Jaewook Shin

Th2 Regulation of Viral Myocarditis in Mice: Different Roles for TLR3 versus TRIF in Progression to Chronic Disease

Viral infections are able to induce autoimmune inflammation in the heart. Here, we investigated the role of virus-activated Toll-like receptor (TLR)3 and its adaptor TRIF on the development of autoimmune coxsackievirus B3 (CVB3) myocarditis in mice. Although TLR3- or TRIF-deficient mice developed similarly worse acute CVB3 myocarditis and viral replication compared to control mice, disease was significantly worse in TRIF compared to TLR3-deficient mice. Interestingly, TLR3-deficient mice developed an interleukin (IL)-4-dominant T helper (Th)2 response during acute CVB3 myocarditis with elevated markers of alternative activation, while TRIF-deficient mice elevated the Th2-associated cytokine IL-33. Treatment of TLR3-deficient mice with recombinant IL-33 improved heart function indicating that elevated IL-33 in the context of a classic Th2-driven response protects against autoimmune heart disease. We show for the first time that TLR3 versus TRIF deficiency results in different Th2 responses that uniquely influence the progression to chronic myocarditis.

Testosterone and interleukin-1β increase cardiac remodeling during coxsackievirus B3 myocarditis via serpin A 3n

Myocarditis and dilated cardiomyopathy (DCM) are often caused by viral infections and occur more frequently in men than in women, but the reasons for the sex difference remain unclear. The aim of this study was to assess whether gene changes in the heart during coxsackievirus B3 (CVB3) myocarditis in male and female BALB/c mice predicted worse DCM in males. Although myocarditis (P = 4.2 × 10(-5)) and cardiac dilation (P = 0.008) were worse in males, there was no difference in viral replication in the heart. Fibrotic remodeling genes, such as tissue inhibitor of metalloproteinase (TIMP)-1 and serpin A 3n, were upregulated in males during myocarditis rather than during DCM. Using gonadectomy and testosterone replacement, we showed that testosterone increased cardiac TIMP-1 (P = 0.04), serpin A 3n (P = 0.007), and matrix metalloproteinase (MMP)-8 (P = 0.04) during myocarditis. Testosterone increased IL-1β levels in the heart (P = 0.02), a cytokine known to regulate cardiovascular remodeling, and IL-1β in turn increased cardiac serpin A 3n mRNA (P = 0.005). We found that 39 of 118 (33%) genes identified in acute DCM patients were significantly altered in the heart during CVB3 myocarditis in mice, including serpin A 3n (3.3-fold change, P = 0.0001). Recombinant serpin A 3n treatment induced cardiac fibrosis during CVB3 myocarditis (P = 0.0008) while decreasing MMP-3 (P = 0.04) and MMP-9 (P = 0.03) levels in the heart. Thus, serpin A 3n was identified as a gene associated with fibrotic cardiac remodeling and progression to DCM in male myocarditis patients and mice.

Single-pixel imaging using compressed sensing and wavelength-dependent scattering

We demonstrate two-dimensional imaging using illumination via a single-mode fiber with a multiply scattering tip and compressed sensing acquisition. We illuminate objects with randomly structured, but deterministic, speckle patterns produced by a coherent light source propagating through a TiO2-coated fiber tip. The coating thickness is optimized to produce speckle patterns that are highly sensitive to laser wavelength, yet repeatable. Images of the object are reconstructed from the characterized wavelength dependence of the speckle patterns and the wavelength dependence of the total light collected from the object using a single photodetector. Our imaging device is mechanically scan-free and insensitive to bending of the fiber, making it suitable for micro-endoscopy.

Compressive fluorescence imaging using a multi-core fiber and spatially dependent scattering

We demonstrate imaging using a multi-core fiber with a scattering distal tip and compressed sensing signal acquisition. We illuminate objects with randomly structured speckle patterns generated by a coherent light source separately coupled through each fiber core to a ground glass diffuser at the distal end. Using the characterized speckle patterns and the total light collected from the object, we computationally recover pixelation-free object images with up to a seven times higher space-bandwidth product than the number of cores. The proposed imaging system is insensitive to bending of the fiber and extremely compact, making it suitable for minimally invasive endomicroscopy.

Compressive temporal focusing microscopy (Conference Presentation)

Multiphoton microscopes are of paramount importance in capturing neural activity with cellular resolution. However, the imaging speed and field-of-view of traditional two-photon microscopes is limited by raster scanning technologies. Temporally-focused two-photon (TFTP) microscopy is a wide-field scan-free approach to increase the speed of two-photon microscopy. In conventional TFTP microscopy, wide-field depth sectioning is obtained by compressing a spatially pre-chirped pulse at the focal plane of the objective. Unfortunately, the greater imaging speed of TFTP microscopes comes at the expense of poor imaging depth in tissue due to scattering of the short-wavelength fluorescence photons en-route to the imaging camera. Here we demonstrate a compressive high-speed two-photon microscope based on wide-field temporally-focused structured illumination, which eliminates the loss of image contrast from scattering of the fluorescence signal by leveraging a single-pixel detector. Specifically, we illuminate the sample with a rapid sequence of randomly structured temporally-focused wide-field illumination pulses and integrate the net two-photon fluorescence response on a single photomultiplier tube (PMT). Notably, the longer wavelength structured illumination is significantly less susceptible to scattering and the use of integrated measurements on a single PMT provides immunity to fluorescence scattering since these measurements are solely concerned with the net fluorescence. Furthermore, our approach provides greater speed than point scanning two-photon microscopes through the use of wide-field illumination and compressive image acquisition. Experimentally we demonstrate this system operating over a 200×250-μm field-of-view and at a compression rate of 10%, which provides an order of magnitude increase in speed over a comparable point scanning architecture.

High-speed imaging using compressed sensing and wavelength-dependent scattering (Conference Presentation)

The process of multiple scattering has inherent characteristics that are attractive for high-speed imaging with high spatial resolution and a wide field-of-view. A coherent source passing through a multiple-scattering medium naturally generates speckle patterns with diffraction-limited features over an arbitrarily large field-of-view. In addition, the process of multiple scattering is deterministic allowing a given speckle pattern to be reliably reproduced with identical illumination conditions. Here, by exploiting wavelength dependent multiple scattering and compressed sensing, we develop a high-speed 2D time-stretch microscope. Highly chirped pulses from a 90-MHz mode-locked laser are sent through a 2D grating and a ground-glass diffuser to produce 2D speckle patterns that rapidly evolve with the instantaneous frequency of the chirped pulse. To image a scene, we first characterize the high-speed evolution of the generated speckle patterns. Subsequently we project the patterns onto the microscopic region of interest and collect the total light from the scene using a single high-speed photodetector. Thus the wavelength dependent speckle patterns serve as high-speed pseudorandom structured illumination of the scene. An image sequence is then recovered using the time-dependent signal received by the photodetector, the known speckle pattern evolution, and compressed sensing algorithms. Notably, the use of compressed sensing allows for reconstruction of a time-dependent scene using a highly sub-Nyquist number of measurements, which both increases the speed of the imager and reduces the amount of data that must be collected and stored. We will discuss our experimental demonstration of this approach and the theoretical limits on imaging speed.