99mTc-labelling of leucocytes with 99mTc-DPO: a complex developed for myocardial imaging

The lipophilic 99mTc-DPO complex, developed as a myocardial imaging radiopharmaceutical, was used to label leucocytes. After an incubation of 0.1 ml 99mTc-DPO (8 micrograms DMPE*2HCl) with mixed leucocytes in plasma, the labelling efficiency was over 70%. During incubation in 5 ml plasma, a loss of activity was found between 20% (1 h) and 35% (3 h) caused by elution. Disturbances of cell viability could not be found with the help of the chemiluminescence test. The in vivo recovery was determined in three dogs and was 45%-50% (0.5 h), 30%-36% (1 h), and 18%-24% (3 h). Autologous 99mTc-DPO-leucocytes were used on seven patients with suspected osteomyelitis, there were four true negative and three true positive results. The target/nontarget ratio determined by ROI in the positive cases was 1.8 to 2.5 at 3 h after injection.     

Technetium-99m pyrophosphate/thallium-201 dual-isotope SPECT imaging predicts reperfusion injury in patients with acute myocardial infarction after reperfusion

Purpose  Microcirculatory failure after reperfusion is clinically indicated to cause reperfusion injury whereas excessive intracellular calcium ion overload is experimentally proved as a key mechanism of reperfusion injury. We hypothesized that technetium-99m (^99mTc) pyrophosphate (Tc-PYP) uptake in injured but viable infarct-related myocardium with preserved myocardial perfusion after reperfusion estimated by thallium-201 (²⁰¹Tl) uptake would be associated with final functional recovery. Methods  Dual-isotope Tc-PYP/²⁰¹Tl single-photon emission computed tomography (SPECT) was performed 2 days after successful reperfusion therapy in patients with first acute myocardial infarction, and 50 patients (63 ± 13 years old, female 22%) with preserved ²⁰¹Tl uptakes of ≥50% in reperfused myocardium was followed for 1 month. Tc-PYP uptake was assessed as the heart-to-sternum (H/S) ratio. Two-dimensional echocardiography was also performed 2 days and 1 month after reperfusion to evaluate functional recovery. Results  High Tc-PYP uptake, defined as the H/S ratio ≥0.81, was predictive of chronic phase no functional recovery (73.7% in 14 of 19 patients with high uptake vs 16.1% in five of 31 patients without those, p < 0.0001). After adjustment for potential confounding variables, including electrocardiographic persistent ST segment elevation at 1 h after reperfusion, high Tc-PYP uptake remained independently predictive of no functional recovery with odds ratio of 8.7 (95% confidential interval = 2 to 38.7; p = 0.005). Conclusion  High Tc-PYP uptake in reperfused but viable infarct-related myocardium was a powerful predictor of no functional recovery, which may reflect excessive intracellular calcium ion overload caused by reperfusion injury. Tc-PYP/²⁰¹Tl dual-isotope SPECT imaging can provide prognostic information after reperfusion.     

Myocardial uptake characteristics of three99mTc-labeled tracers for myocardial perfusion imaging one hour after rest injection

OBJECTIVE: 99mTc-tetrofosmin and 99mTc-sestamibi are approved tracers for myocardial perfusion studies. Recently, a 99mTc-MIBI preparation from a different manufacturer (99mTc-cardiospect-MIBI) has been introduced to the market. Therefore, the aim of this study was the evaluation of 99mTc-tetrofosmin as well as of two different 99mTc-labeled MIBI tracers with regard to differences in imaging quality under resting conditions. METHODS: Sixty patients (mean age 63.8 years +/- 1.25) with known or suspected coronary artery disease but without evidence of rest-ischemia were included. Twenty patients in each group were examined by a two-day-rest-stress protocol using the three 99mTc-labeled tracers. Visual analysis of all images was performed by two experienced physicians blinded with regard to the applied tracer. Regions of interest (ROI) were defined over the heart, lung and whole body only in the rest imaging in order to calculate heart-to-lung, lung-to-whole body-, and heart-to-whole body-ratios. RESULTS: The heart-to-lung ratio was statistically significant higher for 99mTc-cardiospect-MIBI as compared to 99mTc-sestamibi as well as to 99mTc-tetrofosmin. Furthermore, a significantly higher heart-to-lung ratio was found for 99mTc-sestamibi as compared to 99mTc-tetrofosmin. The heart-to-whole body-ratio and the lung-to-whole body-ratio were equivalent between all tracers. Visual analysis revealed only slight differences regarding image quality between all tracers. CONCLUSIONS: ROI analysis surprisingly revealed a significant higher myocardial uptake and consequently a higher heart-to-lung ratio for 99mTc-cardiospect-MIBI. Whether this leads to a better visual image quality has to be evaluated in future studies with larger study populations as well as semiquantitative segmental analysis of the myocardial perfusion images.     

Evaluation of a new leukocyte labeling procedure with 99mTc-HMPAO

A technique for labeling leukocytes with ^99mTc using hexamethylpropylene-amineoxime (HMPAO) was evaluated in vitro. The labeling procedure resulted in a cell bound fraction of radioactivity of 56% after ^99mTc incubation and 96% after 1 washing of cells. In the final cell suspension 84% of the radioactivity was attached to the polymorphonuclear (PMN) leukocytes constituting 94% of white cells. Only 5% was bound to residual red blood cells. The stability evaluated in autologous plasma showed a decline of cell bound activity from 96% to 84% over 3 h. The chemotactic function of PMN leukocytes was unaffected by the labeling procedure. These findings demonstrate that HMPAO, albeit cell unspecific, is efficient for labeling PMN leukocytes with ^99mTc. The stability of the labeling procedure is high and the technique does not affect cell function. No other current ^99mTc leukocyte labeling technique possesses all these qualities.     

The utilization of Tc-99m-TBI as a myocardial perfusion agent in exercise studies: Comparison with Tl-201 thallous chloride and examination of its biodistribution in humans

Twenty-four patients were studied with both ²⁰¹Tl-thallous chloride and ^99mTc-TBI scintigraphy following exercise. Comparison of the two agents in detecting segmental myocardial ischemia and scar was made in 18 patients with evidence of coronary artery disease on ²⁰¹Tl-thallous chloride scintigraphy. Agreement between the two studies was observed in 77% (125 of 162) of left ventricular segments, suggesting that ^99mTc-TBI can be used as a myocardial perfusion agent. Limitations were related to early high background activity from lungs and liver. The high lung activity and early myocardial redistribution within the 1st hour contributed to the failure of ^99mTc-TBI to detect 16 segmental defects seen in the immediate post-exercise thallous chloride scan. Persistently high liver activity additionally affected accurate interpretation in the left ventricular segments close to the diaphargm. Improvement in the accuracy of ^99mTc-TBI stress studies might be achieved with tomographic imaging to reduce the problem of background activity or by the development of ^99mTc-labeled isonitrile analogues with rapid lung and liver clearance.     

Synthesis and biodistribution of a novel [TcN(PNP5)(DMCHDTC)] complex as a potential myocardial perfusion imaging agent

The [^99mTcN(PNP5)(DMCHDTC)]⁺(DMCHDTC: 2,3-dimethyl cyclohexyl dithiocarbamate, PNP5:bis(dimethoxypropylphosphinoethyl)ethoxyethylamine) complex was synthesized through a ligand-exchange reaction. The two-step procedure involved the initial reaction of ^99mTcO₄⁻ with succinic dihydrazide (SDH) as a donor of nitride nitrogen atom (N³⁻) in the presence of stannous chloride dihydrate as reducing agent and propylenediamine tetraacetic acid (PDTA) as complexant, followed by the addition of the PNP5 ligand and the DMCHDTC ligand. The radiochemical purity (RCP) of the product was over 90% as measured by thin layer chromatography (TLC). No decomposition of the complex at room temperature was observed over a period of 6 h. Its partition coefficient indicated that it was a lipophilic complex. The electrophoresis results showed the complex was cationic. The biodistribution results in mice indicated that [^99mTcN(PNP5)(DMCHDTC)]⁺ was significantly retained into the heart. The heart uptake (ID%/g) was 14.47, 12.23 and 8.76 at 5, 30 and 60 min post-injection, respectively. The heart/liver, heart/lung and heart/blood ratios of the complex were 1.24, 3.62 and 23.05 at 60 min post-injection, suggesting it will be a potential myocardial imaging agent.     

A new 99mTc-red blood cell labeling procedure for cardiac blood pool imaging: clinical results

The first clinical results of a new ^99mTc-red blood cell labeling procedure avoiding cell centrifugation are presented. One ml heparinized blood samples were incubated with small amounts of a stannous kit. By titration studies, ideal quantities of sodium hypochlorite for oxidation of extracellular tin and of EDTA as stabilizer of the label were found. The Cl^- concentration and pH of the labeled blood were acceptable, and EDTA increased labeling yield and stability determined in vitro by a few percent. The new procedure gave a slightly higher labeling yield than a current technique using centrifugation of cells. Labeling efficiency expressed as cell bound/total activity was 96.6%±1.3% in healthy subjects and 95.5%±2.2% in cardiac patients and remained high for 2 h after reinjection. The biological halflife of labeled cells following the new procedure was 11–12 h rendering it suitable for serial determinations of radionuclide cardiography.     

Clinical evaluation of the 99mTc-CPI myocardial perfusion imaging

^99mTc-CPI myocardial perfusion scintigraphy including planar images in 35 patients and SPECT images in 16 patients has been studied. Scintigraphic data revealed that high quality ^99mTc-CPI myocardial perfusion images were obtained. The sensitivity and specificity of ^99mTc-CPI planar images in detecting CAD was 92% and 80% respectively. There was no significant difference in sensitivity for detecting CAD between planar and SPECT. However, the specificity of SPECT was much better than that of planar imaging.     

The Clinical Usefulness of 99mTc HMPAO Leukocyte/99mTc Phytate Bone Marrow Scintigraphy for Diagnosis of Prosthetic Knee Infection: A Preliminary Study

Purpose

The preferred radionuclide imaging procedure for diagnosing prosthetic joint infection is combined radiolabeled leukocyte/^99mTc sulfur colloid bone marrow scintigraphy, which has an accuracy of over 90 %. Unfortunately, sulfur colloid is no longer available in South Korea. In this study, we evaluated the usefulness of ^99mTc phytate, a substitute for ^99mTc sulfur colloid, when combined with radiolabeled leukocyte scintigraphy in suspected prosthetic knee infections.

Methods

Eleven patients (nine women, two men; mean age 72 ± 6 years) with painful knee prostheses and a suspicion of infection underwent both ^99mTc HMPAO leukocyte scintigraphy (LS) and ^99mTc phytate bone marrow scintigraphy (BMS). The combined images were interpreted as positive for infection when radioactivity in the LS at the site of clinical interest clearly exceeded that of the BMS (discordant); they were interpreted as negative when the increased activity in the LS was consistent with an increased activity in the BMS (concordant). The final diagnosis was made with microbiological or intraoperative findings and a clinical follow-up of at least 12 months.

Results

Five of eleven patients were diagnosed as having an infected prosthesis. The overall sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of the combined LS/BMS were 100 %, 83 %, 83 %, 100 % and 91 %, respectively.

Conclusion

We find that combined ^99mTc HMPAO LS/^99mTc phytate BMS shows comparable diagnostic performance to other studies utilizing sulfur colloid. Combined ^99mTc HMPAO LS/^99mTc phytate BMS is therefore expected to be an acceptable alternative to combined radiolabeled LS/^99mTc sulfur colloid BMS for diagnosing prosthetic knee infections.     

A micro-autoradiographical study of the localization of 99mTc(Sn)-MDP and 99mTc-MDP in undecalcified bone sections

The localization of ^99mTc(Sn)-MDP in bone tissue was compared with ^99mTc-MDP by means of microautoradiography of undecalcified bone sections. Sections of good histological quality were obtained by a rapid embedding method in methylmethacrylate. No differences were found in the localization of these radiopharmaceuticals in fetal rat calvariae after incubation in vitro or in rat femora after administration in vivo. In the incubation experiment, hydrolyzed ^99mTc was formed. The uptake was high in areas of new bone formation. No uptake was seen in cells or in resorbing areas. In compact bone ^99mTc(Sn)-MDP was predominantly taken up in the vicinity of blood vessels.     

Identification of factors affecting technetium 99m leucocyte labelling by phagocytic engulfment and evelopment of an optimal technique

Human autologous leucocytes can be simply and reproducibly labelled by phagocytic engulfment of technetium 99m stannous colloid with high leucocyte-labelling efficiency (LLE), similar human biodistribution to indium-111-oxine labelled leucocytes and good cell viability. A mean particle size of 2.1 ms is optimal for phagocytosis and the most important parameter in maintaining reproducibly high neutrophil uptake. It is more critical than hitherto appreciated. When such variables as type of colloid, purity of starting materials, speed of mixer rotation when preparing colloid and labelling leucocytes, heparin concentration, freshness of colloid preparation, type of sterilising membrane filter used and incubation time of cells with colloid are rigidly controlled, consistent labelling efficiencies in excess of 90% can be obtained with neutrophil predominance. The lyophilised kit tested produced suboptimal results.     

In vivo labelling of granulocytes with 99mTc anti-NCA monoclonal antibodies for imaging inflammation

We looked for inflammatory lesions in 45 patients using in vivo labelled granulocytes. For the purposes of cell labelling we used a murine, monoclonal antibody reacting with NCA and CEA (BW 250/183) (Bosslet et al. 1985) labelled with ^99mTc-pertechnetate. All abscesses and other inflammatory lesions were visualized with excellent quality scintigrams between 2 and 6 h after the injection. As the antibody can be stored in a freeze-dried form and labelled at any time in any Department of Nuclear Medicine with ^99mTc-pertechnetate, without cell isolation being necessary, the method appears to be suitable even for use in acute diagnosis. No side-effects have so far occurred, even in patients injected up to three times.     

Blood pool ventriculography with a new technetium-labelled complex (99mTc-DEPIC) : a clinical evaluation

A new, single bolus method of in vivo blood pool imaging using a technetium Tc99m phosphine isocyanide complex (DEPIC) which binds to pre-albumin was evaluated in volunteers (n=4) and patients (n=20). DEPIC was assessed for its safety and possible drug interactions. Its duration of action and quality of ventriculography were compared with imaging using standard in vivo red cell labelling (PYP) during two 3-h scanning periods 1 week apart. DEPIC had a mean plasma halflife of 3.3 h. The count rate over the left ventricle was initially 42% higher with DEPIC than with PYP. However, removal of DEPIC by the liver resulted in equivalent count rates by 1 h, and by 3 h PYP count rates were 22% higher than DEPIC. Immediately post injection mean (SD) difference in the left ventricular ejection fraction between the two methods was 2.4% (7.7%). Satisfactory DEPIC scans were obtained up to 2 h post injection, but by 3 h there was a mean difference of 13% (11.3%). DEPIC was found to be a safe alternative to red all labelling for blood pool angiography, suitable for routine work. The single bolus methodology and high initial count rates offer improved efficiency and a capability for truly emergency scanning.     

Dramatic intestinal uptake of99mTc pertechnetate in a pediatric patient with enteritis

A pediatric patient with enteritis, which showed dramatically diffuse uptake of 99mTc pertechnetate in the intestine, is reported. Repeated study after medical treatment exhibited complete disappearance of the intestinal uptake. 99mTc pertechnetate scintigraphy has the potentiality for diagnosing and monitoring active enteritis.     

A (99m)Tc-labeled dual-domain cytokine ligand for imaging of inflammation

Introduction

Interleukin (IL)-1 and IL-18 are potent proinflammatory cytokines in inflammation-related diseases. Their actions are regulated by IL-1 receptor antagonist (IL-1ra) and IL-18 binding protein (IL-18bp). This study was designed to ^99mTc-radiolabel an IL-1ra and IL-18bp dual-domain cytokine ligand, IL-18bp-Fc-IL-1ra, for specific inflammation targeting.

Methods

The ^99mTc-IL-18bp-Fc-IL-1ra was obtained by direct labeling via 2-iminothiolane reduction. Competitive binding of ^99mTc-labeled and unlabeled IL-18bp-Fc-IL-1ra to rat polymorphonuclear leukocytes was assessed in vitro. A mouse ear edema model was used to evaluate specific targeting properties of ^99mTc-IL-18bp-Fc-IL1ra in vivo. The correlation between ^99mTc-IL-18bp-Fc-IL-1ra uptake and ¹¹¹In-labeled polymorphonuclear neutrophil infiltration was studied using ischemic–reperfused rat hearts.

Results

Direct ^99mTc-labeling yielded a stable dual-domain cytokine radioligand with radiochemical purity greater than 95% after gel filtration. Competitive binding studies showed specific targeting of ^99mTc-IL-18bp-Fc-IL-1ra to inflammatory cells. The ^99mTc-IL-18bp-Fc-IL-1ra uptake was 1.80±0.17 % injected dose per gram (%ID/g) in the inflamed ear without blocking, whereas uptake in the presence of IL-18bp-Fc-IL-1ra was 1.09±0.08 %ID/g (P<.05). The amounts of IL-1β and IL-18 were significantly increased in the inflamed ears compared to the vehicle controls. A significant correlation of ^99mTc-IL-18bp-Fc-IL-1ra with ¹¹¹In-labeled neutrophil distribution was observed in the ischemic–reperfused hearts (P<.001).

Conclusion

Targeting proinflammatory cytokines with ^99mTc-IL-18bp-Fc-IL-1ra may provide a suitable approach for specific detection of inflammatory sites.     

Isolation and labelling of human leukocytes with 99mTc

The role of the leukocyte isolation procedure on cell labelling with ^99mTc has been evaluated. Separation of leukocytes was performed by two procedures: (1) sedimentation on methyl cellulose, followed by discontinuous gradient centrifugation; (2) methyl cellulose sedimentation and hypotonic haemolysis of residual red blood cells. After washing the cells in saline and incubation with a stannous pyrophosphate agent, the leukocytes were labelled with 5–10 mCi ^99mTc. Procedure 1 gave a higher purity but lower recovery of polymorphonuclear leukocytes, and a minor contamination of red blood cells. ^99mTc labelling of cells was slightly more efficient with this method, probably due to the presence of red blood cells. Procedure 1 is recommended for in vitro studies on cell kinetics and procedure 2 is recommended for clinical use.     

Technetium-99m mercaptoalbumin as a potential substitute for technetium-99m labelled red blood cells

Technetium-99m labelled red blood cells (^99mTc-RBCs) are far superior to ^99mTc-labelled human serum albumin (^99mTc-HSA) for radionuclide ventriculography, but their labelling is more complex, time consuming and risk bearing (in vitro labelling) or suffers from interference by some medications (in vivo labelling). We have now modified HSA by the introduction of mercapto groups with the purpose of preparing stable and practical ^99mTc-mercaptoalbumin with long retention in the vascular system, that could replace ^99mTc-RBCs. HSA was incubated with N-succinimidyl S-acetylthioacetate (SATA) or N-succinimidyl 2,3-di(S-acetylthio) propionate (SATP) to introduce a chain containing one or two protected sulfhydryl groups on some of the lysine amino groups. After purification by size-exclusion chromatography (SEC), the mercapto groups were deprotected by incubation at alkaline pH or by treatment with hydroxylamine. The reaction products were used with or without SEC purification for direct or exchange labelling experiments with ^99mTc at neutral pH. SEC-HPLC was used to determine labelling yields and to isolate pure ^99mTc-mercaptoalbumin. Stable ^99mTc-mercaptoalbumin complexes could be formed in 90%–95% yield after coupling albumin with SATA or SATP in all molar ratios used followed by deacetylation in one of the mentioned conditions. The most favourable results were obtained after reaction of SATA or SATP with HSA in a 25: 1 ratio and deprotection with NH₂OH. The stability of the resulting ^99mTc-mercaptoacetyl-albumin (^99mTc-MAHSA) and ^99mTc-dimercaptopropionyl-albumin (^99mTcDMP-HSA) and their retention in vivo in plasma of mice and rabbits are clearly higher than that of conventional ^99mTc-HSA preparations. ^99mTc-DMP-HSA approaches the behaviour of ¹²⁵I-HSA quite well in both animal species. A preliminary study with ^99mTc-DMP-HSA in a volunteer showed a retention in the vascular compartment almost identical to that of ^99mTc-RBCs and clearly higher than that of a common ^99mTc-HSA preparation. The results indicate that these ^99mTc-mercaptoalbumins and especially ^99mTc-DMP-HSA are very promising as a practical alternative to ^99mTc-RBCs.K.A. Verbeke is a Research Assistant for the Belgian National Fund for Scientific Research     

99mTc-DTPA and 99mTc-DMSA renal gamma imaging in the surveillance of patients with conduit urinary diversion

We studied renal anatomy and function using ^99mTc-2-3 dimercaptosuccinic acid (DMSA) and ^99mTc-diethylenetriaminepentaacetic acid (DTPA) in 27 patients with conduit urinary diversion. In this condition, free ureteral reflux is often associated with bacteriuria, and these factors are thought to precipitate progressive renal deterioration. Gamma-camera images provided valuable information concerning the structure of the renal parenchyma, the function of individual kidneys and possible ureteral obstruction, thus helping us to decide whether or not to instigate further treatment. The information gained using renal gamma imaging with ^99mTc-DTPA and ^99mTc-DMSA was complementary and partly overlapping. We preferred the use of ^99mTc-DTPA because of its ability to visualise the ureters and the region of ureteroconduit anastomosis. Using diuretic medication, we were able to differentiate true ureteral obstruction from atony in 9 patients using ^99mTc-DTPA.     

Use of pertechnetate 99mTc for abdominal scanning in localising an ileal duplication cyst: case report and review of the literature

A 10-year-old boy presented with periumbilical postprandial pain and some melena. Physical examination was normal. All investigations were negative except a pertechnetate ^99mTc abdominal scan which showed a very large and horn-shaped focus of high activity in the right flank. An ileal duplication was resected. It was lined by antral gastric mucosa with a large ulcer. The patient was treated successfully. The abdominal pertechnetate scan is discussed. Offprint requests to: X. Marchandise     

Osteoid osteoma simulating osteomyelitis: Differentiation with Tc-99m HIG scintigraphy

We present a case of osteoid osteoma with a history of mild pain, local swelling and point tenderness on the right lower leg. The diagnosis of osteoid osteoma was difficult due to the atypical clinical history and misleading radiological and bone scan findings. When it is difficult to differentiate an osteoid osteoma from osteomyelitis using CT, MRI or bone scan; HIG scintigraphy can be used to exclude an infection.