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Photochemical virus inactivation technology is widely used to improve the safety of blood products. However, the process by which this inactivation occurs and the resulting immunogenicity of treated viruses remain to be elucidated. This study aimed to explore the effects of two photochemical inactivation methods (methylene and riboflavin, MP and RP) on hepatitis B virus (HBV) immunogenicity. Inactivated HBV were incubated with PBMC from six healthy donors. Culture supernatants were collected at 0, 24 and 72 h for the analysis of HBsAg and HBeAg expression using ELISA. Cytokine expression was analyzed at 72 h using ELISA. Costimulatory and cell adhesion molecule mRNA expression was analyzed at 24 h by RT-PCR. No significant changes in HBsAg and HBeAg were detected following MP. However, the secretion of TNF-α and IFN-γ was upregulated. Expression of CD80, CD86, ICAM2 and LFA3 mRNA was also upregulated. In contrast, although RP did not significantly alter HBsAg expression, a reduction in HBeAg expression was observed. Furthermore, no upregulation of cytokines and intracellular molecule expression was observed following RP. These data indicate that the immunogenicity of HBV is retained following MP, and the inactivation of HBV could upregulate the Th1-type cellular immune responses, which may play significant roles in the antiviral process. [ABSTRACT FROM AUTHOR]

The mechanism of bacteriophage photoinactivation by methylene blue and light (MB+L) involves genomic RNA damage. In this study, two RNA viruses, Sindbis virus (SINV) and hepatitis C virus were treated by MB+L and their nucleic acids were amplified to show that RNA lesions occurred during inactivation. During MB+L inactivation, the viral load of both viruses was significantly reduced as MB+L exposure increased. The nucleic acid amplification of treated viral RNA was inhibited in a time-dependent manner and the percentage inhibition of amplification reached about 99% after 30 min of treatment. Furthermore, as compared to SINV viral infectivity detected by quantification of the 50% tissue culture infective dose (TCID), the inhibition of SINV RNA amplification strongly correlated with a decrease in in vitro infectivity ( R > 0.94), suggesting that RNA serves as the main target during MB+L inactivation. [ABSTRACT FROM AUTHOR]

Immunocompetent lymphocytes present in blood products are an important cause of immune reactions following blood transfusion such as transfusion-associated graft- versus-host disease (TA-GVHD). In this study the effects of riboflavin photochemical treatment (RPT) on lymphocyte proliferation and cytokine production in vitro were measured to establish whether RPT of blood products can be used to prevent immune reactions resulting from transfused lymphocytes. Lymphocytes and riboflavin were added together in medical PVC transparent bags and then exposed to visible light. Control lymphocytes were exposed to light in the absence of riboflavin. Lymphocytes exposed to riboflavin photochemical treatment (RPT-lymphocytes) and control lymphocytes were tested for the proliferative ability and the production of several cytokines upon stimulation with antigens. Upon stimulation by phytohemagglutinin (PHA) the proliferation of RPT-lymphocytes was inhibited. Using flow cytometry it was shown that RPT-lymphocytes were unable to enter the S-phase of the cell cycle following PHA stimulation. The level of cytokines present in the supernatant of RPT-lymphocytes after stimulation by antigens was significantly lower than those present in the corresponding supernatant of control lymphocytes. RPT with visible light inactivates lymphocytes, inhibits lymphocyte proliferation and the production of cytokines. It appears to be a promising method to prevent immune reactions such as TA-GVHD. [ABSTRACT FROM AUTHOR]