Inactivation of leukocytes in platelet concentrates by photochemical treatment with psoralen plus UVA

A photochemical treatment (PCT) process using a novel psoralen and long wavelength ultraviolet light (UVA, 320-400 nm) has been developed to inactivate bacteria and viruses in platelet concentrates. This study evaluated the efficacy of PCT for inactivation of leukocytes that contaminate platelet preparations. Three psoralens, 8-methoxypsoralen (8-MOP), 4'-aminomethyl 4,5',8-trimethylpsoralen (AMT), and the novel psoralen S-59, were compared using the following four independent but complementary biological and molecular assays. (1) T-cell viability: Treatment with 150 mu mol/L S-59 and 1.0 to 3.0 Joules/cm(2) UVA inactivated >5.4 +/- 0.3 log(10) of T cells in full-sized single-donor plateletpheresis units. Using 1.0 Joule/cm(2) UVA, the lowest dose of S-59, AMT and 8-MOP required to reduce the number of T cells to the limit of detection was 0.05 mu mol/L, 1.0 mu mol/L, and 10.0 mu mol/L, respectively. (2) Cytokine synthesis: Treatment with 1.9 Joules/cm(2) UVA and 150 mu mol/L S-59 or AMT completely inhibited synthesis of the cytokine IL-8 by contaminating leukocytes during 5 days of platelet storage. After treatment with 75 mu mol/L 8-MOP and 1.9 Joules/cm(2) UVA, only low levels of IL-8 were detected. (3) Psoralen-DNA adduct formation: The combination of 1.9 Joules/cm(2) UVA and 150 mu mol/L S-59, AMT, or 8-MOP induced 12.0 +/- 3.0, 6.0 +/- 0.9, and 0.7 psoralen adducts per 1,000 bp DNA, respectively. (4) Replication competence: Polymerase chain reaction (PCR) amplification of small genomic DNA sequences (242-439 bp) after PCT was inhibited. The degree of PCR amplification inhibition correlated with the level of adduct formation (S-59 > AMT > 8-MOP). In contrast, 2,500 cGy gamma radiation, a dose that inactivates >5 log(10) of T cells in blood products, had minimal effect on cytokine synthesis and did not induce sufficient DNA strand breaks to inhibit PCR amplification of the same small DNA sequences. These results demonstrate that leukocytes are sensitive to PCT with psoralens and among the psoralens tested S-59 is the most effective. Therefore, PCT has the potential to reduce the incidence of leukocyte-mediated adverse immune reactions associated with platelet transfusion. (C) 1998 by The American Society of Hematology.