W. Dai

A bivalent virus-like particle based vaccine induces a balanced antibody response against both enterovirus 71 and norovirus in mice

Noroviruses are the main cause of severe viral gastroenteritis, which results in estimated 200,000 deaths each year, primarily in children in the developing world. Genogroup II.4 (GII.4) strains are responsible for the majority of norovirus outbreaks. Enterovirus 71 (EV71), the leading causative agent of hand, foot and mouth disease, has recently been prevalent in Asia-Pacific regions, resulting in significant morbidity and mortality in young children. However, no vaccine is commercially available for either norovirus GII.4 or EV71. Recombinant virus-like particles (VLPs) derived from either GII.4 or EV71 have been shown to be promising monovalent vaccine candidates. In this study, we investigate the possibility to formulate a VLP-based bivalent vaccine for both norovirus GII.4 and EV71. The GII.4- and EV71-VLPs were produced in a baculovirus-insect cell expression system. A bivalent combination vaccine comprised of GII.4 and EV71 VLPs was formulated and compared with monovalent GII.4- and EV71-VLPs for their immunogenicity in mice. We found that the bivalent vaccine elicited durable antibody responses toward both GII.4 and EV71, and the antibody titers were comparable to that induced by the monovalent vaccines, indicating there is no immunological interference between the two antigens in the combination vaccine. More significantly, the bivalent vaccine-immunized mouse sera could efficiently neutralize EV71 infection and block GII.4-VLP binding to mucin. Together, our results demonstrate that the experimental combination vaccine comprised of GII.4 and EV71-VLPs is able to induce a balanced protective antibody response, and therefore strongly support further preclinical and clinical development of such a bivalent VLP vaccine targeting both norovirus GII.4 and EV71.

Thermoacoustically Driven Pulse Tube Coolers with Acoustic Amplifiers

In previous thermoacoustically driven pulse tube coolers, the coolers are directly connected to the thermoacoustic engines with connecting tubes as short as possible for less power dissipation. This coupling method limits the amplitude of the pressure waves available to the coolers, which in turn limits the lowest obtainable temperatures. Through our analysis, if we use a connecting tube with length comparable to one quarter wavelength, the pressure wave coming from the thermoacoustic engine will be greatly amplified and drive the cooler to reach a much lower temperature. Experiments have been done on thermoacoustically driven single‐stage and two‐stage pulse tube cooler and successfully proved the effectiveness of this idea. The pressure ratios of 1.3 and 1.24 were obtained on the single‐stage and two‐stage pulse tube coolers, respectively, which are much higher than the pressure ratios of 1.1 and 1.13 provided by the thermoacoustic engine itself. With the amplified pressure ratio, the no‐load lowest tem...

NUMERICAL SIMULATION OF A THREE-STAGE STIRLING-TYPE PULSE TUBE CRYOCOOLER FOR 4K OPERATION

The thermoacoustically driven two-stage pulse tube cooler has recently reached a lowest temperature of about 18.1K in our lab, and we are now developing a three-stage Stirling-type pulse tube cooler working at about 4K to match the thermoacoustic engine. In this paper, thermoacoustic theory is employed to simulate the three-stage pulse tube cooler. In order to decrease the cooling power losses of the second and third stage cold tips, two thermal bridges are introduced on the first and second stage cold tips to cool down the second and the third stage pulse tubes. Thus, the temperature gradients in the pulse tubes near the cold tips decrease, so do the cooling power losses through the thermal conductivity of the pulse tubes. After numerical optimization, the pulse tube cooler reaches a cooling temperature of 3.87K with 207 Watts of input acoustic power.

Enterovirus D68 virus-like particles expressed in Pichia pastoris potently induce neutralizing antibody responses and confer protection against lethal viral infection in mice

Enterovirus D68 (EV-D68) has been increasingly associated with severe respiratory illness and neurological complications in children worldwide. However, no vaccine is currently available to prevent EV-D68 infection. In the present study, we investigated the possibility of developing a virus-like particle (VLP)-based EV-D68 vaccine. We found that co-expression of the P1 precursor and 3CD protease of EV-D68 in Pichia pastoris yeast resulted in the generation of EV-D68 VLPs, which were composed of processed VP0, VP1, and VP3 capsid proteins and were visualized as ~30 nm spherical particles. Mice immunized with these VLPs produced serum antibodies capable of specifically neutralizing EV-D68 infections in vitro. The in vivo protective efficacy of the EV-D68 VLP candidate vaccine was assessed in two challenge experiments. The first challenge experiment showed that neonatal mice born to the VLP-immunized dams were fully protected from lethal EV-D68 infection, whereas in the second experiment, passive transfer of anti-VLP sera was found to confer complete protection in the recipient mice. Collectively, these results demonstrate the proof-of-concept for VLP-based broadly effective EV-D68 vaccines.

A THERMOACOUSTICALLY-DRIVEN PULSE TUBE CRYOCRYOCOOLER OPERATING AROUND 300HZ

High frequency operation of the thermoacoustic cryocooler system, i.e. pulse tube cryocooler driven by thermoacoustic engine, leads to reduced size, which is quite attractive to small-scale cryogenic applications. In this work, a no-load coldhead temperature of 77.8 K is achieved on a 292 Hz pulse tube cryocooler driven by a standing-wave thermoacoustic engine with 3.92 MPa helium gas and 1750 W heat input. To improve thermal efficiency, a high frequency thermoacoustic-Stirling heat engine is also built to drive the same pulse tube cryocooler, and a no-load temperature of 109 K was obtained with 4.38 MPa helium gas, 292 Hz working frequency and 400W heating power. Ideas such as tapered resonators, acoustic amplifier tubes and simple thin tubes without reservoir are used to effectively suppress harmonic modes, amplify the acoustic pressure wave available to the pulse tube cryocooler and provide desired acoustic impedance for the pulse tube cryocooler, respectively. Comparison of systems with different ther...

Thermoacoustic turbulent-flow model for inertance tubes used for pulse tube refrigerators

Publisher Summary This chapter develops a thermoacoustic turbulent-flow model for describing the inertance tube, by which can both incorporate turbulent-flow effect and heat transfer effect between gas and wall. An inertance tube is supposed to be another powerful shifter, which has the additional function of eliminating DC flow. For typical operating conditions, the flowing of the inertance tube is turbulent. Inertance-tube phase shifter can avoid DC-flow of the double-inlet phase shifter besides. The flow inside the inertance tube is usually turbulent, which makes linear network mode not accurately workable for describing performance of the inertance tube. In addition, relatively extensive experiments on various inertance tubes for low-capacity pulse tube refrigerators are made and only some of the experiment data are shown in this chapter. Comparisons between the simplified turbulent-flow model and the experimental data by hot-wire anemometry are made, which show a good agreement qualitatively, and also provide an acceptable accuracy quantitatively. Thus, a thermoacoustic turbulent-flow model is proposed to solve the problems, which can consider both turbulent-flow effect and heat transfer effect.

A Mouse Model of Enterovirus D68 Infection for Assessment of the Efficacy of Inactivated Vaccine

In recent years, enterovirus D68 (EVD68) has been reported increasingly to be associated with severe respiratory tract infections and acute flaccid myelitis (AFM) in children all over the world. Yet, no effective vaccines or antiviral drugs are currently available for EVD68. Although several experimental animal models have been developed, immunogenicity and protective efficacy of inactivated EVD68 vaccines has not been fully evaluated. To promote the development of vaccines, we established an Institute of Cancer Research (ICR) suckling mouse model of EVD68 infection in this study. The results showed that ICR neonatal mice up to about nine days of age were susceptible to infection with EVD68 clinical strain US/MO/14-18947 by intraperitoneal injection. The infected mice exhibited progressive limb paralysis prior to death and the mortality of mice was age- and virus dose-dependent. Tissue viral load analysis showed that limb muscle and spinal cord were the major sites of viral replication. Moreover, histopathologic examination revealed the severe necrosis of the limb and juxtaspinal muscles, suggesting that US/MO/14-18947 has a strong tropism toward muscle tissues. Additionally, β-propiolactone-inactivated EVD68 vaccine showed high purity and quality and induced robust EVD68-specific neutralizing antibody responses in adult mice. Importantly, results from both antisera transfer and maternal immunization experiments clearly showed that inactivated EVD68 vaccine was able to protect against lethal viral infection in the mouse model. In short, these results demonstrate the successful establishment of the mouse model of EVD68 infection for evaluating candidate vaccines against EVD68 and also provide important information for the development of inactivated virus-based EVD68 vaccines.

A virus-like particle vaccine confers protection against enterovirus D68 lethal challenge in mice

Enterovirus D68 (EV-D68) is increasingly associated with severe acute respiratory infection and acute flaccid myelitis (AFM) in children around the world. However, neither vaccines nor therapeutic drugs are available for EV-D68. Here we report the development of a virus-like particle (VLP) based experimental EV-D68 vaccine. We found that EV-D68 VLPs could be successfully generated in insect cells infected with a recombinant baculovirus co-expressing the P1 precursor and 3CD protease of EV-D68. Biochemical and electron microscopic analyses revealed that EV-D68 VLPs were composed of VP0, VP1, and VP3 capsid proteins derived from precursor P1 and were visualized as spherical particles of ∼30 nm in diameter. Immunization of mice with EV-D68 VLPs resulted in the production of serum antibodies that displayed potent serotype-specific neutralizing activities against EV-D68 virus in vitro. Passive transfer of anti-VLP sera completely protected neonatal recipient mice from lethal EV-D68 infection. Moreover, maternal immunization with these VLPs provided full protection against lethal EV-D68 challenge in suckling mice. Together, these results demonstrate that the recombinant EV-D68 VLP is a promising vaccine candidate against EV-D68 infection.

A virus-like particle-based tetravalent vaccine for hand, foot, and mouth disease elicits broad and balanced protective immunity

Hand, foot, and mouth disease (HFMD) is an infectious disease that mainly affects infants and children, causing considerable morbidity and mortality worldwide. HFMD is commonly caused by enterovirus 71 (EV71) and coxsackieviruses A16 (CVA16), A6 (CVA6), and A10 (CVA10). Formalin-inactivated EV71 vaccines are currently available in China; however, these vaccines fail to confer cross-protection against infections by other HFMD-causing enteroviruses, highlighting the necessity of developing a multivalent HFMD vaccine. Our previous studies demonstrated that recombinant virus-like particles (VLP) of EV71, CVA16, and CVA6 are capable of inducing protective immunity against homologous virus challenges in mice. In this study, we generated CVA10-VLP using a baculovirus-insect cell expression system and then combined CVA10-VLP with EV71-VLP, CVA16-VLP, and CVA6-VLP to formulate a tetravalent VLP vaccine. Immunogenicity and protective efficacy of tetravalent VLP vaccine was compared with that of monovalent VLP vaccines. Mouse immunization studies revealed that the tetravalent vaccine elicited antigen-specific and long-lasting serum antibody responses comparable to those elicited by its corresponding monovalent vaccines. Moreover, tetravalent vaccine immune sera strongly neutralized EV71, CVA16, CVA10, and CVA6 strains with neutralization titers similar to those of their monovalent counterparts, indicating a good compatibility among the four antigens in the combination vaccine. Importantly, passively transferred tetravalent vaccine-immunized sera conferred efficient protection against single or mixed infections with EV71, CVA16, CVA10, and CVA6 viruses in mice, whereas the monovalent vaccines could only protect mice against homotypic virus infections but not heterotypic challenges. These results demonstrate that the tetravalent VLP vaccine represents a promising broad-spectrum HFMD vaccine candidate.

A HIGH FREQUENCY THERMOACOUSTICALLY‐DRIVEN PULSE TUBE CRYOCOOLER WITH COAXIAL RESONATOR

High frequency thermoacoustically‐driven pulse tube cryocoolers are quite promising due to their compact size and high reliability, which can find applications in space use. With continuous effort, a lowest cold head temperature of 68.3 K has been obtained on a 300 Hz pulse tube cryocooler driven by a standing‐wave thermoacoustic heat engine with 4.0 MPa helium gas and 750 W heat input. To further reduce the size of the system, a coaxial resonator was designed and the two sub‐systems, i.e., the pulse tube cryocooler and the standing‐wave thermoacoustic heat engine were properly coupled through an acoustic amplifier tube, which leads to a system axial length of only about 0.7 m. The performance of the system with the coaxial resonator was tested, and shows moderate degradation compared to that with the in‐line resonator, which might be attributed to the large flow loss of the 180 degree corner.