Trannoy L L, Terpstra F G, de Korte D, Lagerberg J W M, Verhoeven A J, Brand A, van Engelenburg F A C
Department of Research and Development, Sanquin Blood Bank Southwest, Leiden, the Netherlands.
Vox Sang. 2006 Aug;91(2):111-8. doi: 10.1111/j.1423-0410.2006.00791.x.
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.
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.
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.
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.
先前已证明，使用阳离子卟啉单苯基三（N - 甲基 - 4 - 吡啶基）氯化卟啉[三 - P（4）]进行光动力治疗（PDT）可有效灭活红细胞浓缩液（RCC）中的水泡性口炎病毒（VSV），且对红细胞（RBC）的损伤有限。本研究的目的是确定使用三 - P（4）进行PDT对更广泛病原体的病原体灭活能力，并确定其对体外RBC质量的相关影响。
将一系列病毒和细菌加入到60%的RCC中。使用25微摩尔三 - P（4）和高达360 kJ/m²的红光进行PDT后，测定病原体灭活情况。评估感染人类免疫缺陷病毒（HIV）的细胞的细胞死亡诱导情况，并分析RCC的溶血诱导情况和ATP含量。
对于包膜病毒牛病毒性腹泻病毒、HIV和伪狂犬病病毒，以及革兰氏阳性菌金黄色葡萄球菌和革兰氏阴性菌铜绿假单胞菌和小肠结肠炎耶尔森菌，使用三 - P（4）进行60分钟的PDT后，测得灭活率≥5 log10。达到该灭活水平所需的治疗时间是VSV所需时间的四倍。对于细胞相关的HIV，尽管明显诱导了HIV感染细胞的细胞死亡，但仅发现1.7 log10的灭活率。无包膜病毒犬细小病毒对该治疗完全耐药。用三 - P（4）对RCC进行60分钟的PDT，随后在AS - 3中储存，储存35天后溶血率为4%。未处理和处理后的RBC在储存期间ATP含量以相似的动力学下降。
用三 - P（4）对RCC进行60分钟的PDT可灭活多种病原体，但不能灭活细胞相关的HIV和一种无包膜病毒，且会损害RBC质量。这降低了用三 - P（4）进行PDT用于红细胞灭菌的适用性。因此，需要进一步改进治疗程序，以增强病原体灭活并保持RBC完整性，从而产生一种有效的RCC灭菌治疗方法。
