EpiVax (United States)

Porcine circovirus type 2 (PCV2) is strongly linked to a group of syndromes referred to as porcine-circovirus-associated diseases (PCVADs), which are controlled through vaccination; however, this does not induce sterilizing immunity but is instead involved in the evolution of the virus and is considered a factor in vaccine failure. This study sampled 84 herds (167 pigs) vaccinated against PCV2 and with clinical signs of PCVADs in five provinces across Colombia. PCV2 was identified and further characterized at the molecular level via genotyping and phylogenetic reconstructions. In addition, PCV2-associated lesions were examined via histopathology. Furthermore, the PCV2-Cap sequences retrieved were compared with three vaccines via the EpiCC tool and T cell epitope coverage. The prevalence of PCV2 was 82% in pigs and 92.9% in herds. The highest viral loads were identified in lymphoid tissue, and PCV2d emerged as the most predominant in pigs and herds (93.4% and 92.3%). Sequences for PCV2-ORF2 (n = 57; 55 PCV2d and 2 PCV2a) were determined, and PCV2d sequences were highly similar. The most common pneumonia pattern was suppurative bronchopneumonia, while the most common lung lesion was exudation in the airways; in lymphoid tissue, there was lymphoid depletion. The bivalent vaccine (PCV2a and PCVb) exhibited a higher EpiCC score (8.36) and T cell epitope coverage (80.6%) than monovalent PCV2a vaccines. In conclusion, PCV2d currently circulates widely in Colombia. Despite vaccination, there are clinical cases of PCV2, and immunoinformatic analyses demonstrate that bivalent vaccines improved the average coverage.

The original article has been updated to correct a statement "The general guidelines for the maximum amount of allowed HCP impurities is 100 ng per mg of HCP." to "The general guidelines for the maximum amount of allowed HCP impurities is 100 ng per mg of product."

Biologics developers are moving beyond antibodies for delivery of a wide range of therapeutic interventions. These non-antibody modalities are often based on ‘natural’ protein scaffolds that are modified to deliver bioactive sequences. Both human-derived and non-human-sourced scaffold proteins have been developed. New types of “non-antibody” scaffolds are still being discovered, as they offer attractive alternatives to monoclonals due to their smaller size, improved stability, and ease of synthesis. They are believed to have low immunogenic potential. However, while several human-sourced protein scaffolds have not been immunogenic in clinical studies, this may not predict their overall performance in other therapeutic applications. A preliminary evaluation of their potential for immunogenicity is warranted. Immunogenicity risk potential has been clearly linked to the presence of T “helper” epitopes in the sequence of biologic therapeutics. In addition, tolerogenic epitopes are present in some human proteins and may decrease their immunogenic potential. While the detailed sequences of many non-antibody scaffold therapeutic candidates remain unpublished, their backbone sequences are available for review and analysis. We assessed 12 example non-antibody scaffold backbone sequences using our epitope-mapping tools (EpiMatrix) for this perspective. Based on EpiMatrix scoring, their HLA DRB1-restricted T cell epitope content appears to be lower than the average protein, and sequences that may act as tolerogenic epitopes are present in selected human-derived scaffolds. Assessing the potential immunogenicity of scaffold proteins regarding self and non-self T cell epitopes may be of use for drug developers and clinicians, as these exciting new non-antibody molecules begin to emerge from the preclinical pipeline into clinical use.

T cell-mediated immunity plays a central role in the control and clearance of intracellular Coxiella burnetii infection, which can cause Q fever. Therefore, we aimed to develop a novel T cell-targeted vaccine that induces pathogen-specific cell-mediated immunity to protect against Q fever in humans while avoiding the reactogenicity of the current inactivated whole cell vaccine. Human HLA class II T cell epitopes from C. burnetii were previously identified and selected by immunoinformatic predictions of HLA binding, conservation in multiple C. burnetii isolates, and low potential for cross-reactivity with the human proteome or microbiome. Epitopes were selected for vaccine inclusion based on long-lived human T cell recall responses to corresponding peptides in individuals that had been naturally exposed to the bacterium during a 2007-2010 Q fever outbreak in the Netherlands. Multiple viral vector-based candidate vaccines were generated that express concatemers of selected epitope sequences arranged to minimize potential junctional neo-epitopes. The vaccine candidates caused no antigen-specific reactogenicity in a sensitized guinea pig model. A subset of the vaccine epitope peptides elicited antigenic recall responses in splenocytes from C57BL/6 mice previously infected with C. burnetii. However, immunogenicity of the vaccine candidates in C57BL/6 mice was dominated by a single epitope and this was insufficient to confer protection against an infection challenge, highlighting the limitations of assessing human-targeted vaccine candidates in murine models. The viral vector-based vaccine candidates induced antigen-specific T cell responses to a broader array of epitopes in cynomolgus macaques, establishing a foundation for future vaccine efficacy studies in this large animal model of C. burnetii infection.

An effective malaria vaccine must prevent disease in a range of populations living in regions with vastly different transmission rates and protect against genetically-diverse Plasmodium falciparum (Pf) strains. The protective efficacy afforded by the currently licensed malaria vaccine, Mosquirix™, promotes strong humoral responses to Pf circumsporozoite protein (CSP) 3D7 but protection is limited in duration and by strain variation. Helper CD4 T cells are central to development of protective immune responses, playing roles in B cell activation and maturation processes, cytokine production, and stimulation of effector T cells. Therefore, we took advantage of recent in silico modeling advances to predict and analyze human leukocyte antigen (HLA)-restricted class II epitopes from PfCSP – across the entire PfCSP 3D7 sequence as well as in 539 PfCSP sequence variants – with the goal of improving PfCSP-based malaria vaccines. Specifically, we developed a systematic workflow to identify peptide sequences capable of binding HLA-DR in a context relevant to achieving broad human population coverage utilizing cognate T cell help and with limited T regulatory cell activation triggers. Through this workflow, we identified seven predicted class II epitope clusters in the N- and C-terminal regions of PfCSP 3D7 and an additional eight clusters through comparative analysis of 539 PfCSP sequence variants. A subset of these predicted class II epitope clusters was synthesized as peptides and assessed for HLA-DR binding in vitro. Further, we characterized the functional capacity of these peptides to prime and activate human peripheral blood mononuclear cells (PBMCs), by monitoring cytokine response profiles using MIMIC® technology (Modular IMmune In vitro Construct). Utilizing this decision framework, we found sufficient differential cellular activation and cytokine profiles among HLA-DR-matched PBMC donors to downselect class II epitope clusters for inclusion in a vaccine targeting PfCSP. Importantly, the downselected clusters are not highly conserved across PfCSP variants but rather, they overlap a hypervariable region (TH2R) in the C-terminus of the protein. We recommend assessing these class II epitope clusters within the context of a PfCSP vaccine, employing a test system capable of measuring immunogenicity across a broad set of HLA-DR alleles.

SummaryHuman antibody responses are established by the generation of combinatorial sequence diversity in antibody variable domains, followed by iterative rounds of mutation and selection via T cell recognition of antigen peptides presented on MHC-II. Here, we report that MHC-II peptide epitope deletion from B cell receptors (BCRs) correlates with antibody developmentin vivo. Large-scale antibody sequence analysis and experimental validation of peptide binding revealed that MHC-II epitope removal from BCRs is linked to genetic signatures of T cell help, and donor-specific antibody repertoire modeling demonstrated that somatic hypermutation selectively targets the personalized MHC-II epitopes in antibody variable regions. Mining of class-switched sequences and serum proteomic data revealed that MHC-II epitope deletion is associated with antibody class switching and long-term secretion into serum. These data suggest that the MHC-II peptide epitope content of a BCR is an important determinant of antibody maturation that shapes the composition and durability of humoral immunity.HighlightsAntibody somatic hypermutation selectively removes MHC-II peptide epitopes from B cell receptors.Antibodies with lower MHC-II epitope content show evidence of greater T cell help, including class-switching.MHC-II peptide epitope removal from a BCR is linked to long-term antibody secretion in serum.MHC-II genotype provides a personalized selection pressure on human antibody development.

Abstract The liver harbors two main innate lymphoid cell (ILC) populations, conventional NK (cNK) cells and tissue-resident NK (trNK) cells also called ILC1s. While the antiviral functions of cNK cells have been well established, trNK cell functions are only beginning to be uncovered. Using the MCMV model of infection, we found that in contrast to liver cNK cells, the trNK cell population initially undergoes a contraction phase prior to a recovery phase to homeostatic levels. The rapid contraction phase is due to apoptosis, while the recovery phase occurs via proliferation in situ. Liver trNK cell apoptosis is not mediated by fratricide, liver lymphocytes, or inflammatory cytokines. Instead, we found that liver trNK cell apoptosis is the consequence of an increased sensitivity to lactic acid. Interestingly, MCMV infection lead to an increase of lactate in the liver, which was supported by changes in lactate dehydrogenase expression. Mechanistic analysis indicates that trNK cell sensitivity to lactate is linked to impaired mitochondrial functions. These findings underscore the unique properties of the liver resident NK cell compartment.

Abstract Clinical studies have highlighted the potential of precision cancer immunotherapy to effectively control the tumor of patients across cancer indications. However, recent studies showcase the difficulty of establishing robust CD8 and CD4 T cell responses. We hypothesize that poor cancer vaccine performance may be due in part to the inadvertent inclusion of suppressive T cell neo-epitopes in neoantigen vaccines that may be recognized by regulatory T cells (Tregs). To test this hypothesis, we used the Ancer™ system to identify and select neo-epitopes from the CT26 syngeneic mouse model. Ancer™ leverages EpiMatrix® and JanusMatrix™, state-of-the-art predictive algorithms that have been extensively validated in prospective vaccine studies for infectious diseases (Moise et al., Hum. Vaccines Immunother 2015; Wada et al., Sci. Rep. 2017). Distinctive features of Ancer™ over other in silico pipelines are its ability to accurately predict CD4 T cell epitopes and to identify tolerated or Treg epitopes. In a first experiment, optimally selected CT26 neoantigen vaccine candidates were identified with Ancer™ and ranked according to tumor expression level and predicted Class I- and Class II-restricted immunogenicity. Self-like, putative Treg epitopes were removed in this process. Naïve Balb/c mice were immunized subcutaneously with a peptide pool comprised of the 20 highest ranking neoantigens delivered with PolyICLC (Oncovir). Immunization with Ancer™-derived neoantigens induced strong IFNg ELISpot responses compared to controls (p < 0.001). Flow cytometry confirmed the vaccine stimulated multifunctional CD4+ and CD8+ T cells. In a follow-on experiment, ten self-like neoantigens, from the same CT26 genome, were selected with Ancer™. These neoantigens may be recognized by Tregs due to their high degree of similarity with self, based on JanusMatrix™. Co-administration of the CT26 self-like neoantigens with our optimally designed neoantigen vaccine in naïve Balb/c mice diminished IFNg ELISpot responses by 5-fold compared to vaccination without the self-like neoantigens (p = 0.003). While it has been well known that Tregs are present in tumors, these results suggest the possibility that tumor-derived neo-epitopes may be recruiting Tregs to the tumor. More importantly, the inadvertent inclusion of Treg driving neoantigens in vaccine formulations may hinder efforts to induce strong T cell-mediated tumor control. In silico screening of neoantigen sequences using specialized tools offers the possibility of enriching and designing new vaccines with higher quality candidates. Efforts are ongoing to determine the effect of Ancer™-derived self-like neoantigens on CD4+ and CD8+ T cells and how the inclusion of self-like neoantigens in vaccines affects their efficacy. Citation Format: Guilhem Richard, Bethany Biron, Christine Boyle, Matthew Ardito, Leonard Moise, William Martin, Gad Berdugo, Anne S. De Groot. Filtering out self-like neoantigens improves immune response to cancer vaccines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 943.

Abstract Next-generation sequencing has opened the door to precision cancer therapies targeting mutations expressed by tumor cells. However, most neo-epitopes selected by traditional T cell epitope prediction algorithms prove to be non-immunogenic. Poor predictive performance may partially be due to inclusion of mutated epitopes cross-conserved with self-epitopes recognized by the T cell receptor of regulatory (Treg), anergic or deleted T cells. Vaccination with self-epitopes can lead to weak effector responses, active immune suppression, and toxicity due to immune-mediated adverse effects. We have developed Ancer, an advanced cancer T cell epitope identification and characterization tool, that streamlines the selection of Class I and Class II T cell neo-epitopes. Ancer leverages EpiMatrix and JanusMatrix, state-of-the-art predictive algorithms that have been extensively validated in prospective vaccine studies for infectious diseases [Moise et al., Hum. Vaccines Immunother 2015; Wada et al., Sci. Rep. 2017]. Distinctive features of Ancer are its ability to accurately predict Class II HLA ligands with EpiMatrix and its 82% positive predictive value, as estimated in previous prospective studies. Additionally, the application of JanusMatrix allows for the prioritization of neo-epitopes with reduced potential for Treg induction, that is responsible for diminished efficacy of current cancer therapies. We validated Ancer's predictive accuracy using datasets of HLA-bound peptides detected by mass spectrometry, which are independent of training sequence data used in model development. Analysis of sequences from Abelin et al., Immunity 2017 shows a 96% agreement between Ancer predictions and peptides eluted from common Class I HLAs, while only 86% of these sequences are accurately predicted by NetMHC or NetMHCpan. An additional retrospective analysis of a cancer immunogenicity study [Strønen et al., Science 2016] demonstrates that Ancer selects immunogenic neo-epitopes with 72% accuracy, as compared to 21% accuracy when using public prediction tools. These results demonstrate that Ancer may focus epitope candidate selection on higher value sequences than conventional algorithms. Class I and Class II neo-epitopes with low Treg activation potential may then be used to support the development of safer and more effective vaccines. Citation Format: Guilhem Richard, Lenny Moise, Matthew Ardito, Frances Terry, Gad Berdugo, William Martin, Anne De Groot. Integrated approaches for design of precision cancer immunotherapies: Selection of Class I and Class II T cell neo-epitopes and removal of Treg epitopes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5311.