The Role of Hypoxia in Brain Tumor Immune Responses

Oxygen is a vital component of living cells. Low levels of oxygen in body tissues, known as hypoxia, can affect multiple cellular functions across a variety of cell types and are a hallmark of brain tumors. In the tumor microenvironment, abnormal vasculature and enhanced oxygen consumption by tumor cells induce broad hypoxia that affects not only tumor cell characteristics but also the antitumor immune system. Although some immune reactions require hypoxia, hypoxia generally negatively affects immunity. Hypoxia induces tumor cell invasion, cellular adaptations to hypoxia, and tumor cell radioresistance. In addition, hypoxia limits the efficacy of immunotherapy and hinders antitumor responses.Therefore, understanding the role of hypoxia in the brain tumor, which usually does not respond to immunotherapy alone is important for the development of effective anti-tumor therapies. In this review, we discuss recent evidence supporting the role of hypoxia in the context of brain tumors.

Monocytes Contribute to IFN-β Production via the MyD88-Dependent Pathway and Cytotoxic T-Cell Responses against Mucosal Respiratory Syncytial Virus Infection

Respiratory syncytial virus (RSV) is the leading cause of respiratory viral infection in infants and children. However, little is known about the contribution of monocytes to antiviral responses against RSV infection. We identified the IFN-β production of monocytes using IFN-β/YFP reporter mice. The kinetic analysis of IFN-β-producing cells in in vivo RSV-infected lung cells indicated that monocytes are recruited to the inflamed lung during the early phase of infection. These cells produced IFN-β via the myeloid differentiation factor 88-mediated pathway, rather than the TLR7- or mitochondrial antiviral signaling protein-mediated pathway. In addition, monocyte-ablated mice exhibited decreased numbers of IFN-γ-producing and RSV Ag-specific CD8 + T cells. Collectively, these data indicate that monocytes play pivotal roles in cytotoxic T-cell responses and act as type I IFN producers during RSV infection.

Single Cell Transcriptomic Re-analysis of Immune Cells in Bronchoalveolar Lavage Fluids Reveals the Correlation of B Cell Characteristics and Disease Severity of Patients with SARS-CoV-2 Infection

The coronavirus disease 2019 (COVID-19) pandemic (severe acute respiratory syndrome coronavirus 2) is a global infectious disease with rapid spread. Some patients have severe symptoms and clinical signs caused by an excessive inflammatory response, which increases the risk of mortality. In this study, we reanalyzed scRNA-seq data of cells from bronchoalveolar lavage fluids of patients with COVID-19 with mild and severe symptoms, focusing on Ab-producing cells. In patients with severe disease, B cells seemed to be more activated and expressed more immunoglobulin genes compared with cells from patients with mild disease, and macrophages expressed higher levels of the TNF superfamily member B-cell activating factor but not of APRIL (a proliferation-inducing ligand). In addition, macrophages from patients with severe disease had increased pro-inflammatory features and pathways associated with Fc receptor-mediated signaling, compared with patients with mild disease. CCR2-positive plasma cells accumulated in patients with severe disease, probably because of increased CCL2 expression on macrophages from patients with severe disease.Together, these results support the hypothesis that different characteristics of B cells might be associated with the severity of COVID-19 infection.

The autophagy Protein Atg5 Plays a Crucial Role in the Maintenance and Reconstitution Ability of Hematopoietic Stem Cells

Hematopoietic stem cells (HSCs) in bone marrow are pluripotent cells that can constitute the hematopoiesis system through self-renewal and differentiation into immune cells and red blood cells. To ensure a competent hematopoietic system for life, the maintenance of HSCs is tightly regulated. Although autophagy, a self-degradation pathway for cell homeostasis, is essential for hematopoiesis, the role of autophagy key protein Atg5 in HSCs has not been thoroughly investigated. In this study, we found that Atg5 deficiency in hematopoietic cells causes survival defects, resulting in severe lymphopenia and anemia in mice. In addition, the absolute numbers of HSCs and multiple-lineage progenitor cells were significantly decreased, and abnormal erythroid development resulted in reduced erythrocytes in blood of Vav_Atg5(−/−) mice. The proliferation of Lin⁻Sca-1⁺c-Kit⁺ HSCs was aberrant in bone marrow of Vav_Atg5(−/−) mice, and mature progenitors and terminally differentiated cells were also significantly altered. Furthermore, the reconstitution ability of HSCs in bone marrow chimeric mice was significantly decreased in the presence of Atg5 deficiency in HSCs. Mechanistically, impairment of autophagy-mediated clearance of damaged mitochondria was the underlying cause of the HSC functional defects. Taken together, these results define the crucial role of Atg5 in the maintenance and the reconstitution ability of HSCs.

Caspase-1 Independent Viral Clearance and Adaptive Immunity Against Mucosal Respiratory Syncytial Virus Infection

Respiratory syncytial virus (RSV) infection is recognized by the innate immune system through Toll like receptors (TLRs) and retinoic acid inducible gene I. These pathways lead to the activation of type I interferons and resistance to infection. In contrast to TLRs, very few studies have examined the role of NOD-like receptors in viral recognition and induction of adaptive immune responses to RSV. Caspase-1 plays an essential role in the immune response via the maturation of the proinflammatory cytokines IL-1beta and IL-18. However, the role of caspase-1 in RSV infection in vivo is unknown. We demonstrate that RSV infection induces IL-1beta secretion and that caspase-1 deficiency in bone marrow derived dendritic cells leads to defective IL-1beta production, while normal RSV viral clearance and T cell responses are observed in caspase-1 deficient mice following respiratory infection with RSV. The frequencies of IFN-gamma producing or RSV specific T cells in lungs from caspase-1 deficient mice are not impaired. In addition, we demonstrate that caspase-1 deficient neonatal or young mice also exhibit normal immune responses. Furthermore, we find that IL-1R deficient mice infected with RSV exhibit normal Th1 and cytotoxic T lymphocytes (CTL) immune responses. Collectively, these results demonstrate that in contrast to TLR pathways, caspase-1 might not play a central role in the induction of Th1 and CTL immune responses to RSV.

Differential Roles of Lung Dendritic Cell Subsets Against Respiratory Virus Infection

Respiratory viruses can induce acute respiratory disease. Clinical symptoms and manifestations are dependent on interactions between the virus and host immune system. Dendritic cells (DCs), along with alveolar macrophages, constitute the first line of sentinel cells in the innate immune response against respiratory viral infection. DCs play an essential role in regulating the immune response by bridging innate and adaptive immunity. In the steady state, lung DCs can be subdivided into CD103+ conventional DCs (cDCs), CD11b+ cDCs, and plasmacytoid DCs (pDCs). In the inflammatory state, like a respiratory viral infection, monocyte-derived DCs (moDCs) are recruited to the lung. In inflammatory lung, discrimination between moDCs and CD11b+ DCs in the inflamed lung has been a critical challenge in understanding their role in the antiviral response. In particular, CD103+ cDCs migrate from the intraepithelial base to the draining mediastinal lymph nodes to primarily induce the CD8+ T cell response against the invading virus. Lymphoid CD8alpha+ cDCs, which have a developmental relationship with CD103+ cDCs, also play an important role in viral antigen presentation. Moreover, pDCs have been reported to promote an antiviral response by inducing type I interferon production rather than adaptive immunity. However, the role of these cells in respiratory infections remains unclear. These different DC subsets have functional specialization against respiratory viral infection. Under certain viral infection, contextually controlling the balance of these specialized DC subsets is important for an effective immune response and maintenance of homeostasis.

Autophagy as an Innate Immune Modulator

Autophagy is a fundamental cellular process in eukaryotic cells for maintaining homeostasis by degrading cellular proteins and organelles. Recently, the roles of autophagy have been expanded to immune systems, which in turn modulate innate immune responses. More specifically, autophagy acts as a direct effector for protection against pathogens, as well as a modulator of pathogen recognition and downstream signaling in innate immune responses. In addition, autophagy controls autoimmunity and inflammatory disorders by negative regulation of immune signaling. In this review, we focus on recent advances in the role of autophagy in innate immune systems.

Autophagy in Innate Recognition of Pathogens and Adaptive Immunity

Autophagy is a specialized cellular pathway involved in maintaining homeostasis by degrading long-lived cellular proteins and organelles. Recent studies have demonstrated that autophagy is utilized by immune systems to protect host cells from invading pathogens and regulate uncontrolled immune responses. During pathogen recognition, induction of autophagy by pattern recognition receptors leads to the promotion or inhibition of consequent signaling pathways. Furthermore, autophagy plays a role in the delivery of pathogen signatures in order to promote the recognition thereof by pattern recognition receptors. In addition to innate recognition, autophagy has been shown to facilitate MHC class II presentation of intracellular antigens to activate CD4 T cells. In this review, we describe the roles of autophagy in innate recognition of pathogens and adaptive immunity, such as antigen presentation, as well as the clinical relevance of autophagy in the treatment of human diseases.

Development of Solenoid RF Coil for Animal Imaging in 3T High Magnetic Field MRI

PURPOSE: The purpose of the present study was to develop and optimize solenoid coil for animal- model in 3 T MRI system and investigate and compare with the birdcage coil concerning the image quality with the various parameters such as SNR and Q-factor. MATERIALS AND METHODS: Solenoid coil for animal-model was made on the acryl structure (diameter 4 cm, length 10 cm) 3 times-winding cooper tape of width 2 cm , thickness 0.05 cm and length 10 cm with 2 cm interval between winded tapes. Capacitors from 2 pF to 100 pF were used, and the solenoid coil was designed for receiver only coil. RESULTS: SNR of the developed solenoid was 985 in CuSO4 0.7 g/L and 995 in rat experiment. Q-factor was 84-89 in unloaded condition and 203-206 in loaded condition. CONCLUSION: The resolution of the image obtained from solenoid was relatively higher than that of the conventional birdcage coil. In addition, the homogeneity of RF field by coil simulation was significantly excellent. The present study demonstrated that the solenoid coil could be useful to obtain small animal images with better contrast, resolution, visibility than images from birdcage.

Quantitative Analysis of Magnetization Transfer by Phase Sensitive Method in Knee Disorder

Magnetization Transfer (MT) imaging generates contrast dependent on the phenomenon of magnetization exchange between free water proton and restricted proton in macromolecules. In biological materials in knee, MT or cross-relaxation is commonly modeled using two spin pools identified by their different T2 relaxation times. Two models for cross-relaxation emphasize the role of proton chemical exchange between protons of water and exchangeable protons on macromolecules, as well as through dipole-dipole interaction between the water and macromolecule protons. The most essential tool in medical image manipulation is the ability to adjust the contrast and intensity. Thus, it is desirable to adjust the contrast and intensity of an image interactively in the real time. The proton density (PD) and T2-weighted SE MR images allow the depiction of knee structures and can demonstrate defects and gross morphologic changes. The PD- and T2-weighted images also show the cartilage internal pathology due to the more intermediate signal of the knee joint in these sequences. Suppression of fat extends the dynamic range of tissue contrast, removes chemical shift artifacts, and decreases motion-related ghost artifacts. Like fat saturation, phase sensitive methods are also based on the difference in precession frequencies of water and fat. In this study, phase sensitive methods look at the phase difference that is accumulated in time as a result of Larmor frequency differences rather than using this difference directly. Although how MT work was given with clinical evidence that leads to quantitative model for MT in tissues, the mathematical formalism used to describe the MT effect applies to explaining to evaluate knee disorder, such as anterior cruciate ligament (ACL) tear and meniscal tear. Calculation of the effect of the effect of the MT saturation is given in the magnetization transfer ratio (MTR) which is a quantitative measure of the relative decrease in signal intensity due to the MT pulse.

High Resolution MR Images from 3T Active-Shield Whole-Body MRI System

PURPOSE: Within a clinically acceptable time frame, we obtained the high resolution MR images of the human brain, knee, foot and wrist from 3T whole-body MRI system which was equipped with the world first 3T active shield magnet. MATERIALS AND METHODS: Spin echo (SE) and Fast Spin Echo (FSE) images were obtained from the human brain, knee, foot and wrist of normal subjects using a homemade birdcage and transverse electromagnetic (TEM) resonators operating in quadrature and tuned to 128 MHz. For acquisition of MR images of knee, foot and wrist, we employed a homemade saddle shaped RF coil. Typical common acquisition parameters were as follows: matrix= 512x512, field of view (FOV) = 20 cm, slice thickness = 3 mm, number of excitations (NEX) = 1. For T1-weighted MR images, we used TR= 500 ms, TE = 10 or 17.4 ms. For T2-weighted MR images, we used TR=4000 ms, TE = 108 ms. RESULTS: Signal to noise ratio (SNR) of 3T system was measured 2.7 times greater than that of prevalent 1.5T system. MR images obtained from 3T system revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter. CONCLUSION: The present results demonstrate that the MR images from 3T system could provide better diagnostic quality of resolution and sensitivity than those of 1.5T system. The elevated SNR observed in the 3T high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the microscopic structural level under in vivo conditions. These images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods.

The Comparison of Susceptibility Changes in 1.5T and 3.0T MRIs due to TE Change in Functional MRI

PURPOSE: The purpose of this study was to find the optimum TE value for enhancing T2* weighting effect and minimizing the SNR degradation and to compare the BOLD effects according to the changes of TE in 1.5T and 3.0T MRI systems. MATERIALS AND METHODS: Healthy normal volunteers (eight males and two females with 24-38 years old) participated in this study. Each volunteer was asked to perform a simple finger-tapping task (sequential opposition of thumb to each of the other four fingers) with right hand with a mean frequency of about 2Hz. The stimulus was initially off for 3 images and was then alternatively switched on and off for 2 cycles of 6 images. Images were acquired on the 1.5T and 3.0T MRI with the FLASH (fast low-angle shot) pulse sequence (TR : 100ms, FA : 20degrees, FOV : 230mm) that was used with 26, 36, 46, 56, 66, 76ms of TE times in 1.5T and 16, 26, 36, 46, 56, 66ms of TE in 3.0T MRI system. After the completion of scan, MR images were transferred into a PC and processed with a home-made analysis program based on the correlation coefficient method with the threshold value of 0.45. To search for the optimum TE value in fMRI, the difference between the activation and the rest by the susceptibility change for each TE was used in 1.5T and 3.0T respectively. In addition, the functional T2* map was calculated to quantify susceptibility change. RESULTS: The calculated optimum TE for fMRI was 61.89+/-2.68 at 1.5T and 47.64+/-13.34 at 3.0T. The maximum percentage of signal intensity change due to the susceptibility effect in activation region was 3.36% at TE 66ms in 1.5T and 10.05% at TE 46ms in 3.0T, respectively. The signal intensity change of 3.0T was about 3 times bigger than that of 1.5T. The calculated optimum TE value was consistent with TE values which were obtained from the maximum signal change for each TE. CONCLUSION: In this study, the 3.0T MRI was clearly more sensitive, about three times bigger than the 1.5T in detecting the susceptibility due to the deoxyhemoglobin level change in the functional MR imaging. So the 3.0T fMRI is more useful than 1.5T.

Development of TEM Head-size Resonator for 3T MRI Head Coil

PURPOSE: To apply a distributed circuit theory, to develop a head-size transverse electromagnetic(TEM) resonator coil for a home-built 3T whole-body MRI system and to report an efficiency of the coil. MATERIALS AND METHODS: The dimension of TEM resonator with 16 cavity element, the diameter of copper rod was 0.63cm and its length was 13.75cm. As raw materials, the purity of copper rod was 98% and the dielectric constant of teflon was 2.08. RESULTS: The TEM head-size resonator with 16cavity elements exhibiting 9-mode resonances was robust to the surrounding influences owing to the self-shielding structure. The isolation of quadrature with a human brain was 364 and the ratio of Q(unloaded/Q(loaded) was 2.9. CONCLUSION: It was successfully demonstrated that the TEM head-size resonator with high Q factor can provide high quality MR images at 3T MRI system. Also, the TEM resonator coil has an advantage for a fine tune with length adjustment of each cavity elements. Thus, it is expected that the TEM resonator at 3T, even higher field could be used in the clinical and research studies in near future.

Metabolic Abnormalities in Patients with Mitochondrial Myopathy Evaluated by In Vivo 31P Magnetic Resonance Spectroscopy

PURPOSE: To investigate the phosphorus metabolic abnormalities in skeletal muscle of patients with mitochondrial myopathy using in vivo 31P magnetic resonance spectroscopy(MRS). MATERIAL AND METHODS: Patients with mitochondrial myopathy(N=10) and normal control subjects (N=10) participated. All in vivo 31P MRS examinations were performed on 1.5T whole-body MRI/MRS system by using an image selected in vivo spectroscopy (ISIS) pulse sequence that provided a 4 X 4 X 4 cm3 volume of interest (VOI) in the right thigh muscle tissue. Peak areas for each phophorus methabolite were measured using a Marquart algorithm. RESULTS: The specific features in patients with mitochondrial myopathy were a significant increase of Pi/PCr ratio (p=0.003) and a significant decrease of ATP/PCr ratio (p=0.004) as compared with normal controls. In particular, the beta-ATP/PCr ratio between controls and patients with mitochondrial myopathy was predominantly altered. CONCLUSIONS: In vivo 31P MRS may be a useful modality in the clinical evaluation of patients with mitochondrial myopathy based on ATP/PCr and Pi/PCr ratios in skeletal muscle tissue and provides a valuable information in further understanding disorders of muscle metabolism.

Funtional MRI of Cerebral Motor Cortex: Comparison between 1.0 T and 1.5 T

PURPOSE: To evaluate the feasibility of functional MR imaging (fMRI) with a 1.0 T scanner, fMRI of normal cerebral motor cortex at 1.0 T was compared with that at 1.5 T. MATERIALS AND METHODS: FMRI of bilateral cerebral motor cortices (left, seven; right, six) was performed in seven healthy male volunteers aged 26-34 (mean 29) years, with BOLD contrast at both 1.0 T and 1.5 T units (Siemens MR scannners). Using both these systems, two-dimensional (2D) FLASH images were obtained with TR/TE of 90/56, flip angle of 40degrees, matrix size 128*128, slice thickness of 5 mm, and FOV 23 cm. A sequence consisting of five-image-off phase (rest phase) followed by five-image-on phase (activation with finger movement) was repeated four times without pause at a single plane. The same study was performed for the contralateral motor cortex in each volunteer. Using the z-test, activation images were obtained for the signal difference between on- and off-phases (p < 0.05) and were then superimposed on 2D FLASH anatomic images at the same plane. Percentage changes of signal intensities (PCSIs) and numbers of activated pixels were compared, using the non-parametric t-test, and periodicity of signal changes was compared, using the Mantel-Haenszel Chi-square test. RESULTS: Mean PCSIs at 1.5 T and 1.0 T in the left motor cortex were 3.13 +/-1.20% and 1.43 +/- 0.56%, respectively (p = 0.009), and in the right, 1.78 +/- 0.95% and 1.34 +/- 0.28%, respectively (p = 0.32). The mean number of activated pixels at 1.5 T and 1.0 T in the left cortex was 21.14 +/- 10.67 and 19.86 +/- 11.36, respectively (p = 0.83), and in the right, 22.5 +/- 6.47 and 16.8 +/- 8.47, respectively (p= 0.22). At 1.5 T, periodicity of signal changes was seen in the left cortex in six of seven volunteers, and in the right cortex, in four of six. At 1.0T, all showed periodicity (left : p = 0.32 ; right : p = 0.14). CONCLUSION: PCSIs in the dominant hemispheres were significantly higher at 1.5 T, but no other indicators showed significant differences between 1.0 T and 1.5 T. Acceptable fMRI can therefore be carried out with a 1.0 T field strength scanner.