Biomaterials created from supramolecular peptides, proteins, and their derivatives have been receiving increasing interest for both immunological applications such as vaccines and immunotherapies, as well as ostensibly non-immunological applications such as therapeutic delivery or tissue engineering. However, simple rules for either maximizing immunogenicity or abolishing it have yet to be elucidated, even though immunogenicity is a prime consideration for the design of any supramolecular biomaterial intended for use in vivo. Here we investigated a range of physicochemical properties of fibrillized peptide biomaterials, identifying negative surface charge as a means for completely abolishing antibody and T cell responses against them in mice, even when they display a competent epitope. The work was facilitated by the modularity of the materials, which enabled the generation of a set of co-assembled fibrillar peptide materials with broad ranges of surface properties. It was found that negative surface charge, provided via negatively charged amino acid residues, prevented T cell and antibody responses to antigen-carrying assemblies because it prevented uptake of the materials by antigen presenting cells (APCs), which in turn prevented presentation of the epitope peptide in the APCs’ major histocompatibility (MHC) class II molecules. Conversely, positive surface charge augmented the uptake of fibrillized peptides by APCs. These findings suggest that some surface characteristics such as extensive negative charge should be avoided in vaccine design using supramolecular peptide assemblies. More importantly, it provides a strategy to switch off potentially problematic immunogenicity for using these materials in non-immunological applications.

Constitutive B-cell receptor signaling leads to overexpression of the anti-apoptotic BCL-2 protein and is implicated in the pathogenesis of many types of B-cell Non-Hodgkin Lymphoma (B-NHL). The BCL-2 small molecule inhibitor venetoclax shows promising clinical response rates in several lymphomas, but is not curative as monotherapy. Radiotherapy (RT) is a rational candidate for combining with BCL-2 inhibition, as DNA damage caused by RT increases the activity of pro-apoptotic BCL-2 pathway proteins, and lymphomas are exquisitely sensitive to radiation. We tested B-NHL responses to venetoclax combined with either external beam RT or radioimmunotherapy (RIT), which joins the selectivity of antibody targeting with the effectiveness of irradiation. We first tested cytotoxicity of cesium-137 irradiation plus venetoclax in 14 B-NHL cell lines representing five lymphoma sub-types. Combination treatment synergistically increased cell death in ten of 14 lines. Lack of synergy was predicted by resistance to single-agent venetoclax and high BCL-XL expression. We then assessed the efficacy of external beam RT plus venetoclax in murine xenograft models of mantle cell (MCL), germinal-center diffuse large B-cell (GCB-DLBCL), and activated B-cell (ABC-DLBCL) lymphomas. In each model, external beam RT plus venetoclax synergistically increased mouse survival time, curing up to 10%. We finally combined venetoclax treatment of MCL and ABC-DLBCL xenografts with a pretargeted RIT (PRIT) system directed against the CD20 antigen. Optimal dosing of PRIT plus venetoclax cured 100% of mice with no detectable toxicity. Venetoclax combined with RT may be a promising treatment for a wide range of lymphomas.

No abstract

<div>Abstract<p>Constitutive B-cell receptor signaling leads to overexpression of the antiapoptotic BCL-2 protein and is implicated in the pathogenesis of many types of B-cell non-Hodgkin lymphoma (B-NHL). The BCL-2 small-molecule inhibitor venetoclax shows promising clinical response rates in several lymphomas, but is not curative as monotherapy. Radiotherapy is a rational candidate for combining with BCL-2 inhibition, as DNA damage caused by radiotherapy increases the activity of pro-apoptotic BCL-2 pathway proteins, and lymphomas are exquisitely sensitive to radiation. We tested B-NHL responses to venetoclax combined with either external beam radiotherapy or radioimmunotherapy (RIT), which joins the selectivity of antibody targeting with the effectiveness of irradiation. We first tested cytotoxicity of cesium-137 irradiation plus venetoclax in 14 B-NHL cell lines representing five lymphoma subtypes. Combination treatment synergistically increased cell death in 10 of 14 lines. Lack of synergy was predicted by resistance to single-agent venetoclax and high BCL-XL expression. We then assessed the efficacy of external beam radiotherapy plus venetoclax in murine xenograft models of mantle cell (MCL), germinal-center diffuse large B-cell (GCB-DLBCL), and activated B-cell (ABC-DLBCL) lymphomas. In each model, external beam radiotherapy plus venetoclax synergistically increased mouse survival time, curing up to 10%. We finally combined venetoclax treatment of MCL and ABC-DLBCL xenografts with a pretargeted RIT (PRIT) system directed against the CD20 antigen. Optimal dosing of PRIT plus venetoclax cured 100% of mice with no detectable toxicity. Venetoclax combined with radiotherapy may be a promising treatment for a wide range of lymphomas <i>Cancer Res; 77(14); 3885–93. ©2017 AACR</i>.</p></div>

<p>Table S1 presents mouse survival data for in vivo experiments</p>

<p>Table S1 presents mouse survival data for in vivo experiments</p>

Sarcocystis falcatula is a well-known cause of fatal pneumonia in some birds, particularly Old World psittacines. Here we describe fatal sarcosystosis due to S. falcatula in 3 penguins (Family Spheniscidae) under managed care, including one African penguin (Spheniscus demersus), and two Southern rockhopper penguins (Eudyptes chrysocome). Randomly distributed foci of necrosis, inflammatory cell infiltrates, edema, and variable numbers of round to elongated protozoal schizonts were observed in sections of lung. Protozoal organisms exhibited strong immunoreactivity for Sarcocystis sp. antigen by immunohistochemistry. Apicomplexan and Sarcocystis genus-specific PCR assays and sequence analysis confirmed S. falcatula as the etiologic agent. These cases of fatal pneumonia attributed to S. falcatula expand the list of aberrant intermediate avian hosts, with particular implications for penguins.

<div>Abstract<p>Constitutive B-cell receptor signaling leads to overexpression of the antiapoptotic BCL-2 protein and is implicated in the pathogenesis of many types of B-cell non-Hodgkin lymphoma (B-NHL). The BCL-2 small-molecule inhibitor venetoclax shows promising clinical response rates in several lymphomas, but is not curative as monotherapy. Radiotherapy is a rational candidate for combining with BCL-2 inhibition, as DNA damage caused by radiotherapy increases the activity of pro-apoptotic BCL-2 pathway proteins, and lymphomas are exquisitely sensitive to radiation. We tested B-NHL responses to venetoclax combined with either external beam radiotherapy or radioimmunotherapy (RIT), which joins the selectivity of antibody targeting with the effectiveness of irradiation. We first tested cytotoxicity of cesium-137 irradiation plus venetoclax in 14 B-NHL cell lines representing five lymphoma subtypes. Combination treatment synergistically increased cell death in 10 of 14 lines. Lack of synergy was predicted by resistance to single-agent venetoclax and high BCL-XL expression. We then assessed the efficacy of external beam radiotherapy plus venetoclax in murine xenograft models of mantle cell (MCL), germinal-center diffuse large B-cell (GCB-DLBCL), and activated B-cell (ABC-DLBCL) lymphomas. In each model, external beam radiotherapy plus venetoclax synergistically increased mouse survival time, curing up to 10%. We finally combined venetoclax treatment of MCL and ABC-DLBCL xenografts with a pretargeted RIT (PRIT) system directed against the CD20 antigen. Optimal dosing of PRIT plus venetoclax cured 100% of mice with no detectable toxicity. Venetoclax combined with radiotherapy may be a promising treatment for a wide range of lymphomas <i>Cancer Res; 77(14); 3885–93. ©2017 AACR</i>.</p></div>