ALTRUINS, NATURAL ANTIBIOTICS OF THE MALE GENITAL TRACT
Robert I Lehrer, Professor
Medicineuniversity Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 5R01AI046015-03 from National Institute Of Allergy And Infectious Diseases IRG: REB
Abstract: Long term objectives. To discover the endogenous antimicrobial effector molecules of humans, and to define their structures and mechanisms of action. This proposal deals with antimicrobial peptides of the male genital tract, an area where remarkably little information currently exists. Hypotheses. Normal human semen contains proteins with latent antimicrobial domains that are activated by serine and aspartyl proteases found in normal semen. The resulting peptides ("altruins") can protect the female from infection by bacteria introduced into the vagina during intercourse. Specific aims. 1). To identify and characterize antimicrobial peptides and their precursors in human semen. 2). To ascertain how the precursors of these peptides are processed and identify the responsible proteases. 3). To test these peptides against C. trachomatis, E. coli, STD pathogens, and normal vaginal flora. 4). To synthesize selected seminal peptides, and elucidate their mechanisms of action. Methods. We will apply various purification procedures, including gel permeation chromatography, preparative electrophoresis and RP-HPLC to purify the antimicrobial components of normal and pH conditioned semen. These fractions will be tested for antimicrobial activity, and the active effector molecules they contain will be purified to homogeneity, sequenced, synthesized, and studied. Health relatedness. Many of the most common infectious diseases in the U.S. are sexually transmitted. According to CDC estimates, STDs other than AIDS add 10 billion dollars to the nation´s annual health care I costs. The proposed studies could lead to novel approaches for STD prophylaxis
Keywords: antibiotic, male reproductive system, semen Chlamydia trachomatis, Escherichia coli, aspartic endopeptidase, protein biosynthesis, protein sequence, serine proteinase, sexually transmitted disease, vagina clinical research, high performance liquid chromatography, human subject, laboratory rabbit, monoclonal antibody, protein purification
Project start date: 1999-03-15
Project end date: 2002-02-28
5R01AI046015-03 (2001): $287609
Sponsored Links Excellgen http://Excellgen.com
ALTRUINS, NATURAL ANTIBIOTICS OF THE MALE GENITAL TRACT
Robert I Lehrer, Professor
Medicineuniversity Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 5R01AI046015-02 from National Institute Of Allergy And Infectious Diseases IRG: REB
Abstract: Long term objectives. To discover the endogenous antimicrobial effector molecules of humans, and to define their structures and mechanisms of action. This proposal deals with antimicrobial peptides of the male genital tract, an area where remarkably little information currently exists. Hypotheses. Normal human semen contains proteins with latent antimicrobial domains that are activated by serine and aspartyl proteases found in normal semen. The resulting peptides ("altruins") can protect the female from infection by bacteria introduced into the vagina during intercourse. Specific aims. 1). To identify and characterize antimicrobial peptides and their precursors in human semen. 2). To ascertain how the precursors of these peptides are processed and identify the responsible proteases. 3). To test these peptides against C. trachomatis, E. coli, STD pathogens, and normal vaginal flora. 4). To synthesize selected seminal peptides, and elucidate their mechanisms of action. Methods. We will apply various purification procedures, including gel permeation chromatography, preparative electrophoresis and RP-HPLC to purify the antimicrobial components of normal and pH conditioned semen. These fractions will be tested for antimicrobial activity, and the active effector molecules they contain will be purified to homogeneity, sequenced, synthesized, and studied. Health relatedness. Many of the most common infectious diseases in the U.S. are sexually transmitted. According to CDC estimates, STDs other than AIDS add 10 billion dollars to the nation´s annual health care I costs. The proposed studies could lead to novel approaches for STD prophylaxis
Keywords: antibiotic, male reproductive system, semen Chlamydia trachomatis, Escherichia coli, aspartic endopeptidase, protein biosynthesis, protein sequence, serine proteinase, sexually transmitted disease, vagina clinical research, high performance liquid chromatography, human subject, laboratory rabbit, monoclonal antibody, protein purification
Project start date: 1999-03-15
Project end date: 2002-02-28
5R01AI046015-02 (2000): $279230
Grants awarded to Robert I Lehrer
Theta-defensins Novel HIV-1 Uptake Inhibitors
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R01AI056921-03 from National Institute Of Allergy And Infectious Diseases IRG: AMCB
Abstract: We propose to study the immune mechanism of defensins in host resistance to human HIV-1 and related retroviruses of non-human primates. Our long-term objectives are 1) to identify innate immune system molecules involved in the pathogenesis of HTV-1 infections in primates. 2) to characterize the carbohydrate-binding specificity of theta- and alpha-defensins, and to correlate this property with their activity against HIV-1. 3) to identify potent theta-defensins for future in vivo studies designed to assess their therapeutic usefulness. No in vivo studies are proposed hi this grant. Theta-defensins are cyclic octadecapeptides expressed by leukocytes of Old World monkeys and some other non human primates. Their ability to recognize certain carbohydrate structures (glycans) allows some 9-defensins to bind both gp120 of HIV-1 and CD4 with high affinity, and to prevent the entry of R5 and X4 strains of HIV-1 into otherwise susceptible human cells. At least 6 0-defensin genes exist in the human genome, and theta-defensin mRNA exists in human bone marrow, thymus, and spleen. However, human theta-defensin genes and their mRNA contain a premature stop codon, and human cells do not produce theta-defensin peptides. Retrocyclins 1-3 are synthetic peptides that represent the theta-defensins that humans could produce if the silencing mutation (the premature stop codon) were absent. Retrocyclin-2 is effective against HIV-1 primary isolates representing subtypes A,B,C and CRF01_AE, which cause most HIV-1 infections worldwide. Because retrocyclins are noncytotoxic and noninjurious to vaginal lactobacilli, they provide intriguing lead compounds for future pharmaceutical development. The studies contained in this proposal are intended to provide a knowledge-platform that will facilitate the development of therapeutic theta-defensins in the future. The specific aims are 1). To synthesize novel theta-defensins, including peptides based on theta-defensin gene sequences in non-human primates and analogues of human retrocyclins, and to test them for activity against HIV-1 and cytotoxicity. 2). To identify the sugars and oligosaccharides that primate theta-defensins bind, and correlate binding with their antiretroviral properties. Aim 1 will be accomplished by solid phase peptide synthesis and in vitro protection assays using peripheral blood mononuclear cells and indicator cell lines. Specific Aim 2 will be realized by a combination of microchemical techniques, including surface plasmon resonance, tandem mass spectrometry, and state-of-the-art lectinomic and peptidomic chemistry.
Keywords: antiAIDS agent, defensin, drug screening /evaluation, human immunodeficiency virus 1, aminoglycoside antibiotic, human immunodeficiency virus 2, lymphocyte, oligosaccharide, simian immunodeficiency virus, Cercocebus, Macaca mulatta, Macaca nemestrina, capillary electrophoresis, cell line, high performance liquid chromatography, human subject, matrix assisted laser desorption ionization, peptide chemical synthesis
Project start date: 2005-05-01
Project end date: 2008-04-30
5R01AI056921-03 (2007): $256365
5R01AI056921-02 (2006): $264022
1R01AI056921-01A2 (2005): $270083
STRUCTURE/FUNCTIONS OF HUMAN TEAR LIPOPHILIN
Robert I Lehrer, Professor
Medicineuniversity Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 5R01EY012080-03 from National Eye Institute IRG: VISA
Abstract: We discovered a new 16.5 kDa molecular (lipophilin) that is a prominent component of normal human tears. Lipophilin molecules are heterodimers whole subunits are linked covalently by 3 intramolecular cystine disulfide bonds. Our preliminary sequences and structure data suggest that lipophilin is homologous to prostatein, a steroid-binding protein secrete by the rate prostate gland. The larger subunit of lipophilin also shows strong homology to certain other steroid-binding and PLA2-inhibitory proteins (mammaglobin, uteroglobin and human CC-10) that are secreted by mammary, uterine, and lung epithelial cells. We have assembled an experienced, multi-disciplinary group of co-investigators to determine the structural and functional properties of lipophilin. Both a consideration of its homologues and the considerable concentration of lipophilin in tears suggest that I may play a crucial role with respect to the nutritional lubricating and anti-inflammatory needs of the cornea and conjunctiva. The Specific Aims are 1) To determine the complete amino acid sequences of both lipophilin components; define how their cysteines pair, and if the heterodimers associate non0covalently into tetramers. 2) To clone each lipophilin component from a lacrimal gland cDNA library; determine if any non-ocular tissues express lipophilin-related peptides; and identify the chromosomal locations of the genes for both lipophilin components. 3) To identify sites of lipophilin storage in the lacrimal gland and other tissues by immunohistochemistry. 4) To establish an ELISA assay for lipophilin and measure its concentration in the tears of normal men and women. 5) To examine the ability of human tear lipophilin to bind biologically significant lipids, including retinoids, selected steroid hormones and phospholipids; to identify endogenous lipids and to examine its ability to inhibit the Type II secretory phospholipase A2 found in normal human tears. The long-term objectives for this research are to learn how lipophilin contributes to corneal and conjuctival health, and to explore its potential for creating new diagnostic and therapeutic modalities
Keywords: protein, protein structure /function, tear lacrimal apparatus, lipid, molecular film, phospholipase A2, phospholipid, protein binding, protein protein interaction, protein sequence, steroid hormone analytical ultracentrifugation, enzyme linked immunosorbent assay, human tissue, immunocytochemistry, molecular cloning, polymerase chain reaction, western blotting
Project start date: 1998-12-01
Project end date: 2002-11-30
5R01EY012080-03 (2001): $206792
5R01EY012080-02 (2000): $200770
ANTIBIOTIC PROTEINS OF THE BLADDER
Robert I Lehrer, Professor
Medicineuniversity Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 5R01AI032930-03 from National Institute Of Allergy And Infectious Diseases IRG: BM
Abstract: Urinary tract infections are important causes of morbidity and mortality. During their lifetime, at least 12% of men and 10-20% of women experience a symptomatic acute urinary tract infection. Each year, over 100,000 patients are hospitalized with renal infections in the United States. The most common cause of these urinary tract infections is E. coli. The initial stage in the pathogenesis of E. coli urinary tract infections involves its adherence to the uroepithelial lining. Once adherent, the organisms encounter local bactericidal mechanisms that can defend the bladder mucosa. Only organisms that survive this host response can go on to produce urinary tract infections. Although much has been learned about the virulence factors of potentially uropathogenic E. coli, virtually nothing is known about the inherent antimicrobial mechanisms of the mammalian bladder. Our long term research objectives are to identify the molecules that enable the mammalian bladder to kill adherent E. coli and to ascertain whether deficiencies in such molecules contribute to the pathogenesis of urinary tract infections in predisposed or vulnerable individuals, such as women and the elderly. Because this general area has not been well explored to date, we have designed a sharply focussed effort to identify and localize antibacterial molecules in the rat bladder. Our preliminary studies have shown that we can recover potently bactericidal molecules from the rat bladder by rinsing it briefly with 1% acetic acid. The microbicidal molecules have been partially purified, and shown to be distinct from defensins, lysozyme and nuclear histones. The Specific Aims of this proposal are 1. To identify and characterize proteins associated with the rat bladder wall mucosa that are bactericidal for E. coli ML-35p. 2. To analyze how various components of urine may influence the bactericidal properties of these endogenous bladder proteins. 3. To test the activity of these antimicrobial bladder proteins against a) E. coli strains freshly recovered from patients with urinary tract infections, b) E. coli strains that differ in attributes potentially associated with uropathogenicity, and against c) other common uropathogens, including, P. mirabilis, P. aeruginosa, S. epidermidis, enterococci, etc. 4. To immunolocalize the microbicidal proteins in the rat bladder. 5. To clone the microbicidal proteins from a rat bladder cDNA library, determine their complete primary amino acid sequences and ascertain their cellular sites of synthesis
Keywords: Escherichia coli, bactericidal immunity, urinary bladder, urinary tract infection Proteus, Streptococcus enterococcus group, protein biosynthesis, urine electron microscopy, in situ hybridization, laboratory rat, molecular cloning, protein purification, protein sequence
Project start date: 1992-08-01
Project end date: 1995-07-31
5R01AI032930-03 (1994): $157047
5R01AI032930-02 (1993): $157049
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 1S10RR019042-01 from National Center For Research Resources IRG: ZRG1
Abstract: This Shared Instrumentation Grant requests a Biacore3000 instrument to perform real-time surface plasmon resonance (SPR) analyses of receptor ligand binding interactions. The core user group consists of five full-time faculty members from the Departments of Medicine (Lehrer, Ganz, Cole), Pathology (Baum, Ganz), Microbiology, Immunology and Molecular Genetics (Lee), and Molecular Biology Institute (Baum, Lehrer). Biamolecular binding interactions are central to the research efforts of all of these investigators, and all will benefit from the exceptional sensitivity and high throughput capacity of the requested instrument. Because three of the users (Baum, Lee and Lehrer) study the interactions of peptides and proteins with carbohydrates and sugars, only the Biacore3000 instrument combines a) the sensitivity necessary to detect oligosaccharide binding, with b) high throughput capacity, and c) the ability to recover analyte samples with minimal dilution. The investigators are committed to sharing the instrument and will be proactive in making it available for other users. This will include providing access to the services of two highly skilled operators for occasional or new users of the instrument. The benefits and health relatedness of the proposal are that it will facilitate accomplishing the research goals of the ten NIH grants or proposals listed in Table 1 within. These include the development of new agents and approaches to prevent and treat infections caused by HIV-1 and other viral pathogens, and fundamental issues related to cell targeting and survival.
Keywords: biomedical equipment purchase, surface plasmon resonance, HIV infection, ligand, microorganism disease chemotherapy, receptor binding, virus disease
Project start date: 2004-04-01
Project end date: 2005-03-31
1S10RR019042-01 (2004): $297688
ANTIMICROBIAL PEPTIDES OF LEUKOCYTES
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R03TW000355-06 from Fogarty International Center IRG: ICP
Abstract: This is a competing renewal application to continue studies to identify endogenous antimicrobial peptides from chicken and porcine leukocytes. Three specific aims are described. 1) Preparative and analytical protein chemistry methods will be used to purify the non-gallinacin antimicrobial peptides of chicken leukocytes and to ascertain the molecular characteristics of these peptides including primary protein sequence and unique structural characteristics (e.g., circular dichroism, disulfide pairing, etc.). Several highly sensitive assays will be applied to measure antibiotic activity against bacterial and fungal pathogens. In particular, extracts of chicken peritoneal exudate cells will be separated by chromatography. Fractions exhibiting antimicrobial activity will be further purified using preparative continuous acid urea gel electrophoresis and/or reversed-phase HPLC. Peptides of interest will be microsequenced. In some instances it may be necessary to derive the entire sequence from peptides produced from the holopeptide by limited proteolysis. 2) Another aspect of this collaboration will be to analyze the structure of prophenin-l, an unusually proline-rich peptide, to examine its post-translational processing and determine the contributions of its reiterated decamer motif to the peptide s tight binding of LPS and to its selective antimicrobial activity. To accomplish this specific aim, well-characterized synthetic peptide fragments will be tested for their ability to protect against LPS-mediated endotoxin shock in mice. 3) Finally, molecular biologic techniques will be used to clone and sequence the genes of avian antimicrobial peptides. Many of the prophenin-related peptides that have been discovered in pigs and cows exhibit highly conserved sequences within the propeptide region. Thus, screening for genes that exhibit homology to the propeptide region may allow the discovery of similar antimicrobial peptides in chickens. Total RNA from chicken bone marrow cells will be used to construct a cDNA library using standard techniques. The library will be screened with synthetic oligonucleotide "guessmers" based on reverse translation of the least degenerate part of the N-terminal sequence of each precursor protein. Hybridizing cDNA clones will then be subcloned, sequenced, and used to screen a commercially obtained chicken genomic library. The cloning of peptide cDNAs and genes will also permit future studies of the regulation of antimicrobial peptide synthesis by microbial substances and cytokines.
Keywords: antibiotic, leukocyte, peptide, protein structure /function, analytical chemistry, neutrophil, peptide structure, posttranslational modification, chicken, gel electrophoresis, genetic library, high performance liquid chromatography, laboratory mouse, molecular cloning, protein purification, protein sequence, swine
Project start date: 1993-09-20
Project end date: 2000-01-31
5R03TW000355-06 (1999): $23991
5R03TW000355-05 (1998): $24191
Sponsored Links Excellgen http://Excellgen.com
TOPICAL PROTEGRINS TO PREVENT SEXUALLY TRANSMITTED DISEASES AND HIV INFECTION
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P01AI037945-040001 from National Institute Of Allergy And Infectious Diseases
Abstract: The long term objective of this Project is to develop protegrins as safe and effective chemoprotective barrier to protect women from the acquisition of HIV, gonorrhea, chlamydia, syphilis, herpes or trichomonas infection during sexual intercourse. Protegrins are recently discovered natural peptide antibiotics that were originally isolated from porcine leukocytes. They contain 16-18 amino acids, including 4 cysteine residues that form 2 intramolecular disulfide bonds. Unlike defensins, to which they show primary sequence homology, and many other antibiotic peptides of animal origin, the antimicrobial activity of protegrins is enhanced by the presence of physiological salt concentrations and serum. The relatively simple structure of protegrins makes them highly amenable to solid phase chemical synthesis, and synthetic and native protegrins show identical antimicrobial activity in vitro. Initial tests of these congeners revealed that they also possessed strong inactivating activity. Despite their unusually broad antimicrobial spectrum, protegrins were not cytotoxic towards mammalian fibroblasts, lymphocytes or macrophages, even when tested at concentrations of 50 mug/ml. The Specific Aims are 1) To design, synthesize and test protegrrins and protegrin congeners that can inactivate HIV and other STD agents of micromolar and sub-micromolar concentrations. 2) To ascertain how specific structural features of protegrins and their congeners contribute to their ability against these STD pathogens. 3) To determine how the structural features affect the interaction of protegrins with host factors, including their binding to serum proteins, the potentiation of their antimicrobial efficacy by factors present in normal serum, their resistance to host proteases, and their cytotoxicity (or lack thereof) and their interactions with normal vaginal flora, as exemplified by lactobacilli. 4) To determine the 3-dimensional structures of protegrins by 2-D NMR and X-ray crystallography.
Keywords: AIDS education /prevention, drug design /synthesis /production, drug screening /evaluation, local antiinfective agent, peptide analog, sexually transmitted disease, HIV infection, antibacterial agent, antiviral agent, peptide chemical synthesis, synthetic peptide, X ray crystallography, nuclear magnetic resonance spectroscopy
TOPICAL PROTEGRINS TO PREVENT STDS AND HIV INFECTION
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P01AI037945-04 from National Institute Of Allergy And Infectious Diseases IRG: SRC
Abstract: The long term objective of this Program Project is to develop protegrins as a safe and effective chemoprotective barrier to protect women from the acquisition of HIV, gonorrhea, chlamydia, syphilis, herpes or trichomonas infection during sexual intercourse. protegrins are recently discovered natural peptide antibiotics that were originally isolated from porcine leukocytes. The contain 16-18 amino acids, including 4 cysteine residues that form 2 intramolecular disulfide bonds. Unlike defensins, to which they show primary sequence homology, and many other antibiotic peptides of animal origin, the antimicrobial activity of protegrins is enhanced by the presence of physiological salt concentrations and serum. The relatively simple structure of protegrins makes them highly amenable to solid phase chemical synthesis, and synthetic and native protegrins show identical antimicrobial activity in vitro. Protegrins rapidly inactivated HIV, C. trachomatis, N. gonorrhoeae and T. pallidum in vitro when added in micromolar concentrations, typically 5 mug/ml or below. Somewhat higher concentrations of protegrins also inactivated herpes simplex type II and Trichomonas vaginalis. Despite their unusually broad antimicrobial spectrum, protegrins were not cytotoxic towards mammalian fibroblasts, lymphocytes or macrophages, even when tested at concentrations of 50 mug/ml. The specific aims of the overall program project include designing protegrin congeners with enhanced efficacy against STD organisms (which will also allow us to examine the structure-function relationships of protegrins), testing the protegrin congeners against laboratory and clinical isolates of these organisms, and ascertaining the ability of topical protegrins to protect the lower genital tracts of female mice from infection by C. trachomatis. This Program Project Grant combines the talents and experiences of a consortium of leading STD scientists in an integrated and interactive program to develop a novel, female-controlled modality to prevent STD.
Keywords: AIDS education /prevention, communicable disease control, local antiinfective agent, peptide analog, sexually transmitted disease, topical drug application
Project start date: 1995-03-01
Project end date: 1999-09-29
5P01AI037945-04 (1998): $862074
5P01AI037945-03 (1997): $759060
DEFENSINS--ANTIMICROBIAL PEPTIDES OF HUMAN NEUTROPHILS
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R37AI022839-15 from National Institute Of Allergy And Infectious Diseases IRG: NSS
Abstract: The ability of neutrophils kill ingested microorganisms is essential for host defense against microbial infection. Our long term goal is to identify the endogenous antibiotic peptides of human neutrophils. The present application has six specific aims. l) To purify "granulins" from human neutrophils, test their antimicrobial, growth-regulatory and cytotoxic properties, and determine their subcellular localization and their mode of release. 2) To determine if human neutrophils contain antimicrobial members of the "four-disulphide-core family" of basic proteins. 3) To establish the prevalence of neutrophil defensin deficiency in inbred strains of mice and to purify and identify the non- defensin antimicrobial proteins of murine neutrophils. 4) To examine selected properties of human defensin HNP-4 relevant to its antimicrobial, cytotoxic and corticostatic functions. 5) To express human defensins in otherwise defensin-deficient murine phagocytes and to determine the consequences for antimicrobial activity against H. capsulatum and C. albicans. 6) To characterize the binding of human defensins to bacteria and human mononuclear cells, and to study the uptake of exogenous defensins by human monocytes and macrophages and determine its consequences for antimicrobial activity against H. capsulatum and C. albicans. The methods used in Specific Aims 1-4 involve cellular and subcellular fractionations, analytical and preparative protein chemistry and various microbiological assays. Specific Aim 5 combines molecular biology and microbiology and will use recently constructed murine cell lines that express and process the human defensin HNP-1. Specific Aim 6 will combine immunoelectron microscopy with measurements of binding and uptake of defensins and quantitative measurements of macrophage-mediated antimicrobial activity. Overall, these studies will define the role of defensins and other cysteine-rich antimicrobial peptides (granulins and four-disulphide-core congeners) in the antimicrobial activity of human neutrophils and assess the potential of defensins to control infections caused by organisms that replicate within macrophages. The work with murine neutrophils, whose content of endogenous antimicrobial polypeptides differs greatly from those of humans, will provide essential information relevant to the interpretation of murine models of human infection.
Keywords: bactericidal immunity, defensin, host organism interaction, microorganism immunology, neutrophil, Candida albicans, Escherichia coli, Histoplasma capsulatum, Listeria, Salmonella typhimurium, adrenocorticotropic hormone, granule, hormone receptor, macrophage, membrane permeability, monocyte, phagocytosis, cell free system, human tissue, immunoelectron microscopy, laboratory mouse, protein purification, tissue /cell culture, transfection
Project start date: 1985-12-01
Project end date: 2002-11-30
5R37AI022839-15 (2000): $362231
5R37AI022839-14 (1999): $377439
4R37AI022839-13 (1998): $339205
5R37AI022839-12 (1997): $289990
5R37AI022839-10 (1995): $268108
5R01AI022839-09 (1994): $257797
Sponsored Links Excellgen http://Excellgen.com
DEFENSINS: ANTIMICROBIAL PEPTIDES OF HUMAN NEUTROPHILS
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2R01AI022839-08 from National Institute Of Allergy And Infectious Diseases IRG: BM
Abstract: The ability of neutrophils kill ingested microorganisms is essential for host defense against microbial infection. Our long term goal is to identify the endogenous antibiotic peptides of human neutrophils. The present application has six specific aims. l) To purify "granulins" from human neutrophils, test their antimicrobial, growth-regulatory and cytotoxic properties, and determine their subcellular localization and their mode of release. 2) To determine if human neutrophils contain antimicrobial members of the "four-disulphide-core family" of basic proteins. 3) To establish the prevalence of neutrophil defensin deficiency in inbred strains of mice and to purify and identify the non- defensin antimicrobial proteins of murine neutrophils. 4) To examine selected properties of human defensin HNP-4 relevant to its antimicrobial, cytotoxic and corticostatic functions. 5) To express human defensins in otherwise defensin-deficient murine phagocytes and to determine the consequences for antimicrobial activity against H. capsulatum and C. albicans. 6) To characterize the binding of human defensins to bacteria and human mononuclear cells, and to study the uptake of exogenous defensins by human monocytes and macrophages and determine its consequences for antimicrobial activity against H. capsulatum and C. albicans. The methods used in Specific Aims 1-4 involve cellular and subcellular fractionations, analytical and preparative protein chemistry and various microbiological assays. Specific Aim 5 combines molecular biology and microbiology and will use recently constructed murine cell lines that express and process the human defensin HNP-1. Specific Aim 6 will combine immunoelectron microscopy with measurements of binding and uptake of defensins and quantitative measurements of macrophage-mediated antimicrobial activity. Overall, these studies will define the role of defensins and other cysteine-rich antimicrobial peptides (granulins and four-disulphide-core congeners) in the antimicrobial activity of human neutrophils and assess the potential of defensins to control infections caused by organisms that replicate within macrophages. The work with murine neutrophils, whose content of endogenous antimicrobial polypeptides differs greatly from those of humans, will provide essential information relevant to the interpretation of murine models of human infection.
Keywords: bactericidal immunity, host organism interaction, microorganism immunology, neutrophil, peptide, Candida albicans, Escherichia coli, Histoplasma capsulatum, Listeria, Salmonella typhimurium, adrenocorticotropic hormone, granule, hormone receptor, macrophage, membrane permeability, monocyte, phagocytosis, cell free system, human tissue, immunoelectron microscopy, laboratory mouse, protein purification, tissue /cell culture, transfection
Project start date: 1985-12-01
Project end date: 1997-11-30
2R01AI022839-08 (1993): $256071
DEFENSINS--ANTIMICROBIAL PEPTIDES OF HUMAN NEUTROPHILS
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R37AI022839-17 from National Institute Of Allergy And Infectious Diseases IRG: NSS
Abstract: The ability of neutrophils kill ingested microorganisms is essential for host defense against microbial infection. Our long term goal is to identify the endogenous antibiotic peptides of human neutrophils. The present application has six specific aims. l) To purify "granulins" from human neutrophils, test their antimicrobial, growth-regulatory and cytotoxic properties, and determine their subcellular localization and their mode of release. 2) To determine if human neutrophils contain antimicrobial members of the "four-disulphide-core family" of basic proteins. 3) To establish the prevalence of neutrophil defensin deficiency in inbred strains of mice and to purify and identify the non- defensin antimicrobial proteins of murine neutrophils. 4) To examine selected properties of human defensin HNP-4 relevant to its antimicrobial, cytotoxic and corticostatic functions. 5) To express human defensins in otherwise defensin-deficient murine phagocytes and to determine the consequences for antimicrobial activity against H. capsulatum and C. albicans. 6) To characterize the binding of human defensins to bacteria and human mononuclear cells, and to study the uptake of exogenous defensins by human monocytes and macrophages and determine its consequences for antimicrobial activity against H. capsulatum and C. albicans. The methods used in Specific Aims 1-4 involve cellular and subcellular fractionations, analytical and preparative protein chemistry and various microbiological assays. Specific Aim 5 combines molecular biology and microbiology and will use recently constructed murine cell lines that express and process the human defensin HNP-1. Specific Aim 6 will combine immunoelectron microscopy with measurements of binding and uptake of defensins and quantitative measurements of macrophage-mediated antimicrobial activity. Overall, these studies will define the role of defensins and other cysteine-rich antimicrobial peptides (granulins and four-disulphide-core congeners) in the antimicrobial activity of human neutrophils and assess the potential of defensins to control infections caused by organisms that replicate within macrophages. The work with murine neutrophils, whose content of endogenous antimicrobial polypeptides differs greatly from those of humans, will provide essential information relevant to the interpretation of murine models of human infection.
Keywords: bactericidal immunity, defensin, host organism interaction, microorganism immunology, neutrophil, Candida albicans, Escherichia coli, Histoplasma capsulatum, Listeria, Salmonella typhimurium, adrenocorticotropic hormone, granule, hormone receptor, macrophage, membrane permeability, monocyte, phagocytosis, cell free system, human tissue, immunoelectron microscopy, laboratory mouse, protein purification, tissue /cell culture, transfection
Project start date: 1985-12-01
Project end date: 2003-11-30
5R37AI022839-17 (2002): $384291
5R37AI022839-16 (2001): $373098
TOPICAL PROTEGRINS TO PREVENT STDS And HIV INFECTION
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P01AI037945-09 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: Our primary goal is to design topical peptide microbicides that will prevent sexually transmitted diseases (STDs) and remedial bacterial vaginosis (BV). A central element of this program involves designing Lactobacillus- sparing protegrin peptides that can inactivate C. trachomatis, kill C. albicans, and eliminate organisms associated with BV (e.g., Gardnerella, Mobiluncus, Prevotella). These peptides will also be tested against HIV-1, herpes simplex virus and Hemophilus ducreyii and Neisseria gonorrhoeae to identify those with broad-spectrum activity. The third project will analyze microbial mechanisms, especially efflux pumps, that allow gonococci to resist endogenous antimicrobial peptides such as LL-37. We will use this knowledge to identify and select protegrin variants that are poor substrates for such pumps. In the fourth project, we will delineate the endogenous antimicrobial polypeptides in vaginal secretions of normal women and subjects with BV. Characterizing the polypeptide effectors of innate resistance in normal vaginal secretions may illuminate the pathogenesis of various bacterial vaginitis/vaginosis syndromes. Protegrins remain the primary focus of this aspect of research. These small beta-sheet peptides were originally isolated from porcine leukocytes their unusually broad antimicrobial spectrum includes the major STD pathogens. During the past four years, we synthesized over 160 protegrin variants and used them to define the structural elements required for activity against bacteria, C. albicans and HIV-1. Although PG-1,our lead molecule, is very active vaginal lactobacilli, we have constructed Lactobacillus-sparing protegrin variants that retain excellent activity against STD agents and BV-associated organisms. Further "fine-tuning" of these protegrin variants will allow us to identify peptides for future in vivo testing in appropriate models. Overall, these studies will facilitate the development of novel, peptide- containing topical microbicides specifically designed for vaginal use. Given the high prevalence of STDs and their serious personal and economic consequences, such topical microbicides are urgently needed.
Keywords: AIDS education /prevention, HIV infection, antibiotic, disease /disorder prevention /control, drug design /synthesis /production, peptide, sexually transmitted disease, topical drug application
Project start date: 1995-03-01
Project end date: 2005-07-31
5P01AI037945-09 (2003): $814680
5P01AI037945-08 (2002): $740923
5P01AI037945-07 (2001): $770122
5P01AI037945-06 (2000): $750048
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P01AI037945-090004 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: Our long term goal is to design protegrin peptides that will be used both as topical microbicides to prevent sexually transmitted diseases (STDs) and as topical therapeutics to remediate bacterial vaginosis (BV). The Specific Aims of this project are 1. To design protegrin-like molecules that inactivate multiple STD agents and the bacteria associated with bacterial vaginosis, without affecting normal vaginal flora. 2. To determine how the beta-sheet and turn regions of protegrins contribute to these activities. 3. To learn how protegrins interact with factors relevant to their use as topical microbicides, including a) host proteins, peptides, peptides and cells; b) host and microbial proteases; c) nonoxynol-9 other surfactants. 4. To study the effects of protegrins on C. albicans a frequent vaginal opportunist. 5. To examine how protegrins assemble into dimers and oligomers, and ascertain if and how such assemble relates to their antimicrobial, cytotoxic and hemolytic properties. Protegrins are small, exceptionally potent, beta-sheet peptides that rapidly inactivate many microbes, including those responsible for most sexually transmitted bacterial infections. We will use solid-phase peptide synthesis and precise methods of antimicrobial testing to "fine tune" protegrins for future intravaginal application. Our intent is to develop protegrin-like peptides that do not affect vaginal lactobacilli (e.g., L. acidophilus and L. crispatus), but are highly active against C. albicans, STD bacteria, and the flora associated with bacterial vaginitis/vaginosis. We can obtain this constellation of properties by introducing one or two amino acid substitutions into protegrins with 15-18 residues and two intramolecular disulfide bonds. We plan to generate a relatively small number of additional protegrin variants and to test their activity against a panel of STD target organisms. Overall, these studies will facilitate the development of novel, peptide- containing topical microbicides that are designed specifically for intravaginal use. Given the prevalence and serious consequences of STDs, topical microbicides that can protect and empower women are urgently needed.
Keywords: Candida albicans, antibiotic, drug design /synthesis /production, peptide, sexually transmitted disease, topical drug application, disease /disorder prevention /control, drug interaction, drug screening /evaluation, endopeptidase, nonoxynol 9, protein structure function, vaginitis, clinical research, human subject
Sponsored Links Excellgen http://Excellgen.com
Theta-defensins: Novel HIV-1 Uptake Inhibitors
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 1R21AI056921-01 from National Institute Of Allergy And Infectious Diseases IRG: AARR
Abstract: Our broad objectives are to ascertain the role of innate immunity in protecting against HIV-infection and disease progression; to explore the antiviral properties of primate theta-defensins and determine their mechanism of activity, and identify promising 0-defensin peptides for future therapeutic development. Theta-defensins are circular octadecapeptides. They are formed in vivo by the post-translational splicing of two nonapeptides, each derived from a truncated, alpha-defensin-like propeptide that is encoded by a DEFT gene. Some 7.5 to 10 million years ago, the DEFT genes of a hominid ancestor of Homo sapiens acquired a premature stop codon in Exon 2, which encodes the signal sequence. Consequently, although human cells still express theta-defensin mRNA, they no longer produce theta-defensin peptides. We synthesized three theta-defensin peptides (retrocyclins- 1, -2 and-3) that human ceils could have produced if their DEFT genes had not been silenced. Retrocyclin-2 was remarkably effective in protecting CD4-positive cells from infection by both T and Mtropic strains of HIV-1. Its wide protective spectrum encompassed the clade B isolates that cause most infections in our country and the Clade C isolates that predominate in southern Africa and India. Our preliminary data revealed that retrocyelins are lectins, and that they bind gp 120, CD4 and galactosylceramide with high affinity. The specific aims of this proposal are a). To synthesize novel theta-defensins, including peptides based on DEFT gene sequences in non-human primates, b). To test theta-defensins and selected alpha- defensins (including human HN 1-3), against laboratory-adapted and wild-type strains of HIV-1, HIV-2, and SIV; c) To identify the sugars and oligosaccharides recognized by primate theta- and alpha-defensins and correlate this with their antiretroviral properties, and d) To determine which human lymphocytes express 0-defensin mRNA, and if arninoglycosides enable human cells to produce 0-defensin peptides. Health relatedness 0- defensins constitute a novel class of HIV-uptake inhibitors with excellent potential for future therapeutic development. The proposed research will allow experienced investigators based at UCLA, The Yerkes Primate Center at Emory University and the CDC to build the required foundation for this development.
Keywords: antiAIDS agent, defensin, drug screening /evaluation, human immunodeficiency virus 1, aminoglycoside antibiotic, human immunodeficiency virus 2, lymphocyte, oligosaccharide, simian immunodeficiency virus, Cercocebus, Macaca mulatta, Macaca nemestrina, capillary electrophoresis, cell line, high performance liquid chromatography, human subject, matrix assisted laser desorption ionization, peptide chemical synthesis
Project start date: 2003-09-01
Project end date: 2004-08-31
1R21AI056921-01 (2003): $381667
Retrocyclin Reinforcement Of Pulmonary Defenses Against Viral Aerosols
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R21AI067327-02 from National Institute Of Allergy And Infectious Diseases IRG: LCMI
Abstract: Introduction In addition to the a and a defensins produced by human leukocytes and epithelial cells, many nonhuman primates produce ?-defensins, representing a third defensin subfamily. ?-defensins lack potent antibacterial or antifungal activity, but have broad antiviral properties. Retrocyclins are ?-defensins whose structures are based on the sequences of expressed (but untranslated) human ?-defensin pseudogenes. Broad, Long term objectives To use retrocyclins to enhance pulmonary defenses to viral aerosols. Specific aims. 1. To test retrocyclins against potential viral bioterrorism agents, including Influenza A and B (Orthomyxoviridae), HPIV3 (Paramyxoviridae), West Nile and Yellow Fever virus (Flaviviridae) SARS-CoV (Coronaviridae), Vaccinia (Poxviridae), VEE, an alphavirus (Togaviridae), and Tacaribe virus (Arenaviridae) 2. To examine the interactions of retrocyclins with surfactant proteins SP-A and SP-D. 3. To test the effects actions of RC2 on monolayers and bilayers of pulmonary surfactant phospholipids. Research Design. Retrocyclin-2 (RC-2) and selected analogs will be prepared by solid phase peptide synthesis. Viruses will be tested in vitro by standard plaque reduction and CPE reduction assays. The effects of retrocyclins on the integrity of surfactant phospholipid films will be tested in monolayer systems by using a Langmuir-Blodgett trough, and in bilayer systems by using vesicles composed of surfactant lipids and associated surfactant proteins SP-B and SP-C. Binding of retrocyclins to SP-A and SP-D will be studied by surface plasmon resonance. Effects of retrocyclins on the intrinsic antiviral activity of SP-A and SP-D (and vice versa) will be studied using proteins purified from abattoir-obtained porcine lungs in standard plaque reduction and CPE reduction assays. Health relatedness and Agency Mission. This application is in direct response to PA 04-119. Lay language summary. Many natural viral infections begin with exposure to air that contains viruses. Viral aerosols may also allow bioterrorists to deliver incapacitating viruses. We propose to test the activity of retrocyclins (newly discovered antiviral peptides) against potential viral bioterrorism agents, and to determine their biocompatability with ELF-the thin fluid layer that covers the surface of the pulmonary air sacs.
Keywords: aerosol, defensin, Alphavirus, Arenaviridae, Baculoviridae, Coronaviridae, Flaviviridae, Gorilla, Orthomyxoviridae, Pan, Paramyxoviridae, Paramyxovirus, Poxviridae, Primate, Togaviridae, alpha globulin, analog, base, behavior, binding protein, bioterrorism /chemical warfare, blood protein, carbohydrate, cell, collagen, cow, crosslink, cytotoxicity, embryo /fetus, experimental design, family, fluid, gene, gene mutation, glycoprotein, head, health, human, infection, influenza, language, lectin, leukocyte, lipid, lung, mannan, membrane, model, molecular film, nucleic acid sequence, particle, peptide, phospholipid, plasma, protein, protein binding, pseudogene, pulmonary respiration, pulmonary surfactant, reduction, serum, surface plasmon resonance, surfactant, swine, virus, yellow fever virus
Project start date: 2006-02-15
Project end date: 2009-01-31
5R21AI067327-02 (2007): $112090
1R21AI067327-01 (2006): $283125
GASTROINTESTINAL AND MYELOID DEFENSINS
Robert I Lehrer, Professor
Adv Prosthodontics, Biomatl, Hosp Dentuniversity Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 5R01AI029595-05 from National Institute Of Allergy And Infectious Diseases IRG: BM
Abstract: Defensins are small, cysteine-rich, antimicrobial peptides (Mr 3,500- 4,000). They are among the principal constituents of mammalian neutrophils (PMN) and certain mammalian macrophages and also appear to be ancestral members of the immune system. Recently, Ouellette and his associates cloned an abundant mRNA species from murine small intestine, and found that it coded for a peptide("cryptdin") whose amino aid sequence identified it as a typical defensin. Although cryptdin mRNA was virtually absent at birth, it was heavily expressed by epithelial cells at the base of intestinal crypts (Paneth cells) in 20d old mice. In older mice, mononuclear cells in the small intestine´s lamina propria also contained mRNA that hybridized with cryptdin cDNA. It is known that certain defensins obtained from myeloid cells can distinguish between mouse- virulent and mouse avirulent strains of S. typhimurium. Although the functions of Paneth cells in man and animals have long been an enigma, the observation that defensin mRNA is abundantly expressed by these long-lived, secretory and phagocytic cells of the small intestine suggests that they may regulate the small intestine´s microbial flora and defend against invasion by microbes. We will isolate cryptdin and other small-intestinal defensins from rodents, determine their amino acid sequences, and examine their antimicrobial activities against phoP-positive (virulent) and phoP- negative (avirulent) isogenic strains of S. typhimurium and selected other microorganisms. Defensins expressed within rodent PMN. will serve as controls in these studies. "Cryptdin" and selected natural or designed congeners will be produced by solid-phase chemical synthesis and used to study defensin congeners will be produced by solid-phase chemical synthesis and used to study defensin structure, function and mechanism(s). Enteric and myeloid defensins will be cloned to enable us to examine their tissue- specific expression, and to identify novel defensins for potential solid- phase synthesis. Benefits that may accrue from this work include new insights into 1) the functions of Paneth cells, 2) the antimicrobial defenses of the gastrointestinal tract, 3) the mechanisms responsible for neonatal susceptibility to enteric infections 4) The distribution and tissue-specific expression of defensins and 5) the mechanisms of defensin- mediated antimicrobial activity. If Paneth cells prove to be the small- intestines´s principal guardians against microorganisms, this work may also lead to increased understanding of gastrointestinal inflammation and infection in humans, and to the development of therapeutic antibiotics specifically designed to work in the small intestine
Keywords: Salmonella typhimurium, bactericidal immunity, gastrointestinal infection, macrophage, neutrophil, protein, small intestine Candida albicans, Escherichia coli, antibody formation, gastrointestinal epithelium, lysozyme, peptide chemical synthesis, protein biosynthesis, protein sequence, protein structure function, secretory immune system, virulence genetic manipulation, infant human (0-1 year), laboratory mouse, laboratory rat, microorganism culture, molecular cloning, nucleic acid sequence
Project start date: 1990-03-01
Project end date: 1995-02-28
5R01AI029595-05 (1994): $246854
5R01AI029595-04 (1993): $244740
ANTIMICROBIAL PEPTIDES OF LEUKOCYTES
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R03TW000355-03 from Fogarty International Center IRG: SRC
Abstract: adapted from investigator s ) This proposal to study the antimicrobial molecules of porcine and chicken neutrophils is based on work being conducted at the Institute of Experimental Medicine (IEM) in St. Petersburg, Russia and is supported by preliminary collaborative studies that have been performed at UCLA. The IEM investigators have identified and partially purified potent antimicrobial peptides from the leukocytes of chickens and pigs. Preliminary studies of the chicken peptides have confirmed their antimicrobial activity, and provided composition and partial sequence data for two of the peptides.Although the peptides were cysteine-rich, they lacked the classical cysteine motif of defensins, more closely resembling "TAP" (tracheal antimicrobial peptide), a recently described endogenous peptide antibiotic of the bovine respiratory tract. This FIRCA proposal has three specific aims 1) to purify the defensin- sized antimicrobial peptides of chicken leukocytes, ascertain their primary structures, test their ability to kill selected bacteria and fungi and compare their structures and activities with previously characterized antimicrobial peptides of other species; 2) to purify the defensin-sized antimicrobial peptides of porcine leukocytes, determine their primary structures, test their ability to kill selected bacteria and fungi and compare them with other previously characterized antimicrobial peptides; 3) to clone the avian antimicrobial proteins from genomic and cDNA libraries and determine their genetic relatedness to defensins and other antimicrobial peptides of leukocytes. This proposal extends the parent grant s scope to encompass species that would not otherwise be tested. The past decade has seen a great expansion of information about endogenous antimicrobial peptides, such as magainins, cecropins and defensins, that could play a central role in innate resistance to infection. Comparative biochemical studies of various animal species have contributed to the identification of homologous peptides in man.For example, the delineation of human defensins was made possible by earlier studies with rabbit neutrophils and macrophages, and antimicrobial granulins were first recognized in equine neutrophils. Thus, the proposed FIRCA study should not only illuminate the evolution of defensin-like peptide antibiotics, it may also provide probes to facilitate the search for novel homologs of these peptides in humans.
Project start date: 1993-09-20
Project end date: 1996-09-19
5R03TW000355-03 (1995): $23263
5R03TW000355-02 (1994): $22273
1R03TW000355-01 (1993): $22228
ALPHA HELICAL CATHELICIDINS AND HOST DEFENSE
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R01AI043934-05 from National Institute Of Allergy And Infectious Diseases IRG: BM
Abstract: adapted from applicant s ) Long term objectives. To define antimicrobial molecules involved in innate immunity, learn how they work, and use this knowledge to create novel ways to prevent and treat infections. Specific aims. 1). To purify human hCAP-18 from leukocytes and secretions and examine its properties. 2) To learn how hCAP-18 and its murine homologue are processed by leukocytes. 3) To study the structures of human LL-37, murine CRAMP-38, and high molecular weight forms of hCAP-18 from plasma and secretions. 4) To define the minimal peptide structure required for LPS-binding and antimicrobial activity against P. aeruginosa. 5) To examine the effects of LPS and other stimuli on in vitro expression of hCAP-18 and examine in vivo synthesis of CRAMP in LPS-treated mice. 6) To measure hCAP-18 levels in secretions, and examine its interactions with other host defense peptides. Methods. These will include peptide synthesis, protein purification by preparative electrophoresis and chromatography (gel permeation, ion exchange, RP-HPLC and affinity), computer modeling, CD and FTIR measurements, antimicrobial testing and Northern and RT-PCR analyses. Health relatedness. The innate immune system is a key element of mucosal immunity that plays a major role in preventing infection. The proposal centers on two newly discovered, pro-antibiotics ("cathelicidins") human hCAP-18 and its murine homologue, CRAMP. These peptides are expressed constitutively by leukocytes, are produced by epithelial cells, and are found in secretions such as human milk, tears and semen. Their C-terminal domains bind LPS avidly and have potent activity against P. aeruginosa and other pathogens, even under high salt condition. The experiments will expand our knowledge about these important peptides, and should provide the information needed to develop them as antibiotics for topical bronchopulmonary use in cystic fibrosis.
Keywords: antibacterial agent, host organism interaction, peptide, protein structure function, antibody, leukocyte, lipopolysaccharide, laboratory mouse, peptide chemical synthesis, polymerase chain reaction, protein purification
Project start date: 1999-06-01
Project end date: 2005-05-31
5R01AI043934-05 (2003): $251526
Sponsored Links Excellgen http://Excellgen.com
5R01AI043934-04 (2002): $244953
5R01AI043934-03 (2001): $237814
5R01AI043934-02 (2000): $230891
ANTIMICROBIAL PEPTIDES OF LEUKOCYTES
Robert I Lehrer, Professor
Medicineuniversity Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 2R03TW000355-04 from Fogarty International Center IRG: ICP
Keywords: antibiotic, leukocyte, peptide, protein structure /function analytical chemistry, neutrophil, peptide structure, posttranslational modification chicken, gel electrophoresis, genetic library, high performance liquid chromatography, laboratory mouse, molecular cloning, protein purification, protein sequence, swine
Project start date: 1993-09-20
Project end date: 2000-01-31
2R03TW000355-04 (1997): $24121
TOPICAL PROTEGRINS TO PREVENT STDS AND HIV INFECTION
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P01AI037945-02 from National Institute Of Allergy And Infectious Diseases IRG: SRC
Project start date: 1995-03-01
Project end date: 1999-02-28
5P01AI037945-02 (1996): $596636
DEFENSINS--ANTIMICROBIAL PEPTIDES OF HUMAN NEUTROPHILS
Robert I Lehrer, Professor
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5R37AI022839-11 from National Institute Of Allergy And Infectious Diseases IRG: BM
Project start date: 1985-12-01
Project end date: 1997-11-30
5R37AI022839-11 (1996): $278834