LAMP-Vax™, A NEW APPROACH TO DNA VACCINES THAT ACHIEVES AN ENHANCED IMMUNE RESPONSE

Lysosomal Associated Membrane Protein (LAMP) is a glycoprotein found on the lysosomal membrane. LAMP-Vax™ DNA vaccines utilize the natural biochemistry of LAMP to intersect with the process that Antigen Presenting Cells, "APCs", use to internalize, digest, and present exogenously derived antigens to the immune system as part of the lysosomal/MHC-II complex. The goal is to result in improved antigen presentation and a greatly enhanced immune system response to a vaccine.

Upon immunization with a LAMP-Vax™ DNA vaccine, APCs take up the DNA and produce the encoded protein sequence inside the cell as part of a fusion protein with LAMP.

In this way, LAMP-DNA vaccines are meant to activate APCs to the immunized antigen(s) through the CD4+ helper T-cell pathway. LAMP-DNA immunization contrasts with the immune response to conventional DNA vaccines, which are processed and primarily presented through MHC-I and elicit a cytotoxic T response. Studies have shown that LAMP-Vax™ DNA plasmids show no decrease in CD8+ cytotoxic T-cell response yet also feature a CD4+ response. This is believed to initiate a more complete immune response including antibody production, cytokine release and immunological memory.


FEATURES & COMPETITIVE ADVANTAGES OF LAMP TECHNOLOGY

MHC-II Targeting Strategy
The addition of LAMP Technology to DNA vaccine design is intended to increase the often limited immune response that occurs when antigen is processed in somatic cells and presented through the generic MHC-I pathway. By accessing the MHC-II antigen presentation pathway, LAMP-Vax™ vaccines are designed to create CD4+T cell responses and elicit antibody responses, while maintaining killer T cell responses.

Lysosomal Targeting for MHC-II Presentation of Antigens
Including the LAMP luminal domain in a LAMP-antigen vaccine is believed to direct antigen accumulation in MHC-II containing lysosomes, thereby enhancing expression. How it's thought to work: The luminal domain of LAMP stabilizes the antigenic-fusion protein and its glycoproteins protect the antigenic sequence from premature proteolysis, while permitting proteolysis in mature lysosomes.

Multiplexing Antigens
LAMP-Vax™ vaccines have been designed to code for multiple antigens simultaneously.

Improved plasmid design
Immunomic Therapeutics’ LAMP-Vax™ is on the cutting edge of improved plasmid design, incorporating Nature Technology Corporation’s high-yield, antibiotic free immunization vector with ITI’s proprietary LAMP-Vax™ sequence which facilities MHC-II targeting. Additionally, ITI’s in-house molecular biology team are experts in optimizing the design of our DNA vaccines through additional methods that are relevant to the state of the art today.

Various delivery systems
Various approaches to delivery of DNA vaccines have arisen over the past several years. These include needle-free injection, microneedles for ID delivery, liposomes, particle mediated (e.g., gold coated particles), and electroporation, among others. Our R&D team continues to advance LAMP in collaboration with various delivery technologies to make sure we are delivering the properly designed vaccines to the right immune cell populations in the body.

Plug & Play Vaccine Design for Rapid Product Line Development When a target antigen or antigens have been identified, ITI has developed the know-how and process for a quick turnaround in design, synthesis and testing of a construct. This versatility enables ITI to quickly expand both its product line and that of its partner companies.

Manufacturing
Further, ITI also has a license to utilize the Nature’s HyperGRO™ process, a proprietary fermentation/manufacturing process designed to produce higher yields of DNA, believed to enable better scale-up manufacturing.

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lmmunomic Therapeutics is developing a new generation of vaccine therapies by incorporating the genetic sequence for the LAMP protein as part of the DNA vaccine. What follows is the proposed mechanism for LAMP-Vax™.
When plasmid DNA encoding for an allergen and LAMP is injected into the skin, it first interacts with innate immune cells such as natural killer cells, as well as dendritic cells.
Since plasmids are derived from a type of bacterial DNA, and bacterial DNA is often naturally found where there is an infection, the plasmid DNA itself makes the immune system respond as if it were an antigen by secreting IFNγ.
 
The plasmid is taken up by the dendritic cell.
As the DC heads to the lymph node, it begins the process of turning the information contained in the vaccine into the desired protein.
The protein is synthesized directly into the endoplasmic reticulum, which, in turn, forms a Golgi complex.
 
Inside the Golgi, the protein is modified with the addition of protective sugars and uses the LAMP trafficking signal to find other proteins that belong in the lysosome.
As these proteins all come together, the vesicle buds off the end of the Golgi, forming a new lysosome. This new lysosome, containing both LAMP and the key MHC-II protein, is now free to begin antigen processing.
In the lysosome the LAMP fusion protein is first anchored in the lysosomal membrane, oriented so that allergen fusion protein is sandwiched between both domains of LAMP: the outward facing cytoplasmic domain and the highly glycosylated luminal domain.
 
Once there, the pH within the lysosome drops and the antigen protein is then processed into small 1-15 amino acid fragments.
These are bound to a special groove on the MHC-II molecules.
The lysosome moves to the DC surface and fuses with the DC membrane, exposing the MHC-II and its cargo to nearby patrolling T cells.
 
As these antigen peptide fragments are presented to these Tcells,
the DC also releases IL-12 and IFNγ, priming the T cells to become ...
... Th 1 cells rather than Th2 cells. This effectively alters the adaptive immune response toward an lgG·oriented response.
 
Th1 cells create even more IFNγ and, over time, out-compete the initial population ofTh2 cells. This turns the tide, remaining Th2 cells gradually fade away, as does the allergic response.
What is expected from this process, based on early stage data, is that the body has learned to treat this antigen protein like an infection, rather than an allergen.
Ultimately, the goal is, when the body encounters this antigen protein, it rapidly disposes of it, and the allergy is no more.