The risk of transmitting cell-associated human immunodeficiency viruses (HIV- 1/-2), human T cell lymphotropic viruses (HTLV-I/-II), and cytomegaloviruses (CMV), and of transmitting cell-free HIV and hepatitis B and C viruses (HBV and HCV) from fully-screened blood can be reduced by in vitro inactivation/removal (IVIR) of blood-borne viruses (BBV). We propose a three-step cascade process for IVIR of BBV carried out on whole blood during routine blood processing: (1) selective removal of white blood cells, which harbor and permit replication of cell-associated BBV; (2) receptor-mediated removal of the cell-free BBV from leukocyte-free whole blood; and (3) biochemical or chemical inactivation of residual BBV by adding pharmacologically-acceptable agents that are biocompatible with blood. For testing this concept, our experimental design employs HIV as a prototype BBV, because HIV occurs both in cell-associated and cell-free forms and can be monitored quantitatively by complementary in vitro amplifications, biochemically by the polymerase chain reaction (PCR) and biologically by co-cultivation with CD4-positive cells. Because unacceptable platelet loss occurs during leukocyte depletion of whole blood, two different polyester filters are currently used for packed red cells and platelet concentrates. The addition of biotinylated receptor proteins (cD4-CD26) or human antibodies to HIV envelope proteins, bound to streptavidin-coated magnetic beads, will enable specific capture of both cell-free HIV and HIV-immune-complexes from the leukocyte-filtered blood. We will investigate multi-target ribozymes for enzymatic inactivation of HIV RNA and/or ascorbic acid for chemical inactivation of residual virions. Results of the studies with HIV will provide an applicable paradigm for analogous IVIR of cell-free HBV and HCV. Although we do not expect significant changes in membrane structure and function of red cells and platelets, we will perform hematological investigations to validate the safety and effectiveness of blood treatment for IVIR of BBV. Ultimately, our research and development efforts could culminate in a collaboration with manufacturer(s) capable of providing an integrated system suitable for routine IVIR of BBV in transfusion practice.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL053384-05
Application #
2771386
Study Section
Special Emphasis Panel (ZHL1-CSR-S (S1))
Project Start
1994-09-30
Project End
2000-08-31
Budget Start
1998-09-01
Budget End
2000-08-31
Support Year
5
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Pathology
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Vyas, Girish N (2008) Participating in the evolution of transfusion medicine from a dispensary into a discipline. Transfus Med Rev 22:162-7
Terrault, Norah A; Vyas, Girish (2003) Hepatitis B immune globulin preparations and use in liver transplantation. Clin Liver Dis 7:537-50
Vyas, G N (2001) Human peripheral blood mononuclear cell substrate for propagating wild type HIV-1. Dev Biol (Basel) 106:345-56; discussion 356-7
Gandhi, M J; Boyd, M R; Yi, L et al. (2000) Properties of cyanovirin-N (CV-N): inactivation of HIV-1 by sessile cyanovirin-N (sCV-N). Dev Biol (Basel) 102:141-8
Vyas, G N (2000) Immunobiology of persistent blood-borne viral infections. Dev Biol (Basel) 102:17-Sep
Gandhi, M J; Yang, G G; McMahon, B J et al. (2000) Hepatitis B virions isolated with antibodies to the pre-S1 domain reveal occult viremia by PCR in Alaska Native HBV carriers who have seroconverted. Transfusion 40:910-6
Yang, G; D'Souza, M P; Vyas, G N (1998) Neutralizing antibodies against HIV determined by amplification of viral long terminal repeat sequences from cells infected in vitro by nonneutralized virions. J Acquir Immune Defic Syndr Hum Retrovirol 17:27-34
Rawal, B D; Vyas, G N (1996) Magnesium-mediated reversal of the apparent virucidal effect of ascorbic acid or congo red reacted in vitro with the human immunodeficiency virus. Biologicals 24:113-6
Prati, D; Rawal, B D; Dang, C et al. (1995) DNA enzyme immunoassay of the PCR-amplified HLA-DQ alpha gene for estimating residual leukocytes in filtered blood. Clin Diagn Lab Immunol 2:182-5