169 Background: Our current tumor lysate, particle-loaded, dendritic cell (TLPLDC) vaccine employs yeast cell wall particles (YCWP) to deliver antigen to DC ex vivo and is being tested in a randomized phase IIb trial. This strategy may be improved by injecting TL-loaded YCWP as an intradermal vaccine, with in vivo DC uptake. Silicate capping should allow YCWP to retain TL longer and stimulate DC uptake. Here, we present preclinical data on the tumor lysate, particle only (TLPO) vaccine concept vs. TLPLDC. Methods: To test the TLPO concept, YCWP were loaded with fluorescence (flr) labeled albumin and capped (cp) vs. uncapped (ucp) particles were compared for flr leak. Next, YCWP were added to cultured macrophages to evaluate DC uptake. Cells were then lysed, centrifuged, and flr in the cytoplasm vs. organelles measured. 3 C57B mice were then injected with 100ml NaCl, 106empty cp YCWP, or 1mcg GM-CSF. 5 hours (hr) post-injection, 100ml NaCl was injected into the same site, withdrawn, and examined via microscope to count monocytes. TLPO was compared to TLPLDC in a B16 murine melanoma survival model. Finally, 4 grey horses with equine melanoma were treated with autologous TLPO, injected biweekly for 4 vaccinations. Target lesions were assessed over 6 months. Results: Compared to ucp YCWP, cp had decreased flr leak at 1 (15.8% vs. 24.7%) and 2 hr (6.7% vs. 16.6%), increased uptake by DC (2hr flr readings 11065 vs. 3928) and higher delivery to DC cytoplasm (68.9% vs. 48.8%). Empty cp YCWP showed increased recruitment of monocytes (276/hpf) vs. GM-CSF (55/hpf) or NaCl alone (18/hpf). In the B16 murine melanoma model, the median survival time in days was 21 for controls (n=5), 42 for TLPLDC (n=10), and 56 for TLPO (n=20). At 6 months, the equine model revealed 1 complete and 3 partial responses (50%, 68%, and 45% tumor reduction). Conclusions: Silicate capping of the YCWP effectively limits leakage of contents, improves uptake by DC and delivery to the cytoplasm without the need for GM-CSF. In early animal studies, TLPO appears to have equivalent efficacy to TLPLDC, but eliminates the need for in vitro DC loading. We plan to perform a bridging study comparing TLPO to TLPLDC after enrollment is complete in our phase IIb TLPLDC melanoma trial.
Comprehensive Evaluation of the Efficiency of Yeast Cell Wall Extract to Adsorb Ochratoxin A and Mitigate Accumulation of the Toxin in Broiler Chickens