TLR4 Agonist Combined with Trivalent Protein JointS of Streptococcus suis Provides Immunological Protection in Animals

Streptococcus suis (S. suis) serotype 2 (SS2) is the causative agent of swine streptococcosis and can cause severe diseases in both pigs and humans. Although the traditional inactive vaccine can protect pigs from SS2 infection, novel vaccine candidates are needed to overcome its shortcomings. Three infection-associated proteins in S. suis—muramidase-released protein (MRP), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and DLD, a novel putative dihydrolipoamide dehydrogenase—have been previously identified by immunoproteomic assays. In this study, the effective immune protection of the recombinant trivalent protein GAPDH-MRP-DLD (JointS) against SS2, SS7, and SS9 was determined in zebrafish. To improve the immune efficacy of JointS, monophosphoryl lipid A (MPLA) as a TLR4 agonist adjuvant, which induces a strong innate immune response in the immune cells of mice and pigs, was combined with JointS to immunize the mice. The results showed that immunized mice could induce the production of a high titer of anti-S. suis antibodies; as a result, 100% of mice survived after SS2 infection. Furthermore, JointS provides good protection against virulent SS2 strain infections in piglets. Given the above, there is potential to develop JointS as a novel subunit vaccine for piglets to prevent infection by SS2 and other S. suis serotypes.

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Adsorption of Toll-Like Receptor 4 Agonist to Alum-Based Tetanus Toxoid Vaccine Dampens Pro-T Helper 2 Activities and Enhances Antibody Responses

Journal of Immunology Research ◽

10.1155/2015/280238 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 9

Author(s):

Juliana Bortolatto ◽

Luciana Mirotti ◽

Dunia Rodriguez ◽

Eliane Gomes ◽

Momtchilo Russo

Keyword(s):

Tetanus Toxoid ◽

Lipid A ◽

Antibody Responses ◽

Aluminum Salts ◽

T Helper 2 ◽

Monophosphoryl Lipid A ◽

Tlr4 Agonist ◽

Specific Antibody Production ◽

Humoral Responses

Aluminum salts gels (alum) are TLR-independent adjuvants and have been used to boost antibody responses in alum-based vaccines such as diphtheria, pertussis, and tetanus toxoid (DPT) triple vaccine. However, the pro-Th2 activity of alum-based vaccine formulations has not been fully appreciated. Here we found that alum-based tetanus toxoid (TT) vaccine was biased toward a Th-2 profile as shown by TT-induced airway eosinophilic inflammation, type 2 cytokine production, and high levels of IgE anaphylactic antibodies. The adsorption into alum of prototypic TLR4 agonists such as lipopolysaccharides (LPS) derived fromEscherichia coliconsistently dampened TT-induced Th2 activities without inducing IFNγor Th1-like responses in the lung. Conversely, adsorption of monophosphoryl lipid A (MPLA) extracted fromSalmonella minnesota, which is a TIR-domain-containing adapter-inducing interferon-β- (TRIF-) biased TLR4 agonist, was less effective in decreasing Th-2 responses. Importantly, in a situation with antigenic competition (OVA plus TT), TT-specific IgG1 or IgG2a was decreased compared with TT sensitization. Notably, LPS increased the production of IgG1 and IgG2a TT-specific antibodies. In conclusion, the addition of LPS induces a more robust IgG1 and IgG2a TT-specific antibody production and concomitantly decreases Th2-cellular and humoral responses, indicating a potential use of alum/TLR-based vaccines.

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Incorporation of the TLR4 Agonist Monophosphoryl Lipid A Into the Bilayer of DDA/TDB Liposomes: Physico-Chemical Characterization and Induction of CD8+ T-Cell Responses In Vivo

Pharmaceutical Research ◽

10.1007/s11095-010-0301-9 ◽

2010 ◽

Vol 28 (3) ◽

pp. 553-562 ◽

Cited By ~ 33

Author(s):

Pernille Nordly ◽

Else Marie Agger ◽

Peter Andersen ◽

Hanne Mørck Nielsen ◽

Camilla Foged

Keyword(s):

T Cell ◽

Lipid A ◽

Chemical Characterization ◽

Cd8 T Cell ◽

Cell Responses ◽

Monophosphoryl Lipid A ◽

Monophosphoryl Lipid ◽

Physico Chemical ◽

Tlr4 Agonist

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Vaccination Route as a Determinant of Protective Antibody Responses against Herpes Simplex Virus

Vaccines ◽

10.3390/vaccines8020277 ◽

2020 ◽

Vol 8 (2) ◽

pp. 277

Author(s):

Clare Burn Aschner ◽

Carl Pierce ◽

David M. Knipe ◽

Betsy C. Herold

Keyword(s):

Herpes Simplex ◽

Neutralizing Antibodies ◽

Lipid A ◽

Low Dose ◽

High Titer ◽

Antibody Responses ◽

Protein Vaccine ◽

Herpes Simplex Viruses ◽

Monophosphoryl Lipid A ◽

Simplex Virus

Herpes simplex viruses (HSV) are significant global health problems associated with mucosal and neurologic disease. Prior experimental vaccines primarily elicited neutralizing antibodies targeting glycoprotein D (gD), but those that advanced to clinical efficacy trials have failed. Preclinical studies with an HSV-2 strain deleted in gD (ΔgD-2) administered subcutaneously demonstrated that it elicited a high titer, weakly neutralizing antibodies that activated Fcγ receptors to mediate antibody-dependent cellular cytotoxicity (ADCC), and completely protected mice against lethal disease and latency following vaginal or skin challenge with HSV-1 or HSV-2. Vaccine efficacy, however, may be impacted by dose and route of immunization. Thus, the current studies were designed to compare immunogenicity and efficacy following different routes of vaccination with escalating doses of ΔgD-2. We compared ΔgD-2 with two other candidates: recombinant gD protein combined with aluminum hydroxide and monophosphoryl lipid A adjuvants and a replication-defective virus deleted in two proteins involved in viral replication, dl5-29. Compared to the subcutaneous route, intramuscular and/or intradermal immunization resulted in increased total HSV antibody responses for all three vaccines and boosted the ADCC, but not the neutralizing response to ΔgD and dl5-29. The adjuvanted gD protein vaccine provided only partial protection and failed to elicit ADCC independent of route of administration. In contrast, the increased ADCC following intramuscular or intradermal administration of ΔgD-2 or dl5-29 translated into significantly increased protection. The ΔgD-2 vaccine provided 100% protection at doses as low as 5 × 104 pfu when administered intramuscularly or intradermally, but not subcutaneously. However, administration of a combination of low dose subcutaneous ΔgD-2 and adjuvanted gD protein resulted in greater protection than low dose ΔgD-2 alone indicating that gD neutralizing antibodies may contribute to protection. Taken together, these results demonstrate that ADCC provides a more predictive correlate of protection against HSV challenge in mice and support intramuscular or intradermal routes of vaccination.

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Lipopolysaccharide directly inhibits bicarbonate absorption by the renal outer medullary collecting duct

Scientific Reports ◽

10.1038/s41598-020-77363-w ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Shuichi Tsuruoka ◽

Jeffrey M. Purkerson ◽

George J. Schwartz

Keyword(s):

Lipid A ◽

Kinase Inhibitor ◽

Collecting Duct ◽

Therapeutic Interventions ◽

E Coli ◽

Monophosphoryl Lipid A ◽

Tlr4 Agonist ◽

Medullary Collecting Duct ◽

Phosphoinositide 3 Kinase

AbstractAcidosis is associated with E. coli induced pyelonephritis but whether bacterial cell wall constituents inhibit HCO3 transport in the outer medullary collecting duct from the inner stripe (OMCDi) is not known. We examined the effect of lipopolysaccharide (LPS), on HCO3 absorption in isolated perfused rabbit OMCDi. LPS caused a ~ 40% decrease in HCO3 absorption, providing a mechanism for E. coli pyelonephritis-induced acidosis. Monophosphoryl lipid A (MPLA), a detoxified TLR4 agonist, and Wortmannin, a phosphoinositide 3-kinase inhibitor, prevented the LPS-mediated decrease, demonstrating the role of TLR4-PI3-kinase signaling and providing proof-of-concept for therapeutic interventions aimed at ameliorating OMCDi dysfunction and pyelonephritis-induced acidosis.

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Toll-like receptor 4 agonist-based nanoparticles orchestrate protection against sepsis

Journal of Materials Science ◽

10.1007/s10853-021-06638-y ◽

2022 ◽

Author(s):

Yongxiang Zhao ◽

Xinjing Lv ◽

Jie Huang ◽

Huiting Zhou ◽

Hairong Wang ◽

...

Keyword(s):

Lipid A ◽

Severe Infection ◽

Plga Nanoparticles ◽

Great Promise ◽

Toll Like Receptor 4 ◽

E Coli ◽

Monophosphoryl Lipid A ◽

Tlr4 Agonist ◽

Life Threatening ◽

Plga Nanoparticle

AbstractSepsis, a life-threatening organ dysfunction induced by severe infection and uncontrolled host immune response, threatens the health of people all over the world. Herein, a type of nanoparticle formulation with simple components is synthesized by encapsulating monophosphoryl lipid A (MPLA), a TLR4 agonist, with poly(lactic-co-glycolic acid) (PLGA) nanoparticle. The obtained nanoparticles (MPLA@PLGA) could provide Escherichia coli (E. coli)-induced sepsis protection by regulating the immune system after sepsis challenge, including promoting the levels of various cytokines, boosting the percentage of natural killer cells and accelerating bacterial clearance. Notably, the survival mice pre-treated with these nanoparticles could resist repeated E. coli-induced sepsis. Our work therefore provides the great promise of MPLA@PLGA nanoparticles as a simple yet effective nano-drug for prevention and protection against E. coli-induced sepsis.

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The TLR4 Agonist Monophosphoryl Lipid A Drives Broad Resistance to Infection via Dynamic Reprogramming of Macrophage Metabolism

The Journal of Immunology ◽

10.4049/jimmunol.1800085 ◽

2018 ◽

Vol 200 (11) ◽

pp. 3777-3789 ◽

Cited By ~ 28

Author(s):

Benjamin A. Fensterheim ◽

Jamey D. Young ◽

Liming Luan ◽

Ruby R. Kleinbard ◽

Cody L. Stothers ◽

...

Keyword(s):

Lipid A ◽

Monophosphoryl Lipid A ◽

Monophosphoryl Lipid ◽

Tlr4 Agonist ◽

Resistance To Infection ◽

Dynamic Reprogramming

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The Use of Translational Modelling and Simulation to Develop Immunomodulatory Therapy as an Adjunct to Antibiotic Treatment in the Context of Pneumonia

Pharmaceutics ◽

10.3390/pharmaceutics13050601 ◽

2021 ◽

Vol 13 (5) ◽

pp. 601

Author(s):

Robin Michelet ◽

Moreno Ursino ◽

Sandrine Boulet ◽

Sebastian Franck ◽

Fiordiligie Casilag ◽

...

Keyword(s):

Lipid A ◽

Bacterial Resistance ◽

Statistical Modelling ◽

Modelling And Simulation ◽

Knowledge Based ◽

Monophosphoryl Lipid A ◽

Tlr4 Agonist ◽

Tract Infections

The treatment of respiratory tract infections is threatened by the emergence of bacterial resistance. Immunomodulatory drugs, which enhance airway innate immune defenses, may improve therapeutic outcome. In this concept paper, we aim to highlight the utility of pharmacometrics and Bayesian inference in the development of immunomodulatory therapeutic agents as an adjunct to antibiotics in the context of pneumonia. For this, two case studies of translational modelling and simulation frameworks are introduced for these types of drugs up to clinical use. First, we evaluate the pharmacokinetic/pharmacodynamic relationship of an experimental combination of amoxicillin and a TLR4 agonist, monophosphoryl lipid A, by developing a pharmacometric model accounting for interaction and potential translation to humans. Capitalizing on this knowledge and associating clinical trial extrapolation and statistical modelling approaches, we then investigate the TLR5 agonist flagellin. The resulting workflow combines expert and prior knowledge on the compound with the in vitro and in vivo data generated during exploratory studies in order to construct high-dimensional models considering the pharmacokinetics and pharmacodynamics of the compound. This workflow can be used to refine preclinical experiments, estimate the best doses for human studies, and create an adaptive knowledge-based design for the next phases of clinical development.

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Small Molecule Potentiator of Adjuvant Activity Enhancing Survival to Influenza Viral Challenge

Frontiers in Immunology ◽

10.3389/fimmu.2021.701445 ◽

2021 ◽

Vol 12 ◽

Author(s):

Tetsuya Saito ◽

Yukiya Sako ◽

Fumi Sato-Kaneko ◽

Tadashi Hosoya ◽

Shiyin Yao ◽

...

Keyword(s):

Immune Responses ◽

Lipid A ◽

High Titer ◽

Adjuvant Activity ◽

Murine Bone Marrow ◽

Virus Challenge ◽

Monophosphoryl Lipid A ◽

Tlr4 Ligand

As viruses continue to mutate the need for rapid high titer neutralizing antibody responses has been highlighted. To meet these emerging threats, agents that enhance vaccine adjuvant activity are needed that are safe with minimal local or systemic side effects. To respond to this demand, we sought small molecules that would sustain and improve the protective effect of a currently approved adjuvant, monophosphoryl lipid A (MPLA), a Toll-like receptor 4 (TLR4) agonist. A lead molecule from a high-throughput screen, (N-(4-(2,5-dimethylphenyl)thiazol-2-yl)-4-(piperidin-1-ylsulfonyl)benzamide, was identified as a hit compound that sustained NF-κB activation by a TLR4 ligand, lipopolysaccharide (LPS), after an extended incubation (16 h). In vitro, the resynthesized compound (2D216) enhanced TLR4 ligand-induced innate immune activation and antigen presenting function in primary murine bone marrow-derived dendritic cells without direct activation of T cells. In vivo murine vaccination studies demonstrated that compound 2D216 acted as a potent co-adjuvant when used in combination with MPLA that enhanced antigen-specific IgG equivalent to that of AS01B. The combination adjuvant MPLA/2D216 produced Th1 dominant immune responses and importantly protected mice from lethal influenza virus challenge. 2D216 alone or 2D216/MPLA demonstrated minimal local reactogenicity and no systemic inflammatory response. In summary, 2D216 augmented the beneficial protective immune responses of MPLA as a co-adjuvant and showed an excellent safety profile.

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Adjuvantation of an Influenza Hemagglutinin Antigen with TLR4 and NOD2 Agonists Encapsulated in Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Enhances Immunogenicity and Protection against Lethal Influenza Virus Infection in Mice

Vaccines ◽

10.3390/vaccines8030519 ◽

2020 ◽

Vol 8 (3) ◽

pp. 519

Author(s):

Amir Tukhvatulin ◽

Alina Dzharullaeva ◽

Alina Erokhova ◽

Anastasia Zemskaya ◽

Maxim Balyasin ◽

...

Keyword(s):

Influenza Virus ◽

Lipid A ◽

Influenza Virus Infection ◽

Muramyl Dipeptide ◽

Plga Nanoparticles ◽

Influenza Hemagglutinin ◽

Monophosphoryl Lipid A ◽

Tlr4 Agonist

Along with their excellent safety profiles, subunit vaccines are typically characterized by much weaker immunogenicity and protection efficacy compared to whole-pathogen vaccines. Here, we present an approach aimed at bridging this disadvantage that is based on synergistic collaboration between pattern-recognition receptors (PRRs) belonging to different families. We prepared a model subunit vaccine formulation using an influenza hemagglutinin antigen incorporated into poly-(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles adjuvanted with monophosphoryl lipid A (TLR4 agonist) and muramyl dipeptide (NOD2 agonist). The efficacy studies were conducted in comparison to control vaccine formulations containing individual PRR agonists. We show that the complex adjuvant based on TLR4 and NOD2 agonists potentiates proinflammatory cell responses (measured by activity of transcription factors and cytokine production both in vitro and in vivo) and enhances the phagocytosis of vaccine particles up to comparable levels of influenza virus uptake. Finally, mice immunized with vaccine nanoparticles containing both PRR agonists exhibited enhanced humoral (IgG, hemagglutination-inhibition antibody titers) and cellular (percentage of proliferating CD4+ T-cells, production of IFNɣ) immunity, leading to increased resistance to lethal influenza challenge. These results support the idea that complex adjuvants stimulating different PRRs may present a better alternative to individual PAMP-based adjuvants and could further narrow the gap between the efficacy of subunit versus whole-pathogen vaccines.

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