Improving Safety of the Blood Supply from Transmission of HIV/AIDS and Other Emerging Retroviral, Blood Borne Viral and Biodefense Agents by Development of Sensitive Diagnostic Tools and Investigations of Disease Pathogenesis

Principal Investigator: Indira Hewlett, PhD
Office / Division / Lab: OBRR / DETTD / LMV

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Overview

Public Health Issue: HIV disease has spread world wide with evolving genetic diversity resulting in numerous recombinant forms of the virus. The increased use of HIV therapies has also resulted in viral variants that harbor drug resistance mutations. These new strains may go undetected by diagnostic and blood screening assays resulting in disease transmission. Similarly smallpox vaccination using current vaccinia virus based vaccines may result in viremia in vaccinated individuals. Early detection of virus is critical to ensuring safety of blood donations from viral transmission. Also tests used to screen blood may be affected by vaccination. Recently other emerging agents such as HHV-8, human papillomaviruses and highly pathogenic avian flu viruses have posed concerns to blood safety since a full risk assessment is not possible without adequate data about viremia and prevalence/incidence of these agents. There is a continual need to evaluate new tools to enhance the detection of multiple pathogens in blood. These approaches could potentially be useful in screening large numbers of samples for multiple pathogens. In order to enhance the detection of multiple agents in small volumes of clinical samples, we are evaluating a new nanotechnology based approach for simultaneous multiplexed detection of the agents. This technique may have the potential to be applicable to point-of-care testing and to simplify current testing strategies.

Regulatory Contribution: The ability of current and new diagnostic/blood screening assays to detect emerging forms of HIV and other related human retroviruses is critical to ensuring patient and blood product safety. CBER should maintain surveillance for new and emerging strains and potentially new recombinant forms of HIV and to determine if they are detected by current assays. Studies on infectivity of vaccinia by blood will help us understand the potential transmissibility of virus through blood products. Also studies on human herpes viruses, human papillomaviruses and new viruses such as highly pathogenic avian flu will help us understand whether these agents are an issue for blood safety.

Research Approach: The goals of the HIV program are to 1) maintain active surveillance for emerging retroviruses in geographic areas of high viral diversity 2) evaluate the significance of variants for diagnostics and blood screening 3) determine the feasibility of new technologies for more rapid and sensitive detection and characterization of emerging strains 4) acquire strains and develop reference reagents to assess the performance and eventual licensure of new assays for sensitive detection of viral variants. In addition to detection, we will also study the pathogenesis of new strains in regard to tropism, co-receptor use and host factors including new apoptotic pathways/cytokine/chemokine regulation in order to understand whether emerging HIV strains have evolved new mechanisms to facilitate enhanced spread of disease. The goals of the vaccinia project are to develop diagnostic tools for vaccinia and to study virus transmission by blood. We have developed methods to isolate virus from blood and to detect viral nucleic acid using a TaqMan assay. We have developed a nanoparticle assay for HIV p24 and antibody detection based on a bio-barcode technique developed by scientists at Northwestern University. The work will be extended to other viruses including HBV, HCV and WNV. Work has also been initiated on a Europium and BCA assay for some major biodefense pathogens.

Mission Relevance & Outcomes: It is FDA's mission to evaluate new and improved assay systems and diagnostic technologies for medical use in order to facilitate their eventual licensure. The development of a comprehensive panel for evaluation of new assays for blood screening would provide a common point of reference for evaluation of multiple, emerging technologies and allow manufacturers to obtain data in a short time frame for rapid approval for such tests. Our studies will also contribute towards defining policy for blood safety through deferral of vaccinated donors and availability due to potential false positive results due to vaccination. Our work on HHV-8 seroprevalence in blood donors should be helpful in defining the need for improved assays to better determine the epidemiology and impact of this virus on blood donation. We will be developing assays for detection of avian and low pathogenic strains of influenza and use these tools to study potential transfusion transmission of highly pathogenic flu. Studies on pathogenesis and transmission will help determine the impact of virus testing on prevention of virus transmission and identify new biomarkers for virus infection.

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Publications

J Infect Dis 2008 Nov 1;198(9):1300-1308
In Vitro Evaluation of the Protective Role of Human Antibodies to West Nile Virus (WNV) Produced during Natural WNV Infection.
Rios M, Daniel S, Dayton AI, Wood O, Hewlett IK, Epstein JS, Caglioti S, Stramer SL

J Acquir Immune Defic Syndr 2007 Oct 1;46(2):231-7
Nanoparticle-Based Biobarcode Amplification Assay (BCA) for Sensitive and Early Detection of Human Immunodeficiency Type 1 Capsid (p24) Antigen.
Tang S, Zhao J, Storhoff JJ, Norris PJ, Little RF, Yarchoan R, Stramer SL, Patno T, Domanus M, Dhar A, Mirkin CA, Hewlett IK

AIDS Res Hum Retroviruses 2007 Oct;23(10):1262-1267
Detection of Emerging HIV Variants in Blood Donors from Urban Areas of Cameroon.
Lee S, Wood O, Tang S, Hu J, Machuca A, Kerby S, Awazi B, Vockley C, Hewlett I

AIDS Res Hum Retroviruses 2007 Aug;23(8):1008-19
Identification of a Novel Circulating Recombinant Form (CRF) 36_cpx in Cameroon That Combines Two CRFs (01_AE and 02_AG) with Ancestral Lineages of Subtypes A and G.
Powell RL, Zhao J, Konings FA, Tang S, Nanfack A, Burda S, Urbanski MM, Saa DR, Hewlett I, Nyambi PN

Clin Infect Dis 2007 Jul 15;45(2):181-6
West nile virus adheres to human red blood cells in whole blood.
Rios M, Daniel S, Chancey C, Hewlett IK, Stramer SL

J Acquir Immune Defic Syndr 2007 Jul 1;45(3):361-363
Increased Genetic Diversity and Intersubtype Recombinants of HIV-1 in Blood Donors From Urban Cameroon.
Machuca A, Tang S, Hu J, Lee S, Wood O, Vockley C, Vutukuri SG, Deshmukh R, Awazi B, Hewlett I

AIDS Res Hum Retroviruses 2007 Jul;23(7):923-33
Circulating recombinant form (CRF) 37_cpx: an old strain in Cameroon composed of diverse, genetically distant lineages of subtypes A and G.
Powell RL, Zhao J, Konings FA, Tang S, Ewane L, Burda S, Urbanski MM, Saa DR, Hewlett I, Nyambi PN

Biochem Biophys Res Commun 2007 May 18;356(4):1017-23
Microarray multiplex assay for the simultaneous detection and discrimination of hepatitis B, hepatitis C, and human immunodeficiency type-1 viruses in human blood samples.
Hsia CC, Chizhikov VE, Yang AX, Selvapandiyan A, Hewlett I, Duncan R, Puri RK, Nakhasi HL, Kaplan GG

Virology 2007 Mar 1;359(1):105-15
A second-site suppressor significantly improves the defective phenotype imposed by mutation of an aromatic residue in the N-terminal domain of the HIV-1 capsid protein.
Tang S, Ablan S, Dueck M, Ayala-Lopez W, Soto B, Caplan M, Nagashima K, Hewlett IK, Freed EO, Levin JG

Peptides 2007 Mar;28(3):496-504
Antibodies against a multiple-peptide conjugate comprising chemically modified human immunodeficiency virus type-1 functional Tat peptides inhibit infection.
Devadas K, Boykins RA, Hewlett IK, Wood OL, Clouse KA, Yamada KM, Dhawan S

J Virol Methods 2006 Nov;137(2):287-91
Development and evaluation of HIV-1 subtype RNA panels for the standardization of HIV-1 NAT assays.
Lee S, Wood O, Taffs RE, Hu J, Machuca A, Vallejo A, Hewlett I

J Acquir Immune Defic Syndr 2006 Nov 1;43(3):304-12
Novel Approach for Differential Diagnosis of HIV Infections in the Face of Vaccine-Generated Antibodies: Utility for Detection of Diverse HIV-1 Subtypes.
Khurana S, Needham J, Park S, Mathieson B, Busch MP, Nemo G, Nyambi P, Zolla-Pazner S, Laal S, Mulenga J, Chomba E, Hunter E, Allen S, McIntyre J, Hewlett I, Lee S, Tang S, Cowan E, Beyrer C, Altfeld M, Yu XG, Tounkara A, Koita O, Kamali A, Nguyen N, Graham BS, Todd D, Mugenyi P, Anzala O, Sanders E, Ketter N, Fast P, Golding H

Transfusion 2006 Oct;46(10):1847-8
Performance of serological assays used to test blood from recent smallpox vaccinees.
Srinivasan K, Lee S, Daniel S, Wood O, Akolkar P, Hewlett I

Transfusion 2006 Sep;46(9):1647-8
Serologic testing for human T-lymphotropic virus-3 and -4.
Switzer WM, Hewlett I, Aaron L, Wolfe ND, Burke DS, Heneine W

Transfusion 2006 Sep;46(9):1589-92
Absence of detectable viremia in plasma and peripheral blood mononuclear cells from smallpox vaccinees: implications for blood safety.
Srinivasan K, Akolkar PN, Taffs RE, Hewlett IK

Peptides 2006 Apr;27(4):611-21
Selective side-chain modification of cysteine and arginine residues blocks pathogenic activity of HIV-1-Tat functional peptides.
Devadas K, Boykins RA, Hardegen NJ, Philp D, Kleinman HK, Osa EO, Wang J, Clouse KA, Wahl LM, Hewlett IK, Rappaport J, Yamada KM, Dhawan S

Transfusion 2006 Apr;46(4):659-667
Monocytes-macrophages are a potential target in human infection with West Nile virus through blood transfusion.
Rios M, Zhang MJ, Grinev A, Srinivasan K, Daniel S, Wood O, Hewlett IK, Dayton AI

Clin Infect Dis 2005 Jun 1;40(11):1673-6
Seroprevalence of human T cell leukemia virus in HIV antibody-negative populations in rural Cameroon.
Machuca A, Wood O, Lee S, Daniel S, Rios M, Wolfe ND, Carr JK, Eitel MN, Tamoufe U, Torimiro JN, Burke D, Hewlett IK

J Immunol 2004 Dec 1;173(11):6735-44
Mechanisms for Macrophage-Mediated HIV-1 Induction.
Devadas K, Hardegen NJ, Wahl LM, Hewlett IK, Clouse KA, Yamada KM, Dhawan S

Transfusion 2004 Jun;44(6):929-33
Window-period human immunodeficiency virus transmission to two recipients by an adolescent blood donor.
Phelps R, Robbins K, Liberti T, Machuca A, Leparc G, Chamberland M, Kalish M, Hewlett I, Folks T, Lee LM, McKenna M