CD40 specific DNA aptamers as vaccine adjuvants

Inventors

AL-OGAILI, Adil Sabr • JIANG, Tieshan • VUONG, Christine N. • Liyanage, Rohana • Lay, Jr., Jackson O. • Thallapuranam, Suresh Kumar • Berghman, Luc R. • Kwon, Young Min • Hargis, Billy

Assignees

Texas A&M University System • University of Arkansas at Little Rock

Abstract

The present invention provides immunostimulatory nucleic acids that have an affinity to a specific target protein. The present invention also provides templates and methods for making and using the immunostimulatory nucleic acids. Further, methods for linking the immunostimulatory nucleic acids to antigens and using the resulting complexes to enhance an immune response are provided.

Core Innovation

The patent describes immunostimulatory nucleic acids comprising protein-binding sequences that bind to CD40. The protein-binding sequences are defined by at least 95% identity to one of SEQ ID NOs: 1-8, and the nucleic acids are used for immunological activity associated with CD40 binding.

The patent further provides multivalent immunostimulatory nucleic acids having a plurality of protein-binding units, each protein-binding unit including a first protein-binding sequence, a different second protein-binding sequence, and a spacer sequence therebetween. The multivalent nucleic acid binds to CD40, and the first and/or second protein-binding sequences have at least 95% identity to one of SEQ ID NOs: 1-8.

The patent also describes circularizable templates that include ordered regions from 5′ to 3′ that enable formation of a primer-binding site upon circularization, supporting production of ordered protein-binding units. The resulting immunostimulatory nucleic acids are amplified using rolling circle amplification to form aptamer-based RCA products, and aptamer constructs are tagged and linked to an antigen to create immunostimulatory complexes.

The patent documents CD40-binding and immunostimulatory activity for CD40-targeting aptamers, including in vitro macrophage activation shown by nitric oxide liberation with aptamer RCA constructs compared to anti-CD40 monoclonal antibody. It also describes limited proteolysis and mass spectrometry to indicate aptamer binding/protection regions on CD40, and in vivo chicken immunization demonstrating IgG induction using an aptamer RCA II–streptavidin–M2e complex with early and sustained antibody responses.

Claims Coverage

The independent claims cover three inventive features: CD40-binding immunostimulatory nucleic acids defined by SEQ ID identity, multivalent CD40-binding immunostimulatory nucleic acids with first and different second protein-binding sequences separated by a spacer, and a circularizable template for producing ordered protein-binding units.

The claims focus on CD40-binding immunostimulatory nucleic acids, multivalent designs built from protein-binding sequences and spacers, and circularizable templates used to generate ordered protein-binding units.

Stated Advantages

Documented Applications