Developing a cell-bound detection system for the screening of oxidase activity using the fluorescent peroxide sensor roGFP2-Orp1.

Accurate yet efficient high-throughput screenings have emerged as essential technology for enzyme engineering via directed evolution. Modern high-throughput screening platforms for oxidoreductases are commonly assisted by technologies such as surface display and rely on emulsification techniques to facilitate single-cell analysis via fluorescence-activated cell sorting. Empowered by the dramatically increased throughput, the screening of significantly larger sequence spaces in acceptable time frames is achieved but usually comes at the cost of restricted applicability. In this work, we tackle this problem by utilizing roGFP2-Orp1 as a fluorescent one-component detection system for enzymatic H2O2 formation. We determined the kinetic parameters of the roGFP2-Orp1 reaction with H2O2 and established an efficient immobilization technique for the sensor on Saccharomyces cerevisiae cells employing the lectin Concanavalin A. This allowed to realize a peroxide-sensing shell on enzyme-displaying cells, a system that was successfully employed to screen for H2O2 formation of enzyme variants in a whole-cell setting.

Measurement of relative lung perfusion with electrical impedance and positron emission tomography: an experimental comparative study in pigs.

BACKGROUND: Electrical impedance tomography (EIT) with indicator dilution may be clinically useful to measure relative lung perfusion, but there is limited information on the performance of this technique. METHODS: Thirteen pigs (50-66 kg) were anaesthetised and mechanically ventilated. Sequential changes in ventilation were made: (i) right-lung ventilation with left-lung collapse, (ii) two-lung ventilation with optimised PEEP, (iii) two-lung ventilation with zero PEEP after saline lung lavage, (iv) two-lung ventilation with maximum PEEP (20/25 cm H2O to achieve peak airway pressure 45 cm H2O), and (v) two-lung ventilation under unilateral pulmonary artery occlusion. Relative lung perfusion was assessed with EIT and central venous injection of saline 3%, 5%, and 10% (10 ml) during breath holds. Relative perfusion was determined by positron emission tomography (PET) using 68Gallium-labelled microspheres. EIT and PET were compared in eight regions of equal ventro-dorsal height (right, left, ventral, mid-ventral, mid-dorsal, and dorsal), and directional changes in regional perfusion were determined. RESULTS: Differences between methods were relatively small (95% of values differed by less than 8.7%, 8.9%, and 9.5% for saline 10%, 5%, and 3%, respectively). Compared with PET, EIT underestimated relative perfusion in dependent, and overestimated it in non-dependent, regions. EIT and PET detected the same direction of change in relative lung perfusion in 68.9-95.9% of measurements. CONCLUSIONS: The agreement between EIT and PET for measuring and tracking changes of relative lung perfusion was satisfactory for clinical purposes. Indicator-based EIT may prove useful for measuring pulmonary perfusion at bedside.

[CMV-enterocolitis as a cause for repeated intestinal intussusceptions in an adult patient after liver transplantation?]. / CMV-Enterokolitis bei einer erwachsenen lebertransplantierten Patientin als Ursache rezidivierender Invaginationen?

Intestinal intussusception in the adult is often idiopathic but also known to be associated with chronic inflammatory bowel disease, coeliac disease, tumours or previous abdominal operations. A 22-year-old women after liver transplantation due to Crigler Najar Syndrome suffered from repeated episodes of abdominal pain. The diagnosis of repeated self-limited intestinal intussusceptions was made by computed tomography and ultrasonography. A laparoscopy revealed no cause for the intussusceptions. During a new episode of abdominal pain caused again by an intussusception a colonoscopy was performed that showed aspects of a discreet colitis. In the biopsies CMV was detected by qualitative PCR, while blood tests for CMV pp65 antigen were negative. A therapy with gancyclovir was initiated which lead to remission of the patient's symptoms. A colonoscopy six weeks later showed a completely normal colon, while in the biopsies CMV was not detectable. After a follow-up of one year the patient has not suffered from any further episodes. This case demonstrates the role of chronic intestinal CMV infection as a possible causative factor for repeated intussusceptions in immunosuppressed patients. Whenever possible a PCR for CMV in colon biopsies should be carried out to detect an intestinal CMV infection because as shown in our case results for immunohistopathology and CMV pp65 can be negative despite a chronic infection.

Assessment of radiation exposure on a dual-source computed tomography-scanner performing coronary computed tomography-angiography.

OBJECTIVE: The radiation exposure of a dual-source-64-channel multi-detector-computed-tomography-scanner (Somatom-Defintion, Siemens, Germany) was assessed in a phantom-study performing coronary-CT-angiography (CTCA) in comparison to patients' data randomly selected from routine scanning. METHODS: 240 CT-acquisitions of a computed tomography dose index (CTDI)-phantom (PTW, Freiburg, Germany) were performed using a synthetically generated Electrocardiography (ECG)-signal with variable heart rates (30-180 beats per minute (bpm)). 120 measurements were acquired using continuous tube-output; 120 measurements were performed using ECG-synchronized tube-modulation. The pulsing window was set at minimum duration at 65% of the cardiac cycle between 30 and 75 bpm. From 90-180 bpm the pulsing window was set at 30-70% of the cardiac cycle. Automated pitch adaptation was always used. A comparison between phantom CTDI and two patient groups' CTDI corresponding to the two pulsing groups was performed. RESULTS: Without ECG-tube-modulation CDTI-values were affected by heart-rate-changes resulting in 85.7 mGray (mGy) at 30 and 45 bpm, 65.5 mGy/60 bpm, 54.7 mGy/75 bpm, 46.5 mGy/90 bpm, 34.2 mGy/120 bpm, 27.0 mGy/150 bpm and 22.1 mGy/180 bpm equal to effective doses between 14.5 mSievert (mSv) at 30/45 bpm and 3.6 mSv at 180 bpm. Using ECG-tube-modulation these CTDI-values resulted: 32.6 mGy/30 bpm, 36.6 mGy/45 bpm, 31.4 mGy/60 bpm, 26.8 mGy/75 bpm, 23.7 mGy/90 bpm, 19.4 mGy/120 bpm, 17.2 mGy/150 bpm and 15.6 mGy/180 bpm equal to effective doses between 5.5 mSv at 30 bpm and 2.6 mSv at 180 bpm. Significant CTDI-differences were found between patients with lower/moderate and higher heart rates in comparison to the phantom CTDI-results. CONCLUSIONS: Dual source CTCA is particularly dose efficient at high heart rates when automated pitch adaptation, especially in combination with ECG-based tube-modulation is used. However in clinical routine scanning for patients with higher heart rates and corresponding enlarged pulsing window a significant different dose resulted.

Is post-mortem CT of the dentition adequate for correct forensic identification?: comparison of dental computed tomograpy and visual dental record.

The gold standard for identification of the dead is the visual dental record. In this context, several authors emphasize computed tomography (CT) as valuable supportive tool for forensic medicine. However, studies focusing on diagnostic accuracy of post-mortem computed tomography (PMCT) are still missing. Therefore, the aim of this study was to compare diagnostic accuracy of the visual dental record and post-mortem computed tomography (PMCT) of the dentition for identification of the dead. Ten whole skulls were included into the study. The entire dentition of each skull was first examined with the visual dental record as a gold standard and second using dental PMCT scans, performed on a 64-multi-detector CT (MDCT). 3D reformations, multi-planar reformations (MPR), and CT-orthopantomography (OPG) were performed in the post-processing. All examinations were analyzed by three independent investigators regarding the criteria for identification of the dead, e.g., in case of disaster. PMCT for the dental identification of the dead was difficult to perform and time consuming. Due to dental overlays and corresponding artifacts, the definite periphery of the dental fillings/inlays was not accurately defined resulting in 2.9% incorrect and 64.1% false negative findings, especially synthetic inlays were hardly or not recognizable at all. For the identification of the dead especially in case of disasters with large numbers of victims, the visual dental record is still to be considered the gold standard. In the identification process itself, there is no room for error at all, although some non-concordant information may occur. Thus, PMCT should only be performed for identification in individual cases due to the relatively high error rate.

[Interventional MRI of the breast. Indications, technique, results and perspectives]. / MR-gestützte Interventionen in der Mammadiagnostik. Indikationen, Techniken, Ergebnisse und Perspektiven.

With the use of diagnostic MR imaging of the breast MR guided minimal invasive interventions are increasingly needed to obtain histologic proof of lesions, that are occult at mammography and ultrasound. MR guided interventions are time consuming and expensive. Therefore it is necessary to insure, that the diagnostic MR examination of the referring institution establishing the indication for biopsy is of high quality.Assuming accurate patient selection percutaneous MR vacuum-assisted biopsy is a safe and accurate method for the evaluation of even small enhancing breast lesions. Up to now the imaging of the biopsy procedure itself is not possible due to a lack of compatibility of the biopsy needle. Careful reviewing of imaging and pathologic findings is therefore recommended. Several newly developed MR compatible devices will allow more precise placement and imaging of the biopsy needle. Procedure times will be reduced. For the evaluation of additional enhancing lesions in patients with breast carcinoma referred for operation the preoperative wire localization is remaining a wide-spread safe and simple method.

Pulmonary MRI--a new approach for the evaluation of febrile neutropenic patients with malignancies.

GOALS OF WORK: Immunocompromised patients with malignant diseases often suffer from pulmonary infections. Early detection of these life-threatening infections is crucial to start effective treatment. The gold standard for the diagnosis of these disorders is high-resolution computed tomography (HR-CT) of the chest. This method, however, has limitations, for instance, in the discrimination of early interstitial infiltrates and the use of X-rays. We conducted a study to determine the feasibility and sensitivity of magnetic resonance imaging (MRI) of the lung compared to HR-CT in immunocompromised patients with persistent fever in neutropenia and suspected pneumonia. MATERIALS AND METHODS: Between January 2003 and July 2004, 50 consecutive neutropenic patients with fever of unknown origin and negative chest X-ray were examined with HR-CT of the lungs. Patients with pulmonary infiltrates were further examined with MRI of the chest within 24 h after HR-CT using a specific lung protocol. In addition, microbiological testing was performed for the characterization of the causative pathogen. RESULTS: Of 50 patients, 35 had pulmonary infiltration according to HR-CT; these were examined with MRI of the lungs. MRI showed a high correlation (91%) with the findings in HR-CT. Both HR-CT and MRI were feasible in 94% of the examined patients. In 12 of 35 patients, fungal pathogens were identified in microbiological testing. CONCLUSIONS: MRI of the lungs is feasible in neutropenic patients with suspected pulmonary infection. Compared to HR-CT, MRI displays a high sensitivity in the detection of pulmonary infiltrates. MRI offers the opportunity of follow-up examinations without repeated X-ray exposure to the patient.

[Chronic pulmonary embolism--radiological imaging and differential diagnosis]. / Chronische Lungenembolie--Radiologische Bildmorphologie und Differenzialdiagnose.

In chronic pulmonary embolism branches of the pulmonary arterial tree remain partially or totally occluded. This may lead to pulmonary hypertension with the development of right ventricular hypertrophy as well as structural changes of pulmonary arteries. Imaging of chronic pulmonary embolism should prove vessel occlusions (pulmonary angiography, MSCT, MRI) and reduction of regional lung perfusion (lung scanning, MSCT, MRI). According to current guidelines ventilation-perfusion lung scanning and pulmonary angiography are still recommended as the methods of choice. MSCT and MRI provide technical alternatives which are helpful in differential diagnosis versus other types of pulmonary hypertension. In spite of medical and surgical measures (in rare cases pulmonary thromboendarterectomy) the prognosis of chronic pulmonary embolism remains unfavourable.

Beam hardening artefacts occur in dental implant scans with the NewTom cone beam CT but not with the dental 4-row multidetector CT.

OBJECTIVES: The aim of this study was to examine beam hardening artefacts of the NewTom 9000 cone-beam computed tomography (CBCT) device compared with the Philips MX 8000 (4-row multidetector CT (MDCT)). METHODS: We modified a SawBone skull to become a standardized model for our study. The skull was used for scans with the NewTom 9000 CBCT device and a standard dental multi-detector CT (MDCT) at a comparable reconstruction resolution with a standard Straumann ITI 4.1 mm implant in four implant positions in the maxilla (first permanent premolar in the right maxilla region, second permanent molar in the right maxilla region, first permanent premolar in the left maxilla region and second permanent molar in the left maxilla region). Results were compared with construction data of the dental implant. An image quality assessment of the images from both devices was performed with four experienced physicians and statistically analysed with the two-tailed Wilcoxon test. RESULTS: Scans with the NewTom 9000 CBCT showed strong beam hardening artefacts in the form of a radiation beam shadow in all reconstructions compared with the MDCT. These imaging artefacts became stronger with greater distance from the centre of the scanned volume. These differences in the imaging quality were proved as significant in a quality evaluation by four experienced physicians (P<0.05). CONCLUSIONS: Visual spatial resolution of the NewTom 9000 CBCT was less accurate than the Philips MX 8000 MDCT in the imaging of metallic dental implants.

Dose reduction in multidetector CT of the urinary tract. Studies in a phantom model.

A novel ureter phantom was developed for investigations of image quality and dose in CT urography. The ureter phantom consisted of a water box (14 cm x 32 cm x 42 cm) with five parallel plastic tubes (diameter 2.7 mm) filled with different concentrations of contrast media (1.88-30 mg iodine/ml). CT density of the tubes and noise of the surrounding water were determined using two multidetector scanners (Philips MX8000 with four rows, Siemens Sensation 16 with 16 rows) with varying tube current-time product (15-100 mAs per slice), voltage (90 kV, 100 kV, 120 kV), pitch (0.875-1.75), and slice thickness (1 mm, 2 mm, 3.2 mm). Contrast-to-noise ratio as a parameter of image quality was correlated with dose (CTDI) and was compared with image evaluation by two radiologists. The CT densities of different concentrations of contrast media and contrast-to-noise ratio were significantly higher when low voltages (90 kV versus 120 kV, 100 kV versus 120 kV) were applied. Smaller slice thickness (1 mm versus 2 mm) did not change CT density but decreased contrast-to-noise ratio due to increased noise. Contrast phantom studies showed favourable effects of low tube voltage on image quality in the low dose range. This may facilitate substantial dose reduction in CT urography.

[Magnetic resonance imaging in the evaluation of pneumonia]. / Nachweis von pneumonischen Infiltraten mit der MRT.

Magnetic resonance imaging (MRI) of the lung is challenging because of substantial drawbacks. However, lung pathologies that are associated with increased attenuation values in CT enhance visualization in MRI: proton density is increased and tissue-air interfaces, resulting in susceptibility artifacts, are reduced in pneumonia, pneumonitis, edema, and carcinoma. On the other hand, many lung diseases result in shortness of breath, so that patients cannot hold their breath for long periods. Therefore, fast imaging techniques are required which should also allow for high spatial resolution so that small lesions can be detected. Calcifications and air pockets within lesions are not readily recognized with MRI. Thin section CT is standard for the diagnosis of pneumonia. With parallel imaging techniques, MRI examination of the lungs can be performed with short periods of breath holding, which allow for sub-centimeter resolution in the z-axis. Especially for follow-up examinations in immunocompromised patients and, in some instances, for the staging of malignant diseases (malignant pleural mesothelioma, lung cancer, respectively), MRI is very promising and may contribute to a decrease in the radiation exposure of the patients.

Aorto-iliac multidetector-row CT angiography with low kV settings: improved vessel enhancement and simultaneous reduction of radiation dose.

The aim of the study was to implement an abdominal CT angiography protocol using 100 kVp and to compare SNR and CNR, as well as subjective image quality, to a standard CT angiography protocol using 120 kVp on a 16 detector-row CT scanner. Forty-eight patients were referred for routine abdominal CT angiography on a 16 detector-row CT scanner. Patients were scanned using either 120 or 100 kVp at constant mAs settings. Vessel opacification was provided by automated contrast injection using similar injection protocols. Density measurements were performed along the aorto-iliac axis with SNR and CNR calculation. In addition, the estimated effective patient radiation dose was calculated. Results of both protocols were compared. The 100-kVp protocol (432+/-80 HU) showed a significantly higher vessel density than the 120-kVp (333+/-90 HU; P<0.001) protocol, corresponding to an average increase in signal intensity of 30.7%. SNR (36.0 vs 37.0) and CNR (31.1 vs 31.7) for the 100-kV protocol were not significantly lower that those for the standard protocol (P=0.79 and P=0.87), whilst the average estimated dose was significantly lower using the 100-kVp protocol (6.7+/-0.4 vs 10.1+/-1.2 mSv; P<0.0001). Tube kVp reduction from 120 to 100 kVp allows for significant reduction of patient dose in abdominal CT angiography, without significant change in SNR,CNR and image quality.

[PET-CT interventional tumour therapy]. / PET-CT und interventionelle Tumortherapie.

Multifocal metastatic spread of solid tumours is often a diagnostic and therapeutic challenge. Those patients often cannot undergo curative surgery. Since several years, various types of interventional tumour therapy have been introduced, which may represent a promising option in those cases. In some cases morphological imaging is not able to assess the viability of a previously treated lesion. The combination of computed tomography and positron emission tomography has the potential to ease planning and guiding of interventional procedures and to improve postinterventional control. Due to its higher sensitivity and specificity, PET/CT enables for a more precise selection of patients for interventional tumor therapy. The success of a particular therapy could be assessed with greater accuracy using PET/CT as compared with the separate modalities. PET/CT was also able to improve planning and guiding interventional therapy. As of yet, no data are available concerning the success rate and long-term survival of PET/CT associated interventional procedures.

[Radiation dose and dose reduction in multidetector row CT (MDCT)]. / Strahlendosis und Möglichkeiten zur Dosisreduktion in der Mehrschicht-CT.

Since its introduction a few years ago, multidetector row CT (MDCT) has become a widely used diagnostic procedure and has been proven to be a valuable tool for various indications. A major issue using this new modality is the inherent risk of applying increased radiation exposure, when compared to single-slice CT or other imaging modalities.However, MDCT offers some valuable options to save radiation exposure, such as choosing optimized exposure parameters or its superior dose efficiency in comparison to single-slice CT. Multi-phasic examinations should be restricted to indications where definitely necessary. Modern scanners offer intelligent tools for further reduction of radiation dose, such as ECG- or bodyshape-based realtime dose modulation. A new field of applications is the low-dose CT for early detection of diseases. While acquiring thin slices with high spatial resolution, the dose can be reduced to similar values as in conventional radiography, especially when examining under high-contrast conditions. Using all these various options available, radiation exposure can sometimes even be lower than using a conventional single-slice helical CT.

[New abdominal CT angiography protocol on a 16 detector-row CT scanner - first results]. / Hochaufgelöste abdominelle Ubersichtsangiographie mit einem 16-Detektorzeilen-CT-System Erste Erfahrungen.

PURPOSE: Purpose of this study was the implementation of a new abdominal CT angiography protocol on a 16 detector-row CT scanner and the comparison to selective catheter angiography. METHODS: 76 patients with various vascular disorders underwent abdominal CT angiography on a recently developed 16 detector-row CT scanner using submillimeter slice collimation (16 x 0.75 mm). Results were compared with mesenteric and/or hepatic angiography in 17 patients performed during tumor embolization. Opacification was provided using individually tailored contrast application with a test bolus setting. To evaluate the contrast injection protocol density measurements within the vessel lumen were performed. RESULTS: Diagnostic image quality was achieved in all patients with angiographic comparison (n = 17). Within the hepatic and mesenteric vasculature up to 4th generation vessels could be identified. Compared to selective angiography CT angiography provided equivalent morphologic information up to the detectable vessel generation. With the applied contrast application regimen there were no significant differences in vessel enhancement along the abdominal aorta and iliac arteries. CONCLUSION: 16 detector-row CT enables whole abdominal angiographic studies with submillimeter resolution in a single breath-hold. The improved spatial resolution enables for high quality 3D visualization. Compared to invasive angiography, 16 detector-row CT reveals equivalent morphologic information.

[CT angiography of the coronary arteries with a 16-row spiral tomograph. Effect of spatial resolution on image quality]. / CT-Angiographie der Koronararterien mit einem 16-Mehrzeilen-Spiral-CT Einfluss der räumlichen Auflösung auf die Bildqualität.

PURPOSE: To evaluate image quality of coronary CT angiography with retrospectively ECG-gated 16 multi-slice spiral CT (MSCT), reconstructed with 0.75 mm slice thickness for optimal spatial resolution and with 1.3 mm slice thickness, to produce spatial resolution comparable to a 4-MSCT. MATERIALS AND METHODS: Ten patients underwent coronary CT angiography with a 16-MSCT (Siemens Sensation 16, Forchheim, Germany) with 0.75 mm detector collimation. Raw helical CT data were retrospectively reconstructed using two different settings. Setting A: B20f smooth kernel, axial MPR with 1.3 mm slice thickness and 0.7 mm increment. Setting B: B35f "HeartView" medium-smooth kernel, 0.75 mm slice thickness, 0.5 mm increment. In the axial slices two regions of interest (ROIs) were placed in the area of the aortic root (AR) and more caudal in the area of the left ventricle (LV). Image noise was determined by the standard deviation of the CT numbers.Two readers determined visibility of coronary arteries by standardized maximum intensity projections (MIP) post-processing in left, right anterior and left anterior oblique projection plane from setting A and B. Each projection was rated on a five point rating scale concerning plaque delineation. Points determined for each data set were summed up and used for comparison. RESULTS: No significant difference between the CT-numbers was found for setting A and B (A: 283.0 in AR/295.9 in LV and B: 282,9 in AR/297.2 in LV; p >0.2). However, the image noise was significantly different for setting A and B (A: 4.46 in AR/1.67 in LV and B: 8.16 in AR/7.38 in LV; p <0.01). Better delineation of the coronary arteries and atherosclerotic lesions could be achieved from MIP projections in setting B compared to setting A. CONCLUSION: Higher image noise is present in coronary 16-MSCT with thin-slice reconstruction compared to simulated 4-MSCT. However the MIP-reconstructions benefit most from the higher spatial resolution.

Multislice CT imaging of pulmonary embolism.

In recent years CT has been established as the method of choice for the diagnosis of central pulmonary embolism (PE) to the level of the segmental arteries. The key advantage of CT over competing modalities is the reliable detection of relevant alternative or additional disease causing the patient's symptoms. Although the clinical relevance of isolated peripheral emboli remains unclear, the alleged poor sensitivity of CT for the detection of such small clots has to date prevented the acceptance of CT as the gold standard for diagnosing PE. With the advent of multislice CT we can now cover the entire chest of a patient with 1-mm slices within one breath-hold. In comparison with thicker sections, the detection rate of subsegmental emboli can be significantly increased with 1-mm slices. In addition, the interobserver correlation which can be achieved with 1-mm sections by far exceeds the reproducibility of competing modalities. Meanwhile use of multislice CT for a combined diagnosis of PE and deep venous thrombosis with the same modality appears to be clinically accepted. In the vast majority of patients who receive a combined thoracic and venous multislice CT examination the scan either confirms the suspected diagnosis or reveals relevant alternative or additional disease. The therapeutic regimen is usually chosen based on the functional effect of embolic vascular occlusion. With the advent of fast CT scanning techniques, also functional parameters of lung perfusion can be non-invasively assessed by CT imaging. These advantages let multislice CT appear as an attractive modality for a non-invasive, fast, accurate, and comprehensive diagnosis of PE, its causes, effects, and differential diagnoses.

Pulmonary embolism: comprehensive diagnosis by using electron-beam CT for detection of emboli and assessment of pulmonary blood flow.

PURPOSE: To comprehensively assess thoracic anatomy and pulmonary microcirculation in pulmonary embolism by using computed tomographic (CT) angiography of the pulmonary arteries combined with functional CT imaging of blood flow. MATERIALS AND METHODS: Twenty-two patients suspected of having acute pulmonary embolism underwent contrast material-enhanced thin-section electron-beam CT angiography of the pulmonary arteries. In addition, in each patient, a dynamic multisection blood flow CT study was performed on a 7.6-cm lung volume with electrocardiographic gating. Pulmonary blood flow was calculated, and perfusion parameters were visualized on color-coded maps. The color-coded maps and CT angiograms were independently evaluated, segment by segment, by two readers for perfusion deficits and the presence of clots, respectively. The results were compared. RESULTS: Mean pulmonary blood flow was 0.63 mL/min/mL in the occluded segments versus 2.27 mL/min/mL in the nonoccluded segments (P: =.001). The sensitivity and specificity of perfusion maps for the presence of segmental pulmonary embolism compared with those of CT angiography were 75.4% and 82.3%, respectively, with positive and negative predictive values of 79.6% and 84.7%, respectively. The false-negative findings were caused mainly by partial occlusion of vessels. In eight patients, a substantial alternative or additional pathologic entity was diagnosed. CONCLUSION: By combining CT angiography and dynamic CT imaging, a comprehensive and noninvasive diagnosis of thoracic structure and function is feasible with a single modality.

Phase 1 trial of subcutaneous IL-6 in patients with refractory cancer: clinical and biologic effects.

The authors evaluated the clinical and biologic effects of human recombinant interleukin-6 (rhIL-6) in patients with refractory cancer. A phase 1 trial using escalating doses of rhIL-6 (1-50 microg x kg(-1) x d(-1), Monday through Friday for 4 weeks) was performed in 30 patients. Toxicity was moderate and the maximum tolerated dose was determined to be 25 microg x kg(-1)x d(-1) based on cardiac and neurocortical toxicity in one patient each and thrombocytosis (platelets > 800,000/microL) in three patients. One patient with non-small-cell lung cancer had a partial response after three cycles of therapy. The biologic effects of rhIL-6 included anemia and dose-related thrombocytosis. Various proinflammatory activities were induced and included dose-related cyclical increases in peripheral blood monocytes and the CD14+/CD45RB+ +/- CD16C+ mononuclear cell populations. These increases were accompanied by increased levels of C-reactive protein, serum neopterin, and type I soluble tumor necrosis factor receptor. In contrast, rhIL-6 did not affect lymphocyte numbers or function (cytotoxicity, cytokine levels, immunoglobulin levels), with the possible exception of IL-2Ralpha mRNA induction in peripheral blood lymphocytes. rhIL-6 has pleiotropic proinflammatory actions in vivo and moderate toxicity when administered as long-term therapy.