DEMIST: A Deep-Learning-Based Detection-Task-Specific Denoising Approach for Myocardial Perfusion SPECT.

There is an important need for methods to process myocardial perfusion imaging (MPI) single-photon emission computed tomography (SPECT) images acquired at lower-radiation dose and/or acquisition time such that the processed images improve observer performance on the clinical task of detecting perfusion defects compared to low-dose images. To address this need, we build upon concepts from model-observer theory and our understanding of the human visual system to propose a detection task-specific deep-learning-based approach for denoising MPI SPECT images (DEMIST). The approach, while performing denoising, is designed to preserve features that influence observer performance on detection tasks. We objectively evaluated DEMIST on the task of detecting perfusion defects using a retrospective study with anonymized clinical data in patients who underwent MPI studies across two scanners (N = 338). The evaluation was performed at low-dose levels of 6.25%, 12.5%, and 25% and using an anthropomorphic channelized Hotelling observer. Performance was quantified using area under the receiver operating characteristics curve (AUC). Images denoised with DEMIST yielded significantly higher AUC compared to corresponding low-dose images and images denoised with a commonly used task-agnostic deep learning-based denoising method. Similar results were observed with stratified analysis based on patient sex and defect type. Additionally, DEMIST improved visual fidelity of the low-dose images as quantified using root mean squared error and structural similarity index metric. A mathematical analysis revealed that DEMIST preserved features that assist in detection tasks while improving the noise properties, resulting in improved observer performance. The results provide strong evidence for further clinical evaluation of DEMIST to denoise low-count images in MPI SPECT.

Can patient-specific acquisition protocol improve performance on defect detection task in myocardial perfusion SPECT?

Myocardial perfusion imaging using single-photon emission computed tomography (SPECT), or myocardial perfusion SPECT (MPS) is a widely used clinical imaging modality for the diagnosis of coronary artery disease. Current clinical protocols for acquiring and reconstructing MPS images are similar for most patients. However, for patients with outlier anatomical characteristics, such as large breasts, images acquired using conventional protocols are often sub-optimal in quality, leading to degraded diagnostic accuracy. Solutions to improve image quality for these patients outside of increased dose or total acquisition time remain challenging. Thus, there is an important need for new methodologies to improve image quality for such patients. One approach to improving this performance is adapting the image acquisition protocol specific to each patient. For this study, we first designed and implemented a personalized patient-specific protocol-optimization strategy, which we term precision SPECT (PRESPECT). This strategy integrates ideal observer theory with the constraints of tomographic reconstruction to optimize the acquisition time for each projection view, such that MPS defect detection performance is maximized. We performed a clinically realistic simulation study on patients with outlier anatomies on the task of detecting perfusion defects on various realizations of low-dose scans by an anthropomorphic channelized Hotelling observer. Our results show that using PRESPECT led to improved performance on the defect detection task for the considered patients. These results provide evidence that personalization of MPS acquisition protocol has the potential to improve defect detection performance, motivating further research to design optimal patient-specific acquisition and reconstruction protocols for MPS, as well as developing similar approaches for other medical imaging modalities.

A task-specific deep-learning-based denoising approach for myocardial perfusion SPECT.

Deep-learning (DL)-based methods have shown significant promise in denoising myocardial perfusion SPECT images acquired at low dose. For clinical application of these methods, evaluation on clinical tasks is crucial. Typically, these methods are designed to minimize some fidelity-based criterion between the predicted denoised image and some reference normal-dose image. However, while promising, studies have shown that these methods may have limited impact on the performance of clinical tasks in SPECT. To address this issue, we use concepts from the literature on model observers and our understanding of the human visual system to propose a DL-based denoising approach designed to preserve observer-related information for detection tasks. The proposed method was objectively evaluated on the task of detecting perfusion defect in myocardial perfusion SPECT images using a retrospective study with anonymized clinical data. Our results demonstrate that the proposed method yields improved performance on this detection task compared to using low-dose images. The results show that by preserving task-specific information, DL may provide a mechanism to improve observer performance in low-dose myocardial perfusion SPECT.

DEMIST: A deep-learning-based task-specific denoising approach for myocardial perfusion SPECT.

There is an important need for methods to process myocardial perfusion imaging (MPI) SPECT images acquired at lower radiation dose and/or acquisition time such that the processed images improve observer performance on the clinical task of detecting perfusion defects. To address this need, we build upon concepts from model-observer theory and our understanding of the human visual system to propose a Detection task-specific deep-learning-based approach for denoising MPI SPECT images (DEMIST). The approach, while performing denoising, is designed to preserve features that influence observer performance on detection tasks. We objectively evaluated DEMIST on the task of detecting perfusion defects using a retrospective study with anonymized clinical data in patients who underwent MPI studies across two scanners (N = 338). The evaluation was performed at low-dose levels of 6.25%, 12.5% and 25% and using an anthropomorphic channelized Hotelling observer. Performance was quantified using area under the receiver operating characteristics curve (AUC). Images denoised with DEMIST yielded significantly higher AUC compared to corresponding low-dose images and images denoised with a commonly used task-agnostic DL-based denoising method. Similar results were observed with stratified analysis based on patient sex and defect type. Additionally, DEMIST improved visual fidelity of the low-dose images as quantified using root mean squared error and structural similarity index metric. A mathematical analysis revealed that DEMIST preserved features that assist in detection tasks while improving the noise properties, resulting in improved observer performance. The results provide strong evidence for further clinical evaluation of DEMIST to denoise low-count images in MPI SPECT.

A list-mode multi-energy window low-count SPECT reconstruction method for isotopes with multiple emission peaks.

BACKGROUND: Single-photon emission computed tomography (SPECT) provides a mechanism to perform absorbed-dose quantification tasks for [Formula: see text]-particle radiopharmaceutical therapies ([Formula: see text]-RPTs). However, quantitative SPECT for [Formula: see text]-RPT is challenging due to the low number of detected counts, the complex emission spectrum, and other image-degrading artifacts. Towards addressing these challenges, we propose a low-count quantitative SPECT reconstruction method for isotopes with multiple emission peaks. METHODS: Given the low-count setting, it is important that the reconstruction method extracts the maximal possible information from each detected photon. Processing data over multiple energy windows and in list-mode (LM) format provide mechanisms to achieve that objective. Towards this goal, we propose a list-mode multi energy window (LM-MEW) ordered-subsets expectation-maximization-based SPECT reconstruction method that uses data from multiple energy windows in LM format and include the energy attribute of each detected photon. For computational efficiency, we developed a multi-GPU-based implementation of this method. The method was evaluated using 2-D SPECT simulation studies in a single-scatter setting conducted in the context of imaging [[Formula: see text]Ra]RaCl[Formula: see text], an FDA-approved RPT for metastatic prostate cancer. RESULTS: The proposed method yielded improved performance on the task of estimating activity uptake within known regions of interest in comparison to approaches that use a single energy window or use binned data. The improved performance was observed in terms of both accuracy and precision and for different sizes of the region of interest. CONCLUSIONS: Results of our studies show that the use of multiple energy windows and processing data in LM format with the proposed LM-MEW method led to improved quantification performance in low-count SPECT of isotopes with multiple emission peaks. These results motivate further development and validation of the LM-MEW method for such imaging applications, including for [Formula: see text]-RPT SPECT.

Development and task-based evaluation of a scatter-window projection and deep learning-based transmission-less attenuation compensation method for myocardial perfusion SPECT.

Attenuation compensation (AC) is beneficial for visual interpretation tasks in single-photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI). However, traditional AC methods require the availability of a transmission scan, most often a CT scan. This approach has the disadvantage of increased radiation dose, increased scanner costs, and the possibility of inaccurate diagnosis in cases of misregistration between the SPECT and CT images. Further, many SPECT systems do not include a CT component. To address these issues, we developed a Scatter-window projection and deep Learning-based AC (SLAC) method to perform AC without a separate transmission scan. To investigate the clinical efficacy of this method, we then objectively evaluated the performance of this method on the clinical task of detecting perfusion defects on MPI in a retrospective study with anonymized clinical SPECT/CT stress MPI images. The proposed method was compared with CT-based AC (CTAC) and no-AC (NAC) methods. Our results showed that the SLAC method yielded an almost overlapping receiver operating characteristic (ROC) plot and a similar area under the ROC (AUC) to the CTAC method on this task. These results demonstrate the capability of the SLAC method for transmission-less AC in SPECT and motivate further clinical evaluation.

A list-mode multi-energy window low-count SPECT reconstruction method for isotopes with multiple emission peaks.

SPECT provides a mechanism to perform absorbed-dose quantification tasks for $\alpha$-particle radiopharmaceutical therapies ($\alpha$-RPTs). However, quantitative SPECT for $\alpha$-RPT is challenging due to the low number of detected counts, the complex emission spectrum, and other image-degrading artifacts. Towards addressing these challenges, we propose a low-count quantitative SPECT reconstruction method for isotopes with multiple emission peaks. Given the low-count setting, it is important that the reconstruction method extract the maximal possible information from each detected photon. Processing data over multiple energy windows and in list-mode (LM) format provide mechanisms to achieve that objective. Towards this goal, we propose a list-mode multi-energy window (LM-MEW) OSEM-based SPECT reconstruction method that uses data from multiple energy windows in LM format, and includes the energy attribute of each detected photon. For computational efficiency, we developed a multi-GPU-based implementation of this method. The method was evaluated using 2-D SPECT simulation studies in a single-scatter setting conducted in the context of imaging [$^{223}$Ra]RaCl${_2}$. The proposed method yielded improved performance on the task of estimating activity uptake within known regions of interest in comparison to approaches that use a single energy window or use binned data. The improved performance was observed in terms of both accuracy and precision and for different sizes of the region of interest. Results of our studies show that the use of multiple energy windows and processing data in LM format with the proposed LM-MEW method led to improved quantification performance in low-count SPECT of isotopes with multiple emission peaks. These results motivate further development and validation of the LM-MEW method for such imaging applications, including for $\alpha$-RPT SPECT.

Need for objective task-based evaluation of deep learning-based denoising methods: A study in the context of myocardial perfusion SPECT.

BACKGROUND: Artificial intelligence-based methods have generated substantial interest in nuclear medicine. An area of significant interest has been the use of deep-learning (DL)-based approaches for denoising images acquired with lower doses, shorter acquisition times, or both. Objective evaluation of these approaches is essential for clinical application. PURPOSE: DL-based approaches for denoising nuclear-medicine images have typically been evaluated using fidelity-based figures of merit (FoMs) such as root mean squared error (RMSE) and structural similarity index measure (SSIM). However, these images are acquired for clinical tasks and thus should be evaluated based on their performance in these tasks. Our objectives were to: (1) investigate whether evaluation with these FoMs is consistent with objective clinical-task-based evaluation; (2) provide a theoretical analysis for determining the impact of denoising on signal-detection tasks; and (3) demonstrate the utility of virtual imaging trials (VITs) to evaluate DL-based methods. METHODS: A VIT to evaluate a DL-based method for denoising myocardial perfusion SPECT (MPS) images was conducted. To conduct this evaluation study, we followed the recently published best practices for the evaluation of AI algorithms for nuclear medicine (the RELAINCE guidelines). An anthropomorphic patient population modeling clinically relevant variability was simulated. Projection data for this patient population at normal and low-dose count levels (20%, 15%, 10%, 5%) were generated using well-validated Monte Carlo-based simulations. The images were reconstructed using a 3-D ordered-subsets expectation maximization-based approach. Next, the low-dose images were denoised using a commonly used convolutional neural network-based approach. The impact of DL-based denoising was evaluated using both fidelity-based FoMs and area under the receiver operating characteristic curve (AUC), which quantified performance on the clinical task of detecting perfusion defects in MPS images as obtained using a model observer with anthropomorphic channels. We then provide a mathematical treatment to probe the impact of post-processing operations on signal-detection tasks and use this treatment to analyze the findings of this study. RESULTS: Based on fidelity-based FoMs, denoising using the considered DL-based method led to significantly superior performance. However, based on ROC analysis, denoising did not improve, and in fact, often degraded detection-task performance. This discordance between fidelity-based FoMs and task-based evaluation was observed at all the low-dose levels and for different cardiac-defect types. Our theoretical analysis revealed that the major reason for this degraded performance was that the denoising method reduced the difference in the means of the reconstructed images and of the channel operator-extracted feature vectors between the defect-absent and defect-present cases. CONCLUSIONS: The results show the discrepancy between the evaluation of DL-based methods with fidelity-based metrics versus the evaluation on clinical tasks. This motivates the need for objective task-based evaluation of DL-based denoising approaches. Further, this study shows how VITs provide a mechanism to conduct such evaluations computationally, in a time and resource-efficient setting, and avoid risks such as radiation dose to the patient. Finally, our theoretical treatment reveals insights into the reasons for the limited performance of the denoising approach and may be used to probe the effect of other post-processing operations on signal-detection tasks.

A task-specific deep-learning-based denoising approach for myocardial perfusion SPECT.

Deep-learning (DL)-based methods have shown significant promise in denoising myocardial perfusion SPECT images acquired at low dose. For clinical application of these methods, evaluation on clinical tasks is crucial. Typically, these methods are designed to minimize some fidelity-based criterion between the predicted denoised image and some reference normal-dose image. However, while promising, studies have shown that these methods may have limited impact on the performance of clinical tasks in SPECT. To address this issue, we use concepts from the literature on model observers and our understanding of the human visual system to propose a DL-based denoising approach designed to preserve observer-related information for detection tasks. The proposed method was objectively evaluated on the task of detecting perfusion defect in myocardial perfusion SPECT images using a retrospective study with anonymized clinical data. Our results demonstrate that the proposed method yields improved performance on this detection task compared to using low-dose images. The results show that by preserving task-specific information, DL may provide a mechanism to improve observer performance in low-dose myocardial perfusion SPECT.

Development and task-based evaluation of a scatter-window projection and deep learning-based transmission-less attenuation compensation method for myocardial perfusion SPECT.

Attenuation compensation (AC) is beneficial for visual interpretation tasks in single-photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI). However, traditional AC methods require the availability of a transmission scan, most often a CT scan. This approach has the disadvantages of increased radiation dose, increased scanner cost, and the possibility of inaccurate diagnosis in cases of misregistration between the SPECT and CT images. Further, many SPECT systems do not include a CT component. To address these issues, we developed a Scatter-window projection and deep Learning-based AC (SLAC) method to perform AC without a separate transmission scan. To investigate the clinical efficacy of this method, we then objectively evaluated the performance of this method on the clinical task of detecting perfusion defects on MPI in a retrospective study with anonymized clinical SPECT/CT stress MPI images. The proposed method was compared with CT-based AC (CTAC) and no-AC (NAC) methods. Our results showed that the SLAC method yielded an almost overlapping receiver operating characteristic (ROC) plot and a similar area under the ROC (AUC) to the CTAC method on this task. These results demonstrate the capability of the SLAC method for transmission-less AC in SPECT and motivate further clinical evaluation.

Need for Objective Task-based Evaluation of Deep Learning-Based Denoising Methods: A Study in the Context of Myocardial Perfusion SPECT.

Artificial intelligence-based methods have generated substantial interest in nuclear medicine. An area of significant interest has been using deep-learning (DL)-based approaches for denoising images acquired with lower doses, shorter acquisition times, or both. Objective evaluation of these approaches is essential for clinical application. DL-based approaches for denoising nuclear-medicine images have typically been evaluated using fidelity-based figures of merit (FoMs) such as RMSE and SSIM. However, these images are acquired for clinical tasks and thus should be evaluated based on their performance in these tasks. Our objectives were to (1) investigate whether evaluation with these FoMs is consistent with objective clinical-task-based evaluation; (2) provide a theoretical analysis for determining the impact of denoising on signal-detection tasks; (3) demonstrate the utility of virtual clinical trials (VCTs) to evaluate DL-based methods. A VCT to evaluate a DL-based method for denoising myocardial perfusion SPECT (MPS) images was conducted. The impact of DL-based denoising was evaluated using fidelity-based FoMs and AUC, which quantified performance on detecting perfusion defects in MPS images as obtained using a model observer with anthropomorphic channels. Based on fidelity-based FoMs, denoising using the considered DL-based method led to significantly superior performance. However, based on ROC analysis, denoising did not improve, and in fact, often degraded detection-task performance. The results motivate the need for objective task-based evaluation of DL-based denoising approaches. Further, this study shows how VCTs provide a mechanism to conduct such evaluations using VCTs. Finally, our theoretical treatment reveals insights into the reasons for the limited performance of the denoising approach.

Ideal-Observer Computation with anthropomorphic phantoms using Markov chain Monte Carlo.

In medical imaging, it is widely recognized that image quality should be objectively evaluated based on performance in clinical tasks. To evaluate performance in signal-detection tasks, the ideal observer (IO) is optimal but also challenging to compute in clinically realistic settings. Markov Chain Monte Carlo (MCMC)-based strategies have demonstrated the ability to compute the IO using pre-computed projections of an anatomical database. To evaluate image quality in clinically realistic scenarios, the observer performance should be measured for a realistic patient distribution. This implies that the anatomical database should also be derived from a realistic population. In this manuscript, we propose to advance the MCMC-based approach towards achieving these goals. We then use the proposed approach to study the effect of anatomical database size on IO computation for the task of detecting perfusion defects in simulated myocardial perfusion SPECT images. Our preliminary results provide evidence that the size of the anatomical database affects the computation of the IO.

Deep-SMOLM: deep learning resolves the 3D orientations and 2D positions of overlapping single molecules with optimal nanoscale resolution.

Dipole-spread function (DSF) engineering reshapes the images of a microscope to maximize the sensitivity of measuring the 3D orientations of dipole-like emitters. However, severe Poisson shot noise, overlapping images, and simultaneously fitting high-dimensional information-both orientation and position-greatly complicates image analysis in single-molecule orientation-localization microscopy (SMOLM). Here, we report a deep-learning based estimator, termed Deep-SMOLM, that achieves superior 3D orientation and 2D position measurement precision within 3% of the theoretical limit (3.8° orientation, 0.32 sr wobble angle, and 8.5 nm lateral position using 1000 detected photons). Deep-SMOLM also demonstrates state-of-art estimation performance on overlapping images of emitters, e.g., a 0.95 Jaccard index for emitters separated by 139 nm, corresponding to a 43% image overlap. Deep-SMOLM accurately and precisely reconstructs 5D information of both simulated biological fibers and experimental amyloid fibrils from images containing highly overlapped DSFs at a speed ~10 times faster than iterative estimators.

Investigating the limited performance of a deep-learning-based SPECT denoising approach: An observer-study-based characterization.

Multiple objective assessment of image-quality (OAIQ)-based studies have reported that several deep-learning (DL)-based denoising methods show limited performance on signal-detection tasks. Our goal was to investigate the reasons for this limited performance. To achieve this goal, we conducted a task-based characterization of a DL-based denoising approach for individual signal properties. We conducted this study in the context of evaluating a DL-based approach for denoising single photon-emission computed tomography (SPECT) images. The training data consisted of signals of different sizes and shapes within a clustered-lumpy background, imaged with a 2D parallel-hole-collimator SPECT system. The projections were generated at normal and 20% low-count level, both of which were reconstructed using an ordered-subset-expectation-maximization (OSEM) algorithm. A convolutional neural network (CNN)-based denoiser was trained to process the low-count images. The performance of this CNN was characterized for five different signal sizes and four different signal-to-background ratio (SBRs) by designing each evaluation as a signal-known-exactly/background-known-statistically (SKE/BKS) signal-detection task. Performance on this task was evaluated using an anthropomorphic channelized Hotelling observer (CHO). As in previous studies, we observed that the DL-based denoising method did not improve performance on signal-detection tasks. Evaluation using the idea of observer-study-based characterization demonstrated that the DL-based denoising approach did not improve performance on the signal-detection task for any of the signal types. Overall, these results provide new insights on the performance of the DL-based denoising approach as a function of signal size and contrast. More generally, the observer study-based characterization provides a mechanism to evaluate the sensitivity of the method to specific object properties, and may be explored as analogous to characterizations such as modulation transfer function for linear systems. Finally, this work underscores the need for objective task-based evaluation of DL-based denoising approaches.

A physics and learning-based transmission-less attenuation compensation method for SPECT.

Attenuation compensation (AC) is a pre-requisite for reliable quantification and beneficial for visual interpretation tasks in single-photon emission computed tomography (SPECT). Typical AC methods require the availability of an attenuation map, which is obtained using a transmission scan, such as a CT scan. This has several disadvantages such as increased radiation dose, higher costs, and possible misalignment between SPECT and CT scans. Also, often a CT scan is unavailable. In this context, we and others are showing that scattered photons in SPECT contain information to estimate the attenuation distribution. To exploit this observation, we propose a physics and learning-based method that uses the SPECT emission data in the photopeak and scatter windows to perform transmission-less AC in SPECT. The proposed method uses data acquired in the scatter window to reconstruct an initial estimate of the attenuation map using a physics-based approach. A convolutional neural network is then trained to segment this initial estimate into different regions. Pre-defined attenuation coefficients are assigned to these regions, yielding the reconstructed attenuation map, which is then used to reconstruct the activity distribution using an ordered subsets expectation maximization (OSEM)-based reconstruction approach. We objectively evaluated the performance of this method using highly realistic simulation studies conducted on the clinically relevant task of detecting perfusion defects in myocardial perfusion SPECT. Our results showed no statistically significant differences between the performance achieved using the proposed method and that with the true attenuation maps. Visually, the images reconstructed using the proposed method looked similar to those with the true attenuation map. Overall, these results provide evidence of the capability of the proposed method to perform transmission-less AC and motivate further evaluation.

Objective Task-Based Evaluation of Artificial Intelligence-Based Medical Imaging Methods:: Framework, Strategies, and Role of the Physician.

Artificial intelligence-based methods are showing promise in medical imaging applications. There is substantial interest in clinical translation of these methods, requiring that they be evaluated rigorously. We lay out a framework for objective task-based evaluation of artificial intelligence methods. We provide a list of available tools to conduct this evaluation. We outline the important role of physicians in conducting these evaluation studies. The examples in this article are proposed in the context of PET scans with a focus on evaluating neural network-based methods. However, the framework is also applicable to evaluate other medical imaging modalities and other types of artificial intelligence methods.

Task-based assessment of binned and list-mode SPECT systems.

In SPECT, list-mode (LM) format allows storing data at higher precision compared to binned data. There is significant interest in investigating whether this higher precision translates to improved performance on clinical tasks. Towards this goal, in this study, we quantitatively investigated whether processing data in LM format, and in particular, the energy attribute of the detected photon, provides improved performance on the task of absolute quantification of region-of-interest (ROI) uptake in comparison to processing the data in binned format. We conducted this evaluation study using a DaTscan brain SPECT acquisition protocol, conducted in the context of imaging patients with Parkinson's disease. This study was conducted with a synthetic phantom. A signal-known exactly/background-known-statistically (SKE/BKS) setup was considered. An ordered-subset expectation-maximization algorithm was used to reconstruct images from data acquired in LM format, including the scatter-window data, and including the energy attribute of each LM event. Using a realistic 2-D SPECT system simulation, quantification tasks were performed on the reconstructed images. The results demonstrated improved quantification performance when LM data was used compared to binning the attributes in all the conducted evaluation studies. Overall, we observed that LM data, including the energy attribute, yielded improved performance on absolute quantification tasks compared to binned data.

A LIST-MODE OSEM-BASED ATTENUATION AND SCATTER COMPENSATION METHOD FOR SPECT.

Reliable attenuation and scatter compensation (ASC) is a pre-requisite for quantification and beneficial for visual interpretation tasks in SPECT. In this paper, we develop a reconstruction method that uses the entire SPECT emission data, i.e. data in both the photopeak and scatter windows, acquired in list-mode format and including the energy attribute of the detected photon, to perform ASC. We implemented a GPU-based version of this method using an ordered subsets expectation maximization (OSEM) algorithm. The method was objectively evaluated using realistic simulation studies on the task of estimating uptake in the striatal regions of the brain in a 2-D dopamine transporter (DaT)-scan SPECT study. We observed that inclusion of data from the scatter window and using list-mode data yielded improved quantification compared to using data only from the photopeak window or using binned data. These results motivate further development of list-mode-based ASC methods that include scatter-window data for SPECT.

A novel vascular targeting strategy for brain-derived endothelial cells using a TCR mimic antibody.

Organ-specific vascular targeting, for example, to the blood-brain barrier, requires the identification of unique molecular addresses on a subset of endothelial cells. The present study describes a crucial step towards tapping the exquisite specificity of the peptide/HLA class I system for this goal. We utilized a novel T-cell receptor (TCR) mimic antibody of high affinity and specificity, which is restricted by HLA-A2 and has been generated to recognize a peptide epitope derived from p68 RNA helicase (YLLPAIVHI). The parent protein is highly expressed by brain endothelial cells. Flow cytometry and confocal imaging showed that the antibody binds to HLA-A2-positive human brain-derived endothelial cells, both immortalized hCMEC/D3 cells and primary cells. The TCR mimic antibody undergoes internalization into vesicles, where significant colocalization occurs with the early endosomal marker EEA-1, but barely with caveolin-1. To our knowledge internalization of neither MHC class I protein nor TCR mimics by brain endothelial cells has been previously observed. Knock down of p68 protein expression by siRNA reduced the presentation of YLLPAIVHI-peptide/HLA-A2 complexes on the cell membrane by half as measured by flow cytometry 48 h later. We also found that brain endothelial cells isolated from HLA-A2 transgenic mouse strains express the A2 transgene, and brain endothelial cells of one of these strains also present YLLPAIVHI-peptide/HLA-A2, making these mouse strains suitable models for studying TCR mimic antibodies in vivo. In conclusion, these data strongly support the notion that TCR mimic antibodies could be a new class of therapeutic targeting agents in a wide variety of diseases.

Changes of visual evoked potential induced by lateral geniculate nucleus kindling in rats.

PURPOSE: The present study was undertaken to investigate the changes of visual evoked potential (VEP) induced by lateral geniculate nucleus (LGN) kindling in comparison with those induced by amygdala (AMG) and hippocampus (HPC) kindling in rats. METHODS: Under pentobarbital anesthesia, the rats were fixed to a stereotaxic apparatus, and stainless steel electrodes were implanted into the LGN, AMG, HPC and the occipital cortex (OCOR). Bipolar stimulation was applied to the LGN, AMG or HPC every day until generalized seizure was obtained. VEP was recorded from the OCOR before the first day of electric stimulation (pre-kindled) and after the development of kindling (post-kindled). RESULTS: After the development of LGN kindling, the amplitudes of early VEP components (P(1)-N(1) and N(1)-P(2)) were significantly augmented in comparison with those of pre-kindled rats. In addition, the duration of the photically evoked after-discharge burst (duration of VEP) was significantly prolonged by LGN kindling. On the other hand, in AMG and HPC kindling, no significant changes were observed in VEP between pre-kindled and post-kindled rats. CONCLUSIONS: The changes of VEP observed in patients with photosensitive epilepsy are also demonstrated in animal study using LGN kindling.