OVERCOMING IMMUNOSENESCENCE BY NANOPARTICLE-MEDIATED ACTIVATION
N Soman
Duke Universitycity: Durham country: United States (us)
Grant 1R01AI096305-01A1 from National Institute Of Allergy And Infectious Diseases
Abstract: One new challenge for vaccine design is the development of effective vaccines for the ever growing geriatric population. Influenza is a highly contagious viral infection of the respiratory tract that afflicts 3-5 million people world-wide with 200,000-500,000 deaths each year. The elderly (>65 years) are especially prone to severe influenza infection. There is now strong evidence that the seasonal H1N1 influenza vaccine is largely ineffective for these elderly subjects, subjecting them to high risk of complication, hospitalization and death. The ineffectiveness of current flu vaccines in the elderly has been associated with immunosenescence, which is a gradual age-associated deterioration of the immune system. There is growing belief that some of the key limitations resulting in immunosenescence can be overcome by improved immunization strategies, such as the use of novel adjuvants to promote effective memory immunity. Current adjuvants typically work by activating dendritic cells (DCs) at the site of vaccine administration and enhancing the trafficking of antigen-loaded DCs to the draining lymph nodes (DLNs), the epicenter of the adaptive immune response and where antigen presentation to T cells occurs. Here, we propose an alternate strategy to accelerate and maximize the immune responses. We recently developed a particle-mediated vaccine system capable of transporting critical immunostimulatory cytokines directly to the DLN during vaccination. This adjuvant technology induced rapid and significant organizational changes to the DLNs of aged mice, animals that are not immunoresponsive, like their human counterparts. The changes in the DLNs induced by targeted delivery of these immunomodulators were consistent with robust immune responses, including vigorous and sustained recruitment of not only DCs but also T cells. The objective of this proposal is to develop an effective immune stimulatory strategy and adjuvant formulation with the potential of overcoming immunosenescence. The Specific aims are 1) Optimize the composition, size and loading of biocompatible and degradable nanoparticles with various combinations of immunodulatory mediators to achieve maximal delivery and controlled release in the DLNs; 2) Demonstrate the safety and efficacy of a vaccine formulation comprising the seasonal Flu vaccine and various cytokine-loaded nanoparticles in promoting maximal immunity in aged mice; 3) Evaluate the prophylactic and therapeutic ability of the flu vaccine in combination with lymph node activating nanoparticles in protecting aged mice against lethal influenza virus infections. If proven successful, this approach could serve as a new paradigm for improving vaccines in general. Relevance to Healthcare Many vaccines are ineffective in elderly patients (>65 years) because of immunosenescence, a gradual age-associated deterioration of the immune system. We have developed a particle- mediated adjuvant technology capable of delivering critical immunostimulatory cytokines directly to the draining lymph node, the epicenter of the adaptive immune response and where antigen presentation to T cells occurs. By accelerating and maximizing the immune responses to previously non-responsive subpopulations, this vaccine technology may significantly improve the healthcare of elderly patients
Keywords: Adjuvant; Adjuvanticity; Affect; Age; age group; aged; Animals; Antigen Presentation; Antigens; base; Belief; Biocompatible; Cells; Cessation of life; Charge; Chitosan; Complex; Complication; controlled release; cytokine; Cytoplasmic Granules; Dendritic Cells; Deterioration; Development; Drug Formulations; Elderly; Encapsulated; Face; flu; Healthcare; Heparin; high risk; Histocompatibility Antigens Class II; Hospitalization; Hour; Human; Immune; Immune response; Immune system; Immunity; Immunization; Immunologic Monitoring; Immunomodulators; immunosenescence; improved; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza virus vaccine; influenzavirus; Interleukin-12; lymph nodes; mast cell; Mediating; Mediator of activation protein; Memory; Morbidity - disease rate; Mortality Vital Statistics; Mus; nanoparticle; novel; older patient; Organizational Change; particle; Particulate; Patients; Pharmaceutical Preparations; Population; prophylactic; Prophylactic treatment; response; Safety; seasonal influenza; Severities; sex; Site; System; T-Lymphocyte; targeted delivery; Technology; Therapeutic; Time; TNF gene; Toxic effect; trafficking; Treatment Efficacy; Vaccination; Vaccine Design; Vaccines; Viral Respiratory Tract Infection; Virus Diseases; Work; young adult
Relevance: Relevance to Healthcare Many vaccines are ineffective in elderly patients (>65 years) because of immunosenescence, a gradual age-associated deterioration of the immune system. We have developed a particle- mediated adjuvant technology capable of delivering critical immunostimulatory cytokines directly to the draining lymph node, the epicenter of the adaptive immune response and where antigen presentation to T cells occurs. By accelerating and maximizing the immune responses to previously non-responsive subpopulations, this vaccine technology may significantly improve the healthcare of elderly patients
Project start date: 2012-02-01
Project end date: 2017-01-31
Budget start date: 1-FEB-2012
Budget end date: 31-JAN-2013
1R01AI096305-01A1 (2012): $392500
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to N Soman
DEVELOPMENT OF EFFICACIOUS VACCINE AGAINST UTI´S
N Soman, Professor
Duke Universitycity: Durham country: United States (us)
Grant 3R21DK077307-02S1 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: UTIs is one of the most frequent bacterial infections of man with over 85% of the outpatient community acquired UTIs caused by E.coli. Since antibiotic resistance among uropathogenic E.coli is growing, prophylactic approaches are increasingly being sought. The virulence of E.coli is attributable to expression of fimbrial adhesion FimH that mediates bacterial adherence and subsequent bacterial invasion of the bladder uroepithelium. Vaccines comprising of FimH or FimH fragments have proven highly effective in protecting mice against UTIs. However, when these FimH vaccines were administered intramuscularly with alum as adjuvant in humans, the levels of FimH specific antibodies in the serum and mucosal surfaces were low and not sustainable. Currently, a major limitation in the use of FimH vaccine in the humans is the absence of effective and safe adjuvants capable of achieving high levels of specific antibodies in serum and mucosal secretions. Recently, we discovered that mast cells (MCs) play a hitherto unrecognized role at sites of infection in orchestrating the trafficking of dendritic cells and B and T cells, Key immune cells critical for the development of adaptive immune responses. These observations led us to investigate whether co-administering antigens along with MC activators would result in elevated antigen-specific immune response. Preliminary studies revealed administering different vaccine antigens along with compound 48/80 in the nasal regions of mice evoked a highly elevated IgG response in the serum as well as IgA responses in various mucosal fluids. These studies revealed that mast cell activators have the potential to serve as potent adjuvants. Here, we hypothesized that if FimH or FimH fragments were nasally co-administered into mice with a mast cell activator, we would evoke highly protective immunity against UTI. The goals of this study are to (1) ) Evaluate the immunogenicity and protective capacity against UTIs of a FimH1-25 peptide vaccine when co administered nasally with compound 48/80. (2) Compare the efficacy and safety of compound 48/80 as a vaccine adjuvant with other known MC activators (3) Confirm that the adjuvant activity demonstrated with mast cell activators acts through specific activation of resident MCs. Vaccines comprising of Fim H or FimH fragments have proven highly effective in protecting mice against UTIs. However, when these FimH vaccines were administered intramuscularly with alum as adjuvant in humans, the levels of FimH antibodies in the serum and mucosal were low and not sustainable. Currently a major limitation in the use of Fim H vaccines in humans is the absence of effective and safe adjuvants capable of achieving high levels of specific antibodies in serum and mucosal secretion. Preliminary studies have revealed that administering different vaccine antigens along with a mast cell activator in the nasal regions of mice evoked a high level of IgG antibodies in the serum as well as IgA antibodies in various mucosal fluids. The primary goal of this study is therefore to evaluate the immunogenisity and protective capacity against UTIs of a FimH peptide vaccine when co-administered nasally with mast cell activator
Keywords: Adherence (attribute); Adhesions; Adjuvant; Adverse effects; Agent 48-80; aluminum sulfate; Antibiotic Resistance; Antibiotics; Antibodies; Antibody Formation; Antigen Targeting; Antigens; Bacterial Adhesins; Bacterial Infections; bacterial resistance; base; Binding (Molecular Function); Bladder; Cells; Cholera Toxin; community-acquired UTI; comparative efficacy; Dendritic Cells; Development; Drug Formulations; Employee Strikes; Failure (biologic function); Genitourinary system; Goals; Human; human subject; Immune; Immune response; Immunity; Immunization; immunogenic; immunogenicity; Immunoglobulin A; Immunoglobulin G; Infection; interest; Liquid substance; lymph nodes; Lymphocyte; Lymphoid Tissue; man; Mannose; mast cell; Mediating; migration; Mus; Needles; nonhuman primate; Nose; Outpatients; Patients; Peptide Vaccines; Play; Prevention strategy; prophylactic; Proteins; public health relevance; Recurrence; Regimen; Research; response; Rodent; Role; Safety; Serum; Site; small molecule; Surface; T-Lymphocyte; Toxic effect; trafficking; Urinary tract infection; Uropathogenic E. coli; Vaccine Adjuvant; Vaccine Antigen; Vaccines; Virulence; Virulence Factors
Project start date: 2009-11-19
Project end date: 2010-10-31
Budget start date: 19-NOV-2009
Budget end date: 31-OCT-2010
PFA/PA: PA-06-181
3R21DK077307-02S1 (2010): $99999
5R21DK077307-02 (2009): $195000
CAVEOLAE MEDIATED BACTERIAL UPTAKE IN THE URINARY TRACT
N Soman, Professor
Duke Universitycity: Durham country: United States (us)
Grant 5R01AI050021-10 from National Institute Of Allergy And Infectious Diseases
Abstract: Uropathogenic E.coli (UPEC) account for over 80% of urinary tract infections (UTIs). It is now known invasion of the bladder epithelial cells (BECs) is a critical initiating step in UTIs. We have demonstrated that UPEC invasion is localized to distinct cellular entities in the BEG membrane loosely- termed caveolae. These caveolae are comprised of lateral assemblies of cholesterol, lipids and select proteins such as caveolin-1. We hypothesized that the entry of UPEC into BECs is a highly dynamic event and therefore the host-cell proteins that mediate bacterial entry could be proteins that are selectively recruited to caveolae or become activated within these microdomains following exposure to UPEC. Our initial studies have revealed that two proteins, flotillin-1 and annexin-ll, were recruited to caveolae whereas the third, caveolin-1, became tyrosine-phosphorylated following exposure to UPEC. Another finding emerging from proteome analysis of BECs is that a significant proportion of the proteins identified as caveolar components are known to be regulated by cAMP, a major second messenger implicated in regulating luminal surface area of the bladder by triggering endo- and exocytosis of BEG vesicles which serve as repositories of luminal membranes. This finding, revealed possible parallels between caveolae mediated bacterial entry nto BECs and regular endocytosis of luminal membrane by BECs following voiding of urine. The goal of this research proposal is to extend and expand this line of investigation. Therefore, we propose to ) Elucidate the mechanism by which flotillin-1, annexin-ll, and caveolin-1 contribute to bacterial entry into 3ECs. (2) Use proteome mining approaches to extend the identification of caveolar components essential for UPEC invasion of BECs.(3) Investigate the regulatory role of cAMP on UPEC invasion of BECs and the mpact of modulators of intracellular cAMP in conferring protection against experimental UTIs. We believe that a systematic approach to the identification of caveolar determinants of bacterial entry will provide a comprehensive picture of the dynamic molecular interactions occurring in the invasion process and also yield candidate proteins that could potentially serve as targets for drug therapy
Keywords: Accounting; Adhesions; Annexin A2; Annexins; Area; Bacteria; Binding (Molecular Function); Bladder; Caveolae; caveolin 1; Cell membrane; Cells; Cholesterol; Cyclic AMP; Development; Endocytosis; Epithelial Cells; Event; Exocytosis; Exposure to; flotillin; Goals; Hair; Invaded; Investigation; Lateral; Lipids; luminal membrane; Maps; Mediating; Mediator of activation protein; Membrane; Membrane Microdomains; Mining; Modeling; Mus; novel strategies; Organ; Pharmacotherapy; Prevention; Process; Proteins; Proteome; Proteomics; Recruitment Activity; Recurrence; repository; Research Proposals; Role; second messenger; Second Messenger Systems; Structure; Surface; Technology; type 1 fimbriae; Tyrosine; uptake; Urinary tract; Urinary tract infection; Urine; Uropathogenic E. coli; Vesicle; Virulence Factors
Project start date: 2001-07-01
Project end date: 2011-11-30
Budget start date: 1-JUN-2010
Budget end date: 30-NOV-2011
5R01AI050021-10 (2010): $367781
MODULATION OF BLADDER CAMP TO COMBAT UTI´S
N Soman, Professor
Duke Universitycity: Durham country: United States (us)
Grant 5R01DK077159-05 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: Urinary tract infections (UTIs) represent one of the most common bacterial infections of humans. Recurrence and persistence of UTIs, in spite of appropriate antibiotic therapry, has been attributed, at least in part, to the pathogen seeking intracellular refuge within bladder cells. Type 1 fimbriated uropathogenic E.coli (UPEC) accounts for over 80% of all UTIs. The virulence of the bacteria is associated with its capacity to invade the superficial bladder epithelium and avoid elimination in the urine. We have recently discovered that during infection in an experimental model of UTI, UPEC were entering and harboring within fusiform vesicles of superficial epithelial cells of the bladder. Remarkably, artificially boosting intracellular levels of cAMP in BECs induced bacterial expulsion from infected BECs, and also impeded bacterial invasion of BECs. Conceivably, use of artificial enhancers of cAMP to reduce entry and to expel bacteria from intracellular niches could be of therapeutic value. In pilot experiments, we have successfully eliminated bacteriurea in UPEC infected mice following intraperitoneal instillation of forskolin, a cAMP booster. Forskolin (a derivative of the plant herb Coleus forskohlii) is unlikely to be toxic to humans considering that Coleus forskohlii extracts have been employed for medicinal purposes for centuries in Asia. In the US, use of forskolin for a variety of ailments is rapidly increasing as interest in alternative medicine has grown. The specific aims are to (i) Employ an ex vivo model of bladder infection, to demonstrate (a), the dependence of UPEC on intracellular cAMP levels during invasion of BECs and (b), the ability to manipulate bacterial burden in infected BECs with modulators of cAMP. (ii) Elucidate the molecular events leading to how intracellular cAMP negatively modulates UPEC invasion into BECs and how artificially increasing cAMP levels triggers bacterial expulsion.(iii) Examine the efficacy of forskolin treatment as an alternative or complement to antibiotic therapy for UTIs. The proposed studies should provide valuable information on the role of cAMP as a potent modulator of bacterial invasion and expulsion from BECs
Keywords: Accounting; Alternative Medicine; Antibiotic Therapy; Antibiotics; Asia; Bacteria; Bacterial Infections; Bladder; Cells; Coleus forskohli; combat; Complement; Cyclic AMP; Dependence; Enhancers; Epithelial Cells; Epithelium; Event; Experimental Models; Forskolin; Herb; Human; Infection; Infective cystitis; interest; intraperitoneal; Invaded; Modeling; Molecular; Mus; novel; pathogen; Plants; Recurrence; research study; Role; Therapeutic; Urinary tract infection; Urine; Uropathogenic E. coli; Vesicle; Virulence
Project start date: 2007-07-15
Project end date: 2012-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
5R01DK077159-05 (2011): $302293
5R01DK077159-04 (2010): $305346
3R01DK077159-03S1 (2009): $103522
SMALL CATIONIC ANTIMICROBIAL PEPTIDES: ACTIVATORS OF INNATE & ADAPTIVE IMMUNITY
N Soman, Professor
Duke Universitycity: Durham country: United States (us)
Grant 1R56AI074751-01A1 from National Institute Of Allergy And Infectious Diseases
Abstract: Cationic antimicrobial peptides, which are 100 amino acids or less in chain size, are produced in all living species. In mammals, they are primarily associated with mucosal epithelial cells and phagocytic cells. These peptides are produced in large amounts at sites of infection and have a broad spectrum of activity not only against gram negative and positive bacteria but also against fungi, viruses and parasites. Seven years ago, it was reported that co-administration of a cationic peptide (human defensin HNP1-3) with ovalbumin (ova), a nonimmunogenic protein, resulted in enhanced production of ova specific IgG antibodies suggesting that these peptides possess intrinsic adjuvant properties. We have extended these studies employing much smaller peptides (~15 amino acids in length) and shown that co-instilling protective antigen (PA) of anthrax with small antimicrobial peptides into the nasal cavities of mice resulted in markedly elevated levels of PA-specific protective antibodies in the serum. Interestingly, the cationic peptides induced immune responses that were markedly superior to those induced by the powerful mucosal adjuvant cholera toxin that cannot be developed for use in humans because of its toxicity. No systemic or local pathologic effects were observed in mice immunized with small peptides. Antibodies specific for the cationic peptides, that may block its adjuvant activity if used repeatedly, were not detected. The adjuvanticity of these cationic peptides was associated with massive migration of dendritic cells from sites of instillation to the draining lymph nodes (DLNs) and large scale sequestration in the DLNs of lymphocytes from the circulation. Our findings support a growing number of studies suggesting that in addition to their antimicrobial functions, these cationic peptides may have the capacity to act on cells of the innate and adaptive immune system and regulate their activity. The specific aims of this proposal are to (i) demonstrate the efficacy and safety of small cationic peptides when used as nasal vaccine adjuvants (ii) elucidate the underlying immunological basis for adjuvant activity of cationic peptides
Keywords: Adjuvant; Adjuvanticity; Amino Acids; anthrax protective factor; Antibodies; Antigens; antimicrobial; Antimicrobial Cationic Peptides; antimicrobial drug; antimicrobial peptide; Bacteria; base; Bioterrorism; Blood Circulation; Cells; Cholera Toxin; Class; Defensins; Dendritic Cells; Development; Epithelial Cells; fungus; Human; Immune; Immune response; Immune system; Immunity; Immunoglobulin G; Infection; Infectious Agent; interest; Length; Life; Ligands; lymph nodes; Lymphocyte; Mammals; Mediator of activation protein; migration; Mus; Nasal cavity; Nose; Numbers; Ovalbumin; Parasites; Pathologic; Peptides; Phagocytes; Production; Property; Proteins; Reporting; Safety; Serum; Site; size; small molecule; synthetic peptide; Toll-like receptors; Toxic effect; Toxin; trafficking; Vaccine Adjuvant; Vaccines; Virus
Project start date: 2008-09-25
Project end date: 2010-08-31
Budget start date: 25-SEP-2008
Budget end date: 31-AUG-2010
PFA/PA: PA-07-070
1R56AI074751-01A1 (2008): $390000