Foxp31 regulatory t cell expression of keratinocyte growth factor enhances lung epithelial proliferation

Repair of the lung epithelium after injury is a critical component for resolution; however, the processes necessary to drive epithelial resolution are not clearly defined. Published data demonstrate that Foxp31 regulatory T cells (Tregs) enhance alveolar epithelial proliferation after injury, and Tregs in vitro directly promote type II alveolar epithelial cell (AT2) proliferation, in part by a contactindependent mechanism. Therefore, we sought to determine the contribution of Treg-specific expression of a growth factor that is known to be important in lung repair, keratinocyte growth factor (kgf). The data demonstrate that Tregs express kgf and that Treg-specific expression of kgf regulates alveolar epithelial proliferation during the resolution phase of acute lung injury and in a model of regenerative alveologenesis in vivo. In vitro experiments demonstrate that AT2 cells cocultured with Tregs lacking kgf have decreased rates of proliferation compared with AT2 cells cocultured with wild-type Tregs. Moreover, Tregs isolated from lung tissue and grown in culture express higher levels of two growth factors that are important for lung repair (kgf and amphiregulin) compared with Tregs isolated from splenic tissue. Lastly, Tregs isolated from human lung tissue can be stimulated ex vivo to induce kgf expression. This study reveals mechanisms by which Tregs direct tissuereparative effects during resolution after acute lung injury, further supporting the emerging role of Tregs in tissue repair

Tracker controls development and control architecture for the Hobby-Eberly Telescope Wide Field Upgrade

To enable the Hobby-Eberly Telescope Wide Field Upgrade, the University of Texas Center for Electromechanics and McDonald Observatory are developing a precision tracker system – a 15,000 kg robot to position a 3,100 kg payload within 10 microns of a desired dynamic track. Performance requirements to meet science needs and safety requirements that emerged from detailed Failure Modes and Effects Analysis resulted in a system of 14 precision controlled actuators and 100 additional analog and digital devices (primarily sensors and safety limit switches). This level of system complexity and emphasis on fail-safe operation is typical of large modern telescopes and numerous industrial applications. Due to this complexity, demanding accuracy requirements, and stringent safety requirements, a highly versatile and easily configurable centralized control system that easily links with modeling and simulation tools during the hardware and software design process was deemed essential. The Matlab/Simulink simulation environment, coupled with dSPACE controller hardware, was selected for controls development and realization. The dSPACE real-time operating system collects sensor information; motor commands are transmitted over a PROFIBUS network to servo amplifiers and drive motor status is received over the same network. Custom designed position feedback loops, supplemented by feed forward force commands for enhanced performance, and algorithms to accommodate self-locking gearboxes (for safety), reside in dSPACE. To interface the dSPACE controller directly to absolute Heidenhain sensors with EnDat 2.2 protocol, a custom communication board was developed. This paper covers details of software and hardware, design choices and analysis, and supporting simulations (primarily Simulink).Center for Electromechanic

Transcriptional analysis of Foxp3+ Tregs and functions of two identified molecules during resolution of ALI

Recovery from acute lung injury (ALI) is an active process. Foxp3+ Tregs contribute to recovery from ALI through modulating immune responses and enhancing alveolar epithelial proliferation and tissue repair. The current study investigates Treg transcriptional profiles during resolution of ALI in mice. Tregs from either lung or splenic tissue were isolated from uninjured mice or mice recovering from ALI and then examined for differential gene expression between these conditions. In mice with ALI, Tregs isolated from the lungs had hundreds of differentially expressed transcripts compared with those from the spleen, indicating that organ specificity and microenvironment are critical in Treg function. These regulated transcripts suggest which intracellular signaling pathways modulate Treg behavior. Interestingly, several transcripts having no prior recognized function in Tregs were differentially expressed by lung Tregs during resolution. Further investigation into 2 identified transcripts, Mmp12 and Sik1, revealed that Treg-specific expression of each plays a role in Treg-promoted ALI resolution. This study provides potentially novel information describing the signals that may expand resident Tregs, recruit or retain them to the lung during ALI, and modulate their function. The results provide insight into both tissue- and immune microenvironment–specific transcriptional differences through which Tregs direct their effects

Bronchus-associated lymphoid tissue in kabuki syndrome with associated hyper-IgM syndrome/common variable immunodeficiency

A 28-year-old woman with a medical history significant for Kabuki syndrome with associated hyper-IgM syndrome/common variable immunodeficiency presented with a 3-month history of dyspnea, pleuritic pain, and nonproductive cough. Imaging demonstrated nodular infiltrates increasing toward the lung bases (Figure 1A). Bronchoscopy with lavage and transbronchial biopsies was performed to evaluate for an infectious or inflammatory etiology; however, results were unrevealing, and no infectious etiology was identified. Therefore, she underwent a video-assisted thoracic surgery biopsy that demonstrated nodular lymphoid hyperplasia with follicles centered on small airways and areas of organizing pneumonia (Figure 1B). Immunostains demonstrated follicles composed of CD31 T cells and CD201 B cells (Figure 1B) and no morphologic evidence of lymphoma or plasma cell neoplasm. Results were consistent with bronchus-associated lymphoid tissue (BALT). High-dose steroids were administered without resolution of symptoms or radiographic findings. A report of effective responses with rituximab and azathioprine in patients with common variable immunodeficiency who had a similar inflammatory lung process containing tertiary lymphoid structures, granulomas, and organizing pneumonia has been published (1). After four doses of rituximab (weekly), together with the initiation of azathioprine, her symptoms and radiographic findings improved (Figure 1C)

Effects of IFN-γ on immune cell kinetics during the resolution of acute lung injury

The immunologic responses that occur early in the acute respiratory distress syndrome (ARDS) elicit immune-mediated damage. The mechanisms underlying the resolution of ARDS, particularly the role of signaling molecules in regulating immune cell kinetics, remain important questions. Th1-mediated responses can contribute to the pathogenesis of acute lung injury (ALI). Interferon-gamma (IFN-γ) orchestrates early inflammatory events, enhancing immune-mediated damage. The current study investigated IFN-γ during resolution in several experimental models of ALI. The absence of IFN-γ resulted in altered kinetics of lymphocyte and macrophage responses, suggesting that IFN-γ present in this microenvironment is influential in ALI resolution. Genetic deficiency of IFN-γ or administering neutralizing IFN-γ antibodies accelerated the pace of resolution. Neutralizing IFN-γ decreased the numbers of interstitial and inflammatory macrophages and increased alveolar macrophage numbers during resolution. Our results underline the complexity of lung injury resolution and provide insight into the effects through which altered IFN-γ concentrations affect immune cell kinetics and the rate of resolution. These findings suggest that therapies that spatially or temporally control IFN-γ signaling may promote ALI resolution. Identifying and elucidating the mechanisms critical to ALI resolution will allow the development of therapeutic approaches to minimize collateral tissue damage without adversely altering the response to injury

Design and development of a high precision, high payload telescope dual drive system

A high precision, dual drive system has been designed and developed for the Wide Field Upgrade to the Hobby-Eberly Telescope* at McDonald Observatory in support of the Hobby-Eberly Telescope Dark Energy Experiment‡. Analysis,design and controls details will be of interest to designers of large scale, high precision robotic motion devices. The drive system positions the 19,000 kg star tracker to a precision of less than 5 microns along its 4-meter travel. While positioning requirements remain essentially equal to the existing HET, tracker mass increases by a factor greater than 5. The 10.5-meter long tracker is driven at each end by planetary roller screws, each having two distinct drive sources dictated by the desired operation: one slowly rotates the screw when tracking celestial objects and the second rotates the nut for rapid displacements. Key results of the roller screw rotordynamics analysis are presented. A description of the complex bearing arrangement providing required degrees of freedom as well as the impact of a detailed Failure Modes and Effects Analysis addressing necessary safety systems is also presented. Finite element analysis results demonstrate how mechanical springs increase the telescope's natural frequency response by 22 percent. The critical analysis and resulting design is provided.Center for Electromechanic

The Development of high-precision hexapod actuators for the Hobby-Eberly Telescope Wide Field Upgrade

Hexapods are finding increased use in telescope applications for positioning large payloads. Engineers from The University of Texas at Austin have been working with engineers from ADS International to develop large, high force, highly precise and controllable hexapod actuators for use on the Wide Field Upgrade (WFU) as part of the Hobby Eberly Telescope Dark Energy Experiment (HETDEX)‡. These actuators are installed in a hexapod arrangement, supporting the 3000+ kg instrument payload which includes the Wide Field Corrector (WFC), support structure, and other optical/electronic components. In addition to force capability, the actuators need to meet the tracking speed (pointing) requirements for accuracy and the slewing speed (rewind) requirements, allowing as many observations in one night as possible. The hexapod actuator stroke (retraction and extension) was very closely monitored during the design phase to make sure all of the science requirements could be met, while minimizing the risk of damaging the WFC optical hardware in the unlikely event of a hexapod actuator or controller failure. This paper discusses the design trade-offs between stiffness, safety, back-drivability, accuracy, and leading to selection of the motor, high ratio worm gear, roller screw, coupling, end mounts, and other key components.Center for Electromechanic

Effect of chronic intermittent hypoxia on triglyceride uptake in different tissues

Chronic intermittent hypoxia (CIH) inhibits plasma lipoprotein clearance and adipose lipoprotein lipase (LPL) activity in association with upregulation of an LPL inhibitor angiopoietin-like protein 4 (Angptl4). We hypothesize that CIH inhibits triglyceride (TG) uptake via Angptl4 and that an anti-Angptl4-neutralizing antibody would abolish the effects of CIH. Male C57BL/6J mice were exposed to four weeks of CIH or intermittent air (IA) while treated with Ab (30 mg/kg ip once a week). TG clearance was assessed by [H 3 ]triolein administration retroorbitally. CIH delayed TG clearance and suppressed TG uptake and LPL activity in all white adipose tissue depots, brown adipose tissue, and lungs, whereas heart, liver, and spleen were not affected. CD146+ CD11b α pulmonary microvascular endothelial cells were responsible for TG uptake in the lungs and its inhibition by CIH. Antibody to Angptl4 decreased plasma TG levels and increased TG clearance and uptake into adipose tissue and lungs in both control and CIH mice to a similar extent, but did not reverse the effects of CIH. The antibody reversed the effects of CIH on LPL in adipose tissue and lungs. In conclusion, CIH inactivates LPL by upregulating Angptl4, but inhibition of TG uptake occurs predominantly via an Angptl4/LPL-independent mechanism

Regulatory T cells reduce acute lung injury fibroproliferation by decreasing fibrocyte recruitment

Acute lung injury (ALI) causes significant morbidity and mortality. Fibroproliferation in ALI results in worse outcomes, but the mechanisms governing fibroproliferation remain poorly understood. Regulatory T cells (Tregs) are important in lung injury resolution. Their role in fibroproliferation is unknown. We sought to identify the role of Tregs in ALI fibroproliferation, using a murine model of lung injury. Wild-type (WT) and lymphocyte-deficient Rag-1-/- mice received intratracheal LPS. Fibroproliferationwascharacterizedby histology and the measurement of lung collagen. Lung fibrocytes were measured by flow cytometry. To dissect the role of Tregs in fibroproliferation, Rag-1-/- mice received CD4 +CD25+ (Tregs) or CD4+ CD25- Tcells (non-Tregs) at the time of LPS injury. To define the role of the chemokine (C-X-C motif) ligand 12 (CXCL12)-CXCR4 pathway in ALI fibroproliferation, Rag-1-/- mice were treated with the CXCR4 antagonist AMD3100 to block fibrocyte recruitment. WT and Rag-1-/- mice demonstrated significant collagen deposition on Day 3 after LPS. WT mice exhibited the clearance of collagen, but Rag-1-/- mice developed persistent fibrosis. This fibrosis was mediated by the sustained epithelial expression of CXCL12 (or stromal cell-derived factor 1 [SDF-1]) that led to increased fibrocyte recruitment. The adoptive transfer of Tregs resolved fibroproliferation by decreasing CXCL12 expression and subsequent fibrocyte recruitment. Blockade of the CXCL12-CXCR4 axis with AMD3100 also decreased lung fibrocytes and fibroproliferation. These results indicate a central role for Tregs in the resolution of ALI fibroproliferation by reducing fibrocyte recruitment along the CXCL12-CXCR4 axis. A dissection of the role of Tregs in ALI fibroproliferation may inform the design of new therapeutic tools for patients with ALI

Baseline and innate immune response characterization of a Zfp30 knockout mouse strain

Airway neutrophilia is correlated with disease severity in a number of chronic and acute pulmonary diseases, and dysregulation of neutrophil chemotaxis can lead to host tissue damage. The gene Zfp30 was previously identified as a candidate regulator of neutrophil recruitment to the lungs and secretion of CXCL1, a potent neutrophil chemokine, in a genome-wide mapping study using the Collaborative Cross. ZFP30 is a putative transcriptional repressor with a KRAB domain capable of inducing heterochromatin formation. Using a CRISPR-mediated knockout mouse model, we investigated the role that Zfp30 plays in recruitment of neutrophils to the lung using models of allergic airway disease and acute lung injury. We found that the Zfp30 null allele did not affect CXCL1 secretion or neutrophil recruitment to the lungs in response to various innate immune stimuli. Intriguingly, despite the lack of neutrophil phenotype, we found there was a significant reduction in the proportion of live Zfp30 homozygous female mutant mice produced from heterozygous matings. This deviation from the expected Mendelian ratios implicates Zfp30 in fertility or embryonic development. Overall, our results indicate that Zfp30 is an essential gene but does not influence neutrophilic inflammation in this particular knockout model