Natural pattern-recognition-receptor agonists as adjuvants for in situ vaccination lymphoma immunotherapy.

2018 ◽  
Vol 36 (5_suppl) ◽  
pp. 123-123 ◽  
Author(s):  
Mark Aleynick ◽  
Paul Peng ◽  
Linda Hammerich ◽  
Ranjan Upadhyay ◽  
Netonia Marshall ◽  
...  
Keyword(s):  
Pattern Recognition ◽  
T Cell ◽  
Immune Activation ◽  
Pattern Recognition Receptor ◽  
Receptor Agonists ◽  
Prophylactic Vaccines ◽  
Recognition Receptor

123 Background: In patients with low-grade lymphoma, in situ vaccination has yielded both partial and complete remissions in clinical trials. Though clinical responses have been observed with multiple pattern recognition receptor agonists (PRRa), the optimal immune stimulant is unknown. We hypothesize that natural PRRa, such as the attenuated pathogens or subunits found in common prophylactic vaccines, could target multiple PRR in a physiologically relevant context and lead to a more robust activation of dendritic cells (DCs) versus synthetic PRRa. Methods: 20 vaccines, including BCG, Typhim Vi, MMR-II, etc. were screened in vitro, where DC phenotype and function were evaluated by flow cytometry. Flt3L-mobilized DC ability to phagocytose, process, present, and cross-present soluble protein or tumor derived antigen, were assessed using CRISPR gene-edited, β2m(-/-) GFP-lymphoma cells and a novel GFP-specific (‘JEDI’) CD8 T cell system. Vaccine mechanism of immune activation was elucidated using a library of PRR-null macrophage cell lines. Potent vaccines were also evaluated in vivo in a Flt3L-primed in situ vaccination using the A20 murine lymphoma model. Results: Several vaccines induced robust DC activation and several showed significant increases in subsequent T cell activation, proliferation, and tumor killing, suggesting increased antigen processing and cross-presentation by DCs. Some vaccines, either as single agents or in combination, were significantly more effective than synthetic PRRa in activating DCs and inducing a T cell response. In vivo, vaccine combination therapies induced tumor regression in a majority of animals, suggesting synergistic immune activation. Conclusions: This data suggests prophylactic vaccines are effective clinical-grade DC activators and can be repurposed for use in the in situ vaccination maneuver, with immediate translation into the clinic. Additionally, by extensive in vitro evaluation in parallel with in vivo studies, this work aims to identify a predictive in vitro molecular immune signature that correlates closely with adjuvant efficacy in vivo.

Natural pattern-recognition-receptor agonists in prophylactic vaccines for in situ vaccination of lymphoma.

2016 ◽  
Vol 34 (15_suppl) ◽  
pp. e14516-e14516
Author(s):  
Mark Aleynick ◽  
Paul Peng ◽  
Linda Hammerich ◽  
Antoine Tanne ◽  
Judith Agudo ◽  
...  
Keyword(s):  
Pattern Recognition ◽  
Pattern Recognition Receptor ◽  
Receptor Agonists ◽  
Prophylactic Vaccines ◽  
Recognition Receptor ◽  
Natural Pattern

Natural Toll-like Receptor Agonists for in Situ Vaccination Lymphoma Immunotherapy

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4002-4002
Author(s):  
Paul Peng ◽  
Joshua Brody ◽  
Linda Hammerich
Keyword(s):  
T Cell ◽  
T Cell Activation ◽  
Cell Activation ◽  
Clinical Grade ◽  
Toll Like Receptor ◽  
Activation Markers ◽  
Prophylactic Vaccines

Abstract Introduction: There is an unmet need for novel and effective treatments for lymphoma, the fifth most common malignancy in the US. In situ vaccination-an immunotherapeutic maneuver involving local irradiation, intratumoral (i.t.) injections of Flt3L and Toll-like receptor (TLR) agonist-has been shown in recent clinical trials (NCT00185965, NCT00880581, NCT00226993) to induce partial and complete remissions in patients with low-grade lymphoma. The strength of anti-tumor response correlates with the potency of immunogenic dendritic cells (DCs) to efficiently uptake and present tumor antigens to T cells. While the latest clinical trial NCT01976585 employs Poly-ICLC-a synthetic TLR3 agonist-to activate DCs, we hypothesize that "natural" TLR agonists (nTLRa) contained within prophylactic vaccines (e.g. live/attenuated bacteria or viruses) could simultaneously target multiple TLRs and be repurposed as clinical-grade DC activators for the in situ vaccination maneuver. Methods: Twenty-four prophylactic vaccines from our hospital's pharmacy were screened using in vitro assays for TLR ligand activity. DC subsets CD11c+ MHC-II+, CD11c+ CD11b+, and CD11c+ CD103+ were gated for flow-cytometric readouts of: (i.) activation markers (e.g. CD40, CD80, CD86, MHC class I and II), (ii.) ability to co-stimulate T cells in the context of TCR activation, as assessed by T cell activation markers (e.g. CD69, intracellular IFNγ), and (iii.) ability of nTLRa-activated DCs to induce T cell proliferation. TLR knock-out cell lines were used for dissecting the mechanistically-distinct properties of each prophylactic vaccine. Best single and combination nTLRa candidates were evaluated with the in situ vaccination maneuver in an in vivo A20 murine lymphoma model. Results: We identify a combination of vaccines Typhim, BCG-TICE, and MMR that demonstrates the ability to induce high expression of costimulatory molecules on DCs (Figure A) in vitro, as well as possessing the ability to co-stimulate in vitro T cell activation and proliferation. DC costimulation of T cell activation and proliferation is highly dependent on the "live" status of several vaccines, including BCG, MMR, Zostavax, which suggests the involvement of other pattern-recognition receptors in addition to TLRs. Cohorts of mice implanted with A20 lymphoma tumors that received i.t. injections of Typhim, BCG, MMR demonstrated slower A20 growth status post in situ vaccination maneuver, as compared to the control cohort receiving only Flt3L and irradiation (XRT). Conclusion: Prophylactic vaccines contain natural ligands to TLRs and are immediately translatable as sources of clinical-grade stimulators of dendritic cells. Combinations of the best nTLRa candidates display synergistic activation of DCs and demonstrate in vivo anti-A20 murine lymphoma immunity. Figure 1. Figure 1. Disclosures No relevant conflicts of interest to declare.

scholarly journals The soluble pattern recognition receptor PTX3 links humoral innate and adaptive immune responses by helping marginal zone B cells

2016 ◽  
Vol 213 (10) ◽  
pp. 2167-2185 ◽  
Author(s):  
Alejo Chorny ◽  
Sandra Casas-Recasens ◽  
Jordi Sintes ◽  
Meimei Shan ◽  
Nadia Polentarutti ◽  
...  
Keyword(s):  
Pattern Recognition ◽  
Immune System ◽  
T Cell ◽  
B Cells ◽  
Marginal Zone ◽  
Fluid Phase ◽  
Pattern Recognition Receptor ◽  
Marginal Zone B Cells ◽  
Adaptive Immune ◽  
Recognition Receptor

Pentraxin 3 (PTX3) is a fluid-phase pattern recognition receptor of the humoral innate immune system with ancestral antibody-like properties but unknown antibody-inducing function. In this study, we found binding of PTX3 to splenic marginal zone (MZ) B cells, an innate-like subset of antibody-producing lymphocytes strategically positioned at the interface between the circulation and the adaptive immune system. PTX3 was released by a subset of neutrophils that surrounded the splenic MZ and expressed an immune activation–related gene signature distinct from that of circulating neutrophils. Binding of PTX3 promoted homeostatic production of IgM and class-switched IgG antibodies to microbial capsular polysaccharides, which decreased in PTX3-deficient mice and humans. In addition, PTX3 increased IgM and IgG production after infection with blood-borne encapsulated bacteria or immunization with bacterial carbohydrates. This immunogenic effect stemmed from the activation of MZ B cells through a neutrophil-regulated pathway that elicited class switching and plasmablast expansion via a combination of T cell–independent and T cell–dependent signals. Thus, PTX3 may bridge the humoral arms of the innate and adaptive immune systems by serving as an endogenous adjuvant for MZ B cells. This property could be harnessed to develop more effective vaccines against encapsulated pathogens.

scholarly journals Signal Transduction by Different Forms of the γδ T Cell–Specific Pattern Recognition Receptor WC1

2014 ◽  
Vol 193 (1) ◽  
pp. 379-390 ◽  
Author(s):  
Chuang Chen ◽  
Haoting Hsu ◽  
Edward Hudgens ◽  
Janice C. Telfer ◽  
Cynthia L. Baldwin
Keyword(s):  
Signal Transduction ◽  
Pattern Recognition ◽  
T Cell ◽  
Specific Pattern ◽  
Pattern Recognition Receptor ◽  
Γδ T Cell ◽  
Recognition Receptor

scholarly journals Mature Dendritic Cells Infiltrate the T Cell-Rich Region of Oral Mucosa in Chronic Periodontitis: In Situ, In Vivo, and In Vitro Studies

2001 ◽  
Vol 167 (8) ◽  
pp. 4693-4700 ◽  
Author(s):  
Ravi Jotwani ◽  
Anna Karolina Palucka ◽  
Montasr Al-Quotub ◽  
Mahyar Nouri-Shirazi ◽  
Jay Kim ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
T Cell ◽  
Oral Mucosa ◽  
Chronic Periodontitis ◽  
In Vitro Studies ◽  
Rich Region

Abstract 15541: Novel Molecular Entity for Effective Cardiac Regeneration: 5’ppp Microrna Induces Reprogramming and Accelerates Cardiomyocyte Maturation

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Jiabiao Hu ◽  
Conrad Hodgkinson ◽  
Syeda S Baksh ◽  
Richard E Pratt ◽  
Victor J Dzau
Keyword(s):  
Pattern Recognition ◽  
Cardiac Regeneration ◽  
Molecular Entity ◽  
Rna Molecules ◽  
Novel Approach ◽  
Nfkb Activation ◽  
Recognition Receptor ◽  
Modest Effect

We have shown that miR combo directly reprograms fibroblasts into cardiomyocytes. However, both in vitro and in vivo, the generated cardiomyocytes are immature. Recently, we discovered that cardiomyocyte maturation is enhanced by NFkB activation. While miR combo initiates reprogramming, the effect on NFkB activity is modest. Therefore, miR combo only has a modest effect on maturation. We have now made an important discovery that 5’triphosphorylation (5’ppp) of RNA molecules markedly enhance NFkB activation; leading to increased and accelerated maturation of reprogramed cardiomyocytes. The addition of 5’ppp-RNA increased the number of mature cardiomyocytes by 4-fold (N=5, P<0.05). Moreover, 5’ppp-RNA also accelerated the appearance of mature cardiomyocyte mRNAs (1 day after transfection vs 4 days for unmodified RNA). Importantly, these effects were lost when the 5’ppp moiety was removed. Knockdown studies suggest that 5’ppp-RNA binds to the Pattern Recognition Receptor Rig1. MNase-seq and ChIP-seq analysis suggests that 5’ppp-RNA activates YY1 (N=4, P<0.001). We next asked if 5’ppp modification of the miRNAs within miR combo would also enhance cardiomyocyte maturation; thus, providing a single molecular entity that can initiate reprogramming and accelerate maturation of cardiomyocytes. Indeed, 5’ppp modification of the miR combo miR-1 increased the expression of the mature cardiomyocyte markers Actn2 (4-fold, N=3, P<0.05) and Myh6 (4-fold, N=3, P<0.05). In conclusion, 5’ppp-miR combo is a novel approach that has the advantage of both initiating reprogramming and accelerating cardiomyocyte maturation and should lead to more effective cardiac regeneration and restore cardiac function.

scholarly journals In situ induction of dendritic cell–based T cell tolerance in humanized mice and nonhuman primates

2011 ◽  
Vol 208 (12) ◽  
pp. 2477-2488 ◽  
Author(s):  
Kyeong Cheon Jung ◽  
Chung-Gyu Park ◽  
Yoon Kyung Jeon ◽  
Hyo Jin Park ◽  
Young Larn Ban ◽  
...  
Keyword(s):  
T Cell ◽  
Humanized Mice ◽  
Intercellular Adhesion Molecule ◽  
T Cell Tolerance ◽  
Cell Tolerance ◽  
Intercellular Adhesion Molecule 1 ◽  
Porcine Islet

Induction of antigen-specific T cell tolerance would aid treatment of diverse immunological disorders and help prevent allograft rejection and graft versus host disease. In this study, we establish a method of inducing antigen-specific T cell tolerance in situ in diabetic humanized mice and Rhesus monkeys receiving porcine islet xenografts. Antigen-specific T cell tolerance is induced by administration of an antibody ligating a particular epitope on ICAM-1 (intercellular adhesion molecule 1). Antibody-mediated ligation of ICAM-1 on dendritic cells (DCs) led to the arrest of DCs in a semimature stage in vitro and in vivo. Ablation of DCs from mice completely abrogated anti–ICAM-1–induced antigen-specific T cell tolerance. T cell responses to unrelated antigens remained unaffected. In situ induction of DC-mediated T cell tolerance using this method may represent a potent therapeutic tool for preventing graft rejection.

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