The Feasibility of Deep Learning-Based Reconstruction for Low-Tube-Voltage CT Angiography for Transcatheter Aortic Valve Implantation.

OBJECTIVE: The purpose of this study is to evaluate the efficacy of deep learning reconstruction (DLR) on low-tube-voltage computed tomographic angiography (CTA) for transcatheter aortic valve implantation (TAVI). METHODS: We enrolled 30 patients who underwent TAVI-CT on a 320-row CT scanner. Electrocardiogram-gated coronary CTA (CCTA) was performed at 100 kV, followed by nongated aortoiliac CTA at 80 kV using a single bolus of contrast material. We used hybrid-iterative reconstruction (HIR), model-based IR (MBIR), and DLR to reconstruct these images. The contrast-to-noise ratios (CNRs) were calculated. Five-point scales were used for the overall image quality analysis. The diameter of the aortic annulus was measured in each reconstructed image, and we compared the interobserver and intraobserver agreements. RESULTS: In the CCTA, the CNR and image quality score for DLR were significantly higher than those for HIR and MBIR ( P < 0.01). In the aortoiliac CTA, the CNR for DLR was significantly higher than that for HIR ( P < 0.01) and significantly lower than that for MBIR ( P ≤ 0.02). The image quality score for DLR was significantly higher than that for HIR ( P < 0.01). No significant differences were observed between the image quality scores for DLR and MBIR. The measured aortic annulus diameter had high interobserver and intraobserver agreement regardless of the reconstruction method (all intraclass correlation coefficients, >0.89). CONCLUSIONS: In low tube voltage TAVI-CT, DLR provides higher image quality than HIR, and DLR provides higher image quality than MBIR in CCTA and is visually comparable to MBIR in aortoiliac CTA.

Virtual monochromatic spectral CT imaging in preoperative evaluations for intraductal spread of breast cancer: comparison with conventional CT and MRI.

PURPOSE: To investigate the efficacy of virtual monochromatic spectral computed tomography imaging (VMI) in the preoperative evaluation for intraductal spread of breast cancer. MATERIALS AND METHODS: Twenty-four women who underwent spectral CT and were pathologically diagnosed with ductal carcinoma with a ≥ 2-cm noninvasive component were retrospectively enrolled in Group 1. Twenty-two women with 22 lesions pathologically diagnosed with ductal carcinoma in situ or microinvasive carcinoma were enrolled in Group 2. We compared the contrast-to-noise ratios (CNRs) of the lesions on conventional 120-kVp CT images and 40-keV VMIs in Group 1. Two board-certified radiologists measured the maximum diameters of enhancing areas on 120-kVp CT, 40-keV VMI, and MRI in Group 2 and compared with histopathological sizes. RESULTS: The quantitative assessment of Group 1 revealed that the mean ± SD of the CNRs in the 40-keV images were significantly greater than those in the 120-kVp images (5.5 ± 1.9 vs. 3.6 ± 1.5, p < 0.0001). The quantitative assessment of Group 2 demonstrated that the lesion size observed in the conventional 120-kVp CT images by both readers was significantly underestimated as compared to the histopathological size (p = 0.017, 0.048), whereas both readers identified no significant differences between the lesion size measured on 40-keV VMI and the histopathological data. In a comparison with MRI, 40-keV VMI provided measurement within a 10-mm error range in more lesions as compared to the conventional 120-kVp CT. CONCLUSION: VMI improves the evaluation of intraductal spread and is useful for the preoperative evaluations of breast cancer.

Efficacy of Dynamic Chest Radiography for Chronic Thromboembolic Pulmonary Hypertension.

Background While current guidelines require lung ventilation-perfusion (V/Q) scanning as the first step to diagnose chronic pulmonary embolism in pulmonary hypertension (PH), its use may be limited by low availability and/or exposure to ionizing radiation. Purpose To compare the performance of dynamic chest radiography (DCR) and lung V/Q scanning for detection of chronic thromboembolic PH (CTEPH). Materials and Methods Patients with PH who underwent DCR and V/Q scanning in the supine position from December 2019 to July 2021 were retrospectively screened. The diagnosis of CTEPH was confirmed with right heart catheterization and invasive pulmonary angiography. Observer tests were conducted to evaluate the diagnostic accuracy of DCR and V/Q scanning. The lungs were divided into six areas (upper, middle, and lower for both) in the anteroposterior image, and the number of lung areas with thromboembolic perfusion defects was scored. Diagnostic performance was compared between DCR and V/Q scanning using the area under the receiver operating characteristic curve. Agreement between the interpretation of DCR and that of V/Q scanning was assessed using the Cohen kappa coefficient and percent agreement. Results A total of 50 patients with PH were analyzed: 29 with CTEPH (mean age, 64 years ± 15 [SD]; 19 women) and 21 without CTEPH (mean age, 61 years ± 22; 14 women). The sensitivity, specificity, and accuracy of DCR were 97%, 86%, and 92%, respectively, and those of V/Q scanning were 100%, 86%, and 94%, respectively. Areas under the receiver operating characteristic curve for DCR and V/Q scanning were 0.92 (95% CI: 0.79, 0.97) and 0.93 (95% CI: 0.78, 0.98). Agreement between the consensus interpretation of DCR and that of V/Q scanning was substantial (κ = 0.79 [95% CI: 0.61, 0.96], percent agreement = 0.9 [95% CI: 0.79, 0.95]). Conclusion Dynamic chest radiography had similar efficacy to ventilation-perfusion scanning in the detection of chronic thromboembolic pulmonary hypertension. © RSNA, 2022 Online supplemental material is available for this article. See also the editorial by Wandtke and Koproth-Joslin in this issue.

Impact of a new deep-learning-based reconstruction algorithm on image quality in ultra-high-resolution CT: clinical observational and phantom studies.

OBJECTIVES: To demonstrate the effect of an improved deep learning-based reconstruction (DLR) algorithm on Ultra-High-Resolution Computed Tomography (U-HRCT) scanners. METHODS: Clinical and phantom studies were conducted. Thirty patients who underwent contrast-enhanced CT examination during the follow-up period were enrolled. Images were reconstructed using improved DLR [termed, New DLR, i.e., Advanced Intelligent Clear-IQ Engine (AiCE) Body Sharp] and conventional DLR (Conv DLR, AiCE Body) algorithms. Two radiologists assessed the overall image quality using a 5-point scale (5 = excellent; 1 = unacceptable). The noise power spectra (NPSs) were calculated to assess the frequency characteristics of the image noise, and the square root of area under the curve (√AUC NPS) between 0.05 and 0.50 cycle/mm was calculated as an indicator of the image noise. Dunnett's test was used for statistical analysis of the visual evaluation score, with statistical significance set at p < 0.05. RESULTS: The overall image quality of New DLR was better than that of the Conv DLR (4.2 ± 0.4 and 3.3 ± 0.4, respectively; p < 0.0001). All New DLR images had an overall image quality score above the average or excellent. The √AUCNPS value of New DLR was lower than that of Conv DLR (13.8 and 14.2, respectively). The median values of reconstruction time required with New DLR and Conv DLR were 5.0 and 7.8 min, respectively. CONCLUSIONS: The new DLR algorithm improved the image quality within a practical reconstruction time. ADVANCES IN KNOWLEDGE: The new DLR enables us to choose whether to improve image quality or reduce the dose.

Evaluation of MR imaging findings differentiating parotid basal cell adenomas from other parotid tumors.

PURPOSE: To evaluate the usefulness of magnetic resonance imaging (MRI) to differentiate basal cell adenomas (BCAs) from other parotid tumors. METHOD: A total of 136 patients with histologically proven parotid gland tumors (13 BCAs, 66 pleomorphic adenomas [PAs], 30 Warthin tumors [WTs], and 27 parotid cancers [PCs]) who underwent a cervical MRI study between December 2011 and March 2019 were retrospectively enrolled. The MRI findings of the tumors were evaluated by two board-certified radiologists. RESULTS: All 13 of the BCAs showed smooth margins, while 19 of the 27 PCs showed irregular margins (p < 0.0001). Eleven BCAs had some cystic components, and five were cyst-dominant. The BCAs had significantly more cystic components than the PAs (p = 0.0077). The mean apparent diffusion coefficient (ADC) value of the BCAs was 1.21 ± 0.20 × 10-3 mm2/sec, which was equivalent to that of the PCs (1.12 ± 0.25 × 10-3 mm2/sec, p = 0.76), significantly lower than that of the PAs (1.61 ± 0.32 × 10-3 mm2/sec, p < 0.0001), and significantly higher than that of the WTs (0.81 ± 0.19 × 10-3 mm2/sec, p = 0.0004). The plateau time-intensity curve (TIC) was the most common type for both BCAs and PCs, seen in 8 of 12 BCAs and 21 of 26 PCs, with no significant difference between these groups (p = 0.34). CONCLUSIONS: BCA should be considered a possibility when a parotid lesion has smooth margins with an entire capsule and includes a cystic component, even if the TIC and diffusion-weighted MR images suggest a malignant pattern.

Right Ventricular Extracellular Volume with Dual-Layer Spectral Detector CT: Value in Chronic Thromboembolic Pulmonary Hypertension.

Background Right ventricular (RV) extracellular volumes (ECVs), as a surrogate for histologic fibrosis, have not been sufficiently investigated. Purpose To evaluate and compare RV and left ventricular (LV) ECVs obtained with dual-layer spectral detector CT (DLCT) in chronic thromboembolic pulmonary hypertension (CTEPH) and investigate the clinical importance of RV ECV. Materials and Methods Retrospective analysis was performed on data from 31 patients with CTEPH (17 were not treated with pulmonary endarterectomy [PEA] or balloon pulmonary angioplasty [BPA] and 14 were) and eight control subjects who underwent myocardial delayed enhancement (MDE) DLCT from January 2019 to June 2020. The ECVs in the RV and LV walls were calculated by using iodine density as derived from spectral data pertaining to MDE. Statistical analyses were performed with one-way repeated analysis of variance with the Tukey post hoc test or the Kruskal-Wallis test with the Steel-Dwass test and linear regression analysis. Results The PEA- and BPA-naive group showed significantly higher ECVs than the PEA- or BPA-treated group and control group in the septum (28.2% ± 2.9 vs 24.3% ± 3.6, P = .005), anterior right ventricular insertion point (RVIP) (32.9% ± 4.6 vs 25.3% ± 3.6, P < .001), posterior RVIP (35.2% ± 5.2 vs 27.3% ± 4.2, P < .001), mean RVIP (34.0% ± 4.2 vs 26.3% ± 3.4, P < .001), RV free wall (29.5% ± 3.3 vs 25.9% ± 4.1, P = .036), and mean RV wall (29.1% ± 3.0 vs 26.1% ± 3.1, P = .029). There were no significant differences between the PEA- or BPA-treated group and control subjects in these segments (septum, P = .93; anterior RVIP, P = .38; posterior RVIP, P = .52; mean RVIP, P = .36; RV free wall, P = .97; and mean RV, P = .33). There were significant correlations between ECV and mean pulmonary artery pressure (PAP) or brain natriuretic peptide (BNP) in the mean RVIP (mean PAP: R = 0.66, P < .001; BNP: R = 0.44, P = .014) and the mean RV (mean PAP: R = 0.49, P = .005; BNP: R = 0.44, P = .013). Conclusion Right ventricular and right ventricular insertion point extracellular volumes could be noninvasive surrogate markers of disease severity and reverse tissue remodeling in chronic thromboembolic pulmonary hypertension. © RSNA, 2021 Online supplemental material is available for this article. See also the editorial by Sandfort and Bluemke in this issue.

Model-based iterative reconstruction for 320-detector row CT angiography reduces radiation exposure in infants with complex congenital heart disease.

PURPOSE: We investigated the impact of model-based iterative reconstruction (MBIR) on 320-detector row computed tomography angiography (CTA) in infants with complex congenital heart disease (CHD). METHODS: Seventy infants with complex CHD who underwent 320-detector row CTA (40 boys and 30 girls; age range, 0-22 months; median age, 60 days) were retrospectively evaluated. First, the images were reconstructed by filtered back projection (FBP), hybrid iterative reconstruction (HIR), or MBIR in 20 cases, and variables were compared among the three iterative reconstruction methods (IR test). Second, the variables were compared between 25 cases scanned using HIR and 25 cases scanned using MBIR, with a 20 standard deviation noise level for both. Attenuation values and contrast-to-noise ratios (CNRs) of the great vessels and heart chambers were calculated. Total dose-length products were recorded for all patients (radiation dose: RD test). RESULTS: In the IR test, the mean CNR values were 4.8±1.3 for FBP, 6.9±1.4 for HIR, and 8.2±1.7 for MBIR (P < 0.0001). The best subjective image qualities in the great vessels and heart chambers were obtained with MBIR. In RD testing, no significant differences between HIR and MBIR in image quality (CNR: HIR, 8.4±2.4; MBIR, 8.3±2.4) were observed. The effective dose was significantly lower for MBIR than for HIR (0.7±0.2 vs. 1.1±0.3 mSv; P < 0.001). CONCLUSION: The MBIR algorithm significantly improved image quality and decreased radiation exposure in 320-row CTA of infants with complex CHD, providing an alternative to FBP or HIR that is both safer and produces better results.

Characterization of parotid gland tumors: added value of permeability MR imaging to DWI and DCE-MRI.

OBJECTIVES: To determine added value of permeability MRI in parotid tumor characterization to T2-weighted imaging (T2WI), semi-quantitative analysis of time-intensity curve (TIC), and intra-voxel incoherent motion diffusion-weighted imaging (IVIM-DWI). METHODS: This retrospective study was approved by the institutional review board, and the informed consent was waived. Sixty-one parotid tumors in 61 patients were examined using T2WI, IVIM-DWI, and permeability MRI. TIC patterns were categorized as persistent, washout, or plateau. Signal intensity ratio of lesion-to-muscle on T2WI, apparent diffusion coefficients (ADCs), D and f values from IVIM-DWI, and Ktrans, kep, Ve, and Vp values from permeability MRI were measured. Multiple comparisons were applied to determine whether any differences among 4 histopathologic types (pleomorphic adenomas, Warthin's tumors, other benign tumors, and malignant tumors) existed. Diagnostic accuracy was compared before and after modification diagnosis referring to permeability MRI. In a validation study, 60 parotid tumors in 60 patients were examined. RESULTS: ADC and D values of malignant tumors were significantly lower than those of benign tumors other than Warthin's tumors, but higher than those of Warthin's tumors. kep and Vp values of Warthin's tumors were significantly higher than those of malignant tumors. Multivariate analyses showed that TIC pattern, D, and kep values were suitable parameters. McNemar's test showed a significant increase of sensitivity (11/12, 92%) and specificity (46/49, 94%) with adding kep. The validation study yielded high sensitivity (14/16, 88%) and specificity (41/44, 93%). CONCLUSION: Permeability MRI offers added value to IVIM-MRI and semi-quantitative TIC analysis of DCE-MRI in characterization of parotid tumors KEY POINTS: • Permeability MR imaging offers added value in the characterization of parotid gland tumors in combination with semi-quantitative TIC analysis and IVIM analyses with D parameter. • The combination of TIC pattern, D, and kep might facilitate accurate characterization of parotid gland tumor, thereby avoiding unnecessary surgery for benign tumors or delayed treatment for malignant tumors. • A combination of permeability and diffusion MR imaging can be used to guide the selection of an appropriate biopsy site.

Amide proton transfer (APT) imaging of parotid tumors: Differentiation of malignant and benign tumors.

PURPOSE: To assess the usefulness of amide proton transfer (APT) imaging in differentiating parotid tumors. MATERIAL AND METHODS: We retrospectively analyzed 43 histopathologically proven parotid solid tumors with diameters ≥2 cm. Twenty-one tumors were benign and 12 tumors were malignant. Two-dimensional APT imaging was performed using a saturation pulse with a duration of 2 s and a saturation power level of 2 µT. For acquiring Z-spectra, the imaging was repeated at 25 saturation frequency offsets from ω = -6 to +6 ppm with a step of 0.5 ppm as well as one scan acquired far off-resonance (-1560 ppm) for signal normalization. For the APT imaging, the asymmetry analysis at 3.5 ppm downfield from the water signal was calculated. The mean APT signal intensity (SI) was compared between the benign and malignant tumors. RESULTS: The mean APT SI was 2.23 ± 0.80 % in the benign tumors and significantly higher at 2.99 ± 0.99 % in the malignant tumors (P = 0.01). A receiver operating curve analysis revealed that the optimal APT SI threshold was 2.40 for distinguishing malignant tumors from benign tumors with an area under the curve of 0.74. The sensitivity, specificity, and accuracy were 83.3%, 61.3%, and 67.4%, respectively. CONCLUSION: The mean APT SI of the malignant parotid tumors was significantly higher than that of the benign parotid tumors.

Balloon pulmonary angioplasty improves right atrial reservoir and conduit functions in chronic thromboembolic pulmonary hypertension.

AIMS: Right atrial (RA) function largely contributes to the maintenance of right ventricular (RV) function. This study investigated the effect of balloon pulmonary angioplasty (BPA) on RA functions in patients with inoperable chronic thromboembolic pulmonary hypertension (CTEPH) using cardiac magnetic resonance imaging (CMRI). METHODS AND RESULTS: CMRI and RV catheterization were performed before BPA sessions and at the follow-up periods in 29 CTEPH patients. Reservoir [RA longitudinal strain (RA-LS)], passive conduit [RA early LS rate (LSR)], and active (RA late LSR) phases were assessed by using cine CMRI and a feature-tracking algorithm. The relationships between the changes in RA functions and in brain natriuretic peptide (BNP) were evaluated in both the dilated and non-dilated RA groups. RA-LS (32.4% vs. 42.7%), RA LSR (6.3% vs. 8.3%), and RA early LSR (-2.3% vs. -4.3%) were improved after BPA, whereas no significant change was seen in RA late LSR. The changes in RA peak LS and in RA early LSR were significantly correlated with the changes in BNP (ΔRA-LS: r = -0.63, ΔRA-early LSR: r = 0.65) and pulmonary vascular resistance (PVR) (ΔRA-LS: r = -0.69, ΔRA-early LSR: r = 0.66) in the nondilated RA group. CONCLUSION: The RA reservoir and passive conduit functions were impaired in inoperable CTEPH, whereas RA active function was preserved. BPA markedly reversed these impaired functions. The improvements in RA reservoir and conduit functions were significantly correlated with the changes in BNP levels and PVR in CTEPH patients with normal RA sizes.

Detectability of the artery of Adamkiewicz on computed tomography angiography of the aorta by using ultra-high-resolution computed tomography.

PURPOSE: To evaluate the detectability of AKA on aortic computed tomography angiography (CTA) using ultra-high-resolution computed tomography (UHRCT). MATERIALS AND METHODS: Twenty-eight patients were enrolled. They underwent aortic CTA with UHRCT (UHRCTA) and had previously undergone aortic conventional CTA (CCTA). The injection protocol of UHRCTA was the same as that of CCTA. The bolus tracking technique was used. UHRCTA images were reconstructed with adaptive iterative dose reduction (strong) and with forward-projected model-based iterative reconstruction solution. The matrix size and slice thickness on UHRCT were 1024 and 0.25 mm, respectively, and those on conventional CT were 512 and 0.5 or 0.67 mm, respectively. The UHRCTA and CCTA images were visually compared by using four scales. A score of 4 or 3 indicated that the AKA was assessable. In this instance, the contrast-to-noise ratios of each UHRCTA were measured. The exposure dose and signal-to-noise ratios were also investigated. RESULTS: The AKA visualization scores obtained with UHRCTA with forward-projected model-based iterative reconstruction solution were significantly higher than those with adaptive iterative dose reduction (p = 0.018) and CCTA (p = 0.0024). CONCLUSION: UHRCT can contribute to the better visualization of the AKA on aortic CTA.

Feasibility and Reproducibility of Fetal Left Ventricular Twist Using Two-Dimensional Speckle-Tracking Analysis in a Japanese Population.

In fetal echocardiography, conventional parameters for assessing cardiac function are limited because of limited echocardiographic windows or the fetus' position. We aimed to evaluate the feasibility and reproducibility of fetal left ventricular (LV) twist by two-dimensional, speckle-tracking echocardiography (2DSTE) in a Japanese population.We included 55 normal fetuses at gestational ages between 21 and 36 weeks. Subjects with adverse maternal health issues were excluded. LV twist was calculated as the net difference between LV basal and apical rotation at end-systole estimated with 2DSTE.We were able to analyze the 2DSTE images in 44 cases (80%). The mean (±SE) apical rotation, basal rotation, and LV twist were 7.88 ± 0.77, -3.68 ± 0.50, and 11.1 ± 0.75 degrees, respectively. We could not analyze 11 cases (20%) because of poor image quality due to fetal position in five cases (45.5%), failure to track the endocardium because of blurred images in five cases (45.5%), and failure to obtain images of the heart due to the presence of the placenta in front of the fetus in one case (9.1%). There were no significant differences in the demographic data between pregnant women in whom LV twist analysis was feasible and not feasible. The intra- and interobserver intraclass correlation coefficients were 0.67 and 0.64, respectively.LV twist analysis by 2DSTE in the fetus was feasible in a substantial population and may provide new insight into cardiac function during the prenatal period. On the other hand, its reproducibility was moderate and needs to be improved.

Clinical usefulness of right ventricular 3D area strain in the assessment of treatment effects of balloon pulmonary angioplasty in chronic thromboembolic pulmonary hypertension: comparison with 2D feature-tracking MRI.

OBJECTIVES: To evaluate the usefulness of right ventricular (RV) area strain analysis via cardiac MRI (CMRI) as a tool for assessing the treatment effects of balloon pulmonary angioplasty (BPA) in inoperable chronic thromboembolic pulmonary hypertension (CTEPH), RV area strain was compared to two-dimensional (2D) strain with feature-tracking MRI (FTMRI) before and after BPA. METHODS: We retrospectively analyzed 21 CTEPH patients who underwent BPA. End-systolic global area strain (GAS), longitudinal strain (LS), circumferential strain (CS), and radial strain (RS) were measured before and after BPA. Changes in GAS and RV ejection fraction (RVEF) values after BPA were defined as ΔGAS and ΔRVEF. Receiver operating characteristic (ROC) analyses were performed to determine the optimal cutoff of the strain at after BPA for detection of improved patients with decreased mean pulmonary artery pressure (mPAP) less than 30 mmHg and increased RVEF more than 50%. RESULTS: ROC analysis revealed the optimal cutoffs of strains (GAS, LS, CS, and RS) for identifying improved patients with mPAP < 30 mmHg (cutoff (%) = - 41.2, - 13.8, - 16.7, and 14.4: area under the curve, 0.75, 0.56, 0.65, and 0.75) and patients with RVEF > 50% (cutoff (%) = - 37.2, - 29.5, - 2.9, and 14.4: area under the curve, 0.81, 0.60, 0.56, and 0.56). CONCLUSIONS: Area strain analysis via CMRI may be a more useful tool for assessing the treatment effects of BPA in patients with CTEPH than 2D strains with FTMRI. KEY POINTS: • Area strain values can detect improvement of right ventricular (RV) pressure and function after balloon pulmonary angioplasty (BPA) equally or more accurately than two-dimensional strains. • Area strain analysis is a useful analytical method that reflects improvements in complex RV myocardial deformation by BPA. • Area strain analysis is a robust method with reproducibility equivalent to that of 2D strain analysis.

Clinical application of radiation dose reduction for head and neck CT.

CT has advantages over MRI including rapid acquisition, and high spatial resolution for detailed anatomical information on the head and neck region. Therefore, CT is the first choice of imaging modality for the larynx, hypopharynx, sinonasal region, and temporal bone. Introduction of multi-detector CT (MDCT) scanning has allowed reduction in scan time, availability of isovoxel image, and relevant 3D image reconstruction; however, it leads to over-ranging due to helical scanning, and increased radiation dose due to 3D-volume imaging, and small detector size. In head and neck CT, reduction and optimization of radiation dose is very important, especially for prevention of the occurrence of cataract development due to radiation to lens, and prevention of the development of malignant tumour development from radiosensitive organs such as the salivary gland, thyroid gland, and retina, especially in children. The goal of dose reduction is "as low as reasonably achievable" (ALARA) level with preservation of appropriate image quality in clinical practice. Reduction of radiation dose per examination is essential; however, indication of repeat examination such as perfusion CT, dynamic contrast-enhanced CT, and follow-up study of malignant tumours should be optimized.

Induction of cytotoxic T lymphocytes against human cancer cell lines using dendritic cell-tumor cell hybrids generated by a newly developed electrofusion technique.

Recently, dendritic cells (DCs) and DC-tumor cell hybrids (DC-tumor hybrids) have been used for cancer vaccine therapy in a clinical trial. DC-tumor hybrids combine the potent antigen-presenting capacity of DCs with the ability to present all tumor antigens expressed on tumor cells to T cells. We used DC-tumor hybrids as stimulator cells to induce tumor-specific cytotoxic T lymphocytes (CTLs) in vitro. DC-tumor hybrids were generated from human monocyte-derived DCs and human cancer-cell lines (GT3TKB, lung cancer; GCIY, gastric cancer) by our newly developed electrofusion technique, established and refined with the use of mouse cells. To evaluate the capacity of DC-tumor hybrids generated by our method to induce tumor antigen-specific CTLs, we performed a cytotoxic assay and an interferon-gamma release assay using CD8-dominant effector lymphocytes induced by them. DC-tumor hybrids more effectively induced tumor-specific primary T-cell response than did stimulation with DCs co-cultured with irradiated tumor cells overnight, irradiated tumor cells alone, or a mixture of DCs and irradiated tumor cells. DC-tumor hybrids were generated at a high fusion rate by our electrofusion technique. When CTLs were induced by DC-tumor hybrids in vitro, the high fusion rate did not contribute to the induction of CTLs with increased tumor-specific cytotoxicity. The addition of interleukin-12 to the culture medium did not augment the cytotoxicity of CTLs. Overall, our results suggest that DC-tumor hybrids effectively induce human tumor-specific CTLs and may thus be applicable for clinical trials of adoptive immunotherapy.

A new strategy using autologous dendritic cells and lymphokine-activated killer cells for cancer immunotherapy: efficient maturation of DCs by co-culture with LAK cells in vitro.

Among a variety of antigen presenting cells (APCs), accumulating results support that the mature dendritic cell (DC) has the potential to induce efficient cytotoxic T lymphocyte (CTL) responses in the context of peptide-based immunotherapy. DCs have been known to assume the mature form by signaling through the CD40-CD40 ligand (CD40L) interaction, which may be provided by activated CD4+ T cells expressing abundant CD40L molecules on their surfaces. Here, we report that DCs generated from peripheral blood monocytes obtained from patients with advanced cancer exhibit a mature phenotype after co-culturing with autologous lymphokine-activated killer (LAK) cells generated by the stimulation of peripheral blood mononuclear cells with anti-CD3 monoclonal antibody (mAb) and interleukin (IL)-2. Part of this process appeared to be dependent on the expression of CD40L on the surface of LAK cells, although it was also suggested that some other humoral factors produced by LAK cells may be involved in this effect as well. DCs derived from the donors, of which LAK cells demonstrated a higher Th1/Th2 ratio upon activation determined by the intracellular detection of interferon-gamma and IL-4, showed more efficient maturation upon co-culture with LAK cells than DCs from donors with a low Th1/Th2 ratio. Importantly, these matured DCs induced a two-times stronger antigen-presenting capacity measured by an allo-reactive mixed lymphocytes reaction assay as compared to immature DCs. These results imply the use of the combination of DCs and LAK cells for immunotherapy against cancer.

[Immunogenic reactivity of CTLs induced by electrofusion cells of human dendritic cells and gastric cancer cells].

In tumor immunotherapy, there were several reports of attempts to induce anti-tumor immunity by fusion hybrid cells generated with dendritic and tumor cells. One of them reported that vaccination of hybrid cells resulted in a remarkable reduction of tumor cells in a lab mouse experiment. In our study, fusion cells were generated successfully with human matured dendritic and human gastric cancer cells by electrofusion technique and employed to induce CTLs. The evaluated fusion rate was 47.8% by FACS analysis. We tried to induce CTLs by co culture of effector and stimulator cells in the presence of IL-2, IL-7 and IL-12 for 4 weeks. Although it was not statistically significant in tumor cytotoxic assay, effector cells induced by the fusion cells as stimulator cells showed a few cytotoxic responses in an immunological tumor specific manner. Our data suggest that fusion hybrid cells may facilitate stimulation and expansion of tumor-specific T cells, but further investigation is required for clinical application of fusion cells in adoptive immunotherapy.