Abstract: The present invention relates to an extracellular vesicle (EV)-based nucleic acid composition or vaccine (EV-NAV), comprising EVs loaded with polynucleotides each encoding, e.g., the SARS-CoV-2 spike protein, and polynucleotides each encoding, e.g., SARS-CoV-2 nucleocapsid protein, wherein said polynucleotides are designed to be simultaneously expressed, and to induce a humoral immune response and/or a cellular immune response, in a subject. The present invention also relates to compositions and methods for the design, preparation, manufacture, formulation, and therapeutic or prophylactic use of said EV-NAVs, e.g., exosomes loaded with mRNAs encoding multiple surface and cytoplasmic antigens derived from, e.g., SARS-CoV-2, to elicit strong humoral and cellular immune responses.

Abstract: Coronavirus egress is mediated by lysosomal exocytosis. It is demonstrated herein that the D614G mutation enhances Spike trafficking to lysosomes and the lysosomal accumulation of newly synthesized virus particles, augments Spike-mediated disruption of endomembrane homeostasis, and causes a 3-fold reduction in cell surface Spike expression. Moreover, it is shown that the D614G mutation is an intragenic suppressor of the 12 nucleotide-long furin cleavage site (FCS) insertion, restoring Spike trafficking to lysosomes and TMPRSS2-independent infectivity, both of which had been impaired by the prior FCS insertion mutation. This data identifies enhanced lysosomal sorting as the earliest known manifestation of the D614G mutation, have implications for virus evolution, immunity, and vaccine design, and support a lysosomal model of coronavirus biogenesis and entry.

Abstract: The present disclosure provides methods for the detection of target specific antibodies in samples. The methods include the detection of pathogen-specific antibodies, such as SARS-CoV-2 specific antibodies. Also included is a kit for use for the detection of pathogen specific antibodies in a sample. In one aspect, the disclosure provides a method of detecting the presence of a target specific antibody in a sample by (i) contacting the sample with a test cell comprising one or more exogenous nucleic acid sequences encoding one or more target proteins; and (ii) detecting the presence of the target specific antibody in the sample by contacting the immune complex of (i) with an anti-immunoglobulin (Ig) antibody, and detecting the anti-immunoglobulin (Ig) antibody, thereby detecting the presence of a target specific antibody.

Abstract: The present invention relates to the field of extracellular vesicles. More specifically, the present invention provides methods and compositions for using extracellular vesicles as a vector for nucleic acid treatment in vivo of various diseases. In a specific embodiment, the present invention provides an extracellular vesicle isolated from a cell comprising one or more microRNAs (miRNAs) that have been loaded ex vivo into the vesicle so that the miRNAs are present in a higher concentration than when measured in the same extracellular vesicle isolated directly from the cell. In another embodiment, the present invention provides a method for treating cholangiocarcinoma in a subject comprising the step of administering to the subject a plurality of exosomes comprising miR-195.

Abstract: The present invention relates to the field of extracellular vesicles. More specifically, the present invention provides methods and compositions for using extracellular vesicles as a vector for nucleic acid treatment in vivo of various diseases. In a specific embodiment, the present invention provides an extracellular vesicle isolated from a cell comprising one or more microRNAs (miRNAs) that have been loaded ex vivo into the vesicle so that the miRNAs are present in a higher concentration than when measured in the same extracellular vesicle isolated directly from the cell. In another embodiment, the present invention provides a method for treating cholangiocarcinoma in a subject comprising the step of administering to the subject a plurality of exosomes comprising miR-195.

Abstract: The present invention relates to the field of extracellular vesicles. More specifically, the present invention provides methods and compositions for using extracellular vesicles as a vector for nucleic acid treatment in vivo of various diseases. In a specific embodiment, the present invention provides an extracellular vesicle isolated from a cell comprising one or more microRNAs (miRNAs) that have been loaded ex vivo into the vesicle so that the miRNAs are present in a higher concentration than when measured in the same extracellular vesicle isolated directly from the cell. In another embodiment, the present invention provides a method for treating cholangiocarcinoma in a subject comprising the step of administering to the subject a plurality of exosomes comprising miR-195.