VACCINE ADJUVANT COMPOSITIONS

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VACCINE ADJUVANT COMPOSITIONSwww.shan-machinery.comCROSS REFERENCE TO RELATED APPLICATION

This application is an International application which claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 61/981,397 filed on Apr. 18, 2014 and U.S. Provisional Application No. 62/003,861 filed on May 28, 2014, the contents of each application are incorporated herein by reference in its entirety.

FIELD OF THE DISCLOSURE

This invention relates to compositions for vaccine adjuvants and uses thereof for enhancing immune response to antigens and infection.

BACKGROUND

The purpose of vaccination is to generate a strong and lasting immune response providing long-term protection against infection. However, many licensed vaccines currently induce only suboptimal immunity, requiring multiple boosts to generate a robust protective response. Adjuvantation of vaccines (i.e., addition of adjuvant) enhances both antibody and cell-mediated immune responses to antigens without the required multiple boots typical of these inoculations. An immunologic adjuvant is any substance that when incorporated into a vaccine formulation acts generally to accelerate, prolong, or enhance the quality of specific immune responses to vaccine antigens. Adjuvants can be used for various purposes: (1) to enhance the immunogenicity of highly purified or recombinant antigens; (2) to reduce the amount of antigen or the number of immunizations needed for protective immunity; (3) to improve the efficacy of vaccines in newborns, the elderly or immuno-compromised persons; or (4) as antigen delivery systems for the uptake of antigens by the mucosa.

Due to impairments in cell-mediated immunity, newborns and infants are markedly susceptible to infection with intracellular pathogens including bacteria such as Listeria spp. and viruses such as Respiratory Syncytial Virus (RSV). Prior efforts to immunize against RSV have been hampered by formalin-inactivated vaccines that induced an inadequate T-helper 1 (Th1) cell response, resulting in enhancement of the disease in newborns and infants. Indeed, inclusion of adjuvants to induce a Th1 response is associated with beneficial protection with RSV vaccines. Impaired newborn immunity, including reduced function of dendritic cells (DCs), which are key antigen-presenting cells (APCs), puts them at risk for infection and limits their Th1 responses to many vaccines. In this context, there is an unmet need for adjuvants that activate newborn DCs to facilitate the production of Th1-polarizing cytokines, resulting in T-cell activation.

Adjuvants have been used since the early 20th Century to enhance an immune response to an antigen. The concept of adjuvants arose from observations that an abscess at the inoculation site assisted the generation of higher specific antibody titers. Adjuvant activity was first demonstrated in 1926 with aluminum, when diphtheria toxoid absorbed to alum. Despite the discovery of many more potent adjuvants, such as Freund's complete adjuvant or lipolysaccharide, aluminum-based adjuvants remain the most prominent of the vaccine enhancers. Many of the newly discovered adjuvants have proven to be unsuitable for human use, as they result in local and systemic toxicity and do not meet the rigorous standards of pre-clinical or clinical trials. The need for adjuvants as a component of vaccines is still acute, especially as newer antigens may be weak immunogens or have limited availability. More recently, approvals have been obtained in Europe for MF59, a detergent-stabilized oil-in-water emulsion, as an adjuvant component of flu vaccine for elderly patients (Fluad®, Novartis Vaccines) and AS04 (combination of alum and monophosphoryl lipid A (MPLA), GlaxoSmithKline) as the adjuvant for a viral vaccines (hepatitis B, HPV).

Other categories of vaccine adjuvants in development, testing, or use include the following: Mineral salts—e.g., aluminum hydroxide (“alum”), aluminum phosphate, calcium phosphate; Oil emulsions—e.g., MF59; Particulate adjuvants—e.g., virosomes, ISCOMS (structured complex of saponins and lipids); Microbial derivatives—e.g., MPLA™ (monophosphoryl lipid A), CpG motifs, modified toxins; Plant derivatives—e.g., saponins (QS-21); Endogenous immunostimulatory adjuvants—e.g., cytokines.

SUMMARY

Embodiments of the present disclosure are combinatorial compositions that improve or stimulate the immune response. When used in conjunction with antigens as combination adjuvant compositions, the combination adjuvants to improve vaccine efficacy. The combinatorial compositions are also useful for preventing or treating infections. The inventors have discovered that dual synergy stimulation with Toll-like receptor agonists (TLRAs) and C-type lectin agonists (CLRAs) can overcome the reduced response of newborn DCs to common vaccine formulations and thereby enable vaccinations at a younger age. The inventors also found that dual synergy activation through certain TLRs and CLRs can induce enhanced activation of in adult DCs, but the specific combinations of TLR(s)/CLR(s) are different for the different age group. For newborns particularly, dual stimulation through the receptors Mincle and TLR8, or through the receptors Dectin-1 and TLR4 is optimal for the induction of Th1-polarizing cytokines, IFN-γ and IL-12p70 cytokines, and for the priming of naïve T cells to differentiate into IFN-γ-producing T cells. The DCs that make these Th1 cytokines actually induce a greater amount of Th1 differentiation from T cells, which is signified by the production of IFN-γ by these T cells. These cytokines and the induction of naïve T cells differentiation are important for adaptive immune responses and host defense against intracellular pathogens, and are usually minimally produced by neonatal cells. The synergistic effects of the TLRs with CLRs were associated with enhanced activation of the NF-kB and NLRP3 inflammasome pathways.

Accordingly, combinatorial activation of TLRAs and the endocytic receptors Mincle or Dectin-1 (CLRAs) is a powerful approach to induce Th1-mediated immunity in newborns, infants and also adults.

In addition, because both the TLR and CLR pathways are activated, these combinatorial compositions comprising TLR and CLR agonists are also ideal as “stand alone” immunomodulators to prevent or treat infections in newborns, infants and also adults. For example, in an immunocompromised individual.

Accordingly, it is the objective of this disclosure to provide combinatorial compositions for the purpose of inducing and/or enhancing the Th1-mediated immunity in a subject. The subject can be a newborn, an infant, a child younger than 12 years old, an elderly subject over 65 years old, or any subject in need of inducing and/or enhancing the Th1-mediated immunity. When used in conjunction with antigens, the combinatorial compositions are referred to as combinatorial adjuvant compositions or vaccine adjuvant compositions.

It is also the objective of this disclosure to provide combinatorial adjuvant compositions for the purpose of inducing and/or enhancing the Th1-mediated immunity in a subject to an antigen.

It is also the objective of this disclosure to provide combinatorial compositions for the purpose of priming and/or inducing the priming of naïve T cells to differentiate into IFN-γ-producing T cells.

It is also the objective of this disclosure to provide combinatorial compositions for the purpose of inducing and/or enhancing the Th1-mediated immunity in newborns. When used in conjunction with antigens, the combinatorial compositions are referred to as combinatorial adjuvant compositions or vaccine adjuvant compositions.

It is also the objective of this disclosure to provide combinatorial compositions for the purpose of inducing and/or enhancing the Th1-mediated immunity in newborns to an antigen.

It is also the objective of this disclosure to provide combinatorial TLR/CLR agonists compositions for the purpose of preventing infection in a subject, for example, in a subject who is immune compromise, or has weakened immune system or response, or has a defect in the immune system. For example, the very young or very old, one who is diabetic, one with a weakened immune system, one prone to infections, or one who has HIV.

It is also the objective of this disclosure to provide combinatorial TLR/CLR agonists compositions for the purpose of treating infection in a subject.

It is also the objective of this disclosure to provide combinatorial adjuvant compositions for the purpose of enhancing the immune response in a subject. The subject can be newborns, infants, the elderly or any subject in need of enhancing the immune system. For example, in a subject who is immune compromise, or has weakened immune system or response, or has a defect in the immune system.

It is also the objective of this disclosure to provide combinatorial adjuvant compositions for the purpose of enhancing the immune response in a subject to an antigen.

It is also the objective of this disclosure to provide a method of inducing and/or enhancing the Th1-mediated immunity in a subject, e.g., newborns.

It is also the objective of this disclosure to provide a method of inducing and/or enhancing the Th1-mediated immunity in a subject to an antigen.

It is also the objective of this disclosure to provide a method of enhancing the immune response in a subject.

It is also the objective of this disclosure to provide a method of enhancing the immune response in a subject to an antigen.

Accordingly, in one embodiment, provided herein is a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1A agonist, wherein the at least TLR4 agonist is selected from monophosphoryl lipid A (MPLA) or glycopyranosyl lipid A (GLA).

In one embodiment, provided herein is a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist.

In some embodiments, when the combinatorial compositions are used in conjunction with antigens, the combinatorial compositions are referred to as combinatorial adjuvant compositions or vaccine adjuvant compositions.

In one embodiment, provided herein is a vaccine composition comprising at least one antigen and a vaccine adjuvant composition described herein.

In one embodiment, provided herein is a use of at least one TLR4 agonist and at least one Dectin-1A agonist for the manufacture of a combination adjuvant composition wherein the at least TLR4 agonist is monophosphoryl lipid A (MPLA) or glycopyranosyl lipid A (GLA).

In one embodiment, provided herein is a use of at least one TLR7/8 agonist and at least one Mincle agonist for the manufacture of a combination adjuvant composition.

In one embodiment, provided herein is a use of at least one TLR4 agonist and at least one Dectin-1A agonist for (i) enhancing an immune response in a subject; (ii) enhancing an immune response to a commercial vaccine in a subject, (iii) for inducing and/or enhancing the Th-1 mediated immunity in a subject; (iv) for preventing an infection in a subject; or (v) for treating an infection in a subject, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a use of a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1A agonist for (i) enhancing an immune response in a subject; (ii) enhancing an immune response to a commercial vaccine in a subject, (iii) for inducing and/or enhancing the Th-1 mediated immunity in a subject; (iv) for preventing an infection in a subject; or (v) for treating an infection in a subject, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a use of at least one TLR7/8 agonist and at least one Mincle agonist for (i) enhancing an immune response in a subject; (ii) enhancing an immune response to a commercial vaccine in a subject, (iii) for inducing and/or enhancing the Th-1 mediated immunity in a subject; (iv) for preventing an infection in a subject; or (v) for treating an infection in a subject.

In one embodiment, provided herein is a use of at least one TLR4 agonist and at least one Dectin-1A agonist for the synergistic enhanced activation of the NF-kB and NLRP3 inflammasome pathways, and/or the priming or induction of naïve T cells to differentiate into IFN-γ-producing T cells.

In one embodiment, provided herein is a use of a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist for (i) enhancing an immune response in a subject; (ii) enhancing an immune response to a commercial vaccine in a subject, (iii) for inducing and/or enhancing the Th-1 mediated immunity in a subject; (iv) for preventing an infection in a subject; or (v) for treating an infection in a subject.

In one embodiment, provided herein is a use of at least one TLR7/8 agonist and at least one Mincle agonist for the synergistic enhanced activation of the NF-kB and NLRP3 inflammasome pathways, and/or the priming or induction of naïve T cells to differentiate into IFN-γ-producing T cells.

In one embodiment, provided herein is a method of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in dendritic cells (DCs), and/or priming or inducing naïve T cells to differentiate into IFN-γ-producing T cells comprising contacting dendritic cells with a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a method of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in DCs, and/or priming or inducing naïve T cells to differentiate into IFN-γ-producing T cells comprising contacting the dendritic cells with a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in a subject, the method comprising administering to the subject a vaccine adjuvant composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist, wherein the at least TLR4 agonist is selected from MPLA or GLA. In some embodiments, the subject is a newborn, an infant, an elderly subject or an immune compromised subject.

In some embodiments, inducing and/or enhancing the Th1-mediated immunity in a subject comprises priming or inducing naïve T cells to differentiate into IFN-γ-producing T cells.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in a subject to an antigen, the method comprising administering to the subject a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist with the antigen, wherein the at least TLR4 agonist is selected from MPLA or GLA. In some embodiments, the subject is a newborn, an infant, an elderly subject or an immune compromised subject.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in newborns, the method comprising administering to the newborn a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in newborns to an antigen, the method comprising administering to the newborn a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist with the antigen, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a method of enhancing an immune response in a subject comprising administering to the subject a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a method of enhancing immune response to an antigen in a subject comprising administering to the subject a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist with the antigen, wherein the at least TLR4 agonist is selected from MPLA or GLA.

In one embodiment, provided herein is a method of preventing or treating an infection in a subject comprising enhancing an immune response and/or enhancing the Th1-mediated immunity in the subject. In some embodiments, the subject is a newborn, an infant, a diabetic, an elderly subject or an immune compromised subject. For example, in a subject who is immune compromise, or has weakened immune system or response, or has a defect in the immune system. In one embodiment, the enhancing the immune response and/or enhancing the Th1-mediated immunity comprises administering a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist with the antigen, wherein the at least TLR4 agonist is selected from MPLA or GLA. In some embodiments, inducing and/or enhancing the Th1-mediated immunity in a subject comprises priming or inducing naïve T cells to differentiate into IFN-γ-producing T cells. In one embodiment, inducing and/or enhancing the Th1-mediated immunity in a subject inducing the production of Th-1 polarization cytokines such as INF-γ and IL-12p70.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in a newborn or an infant, the method comprising administering to the newborn or infant a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in a newborn or an infant to an antigen, the method comprising administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist with the antigen to the newborn or infant.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in a subject, the method comprising administering to the subject a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist. The subject can be newborns, infants, an elderly subject over 65 years old or any subject in need of enhancing the immune system, e.g., an immune compromised subject.

In one embodiment, provided herein is a method of inducing and/or enhancing the Th1-mediated immunity in a subject to an antigen, the method comprising administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist with the antigen to the subject.

In one embodiment, provided herein is a method of enhancing immune response in a subject comprising administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist to the subject. In one embodiment, the subject is a newborn or an infant.

In one embodiment, provided herein is a method of enhancing immune response to an antigen in a subject comprising administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist with the antigen to the subject. In one embodiment, the subject is a newborn or an infant.

In one embodiment, provided herein is a method of preventing or treating an infection in a subject comprising enhancing the immune response and/or enhancing the Th1-mediated immunity in the subject. In some embodiments, the subject is a newborn, an infant, an elderly subject or an immune compromised subject. In one embodiment, the enhancing the immune response and/or enhancing the Th1-mediated immunity comprises administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist to the subject.

In one embodiment, provided herein is a method of preventing an infection in a subject comprising administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist to the subject. In one embodiment, the subject is a newborn or an infant. In another embodiment, the subject is an elderly subject over 65 years old or an immune compromised subject.

In one embodiment, provided herein is a method of treating an infection in a subject comprising administering a combinatorial composition comprising at least one TLR7/8 agonist and at least one Mincle agonist to the subject. In one embodiment, the subject is a newborn or an infant. In another embodiment, the subject is an elderly subject over 65 years old or an immune compromised subject.

In one embodiment, provided herein is a method of preventing an infection in a subject comprising administering a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist to the subject, wherein the at least TLR4 agonist is selected from MPLA or GLA. In one embodiment, the subject is a newborn or an infant. In another embodiment, the subject is an elderly subject over 65 years old or an immune compromised subject.

In one embodiment, provided herein is a method of treating an infection in a subject comprising administering a combinatorial composition comprising at least one TLR4 agonist and at least one Dectin-1 agonist to the subject, wherein the at least TLR4 agonist is selected from MPLA or GLA. In one embodiment, the subject is a newborn or an infant. In another embodiment, the subject is an elderly subject over 65 years old or an immune compromised subject.

In one embodiment of the vaccine adjuvant composition, vaccine or method described, the Dectin-1 agonist is a Dectin 1A agonist or a Dectin 1B agonist.

In one embodiment of the combinatorial composition or vaccine adjuvant composition, vaccine or method described, the at least one TLR7/8 agonist is any agent that would activate the receptor TLR7 and/or TLR8. In some embodiments, the at least one TLR7/8 agonist includes but is not limited to gardiquimod, imiquimod, imidazoquinoline compound R848 (resiquimod), a benzazepine TLR8 agonist VTX-294, 3M-052, 3M compounds listed U.S. Pat. No. 7,799,800, compounds CL087, CL097, and CL075 of INVITROGEN™. VTX-294 is a benzazepine, provided by VentiRx Pharmaceuticals. 3M-052 is an imidazoquinoline compound from 3M, N-(4-{[4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl]oxy}butyl)octadecanamide. See “Vaccine adjuvant activity of 3M-052: An imidazoquinoline designed for local activity without systemic cytokine induction,” by Dmitri Smirnov et al., in Vaccine, Volume 29, Issue 33, 26 Jul. 2011, Pages 5434-5442; and in “Intratumoral immunotherapy with the TLR7/8” by Dmitri Smirnov et al., in Journal for ImmunoTherapy of Cancer 2013, 1(Suppl 1):P138.

In one embodiment of any one of the combinatorial composition or vaccine adjuvant composition, vaccine or method described, the at least one TLR7/8 agonist is selected from the group consisting of gardiquimod, imiquimod, imidazoquinoline compound R848 (resiquimod), CL087, CL097, and CL075.

In one embodiment of any one of the combinatorial composition or vaccine adjuvant composition, vaccine or method described, the at least one Mincle agonist includes but is not limited to heat-killed Mycobacterium tuberculosisis (HKMT), trehalose-6,6-dibehenate (TDB), Trehalose-6,6-dibehenate formulated with Kolliphor® HS 15 (TDB-HS15), and trehalose-6,6′-dimycolate (TDM).

In one embodiment of any one of the combinatorial composition or vaccine adjuvant composition, vaccine or method described, the at least one Mincle agonist is selected from a group consisting of HKMT, TDB, TDB-HS15, and TDM.

In one embodiment of any one of the combinatorial composition or vaccine adjuvant composition, vaccine or method described, the vaccine adjuvant composition further comprises a pharmaceutically acceptable carrier.

In one embodiment of any one of the vaccine composition described, the at least one antigen includes but is not limited to an antigen that is a live attenuated micro-organism that causes known diseases, an antigen that is an inactivated or killed micro-organism that causes known diseases, an antigen that is an inactivated toxin that is produced by a micro-organism that causes known diseases, or an antigen that is a subunit or a conjugate of a subunit of a micro-organism that causes known diseases.

In one embodiment of any one of the vaccine composition described, the combination adjuvant can be paired with a broad range of antigens, including but not limited to all the antigens that are known in the art for vaccination against known diseases, that is, all antigens that the combination adjuvant can pair with. For example, respiratory syncytial virus (RSV) F protein, RSV pre-fusion (F) protein, RSV Nucleoprotein N, HIV-GAG, HIV-gp140, or various outer membrane proteins from Gram-positive, Gram-negative or myco-bacteria.

In one embodiment of any one of the vaccine composition described, the at least one antigen includes but is not limited to an antigen against measles, mumps, rubella, chicken pox (Varicella), shingles (Zoster), Influenza (e.g. Haemophilus influenza type b), pneumonia (Pneumococcal diseases caused by Streptococcus pneumoniae), pneumococcal bacteremia, meningitis (Meningococcal diseases caused by the bacterium, Neisseria meningitidis), Rotavirus, diphtheria, tetanus, pertussis (whooping cough), polio (IPV), smallpox, HIV/AIDS, malaria, and Leishmaniasis, Hepatitis A, Hepatitis B, Hepatitis C, Anthrax, Yellow fever, rabies, Human papillomavirus (HPV), Clostridium tetani bacterium neurotoxin (tetanospasmin), tuberculosis, Dengue, typhoid, and Japanese encephalitis.

In one embodiment of any one of the vaccine composition described, the at least one antigen is selected from a group consisting of an antigen against measles, mumps, rubella, chicken pox (Varicella), shingles (Zoster), Influenza (e.g. Haemophilus influenza type b), pneumonia (Pneumococcal diseases caused by Streptococcus pneumoniae), pneumococcal bacteremia, meningitis (Meningococcal diseases caused by the bacterium, Neisseria meningitidis), Rotavirus, diphtheria, tetanus, pertussis (whooping cough), polio (IPV), smallpox, HIV/AIDS, malaria, and Leishmaniasis, Hepatitis A, Hepatitis B, Hepatitis C, Anthrax, Cholera, Yellow fever, rabies, Human papillomavirus (HPV), Clostridium tetani bacterium neurotoxin (tetanospasmin), typhoid, and Japanese encephalitis.

In one embodiment of any one of the vaccine composition described, the combinatorial composition or vaccine adjuvant composition is in an amount of about 85 to 99% of the mass of the vaccine.

In one embodiment of any one of the vaccine composition described, the at least TLR4 agonist is in the range of about 0.1 to about 5% of the mass of the fraction.

In one embodiment of any one of the vaccine composition described, the at least TLR4 agonist is in the range of about 0.1 to about 3% of the mass of the fraction.

In one embodiment of any one of the vaccine composition described, the at least Dectin-1 agonist is in the range of about 0.1 to about 5% of the mass of the fraction.

In one embodiment of any one of the vaccine composition described, the at least Dectin-1 agonist is in the range of about 0.1 to about 3% of the mass of the fraction.

In one embodiment of any one of the vaccine composition described, the vaccine composition further comprises alum-hydroxide as a co-adjuvant.

In one embodiment of any one of the vaccine composition described, the concentration of the adjuvant composition is about 150 μg-about 150 mg/single dose, wherein about 150 μg-about 150 mg is the dosage for the combined agonists in the composition.

In some embodiments of any one of the compositions described, the dose of TDB is about 10-100 microgram/ml, the dose of R848 is about 1-50 micromolar, the dose of Zymosan/Dectin is about 10-100 microgram/ml, and the dose of MPLA or GLA is about 10 nanogram/ml-1 microgram/ml.

In one embodiment of any one of the vaccine compositions or combination adjuvant composition described, the vaccine composition or combination adjuvant composition is formulated in an oil-in-water emulsion. In one embodiment of any one of the vaccine compositions or combination adjuvant composition described, the vaccine composition or combination adjuvant composition is formulated as a nanoparticle, such as a polymersome,

In one embodiment of any one of the methods described, the subject is a human adult. In another embodiment of any one of the method described, the subject is a human newborn. In another embodiment of any one of the method described, the subject is a non-human mammal. The subject can be newborns, infants, the elderly or any subject in need of enhancing the immune system. For example, in a subject who is immune compromise, or has weakened immune system or response, or has a defect in the immune system.

In one embodiment of any one of the methods described, there is an induction of Th1-polarizing cytokines in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods described, the Th1-polarizing cytokines are TNF-α, IFN-γ and the T-helper 17 (Th17)-polarizing cytokine IL-1β.

In one embodiment of any one of the methods described, there is an induction of IFN-γ cytokine in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods described, there is an induction of TNF-α cytokine in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods described, there is an induction of IL-1β cytokine in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods described, there is an induction of IL-12p70 cytokine in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods described, there is an induction of differentiation of naïve T cells to differentiate into IFN-γ-producing T cells in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods described, there is a synergistic effect associated with an enhanced activation of the NF-kB and NLRP3 inflammasome pathways in the subject upon administration of the combination composition described.

In one embodiment of any one of the methods of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in dendritic cells, the Th1-polarizing cytokines are TNF-α, IFN-γ and the T-helper 17 (Th17)-polarizing cytokine IL-1β.

In one embodiment of any one of the methods of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in dendritic cells, the dendritic cells are immature dendritic cells.

In one embodiment of any one of the methods of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in dendritic cells, the dendritic cells are adult DCs, newborn DCs, or infant DCs.

In one embodiment of any one of the methods of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in dendritic cells, the contacting is in vivo, ex vivo or in vitro.

In one embodiment of any one of the methods of inducing the production of Th1-polarizing cytokines, IFN-γ and/or IL-12p70 cytokines in dendritic cells, the contacting is in vivo in the subject.

In one embodiment of any one of the methods, the method further comprises selecting a subject for treatment of infection, for prevention of infection, for enhancing the immune system or response in the subject, for inducing Th-1 meditate immunity and/or vaccination. For example, the subject selected for any one of the procedures is a newborn, an infant, a child, an adolescent, an elderly subject or an immune compromised or immune deficient subject. It is envisioned that the combinatorial compositions described herein would enhance the immune system in the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows that neonatal MoDCs demonstrate a distinct MPLA-induced innate immune transcriptome. Messenger RNA ΔcT levels of newborn and adult MoDCs are shown. MoDCs were stimulated with MPLA (1 μg/ml) for 24 hours. Total RNA was isolated from lysates of cells and cDNA was synthesized from RNA and analyzed by quantitative RT-PCR. Delta-cT levels of adult cells are plotted against their newborn counterparts (dashed lines indicate 3-fold change). Newborn MoDCs expressed greater levels of mRNA encoding IL-6 (Th2/Th17) and IL-10 (anti-inflammatory) but less of mRNAs encoding CCR4, IRAK1 and IRF7, amongst others. IRAK1 is an important signaling component downstream of TLR activation. N=2 per group.

FIGS. 2A-2F show that TLRAs and CLRAs synergistically induce TNF production by Human newborn and adult MoDCs. Newborn and adult MoDCs were generated in the presence of 10% autologous plasma and incubated for 18 hours with combinations of indicated TLRAs and CLRAs. Supernatants were collected and analyzed by TNF-α ELISA (n=3).

FIG. 2A shows adult MoDCs were incubated with different combinations of TLRAs and CLRAs. The black lines indicate the amount of secreted TNF-α after stimulation with the TLRA alone, as indicated on the x-axis. The other shades of gray lines indicate TNF-α secretion after stimulation with the TLRA (x-axis) in combination with a CLRA (other shades of gray indicated in the legend).

FIG. 2B shows newborn MoDCs were incubated with different combinations of TLRAs and CLRAs. The black lines indicate the amount of secreted TNF-α after stimulation with the TLRA alone, as indicated on the x-axis. The other shades of gray lines indicate TNF-α secretion after stimulation with the TLRA (x-axis) in combination with a CLRA (other shades of gray indicated in the legend).

FIG. 2C shows the combination of MPLA with zymosan induces synergistic activation. Dual stimulation of MoDCs with a combination of MPLA and Zymosan induces TNF-α to adult-like levels in newborns (MPLA low, 10 ng/ml; MPLA high, 1 μg/ml; Zymosan, 100 μg/ml).

FIG. 2D shows the combination of R848 with TDB induces synergistic activation. Newborn and adult MoDCs incubated with a combination of R848 and TDB (R848 low, 1 μM; R848 high, 50 μM; TDB, 10 μg/ml) show increased production of TNF-α.

FIG. 2E shows the secretion of TNF-α in response to single TLRAs and CLRAs. Newborns are impaired in the production of TNF-α. (Students' t-test; *phttps://www.shan-machinery.com