HIV INHIBITORY FACTORS IN HUMAN SALIVA
(1994)
Abstract :
HIV, the causative agent of AIDS, has been infrequently recovered from salivary secretions. Preliminary reports support the contention that human saliva contains inhibitory activity directed against HIV> The specific aims of this project are to (1) verify and amplify the anti-HIV effects of saliva, (2) fractionate the salivas (parotid, submandibular/sublingual and/or labial) with the activity to isolate the component(s), (3) identify and characterize the biologic and molecular nature of the substance(s), and (4) determine the mechanism of anti-HIV action of the active component(s). Human whole and individual gland salivary secretions will be obtained from normal human subjects and tested for anti-HIV activity utilizing a sensitive, standardized, and reproducible human host-cell in vitro screening assay (MT-2 syncytium-formation TCID50 assay). Those glandular secretions showing inhibitory activity toward HIV infectivity will be fractionated and individual fractions possessing the inhibitory activity will be isolated and characterized. The isolated inhibitory factors will also be utilized in studies designed to elucidate the mechanism of action of this inhibition towards HIV. The elucidation of the molecular structure and mechanism of action of such a factor will not only allow an assessment of the protective effects of saliva against HIV infection, but could lead to strategies for the development of potent anti-HIV agents.
| Project Number : | 5R01DE009563-03 |
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| ICD : | NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH |
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| IRG : | SRC |
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| Project Terms : | antiviral agent, chemical structure function, human immunodeficiency virus, saliva glycoprotein, mucin, parotid gland, protein structure function, sublingual gland, submandibular gland, virus host interaction bioassay, human tissue, physical separation |
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NEUROSYPHILIS AND AIDS--PCR METHOD TO DETECT T. PALLIDUM
(1993)
Abstract :
The clinical diagnosis of neurosyphilis, especially in HIV infected patients, can be extremely difficult. Serologic assays are the accepted methods for laboratory diagnoses. However, these assays lack sensitivity or specificity, especially in the diagnosis of neurosyphilis. In addition, individuals with HIV infection have been documented to lose their treponemal test reactivity to MHA-TP and FTA-ABS. Therefore, the need for the development of more effective methods to detect T. pallidum exists. The specific aims of the "pilot" project are to establish a sensitive and specific diagnostic test for the detection of T. pallidum infection based on the polymerase chain reaction (PCR) method, and to evaluate the performance of the PCR assay on well-characterized archived clinical specimens from patients with a clinical or histopathological diagnosis of neurosyphilis. Preliminary studies indicate that a highly sensitive and specific assay to detect T. pallidum based upon the PCR can be developed. A 210-bp sequence of the gene encoding the 47kDa membrane immunogen was amplified and the PCR products probed by DNA hybridization with a 40-bp fragment internal to the amplified DNA. The assay sensitivity is capable of detecting five or more organisms consistently and, in some cases, a single organism when calculated by sequential dilutions of suspensions of treponemes. The specificity of the 47kDa primer pairs/probe was assessed using a panel of pathogenic and non-pathogenic spirochetes. Positive amplification products were consistently detected only with DNA obtained from either T. pallidum or T. pertenue. Similar results were obtained when a second set of primers/probe from the T. pallidum 4D antigen were implemented. The PCR method will be optimized to accommodate the highly sensitive and specific detection of T. pallidum from clinical specimens. Well-characterized archived post-mortem clinical specimens with a clinical/histopathologic diagnosis of neurosyphilis will be processed to validate the PCR method. This approach will allow for a comparison of the sensitivity/specificity of PCR to the currently utilized serologic studies in the diagnosis of neurosyphilis. It is hoped that these pilot studies will provide a valuable diagnostic technique that can be utilized in the investigation of the pathogenesis of neurosyphilis.
| Project Number : | 1R01NS029924-01A1 |
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| ICD : | NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE |
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| IRG : | ARRE |
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| Project Terms : | AIDS, Treponema pallidum, communicable disease diagnosis, diagnosis design /evaluation, human immunodeficiency virus 1, nervous system infection, syphilis diagnosis quality /standard human tissue, polymerase chain reaction |
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NEUROSYPHILIS AND AIDS--PCR METHOD TO DETECT T. PALLIDUM
(1993)
Abstract :
The clinical diagnosis of neurosyphilis, especially in HIV infected patients, can be extremely difficult. Serologic assays are the accepted methods for laboratory diagnoses. However, these assays lack sensitivity or specificity, especially in the diagnosis of neurosyphilis. In addition, individuals with HIV infection have been documented to lose their treponemal test reactivity to MHA-TP and FTA-ABS. Therefore, the need for the development of more effective methods to detect T. pallidum exists. The specific aims of the "pilot" project are to establish a sensitive and specific diagnostic test for the detection of T. pallidum infection based on the polymerase chain reaction (PCR) method, and to evaluate the performance of the PCR assay on well-characterized archived clinical specimens from patients with a clinical or histopathological diagnosis of neurosyphilis. Preliminary studies indicate that a highly sensitive and specific assay to detect T. pallidum based upon the PCR can be developed. A 210-bp sequence of the gene encoding the 47kDa membrane immunogen was amplified and the PCR products probed by DNA hybridization with a 40-bp fragment internal to the amplified DNA. The assay sensitivity is capable of detecting five or more organisms consistently and, in some cases, a single organism when calculated by sequential dilutions of suspensions of treponemes. The specificity of the 47kDa primer pairs/probe was assessed using a panel of pathogenic and non-pathogenic spirochetes. Positive amplification products were consistently detected only with DNA obtained from either T. pallidum or T. pertenue. Similar results were obtained when a second set of primers/probe from the T. pallidum 4D antigen were implemented. The PCR method will be optimized to accommodate the highly sensitive and specific detection of T. pallidum from clinical specimens. Well-characterized archived post-mortem clinical specimens with a clinical/histopathologic diagnosis of neurosyphilis will be processed to validate the PCR method. This approach will allow for a comparison of the sensitivity/specificity of PCR to the currently utilized serologic studies in the diagnosis of neurosyphilis. It is hoped that these pilot studies will provide a valuable diagnostic technique that can be utilized in the investigation of the pathogenesis of neurosyphilis.
| Project Number : | 5R01NS029924-02 |
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| ICD : | NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE |
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| IRG : | ARRE |
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| Project Terms : | AIDS, Treponema pallidum, communicable disease diagnosis, diagnosis design /evaluation, human immunodeficiency virus 1, nervous system infection, syphilis diagnosis quality /standard human tissue, polymerase chain reaction |
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HIV INHIBITORY FACTORS IN HUMAN SALIVA
(1993)
Abstract :
HIV, the causative agent of AIDS, has been infrequently recovered from salivary secretions. Preliminary reports support the contention that human saliva contains inhibitory activity directed against HIV> The specific aims of this project are to (1) verify and amplify the anti-HIV effects of saliva, (2) fractionate the salivas (parotid, submandibular/sublingual and/or labial) with the activity to isolate the component(s), (3) identify and characterize the biologic and molecular nature of the substance(s), and (4) determine the mechanism of anti-HIV action of the active component(s). Human whole and individual gland salivary secretions will be obtained from normal human subjects and tested for anti-HIV activity utilizing a sensitive, standardized, and reproducible human host-cell in vitro screening assay (MT-2 syncytium-formation TCID50 assay). Those glandular secretions showing inhibitory activity toward HIV infectivity will be fractionated and individual fractions possessing the inhibitory activity will be isolated and characterized. The isolated inhibitory factors will also be utilized in studies designed to elucidate the mechanism of action of this inhibition towards HIV. The elucidation of the molecular structure and mechanism of action of such a factor will not only allow an assessment of the protective effects of saliva against HIV infection, but could lead to strategies for the development of potent anti-HIV agents.
| Project Number : | 1R01DE009563-01 |
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| ICD : | NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH |
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| IRG : | SRC |
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| Project Terms : | CHEMICAL STRUCTURE--BIOLOGICAL ACTIVITY, COMMUNICABLE DISEASE CONTROL AGENTS, ANTIVIRAL, ORAL-PHARYNGEAL, SALIVA, VIRUSES, RETROVIRIDAE, HUMAN IMMUNODEFICIENCY VIRUSES (HIV) ORAL-PHARYNGEAL, SALIVARY GLANDS, PAROTID, ORAL-PHARYNGEAL, SALIVARY GLANDS, SUBLINGUAL, ORAL-PHARYNGEAL, SALIVARY GLANDS, SUBMANDIBULAR, PROTEIN STRUCTURE AND FUNCTION, PROTEINS, GLYCOPROTEINS, PROTEOGLYCANS, MUCIN, VIRUS DISEASE CHARACTERISTICS, HOST-VIRUS HUMAN, HUMAN TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PR OJECT, PHYSICAL SEPARATION, FRACTIONATION, bioassay |
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HIV INHIBITORY FACTORS IN HUMAN SALIVA
(1993)
Abstract :
HIV, the causative agent of AIDS, has been infrequently recovered from salivary secretions. Preliminary reports support the contention that human saliva contains inhibitory activity directed against HIV> The specific aims of this project are to (1) verify and amplify the anti-HIV effects of saliva, (2) fractionate the salivas (parotid, submandibular/sublingual and/or labial) with the activity to isolate the component(s), (3) identify and characterize the biologic and molecular nature of the substance(s), and (4) determine the mechanism of anti-HIV action of the active component(s). Human whole and individual gland salivary secretions will be obtained from normal human subjects and tested for anti-HIV activity utilizing a sensitive, standardized, and reproducible human host-cell in vitro screening assay (MT-2 syncytium-formation TCID50 assay). Those glandular secretions showing inhibitory activity toward HIV infectivity will be fractionated and individual fractions possessing the inhibitory activity will be isolated and characterized. The isolated inhibitory factors will also be utilized in studies designed to elucidate the mechanism of action of this inhibition towards HIV. The elucidation of the molecular structure and mechanism of action of such a factor will not only allow an assessment of the protective effects of saliva against HIV infection, but could lead to strategies for the development of potent anti-HIV agents.
| Project Number : | 5R01DE009563-02 |
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| ICD : | NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH |
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| IRG : | SRC |
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| Project Terms : | antiviral agent, chemical structure function, human immunodeficiency virus, saliva glycoprotein, mucin, parotid gland, protein structure function, sublingual gland, submandibular gland, virus host interaction bioassay, human tissue from nonrelated source, physical separation |
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POLYMERASE CHAIN REACTION TO ID MYCOBACTERIAL ORGANISMS
(1991)
| Project Number : | 2S07RR005662-180151 |
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| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
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POLYMERASE CHAIN REACTION TO IDENTIFICATION OF PATHOGENIC TREPONEMES
(1990)
| Project Number : | 2S07RR005662-170522 |
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| ICD : | NATIONAL CENTER FOR RESEARCH RESOURCES |
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