Contracts Management Section, National Institute of Dental Research, NIH, 45 Center Drive, Room 4AN-44D, Bethesda, Maryland 20892

A -- IMMUNOLOGIC FACTORS IN SALIVA AND BLOOD AS DETERMINANTS OF HIV-RELATED DISEASES POC Marilyn R. Zuckerman, Contracting Officer, (301) 594-5569. The National Institute of Dental Research (NIDR) is conducting a market survey to determine the availability and potential technical capability of small businesses; the applicable SIC code is 8731 and the size standard is 500 employees. The NIDR has a requirement to develop methods and assay for selected immunological and viral characteristics in the saliva and serum of members of a cohort of HIV positive U.S. military personnel. The Contractor will test approximately 2000 serum and 2000 saliva samples (to be supplied by the Government) for total IgG, IgM, and IgA; HIV-specific IgG, IgM, and IgA; nonspecific immune factors (lactoferrin, lysozyme and lactoperoxidase); levels of secretory leukocyte protease inhibitor; quantitative HIV virions; and infectivity of human peripheral blood mononuclear cells (PBMC). Special features of this project are the large number of samples and the need for the development of procedures to apply to saliva as the matrix and to conserve volume of specimen as much as possible. Subsequent to the assaying of the samples, the laboratory findings will be linked to the demographic, medical, behavioral, oral examination and follow-up data available on this cohort. This project will entail two phases. Phase I will consist of all activities undertaken prior to commencement of production assays. During Phase I the contractor will develop, test, and prepare the final protocol for the study, including plans for quality control, for approval by the NIDR. Phase II will include performing the assays on the serum and saliva. The Government estimates that three years with a total of 19,200 hours will be required to carry out the study. It is estimated that 2000 professional, 1400 technical, and 3000 support hours (6400 hours total) will be required in each of the three years. General objectives include the following: 1. To quantify in all saliva and serum samples (a) total IgM, IgA (IgA1 and IgA2) and IgG; (b) HIV-specific IgM, IgA (IgA1 and IgA2) and IgG, including nonspecific immune factors including SLPI, mucins, lactoferrin, lysozyme and lactoperoxidase, as well as HIV-1 viral load as measured by culture and nucleic acid detection HIV (RNA PCR). 2. To characterize the phenotype of HIV-1 isolated from saliva and blood according to syncytium-inducing ability (SI vs NSI), cell tropism (macrophage vs T cell), replication kinetics and strain variability. Specific technical requirements include the following: 1. SPECIMEN MANAGEMENT: (a) Maintain and update at all times, an electronic receipt and control system that keeps track of all specimens received and returned at all times; (b) Ensure proper disposal of specimen containers; (c) Ensure proper handling of specimens to prevent degradation and to ensure proper infection control; (d) Return unused specimens to a repository designated by NIDR in the Washington, DC, metropolitan area. 2. DEVELOPMENT OF ASSAYS: Due to the presence of proteinases, inhibitors, bacteria, and other biological variables in saliva, preliminary experiments are required to assess the best method of viral detection and/or culture of virus in these preparations (e.g., centrifuged vs uncentrifuged; cell vs supernatant). Assays must be standardized to enable direct comparison and correlations between saliva specimens (cells and saliva) and blood specimens for HIV and other parameters. Specifically, the contractor must use the following techniques for the listed assays: quantitative HIV RNA PCR assessment for plasma, use of specified commercially available assays may require adjustment for saliva (unspun/cells). 3. REQUIREMENTS OF ASSAYS IN PRODUCTION RUNS: (a) Total and HIV-specific immunoglobulins; use ELISA techniques for total and HIV-specific immunoglobulins IgM, IgA (IgA~ and IgA2) and IgG; additional assays may include neutralizing antibodies and antibody dependent cellular cytotoxicity (ADCC); (b) HIV-viral load: HIV RNA PCR assay (quantitative RT-PCR or another commercially available method); use cell microculture techniques for PBMC and saliva pellet cell associated HIV; use p24 antigen ELISA on serum and saliva supernatant to assess HIV levels; in situ hybridization from smears of cells; (c) Non-specific immune factors; (d) Syncytia-inducing ability: Use cell microculture techniques to determine cell-associated syncytia inducing (SI) HIV phenotypes and non-syncytia inducing (NSI) macrophage tropism (isolates from saliva cell pellet); and (e) Replication kinetics and heteroduplex mobility assay (HMA). This is NOT a Request for Proposal; there is no solicitation. The Government is not committed to award a contract pursuant to this announcement. Responses should not include cost or pricing information. Sources believing they have the required capabilities to undertake this project should submit four (4) copies of the requested material addressing experience and capabilities by 4:00 p.m., local time, on June 16, 1997. (0134)

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