Recombinant ATV Mammalian Expression System for Airway Immunity
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Solution Overview
Problem
Current viral-vector protein expression platforms for vaccines, such as those based on mammalian viruses, pose safety concerns due to pre-existing host immunity and lack an effective solution for generating antiviral airway immunity against respiratory viral infections like rhinovirus, which are common triggers for asthma attacks and have significant healthcare burdens.
Innovation Solution
A recombinant, attenuated Ambystoma tigrinum virus (ATV) is engineered to express foreign proteins in mammalian cells without replicating, using unique replication events in both the nucleus and cytoplasm, and incorporating mammalian transcription and translation enhancement elements, ensuring temperature-sensitive expression in airway mucosa and safety from human infection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If mammalian viruses are used as viral-vector protein expression platforms, then protein expression capability is improved, but safety concerns arise due to pre-existing host immunity
Solution Approach 1:
The patent segments the viral vector system by using a non-mammalian virus (Ambystoma tigrinum virus, an amphibian ranavirus) instead of mammalian viruses. This segmentation separates the expression system from the human immune system, eliminating pre-existing immunity issues while maintaining protein expression capability in mammalian cells through engineered promoters and transcriptional elements
Solution Approach 2:
The patent introduces an intermediary organism (amphibians) as the viral vector source. The Ambystoma tigrinum virus naturally infects amphibians but has been engineered to express mammalian proteins in mammalian cells without causing disease, serving as a safe intermediary that bridges protein expression needs with safety requirements
2Reliability
If conventional viral vectors are used to generate antiviral immunity, then immune response is improved, but inability to generate specific antiviral airway immunity against respiratory viruses persists
Solution Approach 1:
The patent applies local quality by engineering the Ambystoma tigrinum virus to specifically target and express antigens in the airway mucosa. The virus is modified with mammalian transcriptional enhancement elements and specific promoters that direct localized expression in respiratory epithelial cells, generating site-specific antiviral IgA immunity where it is most needed
Solution Approach 2:
The patent changes key parameters of the viral vector system: using a temperature-sensitive amphibian virus (replicates at 28°C but not at 37°C), engineering mammalian transcriptional control elements, and selecting for airway-specific antigen expression. These parameter changes enable the virus to safely express respiratory virus antigens and generate targeted airway immunity
3Reliability
If amphibian-based viral expression system is used, then safety from human infection is improved, but protein expression efficiency in mammalian cells must be enhanced
Solution Approach 1:
The patent applies preliminary action by pre-engineering the Ambystoma tigrinum virus with mammalian transcriptional enhancement elements, promoters, and polyadenylation signals before use. These modifications are incorporated into the viral genome in advance, ensuring efficient mammalian protein expression without requiring post-infection optimization
Solution Approach 2:
The patent creates a composite viral system by combining the Ambystoma tigrinum virus backbone with mammalian transcriptional control elements and enhancement sequences. This hybrid construct merges the safety of an amphibian virus with the expression efficiency of mammalian regulatory elements, achieving both safety and productivity
Data Source
AI summary
A mammalian expression system comprising an attenuated, recombinant Ranavirus that has at least one foreign expression element using a unique combination of mammalian transcriptional and translational enhancement elements is disclosed. In other contemplated embodiments, a mammalian expression system comprising a virus, wherein the virus is engineered to express at least two vaccine antigens is disclosed. In addition, methods of delivering human antigens to a mammal are disclosed that include: providing a non-mammalian virus, engineering a recombinant virus that can express at least one foreign molecule by modifying the non-mammalian virus, and using the recombinant Ranavirus to express and deliver foreign antigens to a mammal.