Inactivation of parvovirus B19 in coagulation factor concentrates by UVC radiation: assessment by an in vitro infectivity assay using CFU-E derived from peripheral blood CD34+ cells.

BACKGROUND: Nonenveloped and thermostable viruses such as parvovirus B19 (B19) can be transmitted to patients who are receiving plasma-derived coagulation factor concentrates treated by the S/D method for inactivating enveloped viruses. Therefore, it is important to develop and validate new methods for the inactivation of nonenveloped viruses. STUDY DESIGN AND METHODS: Suspensions of B19 in coagulation factor concentrates (FVIII) were irradiated with UVC light. B19 infectivity was determined by an indirect immunofluorescence assay using CFU-E, as a host cell, derived from peripheral blood CD34+ cells. The effects of catechins on B19 infectivity and on FVIII activity after UVC illumination were also examined. RESULTS: The indirect immunofluorescence assay estimated the B19 infectivity of samples containing virus copies of 10(5) to 10(11) per 10 microL to be a median tissue culture-infectious dose of 10(0.3) to 10(5.4) per 10 microL. B19 was inactivated by 3 log at 750 J per m(2) of UVC radiation and was undetectable after 1000 or 2000 J per m(2) of irradiation. However, FVIII activity decreased to 55 to 60 percent of pretreatment activity after 2000 J per m(2) of UVC radiation. This was inhibited in the presence of rutin or catechins. Epigallocatechin gallate could maintain FVIII activity at almost 100 percent of pretreatment activity after 2000 J per m(2) of UVC radiation, while B19 infectivity was decreased to undetectable levels, which resulted in >3.9 log inactivation. CONCLUSION: UVC radiation in the presence of catechins, especially epigallocatechin gallate, appears to be an effective method of increasing the viral safety of FVIII concentrates without the loss of coagulation activity.

Elimination of both cell-free and cell-associated HIV infectivity in plasma by a filtration/methylene blue photoinactivation system.

BACKGROUND: Methylene blue phototreatment effectively inactivates cell-free viruses in plasma while maintaining coagulation activities. However, this treatment is considered to be less effective for cell-associated virus inactivation. This report describes a new virus elimination system designed to eliminate cell-associated viruses with a cell-removal filter followed by methylene blue photoinactivation of cell-free viruses in plasma. STUDY DESIGN AND METHODS: Fresh plasma was inoculated with HIV or HIV-infected Molt4 cells (Molt4(IIIB)). The plasma was transferred to a bag containing methylene blue by passing it through a cell-removal filter and was irradiated with white fluorescent light. HIV infectivity was detected by indirect fluorescence assay. In parallel studies, coagulation activities in identically treated plasma were measured during 1 year of storage at -80 degrees C. RESULTS: Initial cell-free HIV titer of 10(6.2) TCID(50) per 0.1 mL dropped to 10(-0. 3) and <10(-0.5) TCID(50) per 0.1 mL after 10 or 20 J per cm(2) radiation, respectively. Cellular components were not detectable in plasma after filtration. The cell-free state of the plasma was ascertained from the observation that the DNase-resistant beta-globin gene, as a marker of intact WBCs, was not detected in the filtrates by PCR. The infectivity of Molt4(IIIB) was reduced to below the detection limit after filtration and radiation, and proviral HIV DNA was not detected in the filtrates by PCR. Coagulation activities including factor VIII in the treated plasma were maintained at more than 76 percent compared with the percentage in untreated plasma after 1 year of storage. CONCLUSION: The filtration/methylene blue photoinactivation system eliminated both cell-free and cell-associated HIV infectivities from plasma while maintaining coagulation activities for 1 year at -80 degrees C storage.