15 karma · joined January 10, 2014
That was a complex project that required a lot of special technology, much of which we developed ourselves, to make and characterize micro-particles.
CV Herst, Pete Lloyd and I (all part of the Aradigm founding team) have been working together at Immunity Project since the beginning - Pete is an ME (UC Berkeley and Davis) who built and operates our precision spray drying system for sphere manufacturing. CV got his MPH at Berkeley in Med Micro and his PhD from North Western in Mo Bio later doing a post doc at MD Anderson. He has extensive experience working with HIV in both the diagnostic and lab research setting requiring handling the virus in cell culture.
Aston S, Rees T. Vicryl Sutures. Aesthetic Plastic Surgery 1977;1:5.
The "threatening point" is just a way of describing what TLR-4 (MPLA) and TLR-9 (CpG) do. In particular, MPLA "looks like" bacterial cell wall and CpG "looks like" bacterial DNA to the immune system.
Steinhagen F. TLR-based immune adjuvants. Vaccine, 2011: 12.
We did not consider using nano spheres because we wanted to get as much payload as possible into the antigen presenting cells without requiring multiple phagocytosis events per cell.
1. We have successfully gotten an immune response with memory (by elispot) after a single dose of the PLGA microspheres containing one or two peptides when combined with TLR-4 (MPLA) and TLR-9 (CpG) agonists. We used H2d restricted epitopes to do the study in C57BL/6 mice – this study is done and in the PRJ review process. We are being careful about posting the results in the event we need to re-submit to a journal who would get heartburn if they saw the data posted.
2. We are raising money now to essentially repeat the experiment (on a smaller scale because we think we understand the optimal formulation configuration now) in C57BL/6 NOG mice (with human PBMCs grafted in from a donor with known HLA type). The new experiment starts off looking like the experiment in (1) above: intradermal tail injection, wait 14 days, sacrifice, get the spleen, confirm immune response to peptide target by flurospot. The next part is new: separate CD8 from CD4 cells (from the mouse spleen), infect the CD4 cells with HIV, see if the vaccination attempt was good enough to see p24 antigen suppression and/or a change in the CD4 counts (all in-vitro). We can finish this study in a couple months, assuming the crowd funding keeps going well and we start in a couple weeks. We will use a different mechanism to publish our paper around this experiment so that we can get it out right away in a fashion that will lead to peer review and open access (We just learned about peerj.com – thoughts on that?).
Looking forward to hearing your thoughts on our approach.