title: Differential sensitivities of pathogens in red cell concentrates to Tri-P(4)-photoinactivation.
abstract: OBJECTIVE
  Photodynamic treatment (PDT) with the cationic porphyrin, mono-phenyl-tri-(N-methyl-4-pyridyl)-porphyrin chloride [Tri-P(4)], has previously been shown to be effective at inactivating vesicle stomatitis virus (VSV) in red cell concentrates (RCC) with limited damage to red blood cells (RBC). The aim of this study was to determine the pathogen-inactivating capacity of PDT with Tri-P(4) for a broader range of pathogens and to establish the associated effect on in vitro RBC quality.
  
  
  METHODS
  A series of viruses and bacteria was spiked into 60% RCC. Pathogen inactivation was determined after PDT with 25 microm Tri-P(4) and red light up to 360 kJ/m2. Human immunodeficiency virus (HIV)-infected cells were evaluated for cell death induction, and RCC were analysed for the induction of haemolysis and ATP content.
  
  
  RESULTS
  For the lipid-enveloped viruses bovine viral diarrhoea virus, HIV and pseudorabies virus, and for the Gram positive bacterium, Staphylococcus aureus, and the Gram-negative bacteria, Pseudomonas aeruginosa and Yersinia enterolitica, inactivation of > or = 5 log10 was measured after 60 min of PDT with Tri-P(4). The required treatment time to achieve this level of inactivation was four times longer than required for VSV. For cell-associated HIV, only 1.7 log10 of inactivation was found, despite clear induction of cell death of HIV-infected cells. The non-enveloped virus, canine parvovirus, was completely resistant to the treatment. PDT of RCC with Tri-P(4) for 60 min, and subsequent storage in AS-3, resulted in 4% haemolysis after 35 days of storage. The ATP content of untreated and treated RBC declined with similar kinetics during storage.
  
  
  CONCLUSIONS
  PDT of RCC with Tri-P(4) for 60 min inactivates a wide range of pathogens, but not cell-associated HIV and a non-enveloped virus, and compromises RBC quality. This reduces the suitability of PDT with Tri-P(4) for red cell sterilization. Therefore, further improvements in the treatment procedures to potentiate pathogen inactivation and to preserve RBC integrity will be required to generate an effective treatment for sterilizing RCC.
authors.affiliation: Department of Research and Development, Sanquin Blood Bank Southwest, Leiden, the Netherlands. l.trannoy@zonnet.nl
authors.forename: L L
authors.initials: LL
authors.lastname: Trannoy
authors.forename: F G
authors.initials: FG
authors.lastname: Terpstra
authors.forename: D
authors.initials: D
authors.lastname: de Korte
authors.forename: J W M
authors.initials: JW
authors.lastname: Lagerberg
authors.forename: A J
authors.initials: AJ
authors.lastname: Verhoeven
authors.forename: A
authors.initials: A
authors.lastname: Brand
authors.forename: F A C
authors.initials: FA
authors.lastname: van Engelenburg
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D001793:Blood Preservation / Q000379:methods; D016923:Cell Death; D017707:Erythrocyte Transfusion / Q000009:adverse effects; D004912:Erythrocytes / Q000187:drug effects / Q000382:microbiology / Q000821:virology; D006414:Hematoporphyrin Photoradiation / Q000379:methods*; D006801:Humans; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011166:Porphyrins / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D013211:Staphylococcus aureus / Q000187:drug effects; D013242:Sterilization / Q000379:methods*; D038301:Virus Inactivation; D014780:Viruses / Q000187:drug effects; D015008:Yersinia enterocolitica / Q000187:drug effects
chemical_list: C508221:5-phenyl-10,15,20-tris(N-methyl-4-pyridyl)porphyrin chloride; D000900:Anti-Bacterial Agents; D000998:Antiviral Agents; D017319:Photosensitizing Agents; D011166:Porphyrins