent and G-CSF. The gold nanocarrier consisted of nanoparticles of 2-3 nm diameter elaborated with DTPA groups on the surface and functionalized with G-CSF. We studied the particle biodistribution in mice with special attention to organs involved in the immune system. The two nanocarriers with similar functionalization of surface showed different pathways in mice, probably due to their difference in size. Considering the biodistribution after G-CSF functionalization, we confirmed that the protein was capable of modifying the pharmacokinetics by increasing the nanocarrier concentration in the spleen, a reservoir of G-CSF receptor expressing cells.

Sol-Gel Derived Adsorbents with Enzymatic and Complexonate Functions for Complex Water Remediation.

Sol-gel technology is a versatile tool for preparation of complex silica-based materials with targeting functions for use as adsorbents in water purification. Most efficient removal of organic pollutants is achieved by using enzymatic reagents grafted on nano-carriers. However, enzymes are easily deactivated in the presence of heavy metal cations. In this work, we avoided inactivation of immobilized urease by Cu (II) and Cd (II) ions using magnetic nanoparticles provided with additional complexonate (diethylene triamine pentaacetic acid or DTPA) functions. Obtained nanomaterials were characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). According to TGA, the obtained Fe3O4/SiO2-NH2-DTPA nanoadsorbents contained up to 0.401 mmol/g of DTPA groups. In the concentration range Ceq = 0–50 mmol/L, maximum adsorption capacities towards Cu (II) and Cd (II) ions were 1.1 mmol/g and 1.7 mmol/g, respectively. Langmuir adsorption model fits experimental data in concentration range Ceq = 0–10 mmol/L. The adsorption mechanisms have been evaluated for both of cations. Crosslinking of 5 wt % of immobilized urease with glutaraldehyde prevented the loss of the enzyme in repeated use of the adsorbent and improved the stability of the enzymatic function leading to unchanged activity in at least 18 cycles. Crosslinking of 10 wt % urease on the surface of the particles allowed a decrease in urea concentration in 20 mmol/L model solutions to 2 mmol/L in up to 10 consequent decomposition cycles. Due to the presence of DTPA groups, Cu2+ ions in concentration 1 µmol/L did not significantly affect the urease activity. Obtained magnetic Fe3O4/SiO2-NH2-DTPA-Urease nanocomposite sorbents revealed a high potential for urease decomposition, even in presence of heavy metal ions.

Trastuzumab Labeled to High Specific Activity with (111)In by Site-Specific Conjugation to a Metal-Chelating Polymer Exhibits Amplified Auger Electron-Mediated Cytotoxicity on HER2-Positive Breast Cancer Cells.

Our objective was to evaluate the cytotoxicity toward HER2-positive human breast cancer (BC) cells of trastuzumab modified site-specifically with a metal-chelating polymer (MCP) that presents multiple DTPA chelators for complexing (111)In. (111)In emits subcellular range Auger electrons that induce multiple lethal DNA double-strand breaks (DSBs) in cells. MCPs were synthesized with a polyglutamide backbone with 24 or 29 pendant DTPA groups, with or without nuclear translocation sequence (NLS) peptide modification and a terminal hydrazide group for reaction with aldehydes generated by sodium periodate (NaIO4)-oxidation of glycans on the Fc-domain of trastuzumab. Trastuzumab was site-specifically modified with two DTPA and labeled with (111)In for comparison (trastuzumab-NH-Bn-DTPA-(111)In). The maximum specific activity (SA) for labeling trastuzumab-Hy-MCP with (111)In was 90-fold greater than for trastuzumab-NH-Bn-DTPA-(111)In [8.9 MBq/μg (1.5 × 10(6) MBq/μmol) vs 0.1 MBq/μg (1.2 × 10(4) MBq/μmol)]. Trastuzumab-Hy-MCP-(111)In was bound, internalized, and imported into the nucleus of SK-BR-3 cells. NLS peptide modification of MCPs did not increase nuclear importation. A greater density of DNA DSBs was found for BC cells exposed to high SA (5.5 MBq/μg) than low SA (0.37 MBq/μg) radioimmunoconjugates. At 20 nmol/L, high SA trastuzumab-Hy-MCP-(111)In was 6-fold more effective at reducing the clonogenic survival (CS) of HER2 overexpressed and HER2 gene-amplified SK-BR-3 cells (1.3 × 10(6) receptors/cell) than low SA MCP-radioimmunoconjugates (CS = 1.8 ± 1.3 vs 10.9 ± 0.7%; P = 0.001). Low SA trastuzumab-NH-Bn-DTPA-(111)In (20 nmol/L) reduced the CS of SK-BR-3 cells to 15.8 ± 2.1%. The CS of ZR-75-1 cells with intermediate HER2 density (4 × 10(5) receptors/cell) but without HER2 gene amplification was reduced to 20.5 ± 4.3% by high SA trastuzumab-Hy-MCP-(111)In (20 nmol/L). The CS of HER2-overexpressed (5 × 10(5) HER2/cell) but trastuzumab-resistant TrR1 cells was decreased to 17.1 ± 1.6% by high SA trastuzumab-Hy-MCP-(111)In. Unlabeled trastuzumab (20 nmol/L) was 18-fold less potent than high SA trastuzumab-Hy-MCP-(111)In at reducing the CS of SK-BR-3 cells (CS = 37.0 ± 5.3%) and 3-fold less effective against Zr-75-1 cells (CS = 53.1 ± 9.8%). Unlabeled trastuzumab had no effect on the survival of TrR1 cells. We conclude that increasing the SA for labeling with (111)In by site-specific conjugation of MCPs to trastuzumab greatly amplified the cytotoxic potency against HER2-overexpressed and gene-amplified BC cells and extended its cytotoxicity to cells with intermediate HER2 expression but without gene amplification and to cells that are HER2 overexpressed but trastuzumab-resistant.

Multifunctional pH-sensitive polymeric nanoparticles for theranostics evaluated experimentally in cancer

A multifunctional pH-sensitive polymeric nanoparticle system was developed for simultaneous tumor magnetic resonance imaging (MRI) and therapy. The nanoparticles were self-assembled using the multi-block polymer poly(lactic acid)-poly(ethylene glycol)-poly(l-lysine)-diethylenetriamine pentaacetic acid (PLA-PEG-PLL-DTPA) and the pH-sensitive material poly(l-histidine)-poly(ethylene glycol)-biotin (PLH-PEG-biotin). The anti-hepatocellular carcinoma (HCC) drug sorafenib was encapsulated inside the nanoparticles. Gd ions were chelated to the DTPA groups which were distributed on the nanoparticle surface. Biotinylated vascular endothelial growth factor receptor (VEGFR) antibodies were linked to the surface biotin groups of nanoparticles through the avidin linker to form the target pH-sensitive theranostic nanoparticles (TPTN). TPTN exhibited spherical or ellipsoidal shapes, uniform particle size distribution (181.4 ± 3.4 nm), positive zeta potential (14.95 ± 0.60 mV), high encapsulation efficiency (95.02 ± 1.47%) and drug loading (2.38 ± 0.04%). The pH-sensitive sorafenib release from TPTN was observed under different pH values (47.81% at pH = 7.4 and 99.32% at pH = 5.0, respectively). In cell cytotoxicity studies, TPTN showed similar antitumor effect against HepG2 cells compared to solubilized sorafenib solution after pre-incubation in acid PBS (pH = 5.0) for 1 h in vitro (P > 0.05). In in vivo anti-tumor studies, TPTN showed significantly higher antitumor effect in H22 tumor (VEGFR overexpressed cell line) bearing mice compared to the solubilized sorafenib solution (oral or i.v. administration) group (P < 0.05). In the MRI test, the T1 relaxivity value of TPTN was 17.300 mM(-1) s(-1) which was 3.6 times higher than Magnevist® (r1 = 4.8 mM(-1) s(-1)). As a positive contrast agent, TPTN exhibited higher resolution and longer imaging time (more than 90 min) in the MRI diagnosis of tumor-bearing mice compared to Magnevist® (more than 60 min). Furthermore, histological examination of TBN (blank TPTN, without sorafenib loaded) showed no visible tissue toxicity compared to normal saline. Thus, TPTN possessed dual-loading drugs and imaging agents, active targeting and pH-triggered drug release properties in one platform with good biocompatibility. All of these results indicated that TPTN was a promising theranostic carrier which could be a platform for the development of novel multifunctional theranostic agents.

Abstract P4-12-14: Metal chelating polymer-conjugated trastuzumab labeled to high specific activity with indium-111 is cytotoxic to trastuzumab-sensitive and resistant breast cancer cells through the emission of auger electrons

Abstract Purpose: Auger electrons are very low energy electrons emitted by Indium-111(111In) that cause lethal DNA damage when emitted in close to the cell nucleus. Our laboratory reported previously that trastuzumab modified with nuclear localizing sequence (NLS) peptides and labeled with 111In(111In-NLS-DTPA-trastuzumab) was internalized and transported to the nucleus of HER2-overexpressing breast cancer (BC) cells, reducing their clonogenic survival (CS) to ≤10%. However, a limitation was that at the specific activity (SA) achieved (0.24 MBq/μg) for 111In-DTPA-trastuzumab without or with NLS, only 1 in 50 molecules was radiolabeled. This reduced the potency for cells with low or intermediate HER2 density. Our purpose was to dramatically increase the SA of 111In-labeled trastuzumab by conjugation to a metal chelating polymer (MCP) that displays multiple DTPA chelators for 111In and compare its potency to 111In-NLS-trastuzumab on BC cells with a wide range of HER2 density including cells resistant to trastuzumab. Methods: An MCP with a polyglutamide backbone, 29 DTPA groups, and a hydrazide reactive end-group was site-specifically linked to periodate-oxidized carbohydrates on the Fc domain of trastuzumab. HER2 immunoreactivity, internalization and nuclear importation were evaluated by cell binding and fractionation studies using HER2-overexpressing SK-Br-3 human BC cells. The SA achievable was assessed by radiolabeling 1, 5, 50 and 100 μg of the immunoconjugates with 9.25 MBq of 111In. The effect of high SA (5.5 MBq/μg) 111In-MCP-trastuzumab to on the CS of human BC cells with a wide range of HER2 density (5×105-1.3×106 HER2 receptors/cell) that were sensitive or resistant to trastuzumab was compared to low SA (0.37 MBq/μg) 111In-labeled trastuzumab conjugates. Studies are also in progress to assess DNA double-strand breaks caused by the Auger electrons emitted by 111In using the γH2AX assay. Results: MCP-trastuzumab bound specifically to HER2 with high affinity (KD = 10.7±1.5nM) and was internalized and imported into the nucleus of HER2-positive BC cells. The smallest mass that could be labeled (labeling efficiency&amp;gt;80%) with 111In for MCP-trastuzumab was 1 μg (SA = 8.9 MBq/μg) compared to 50 μg for DTPA-trastuzumab (SA = 0.08 MBq/μg). High SA 111In-MCP-trastuzumab reduced the CS of SK-Br-3 cells with high HER2 density to 1.8±1.3% and the CS of TrR1 trastuzumab-resistant BC cells with intermediate HER2 density to 17.1±1.6%. These represent a 4-5 and 5-6 fold greater potency compared to low SA 111In-trastuzumab conjugates on cells with high or intermediate HER2 expression, respectively. Conclusion: 111In-MCP-trastuzumab was labeled to 37-fold higher SA than 111In-DTPA-trastuzumab. High SA increased its cytotoxicity potency for HER2-positive BC cells that were sensitive or resistant to trastuzumab. Auger electron radioimmunotherapy is a promising strategy for HER2-positive BC which may extend to trastuzumab-resistant tumours. Supported by a grant from the Canadian Breast Cancer Research Alliance with funds from the Canadian Cancer Society. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-12-14.

Curious Results with Palladium- and Platinum-Carrying Polymers in Mass Cytometry Bioassays and an Unexpected Application as a Dead Cell Stain

We describe the synthesis of metal-chelating polymers (MCPs) with four different pendant polyaminocarboxylate ligands (EDTA, DTPA, TTHA, DOTA) and an orthogonal end-group, either a fluorescein molecule or a bismaleimide linker for antibody attachment. Polymer characterization by a combination of (1)H NMR, UV/vis absorption measurements, and thermal gravimetric analysis (TGA) indicated that each chain of the fluorescein-terminated polymers contained one dye molecule. These polymer samples were loaded with three different types of lanthanide ions as well as palladium and platinum ions. The numbers of metal atoms per chain were determined by a combination of UV/vis and conventional ICP-MS measurements. The experiments with lanthanide ions demonstrated that a net anionic charge on the polymer is important for water solubility. These experiments also showed that at least one type of lanthanide ion (La(3+)) is capable of forming a bimetallic complex with pendant DTPA groups. Conditions were developed for loading these polymers with palladium and platinum ions. While these polymers could be conjugated to antibodies, the presence of Pd or Pt ions in the polymer interfered with the ability of the antibody to recognize its antigen. For example, a goat anti-mouse (secondary) antibody labeled with polymers that contain Pd or Pt no longer recognized a primary antibody in a sandwich assay. In mass cytometry assays, these Pd- or Pt-containing MCPs were very effective in recognizing dead cells and provide a new and robust assay for distinguishing live cells from dead cells.

Recurrence of extensive injection site reactions following DTPa or dTpa vaccine in children 4–6 years old

Objectives The aim of this study was to compare the immunogenicity and reactogenicity of a lower dose diphtheria, tetanus and pertussis vaccine (dTpa) with the recommended vaccine (DTPa) given as a fifth dose to 4–6-year old children who previously experienced an extensive injection site reaction (ISR). Material and methods Children aged 4–6 years who had experienced an extensive ISR following a 4th dose of DTPa were recruited and randomly assigned to receive either the recommended DTPa or the lower dose dTpa vaccine. Parents recorded local reactions and systemic events for 15 days following vaccination. Immunogenicity was assessed pre and post vaccination by ELISA for diphtheria (D), tetanus (T), pertussis toxin (PT), filamentous haemagglutinin (FHA), and pertactin (PRN). Results A total of 53 participants were vaccinated. There was a 72% recurrence rate of ISR, with a trend ( p = 0.055) towards fewer ISR in the dTpa (61.5%) compared with the DTPa group (85.2%). There was no difference in reports of pain or irritability between groups. All participants had seroprotective levels of antibody to D and T and seroresponse to each of the 3 pertussis antigens following vaccination with higher GMCs in DTPa vs dTpa group. There was no increase in antibody avidity observed post vaccination, regardless of vaccine given. Conclusion Recurrence of ISR with the 5th dose of diphtheria, tetanus and pertussis vaccination in children who have previously experienced an extensive ISR is high. Vaccination with a dTpa vaccine may reduce the risk of fifth dose ISR.

Preparation of 111In-DTPA morpholino oligomer for low abdominal accumulation

An ability to quantitate the beta cell mass by noninvasive nuclear imaging will be very useful in the prevention, diagnosis, and treatment of diabetes. However, to be successful, radioactivity from the pancreas must not be obscured by the background radioactivity in the abdomen. Pretargeting offers the promise of achieving high target organ to normal tissue ratios. In preparation for pancreas imaging studies by pretargeting using morpholino oligomers (MORF/cMORF), it was necessary to develop a simple and efficient method to radiolabel the cMORF effector. Because we have shown that labeling the cMORF with (111)In via DTPA reduces excretion into the intestines compared to labeling with (99m)Tc via MAG(3), the conjugation of DTPA to cMORF was investigated for (111)In labeling. The amine-derivatized cMORF was conjugated with DTPA using 1-ethyl-3(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) as an alternative to the conventional cyclic anhydride. The conjugation efficiency (represented by the number of DTPA groups attached per cMORF) was investigated by changing the EDC, DTPA, and cMORF molar ratios. Different open columns were considered for the purification of DTPA-cMORF. Before conjugation, each cMORF molecule was confirmed to have an amine by trinitrobenzene sulfonic acid (TNBS) assay using the omega-amino butyric acid as positive standard and the non-amine derivatized cMORF as negative standard. The average number of DTPA groups per cMORF was 0.15-0.20 following the conjugation over a cMORF/DTPA molar ratio of 0.5-5 and over a cMORF/EDC molar ratio of 20-60. The conjugation efficiency was lower than expected probably due to steric hindrance. A 1 x 50cm P-4 column using ammonium acetate as eluting buffer provided an adequate separation of DTPA-cMORF from free DTPA. The (111)In labeling efficiency by transchelation from acetate exceeded 95%, thus avoiding the need for postlabeling purification. Despite the lower than expected conjugation efficiency in which no more than one-fifth of the cMORF were DTPA-derivatized, a specific radioactivity of at least 300microCi/microg or 1.90Ci/micromol of cMORF was achieved. In conclusion, a protocol is described for (111)In-DTPA-cMORF that provides the high specific activity favorable to beta cell imaging because of the low mass fraction of beta cells in pancreas (1-2%) and obviates the need for postlabeling purification.

High-Affinity Recognition of Lanthanide(III) Chelate Complexes by a Reprogrammed Human Lipocalin 2

Human lipocalin 2 (Lcn2), also known as neutrophil gelatinase-associated lipocalin (NGAL), which naturally scavenges bacterial ferric siderophores, has been engineered to specifically bind rare-earth and related metal ions as chelate complexes with [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diaminepentaacetic acid (p-NH(2)-Bn-CHX-A''-DTPA). To this end, 12 amino acid residues in the ligand pocket of Lcn2, which is formed by four loops at the open end of an eight-stranded beta-barrel, were subjected to targeted random mutagenesis, and from the resulting library, variants with binding activity for the Me x DTPA group were selected using the method of bacterial phage display. One promising candidate was further developed in several cycles of in vitro affinity maturation using partial random mutagenesis and selection (via phage display and/or Escherichia coli colony screening) under conditions of increasing stringency. As result, an Lcn2 variant was obtained that binds Y x DTPA with a dissociation constant as low as 400 pM. The Lcn2 variant specifically recognizes the artificial ligand, as exemplified in (competitive) ELISA and real-time surface plasmon resonance analyses. DTPA-complexed Y(3+), Tb(3+), Gd(3+), and Lu(3+) are most tightly bound, comprising metal ions whose isotopes are in common use for radiotherapy and imaging. All of the Lcn2 variants are stably folded and can be functionally produced in high yield in E. coli. X-ray crystallographic analyses show that the new ligand is well-accommodated in the central cavity of the engineered lipocalin, whose fold is largely preserved, but that the mode of binding differs from the one seen with the natural ligand Fe x enterobactin. This structural study reveals analogies but also differences with respect to previously described antibody-metal chelate complexes. Notably, the functionalized side chain of DTPA protrudes from the ligand pocket of the lipocalin in such a way that its conjugates (with other haptens, for example) are recognized too. With their small sizes and robust fold based on a single polypeptide chain, the engineered Lcn2 variants provide novel modules and/or fusion partners for radionuclide-chelate capturing strategies that bear promise for medical diagnostics and therapy.

High levels of antibody in adults three years after vaccination with a reduced antigen content diphtheria-tetanus-acellular pertussis vaccine

There is increasing interest in prevention of pertussis in adults by vaccination, but little is known about the duration of the antibody response to pertussis, diphtheria or tetanus in reduced antigen content vaccines formulated for adult use. Follow-up of a clinical trial including 550 adults comparing responses to reduced antigen content diphtheria-tetanus-acellular pertussis (dTpa) vaccine, or a licensed Td vaccine, provided the opportunity to evaluate this. Blood samples were collected at 0, 1, 12, 24 and 36 months following vaccination; of the original cohort of 550, 387 subjects (dTpa group N = 310, Td + pa group N = 77) were tested at month 36. Following a decrease in antibody levels against all vaccine antigens between one and 24 months following vaccination, levels stabilized during the third year, remaining higher at 36 months than pre-vaccination for all vaccine antigens. In particular, more than 90% of subjects remained seropositive for pertussis toxin and pertactin antibodies at 36 months after vaccination, suggesting ongoing protection against pertussis. Adult-formulated dTpa vaccines could replace Td for routine booster vaccination of older individuals.

Reactogenicity and immunogenicity of reduced antigen content diphtheria–tetanus–acellular pertussis vaccine (dTpa) administered as a booster to 4–6 year-old children primed with four doses of whole-cell pertussis vaccine

A trial to compare the reactogenicity and immunogenicity of a reduced antigen content diphtheria–tetanus–acellular pertussis (dTpa) vaccine with diphtheria–tetanus–whole-cell pertussis (DTPw) vaccine was conducted in Thailand. Three hundred and thirty children aged 4–6 years, primed with four doses of DTPw, received a single injection of either dTpa or DTPw. There was a significantly lower incidence of local and general reactions following dTpa than DTPw ( P<0.001). One month after vaccination, 99.4 and 100% of all subjects had protective anti-diphtheria and -tetanus titers, respectively. The vaccine response rate to pertussis antigens was similar in both groups, with 96.9% versus 92.5% for anti-pertussis toxin (PT), 96.9% versus 97.5% for anti-filamentous hemagglutinin (FHA) and 95.1% versus 90.8% for anti-pertactin (PRN) in the dTpa and DTPw groups, respectively. For anti-BPT, the vaccine response in the dTpa group was 29.6% versus 94.4% for DTPw. In conclusion, the dTpa vaccine was as immunogenic and significantly better tolerated than DTPw. The new dTpa vaccine could improve coverage for routine booster vaccination in children and provide a good replacement for DTP vaccines at 4–6 years of age.

A Molecular CT Blood Pool Contrast Agent

Rationale and Objectives A molecular-based computed tomographic (CT) contrast agent with prolonged vascular residence time is needed for vascular and tumor imaging. No particulate agents have reached clinical practice due to nonspecific macrophage activation. The authors' objective was to synthesize a water-soluble macromolecular agent. Materials and Methods Dysprosium-DTPA-dextran was synthesized through activation of the hydroxyl units of dextran PM40 with allylbromine and subsequent reaction with amino ethanethiol to produce amino-terminated leashes. These leashes were then coupled to DTPA by means of the mixed anhydride method. Complexation of dysprosium by DTPA-dextran was achieved in an acidic solution of 0.2 M dysprosium chloride. One rabbit with a VX2 tumor was imaged with [Dy]DTPA-dextran (0.5 mL, 3.1 g, 1.15 mmol of dysprosium per kilogram). Transaxial scans were acquired through the liver and tumor for 45 minutes. A second healthy rabbit was imaged with Optiray-320 (6.0 mL, 5.0 mmol of iodine per kilogram) at 1-minute intervals for 10 minutes and again at 20 minutes. Results Each dextran PM40 molecule (diameter, 8.8 nm) contained 95 [Dy]DTPA groups, increasing its average molecular weight from 40,500 to 101,537 g/mol. The baseline-corrected inferior vena cava (IVC) enhancement for [Dy]DTPA-dextran decreased, with an 8-minute half-time during the first 15 minutes followed by a nearly zero slope for the rest of the observation period. The IVC remained brighter than liver throughout the observation period. The solid portion of the tumor was enhanced by 5–10 CT numbers, rendering areas of necrosis more apparent. The baseline-corrected IVC curve for Optiray-320 also demonstrated two phases, with half-times of 2.5 and 45 minutes. The IVC became less dense than liver within 5–8 minutes. Conclusion [Dy]DTPA-dextran is water soluble and can be synthesized without intermolecular cross-linking to carry a high load of dysprosium. It provides blood pool enhancement characteristics with a long intravascular dwell time.

Evaluation of three rhenium-188 candidates for intravascular radiation therapy with liquid-filled balloons to prevent restenosis

Background. Intravascular brachytherapy is an effective method for inhibiting coronary restenosis after percutaneous transluminal coronary angioplasty. A new concept for preventing restenosis is the use of a liquid-filled balloon containing a beta-ray-emitting radioisotope. Generator-produced rhenium-188 (Re-188) is a good candidate for intravascular brachytherapy. However, in the unlikely event of balloon rupture, release of Re-188 perrhenate may cause a high radiation dose to the thyroid and stomach. In this study, we compared the biodistributions of three Re-188 preparations (Re-188 perrhenate, Re-188 pentetic acid [DTPA], and Re-188 MAG3) to assess the radiation dose to organs in a rat model that mimicked balloon rupture. Methods and Results. After injection of Re-188 preparations intravenously, rats were killed at 10 minutes, 30 minutes, 60 minutes, 2 hours, and 6 hours (n = 5/group). Tissue concentrations were calculated and expressed as percent injected dose per gram or per milliliter. In addition, urine excretion and thyroid gland uptake were evaluated in rats (n = 5/group) with a gamma camera after administration of 37 MBq (1 mCi) of each agent. Our data showed all 3 agents were excreted primarily via urine. In the Re-188 MAG3 group, 82% was excreted within 1 hour, but in the Re-188 perrhenate group, only 28% was excreted. The biodistribution data for these agents revealed that radioactivity levels in the stomach and the thyroid gland were high in the perrhenate group but low in the Re-188 DTPA and Re-188 MAG3 groups. The concentration levels in other tissues including lung, liver, testis, muscle, and blood were low throughout this study for all 3 agents. The thyroid radiation values were 0.163, 0.0167, and 0.00728 mGy/MBq for Re-188 perrhenate, Re-188 DTPA, and Re-188 MAG3, respectively. The stomach radiation values were 0.127 mGy/MBq for Re-188 perrhenate, 0.013 mGy/MBq for Re-188 DTPA, and 0.0104 mGy/MBq for Re-188 MAG3. Conclusions. In the event of balloon rupture, the release of Re-188 MAG3 or Re-188 DTPA results in lower radiation doses than release of Re-188 perrhenate, especially to the thyroid gland and the stomach. (J Nucl Cardiol 2000;7:37–42.)

Effect of Local Massage on Vaccination: DTP and DTPa

We previously demonstrated that local massage for one minute can enhance immunogenicity of diphtheria, tetanus, and whole-cell pertussis (DTPw) vaccination. This study further analyzes the effects of more intense local manipulation on infants after DTPw and DTPa (diphtheria, tetanus, and acellular pertussis) vaccination. A total of 808 infants aged two months were recruited to be vaccinated with either DTPw or DTPa. Vaccinees in both groups were further divided into two groups; those receiving local manipulation (massage and hot packing after vaccinations) and those receiving only vaccinations. Results showed that safety profiles were largely similar between those who had local manipulation following vaccination and those without. The only significant difference was more frequent local reactions including pain and swelling following the first two doses in both the DTPa and DTPw groups receiving manipulation compared with the groups not receiving manipulation. Serologic tests revealed that local manipulation had no significant effect on antibody response to pertussis toxin and filamentous hemagglutinin, and diphtheria and tetanus toxins. The effect of local massage on DTPw was related to the intensity of local massage. Too vigorous a local manipulation caused adverse local reactions and no beneficial effect on antibody response. As for the infants receiving DTPa and local massages for two minutes with hot packing, no significant effect on either the reactogenicity or immunogenicily was found.

Labelling of β2-microglobulin with 111In

The modification of β 2-microglobulin by coupling of DTPA groups and subsequent labelling with 111In is described. Purification steps are an ultrafiltration following reaction of the protein with DTPA dianhydride and a gel chromatography of the labelled product. [ 111 In]DTPA-β 2- microglobulin is obtained with a radiochemical yield of 37%.

Simple and rapid methods for labelling of human IgG by four radionuclides

Labelling of DTPA bicyclic anhydride coupled antibodies were investigated by determining the effect of DTPA: antibodies, DTPA: Sn molar ratios, pH, dimer and polymer formation of antibodies coupled with DTPA, using three different radionuclides, [111In,90Y and99mTc]. Analyses were performed with by Whatman No. 1 paper strips. Under optimal conditions we have achieved specific activities of111In or90Y labelled antibodies of about 37 kBq/1 μg for IgG coupled with about 2 DTPA groups per molecule and protein concentration of 15 mg/ml.

The influence of free DTPA on the dose distribution of yttrium-90 radiolabelled antibodies

Yttrium-90 ( 90Y) bound to antibody via a DTPA group, was stable in-vitro until exposed to buffers below pH 5.4. It also exhibited minimal transchelation to transferrin during serum incubation. In-vivo, two 90Y labelled antibodies of the same isotype but different specificities, showed similar distributions in tumour-bearing animals. Subsequent administration of a DTPA solution removed activity from liver and bone. This suggests that these tissues contain an environment in which a proportion of yttrium is released from the antibody, in a form suitable for removal with DTPA.