Alpha-1 antitrypsin limits neutrophil extracellular trap disruption of airway epithelial barrier function.

Neutrophil extracellular traps contribute to lung injury in cystic fibrosis and asthma, but the mechanisms are poorly understood. We sought to understand the impact of human NETs on barrier function in primary human bronchial epithelial and a human airway epithelial cell line. We demonstrate that NETs disrupt airway epithelial barrier function by decreasing transepithelial electrical resistance and increasing paracellular flux, partially by NET-induced airway cell apoptosis. NETs selectively impact the expression of tight junction genes claudins 4, 8 and 11. Bronchial epithelia exposed to NETs demonstrate visible gaps in E-cadherin staining, a decrease in full-length E-cadherin protein and the appearance of cleaved E-cadherin peptides. Pretreatment of NETs with alpha-1 antitrypsin (A1AT) inhibits NET serine protease activity, limits E-cadherin cleavage, decreases bronchial cell apoptosis and preserves epithelial integrity. In conclusion, NETs disrupt human airway epithelial barrier function through bronchial cell death and degradation of E-cadherin, which are limited by exogenous A1AT.

Evaluation of a semiautomatic 3D fusion technique applied to molecular imaging and MRI brain/frame volume data sets.

A generally applicable 3D fusion method was evaluated using molecular imaging and MRI volumetric data sets from 15 brain tumor patients with stereotactic frames attached to their skull. Point pairs, placed on the frame only, were chosen, polynomial warping coefficients were generated to map voxels from one coordinate space to the other. The MRI frame was considered the reference structure and the standard for "correct" registration. An ANOVA test (p > 0.05) confirmed the point pair choice to be consistent. The 95% confidence interval for the t-test showed the measured distance difference between the registered volumes was within one MRI voxel. A further experiment was conducted to independently evaluate the brain registration based on testing for consistency of randomly selected interior/exterior points. A t-test result (p < 0.05) showed that the consistency (i.e., both interior or both exterior) before and after volume registration were significantly different. This fusion method may be a viable alternative when other methods fail.

A versatile functional-anatomic image fusion method for volume data sets.

We describe and validate a volumetric three-dimensional registration method, and compare it to our previously validated two-dimensional/three-dimensional method. CT/MRI and SPECT data from 14 patients were interactively fused using a polynomial warping technique. Registration accuracy was confirmed visually and by a nonsignificant F value from multivariate analysis of the transformed landmarks, a significant difference of the squared sum of intensity differences between the transformed/untransformed and the reference volume both at the 0.05 (p > 0.05) confidence level and an average 31% improvement of the correlation coefficient and cross correlation. For the two-dimensional/three-dimensional method, ROI center-to-center distance ranged from 1.42 to 11.32 mm (for liver) with an average of 6.13 mm +/- 3.09 mm. The average ROI overlap was 92.51% with a 95% confidence interval of 90.20-96.88%. The new method is superior because it operates on the true three-dimensional volume. Both methods give good registration results, take 10 to 30 min, and require anatomic knowledge.

Ictal SPECT during autonomic crisis in familial dysautonomia.

The authors report results of SPECT cerebral perfusion studies in two patients with familial dysautonomia (FD) during dysautonomic crises and when clinically stable. SPECT imaging studies used 99mTc ethylene cysteine dimer. During dysautonomic crises, regions in the temporoparietal and frontal lobes had increased uptake. Uptake in these areas was less during asymptomatic periods. Episodic asymmetric cerebral perfusion during crises especially affecting the frontal and temporal lobes is suggestive of ictal activity.

Initial clinical evaluation of radiolabeled MX-DTPA humanized BrE-3 antibody in patients with advanced breast cancer.

To evaluate radiometal-labeled humanized BrE-3 (huBrE-3) monoclonal antibody as a radioimmunolocalization and therapeutic agent in breast cancer patients, tumor localization, pharmacokinetics, radiation dosimetry, and immunogenicity of (111)In-labeled combined 1-p-isothiocyanatobenzyl 3-methyl- and 1-p-isothiocyanatobenzyl 4-methyldiethylenetriamine pentaacetic acid (MX-DTPA) huBrE-3 were studied. Seven women with BrE-3 antigen-positive, metastatic breast carcinoma underwent (111)In huBrE-3 infusion (5 mCi; 50 mg), followed by serial gamma camera imaging and plasma sampling. Region of interest analysis of images was used to make radiation absorbed dose estimates for (111)In huBrE-3. Data were extrapolated to 90Y huBrE-3. Human anti-human antibody (HAHA) response was measured in serum samples obtained up to 3 months after infusion. Patients tolerated infusions well. Seventy-six percent of 105 known sites of disease were identified on planar and single-photon emission computed tomography scans. For six of seven patients, a biexponential model fit the plasma time-activity curve best with an average T1/2alpha=10.6+/-8.5 (SD) h and average T1/2beta=114.2+/-39.2 h. Radiation absorbed dose estimates for (111)In huBrE-3 for whole body averaged 0.53+/-.08 rads/mCi. Dose estimates for 90Y huBrE-3 for marrow averaged 8.4+/-11.9 rads/mCi, and for tumors, 70+/-31.5 rads/mCi. Liver radioactivity uptake averaged 19.7+/-8.8% injected dose at 24 h after infusion, translating into an average radiation absorbed dose 21.1+/-12 rads/90Y mCi administered. Only one of seven patients demonstrated a low level of HAHA response. Although the plasma half-lives are longer and marrow dose higher for radiolabeled huBrE-3 compared with the murine construct, the excellent tumor localization, good tumor dosimetry, and low immunogenicity support the use of 90Y-huBrE-3 antibody for radioimmunotherapy of breast cancer.

Tc-99m LL-2 Fab' monoclonal antibody imaging in acquired immune deficiency syndrome-related lymphoma.

BACKGROUND: Both systemic and primary central nervous system (CNS) non-Hodgkin's lymphomas (NHL) occur in people with acquired immune deficiency syndrome (AIDS). The radiographic manifestations may be similar to other neoplasms and opportunistic infections that are also found frequently in AIDS. Furthermore, these diseases may coexist with NHL in the AIDS patient. METHODS: To evaluate the use of Tc-99m Lymphoscan (the Fab' fragment of the anti-CD-22 antibody LL-2; Immunomedics, Inc., Morris Plains, NJ) in patients with suspected AIDS lymphoma, we studied 7 patients with 35 sites of suspected disease. Six had CNS lesions suspicious for parenchymal brain lymphoma. Each patient underwent planar and single photon emission computed tomography imaging at 3-5 and 18-24 hours after administration of Lymphoscan. Scintigraphic results were compared with results of conventional diagnostic modalities. RESULTS: Overall, the sensitivity of Lymphoscan was 92% and the specificity was 86%. In brain lesions, there was 100% sensitivity and 100% specificity. Lymphoscan also had 100% sensitivity for sites of lymphomatous lymphadenopathy and for liver involvement. Although less specific in extracranial sites, Lymphoscan was correctly negative in sites of coexisting adenocarcinoma and pneumonia. Two patients had both parenchymal CNS and systemic lymphoma proven by biopsy. CONCLUSIONS: Lymphoscan appears to be a sensitive and specific method for diagnosing CNS lymphoma in AIDS patients. Although slightly less specific in extracranial sites, it may be helpful in differentiating lymphoma from other etiologies in these patients at risk for multiple neoplasms and opportunistic infections.

Thallium-201 uptake in cytomegalovirus encephalitis.

A 36-yr-old man with AIDS exhibited intense 201Tl uptake (lesion-to-brain uptake ratio 5.38) in a brain lesion previously detected by MRI and CT. The lesion was biopsied and found to contain cells with viral inclusions diagnostic of cytomegalovirus infection, not tumor as the thallium SPECT results suggested. Thallium-201 SPECT may be less specific than previously reported for differentiating neoplastic disease from opportunistic infections in AIDS patients.

Radioimmunotherapy for breast cancer using indium-111/yttrium-90 BrE-3: results of a phase I clinical trial.

UNLABELLED: BrE-3 is a murine IgG1 monoclonal antibody that binds to 97% of human ductal breast cancer specimens. A previous study documented the ability of 111In-labeled 1,4-methyl-benzyl isothiocyanate diethylenetriamine pentaacetic acid (111In-MX-DTPA) BrE-3 to specifically target breast cancer tissue in patients, and the dosimetry derived from the pharmacokinetics suggested that a useful therapeutic index could be obtained with 90Y-MX-DTPA BrE-3. A Phase I maximum tolerated dose study was, therefore, initiated. METHODS: Six patients received 111In/90Y-MX-DTPA BrE-3, three of them receiving 6.25 and the other three receiving 9.25 mCi/m2 of 90Y. Pharmacokinetics, dosimetry, human anti-mouse antibody (HAMA), toxicity and clinical responses were evaluated. RESULTS: Three of six patients demonstrated a minor and transient, but objective tumor response, and none of the patients had significant toxicity. Tumor dosimetry ranged from 39 to 167 rad/mCi of 90Y (442-1887 rad/ dose). HAMA response occurred in five of six patients. CONCLUSION: Minimal toxicity, dosimetric calculations and clinical assessment indicate that a useful therapeutic index can be achieved with this therapy. Indium-111/yttrium-90-MX-DTPA BrE-3 can be safely administered to patients with metastatic breast cancer, and therapy doses yielded pharmacokinetics similar to those of tracer doses. Clinical responses, albeit transient, were achieved with single-dose therapy. Rapid onset of the HAMA response will hinder multicycle therapy, unless it is prevented with immunosuppressive drugs or the use of a "humanized" antibody. Further studies are needed to determine the optimal use of BrE-3 for radioimmunotherapy.

Graphical 3D medical image registration and quantification.

We present a graphical three-dimensional method that facilitates image registration and fusion, and provides quantitative geometric and volume information. In particular it enhances the use of functional (radiopharmaceutical) imaging (SPECT, PET) which, though a powerful clinical tool, has the disadvantage of low spatial resolution and ill-defined boundaries. Registration between functional images and structural images (MRI, CT) can augment the anatomical context of these functional images.

Fusion of immunoscintigraphy single photon emission computed tomography (SPECT) with CT of the chest in patients with non-small cell lung cancer.

In non-small cell lung cancer (NSCLC), accurate staging is critical in deciding between potentially curative surgery and palliative treatment. Image registration, or fusion, combines the unique functional information provided by SPECT imaging with the excellent anatomic detail offered by computed tomography (CT) or magnetic resonance imaging to better characterize the information provided by each separate modality. In this study, we explored the role of fusion of immunoscintigraphy SPECT with CT in the staging of NSCLC. We fused chest CT with 99mTc-labeled IMMU-4 anti-carcinoembryonic antigen Fab' antibody fragment SPECT in 14 patients with NSCLC using a landmark-based algorithm. The algorithm's accuracy was a measure from the center-to-center distance and the percentage overlap of two regions of interest: one drawn on CT and warped onto SPECT, the other drawn directly on the SPECT. We found that the average center-to-center distance was 1.3 +/- 0.8 pixels. Average overlap was 46 +/- 20%. CT-SPECT fusion helped differentiate tumor from normal blood pool, necrotic areas within viable tumor, tumor recurrence from scar, and malignant lymphadenopathy from hyperplasia. We conclude that fusion of CT and SPECT augments the information provided by each separate modality. Future clinical applications of fusion in NSCLC staging using immunoscintigraphy appear promising.

Radioimmunodetection of non-small cell lung cancer using technetium-99m-anticarcinoembryonic antigen IMMU-4 Fab' fragment. Preliminary results.

BACKGROUND: Although computed tomography and magnetic resonance imaging have improved the staging and evaluation of non-small cell lung cancer (NSCLC), mediastinal staging lacks adequate specificity and sensitivity. Radioimmunodetection may augment computed tomography and magnetic resonance imaging. The authors evaluated the ability of the technetium 99m-anticarcinoembryonic antigen IMMU-4 Fab' fragment to localize NSCLC in vivo, measured its pharmacokinetics, and estimated its radiation dose. METHODS: Seventeen patients with carcinoembryonic antigen-positive NSCLC received 16-30 mCi of technetium 99m IMMU-4 Fab'. Planar imaging was performed at 1-7 hours and 20-24 hours. Single-photon emission computed tomography (SPECT) was performed within 8 hours after injection. In 10 patients, blood sampling, urine collection, and quantitative imaging were performed to determine blood and urine pharmacokinetics and radiation dose estimates. Human anti-mouse antibody response was measured for as long as 3 months after administration. RESULTS: Planar and/or SPECT imaging detected 72% of 32 known lesions. SPECT was more sensitive than planar imaging. T1/2 alpha averaged 0.18 +/- 0.33 hours; T1/2 beta averaged 8.02 +/- 5.53 hours. The mean concentration versus time value was 1.11 +/- 0.56 mg.h. The average whole body dose estimated for administration of 30 mCi was 0.45 +/- 0.08 rads. No human anti-mouse antibody responses were detected. CONCLUSION: The tumor detection rate was high, but the persistent blood pool at < 8 hours complicated image interpretation. An intermediate imaging time point (12-16 hours) might be preferable. SPECT is an important adjunct to imaging with this radioimmunoconjugate. The acceptable dosimetry estimated for 30 mCi Technetium 99m IMMU-4 Fab' and the lack of human anti-mouse antibody responses suggest this is a promising localizing tool for NSCLC:

Radioimmunolocalization of metastatic breast carcinoma using indium-111-methyl benzyl DTPA BrE-3 monoclonal antibody: phase I study.

Pharmacokinetics of radiolabeled BrE3 monoclonal antibody (Mab), reactive against a breast mucin epitope, were assessed in 15 patients with advanced breast cancer. Patients received 5 mCi (185 MBq) of 111In-methyl benzyl isothiocyanate DTPA (MX-DTPA) conjugated BrE-3 Mab intravenously with total antibody doses of 10, 50 or 100 mg. Serial quantitative imaging, blood and urine clearance were obtained to measure pharmacokinetics, assess tumor localization and estimate radiation dose. Organ function was followed to determine toxicity. Mild allergic reactions occurred in four patients. Eighty-six percent of 70 known lesions and 5 unsuspected lesions were detected by antibody imaging. Biexponential modeling of radiolabeled antibody in serum showed a T1/2 alpha = 9.5 +/- 2.7 hr and T1/2 beta = 56 +/- 25.4 hr. Total urinary excretion averaged 35.5% +/- 19.3% injected dose (ID) by Day 8. Quantitative imaging showed that 0.02-2.56% ID localized in tumors. Extrapolating dosimetry from 111In-MX-DTPA-BrE-3 to 90Y-MX-DTPA-BrE-3, we estimate therapeutic radiation doses could be delivered to some tumors with tolerable toxicity.

Graphical interface for medical image processing.

We have developed a graphical interface which allows users of varying levels of computer experience and proficiency to manipulate medical image-processing data with "point-and-click" ease. The power which had formerly been associated with protocols and shell scripts has been combined with the flexibility and "user-friendliness" of buttons and dialog boxes.

An integrated approach to biodistribution radiation absorbed dose estimates.

An integrated approach to existing methods of extracting biodistribution data, pharmacokinetics and radiation absorbed dose estimates from serial scintigraphic images is described. This approach employs a single computer-generated user interface to reformat planar scans into a standard file type, align conjugate (anterior and posterior) images, draw regions of interest (ROIs) over selected organs and lesions and generate count data for anterior and posterior views and calculated geometric means. Using standard correction methods, the fraction injected activity is obtained for all ROIs and total body. This methodology has been applied to the analysis of indium-III-labelled breast-cancer-directed antibodies and technetium-90m-labelled CEA-specific antibody fragments in non-small-cell lung cancer. It is anticipated that this approach will be useful for evaluating the dosimetry of other radiolabelled monoclonal antibodies, as well as other radiopharmaceuticals.

Radionuclide quantitation of renal function.

Quantitation of renal function may be performed with a variety of radiopharmaceuticals which reflect slightly different renal functions. Plasma sampling techniques and imaging techniques have been used to derive absolute measurements of renal function. The addition of imaging permits the determination of relative or "split" function. Time-activity curves from renal studies provide other quantitative parameters of function reflecting arterial supply, renal cortical function, and patency of the renal collecting system. Quantitative radionuclide studies of the kidneys provide comprehensive, reproducible, and objective assessments of renal function.

Hepatic hemangiomas: diagnosis with fusion of MR, CT, and Tc-99m-labeled red blood cell SPECT images.

A method of image analysis was developed for correlation of hemangiomas detected at computed tomography (CT) and/or magnetic resonance (MR) imaging with increased blood pool activity evident at single photon emission CT (SPECT) performed after labeling of red blood cells with technetium-99m. Image analysis was performed in 20 patients with 35 known hepatic hemangiomas. After section thickness and pixel sizes of the different studies were matched, intrinsic landmarks were chosen to identify anatomically corresponding locations. Regions of interest (ROIs) drawn on the CT and/or MR images were translated, rotated, and reprojected to match the areas of interest on the corresponding SPECT images by means of a two-dimensional polynomial-based warping algorithm. Analysis of ROIs on 30 SPECT-MR and 20 SPECT-CT pairs of registered images provided absolute confirmation that 34 suspected hemangiomas identified on SPECT images correlated exactly with lesions seen on CT and/or MR images. Accuracy of fusion was within an average of 1.5 pixels +/- 0.8 (+/- 1 standard deviation). The technique enabled diagnostic confirmation of hemangiomas as small as 1.0 cm and proved useful for evaluating lesions located adjacent to intrahepatic vessels.

Pulmonary applications of nuclear medicine.

Nuclear medicine techniques have a long history in pulmonary medicine, one that has been continually changing and growing. Even longstanding methods, such as perfusion scanning for embolic disease or for pretherapy pulmonary function evaluation, have largely withstood the test of recent careful scrutiny. Not only have these techniques remained an important part of the diagnostic armamentarium, but we have learned how to use them more effectively. Furthermore, because of technical advances, we are in a phase of expanding roles for nuclear imaging. Gallium citrate scanning for the mediastinal staging and follow-up of lymphoma has been recognized as a valuable adjunct to the anatomic information provided by CT and MRI. With the growth of PET technology in areas that have been explored in a limited fashion until now, such as noncardiogenic pulmonary edema and lung carcinoma, evaluation and management of these patients may substantially improve. Finally, in the field of radiolabeled monoclonal antibodies, attention is now being turned to both the diagnostic and the therapeutic problems presented by lung carcinoma. As radiolabeling methods are refined and as new and better antibodies are developed, radioimmunodetection and therapy in lung carcinoma may begin to make inroads on this common and hard to control disease.

CT-SPECT fusion for analysis of radiolabeled antibodies: applications in gastrointestinal and lung carcinoma.

Fusing or image registration improves the information obtained by correlating images from various imaging modalities. We "fused" radiolabeled antibody SPECT with CT in patients with colorectal or lung cancer. We identified corresponding landmarks on cross-sectional images and used standard graphics algorithms for untilting to match planes of reconstruction and for two-dimensional warping or transformation of images or regions of interest. Fusing localizes activity on SPECT to specific anatomic structures and decreases SPECT false positives and CT false negatives.