Blood sterilisation by PDT: study of photosensitiser toxicity on red blood cells (original) (raw)
Related papers
Transfusion, 1999
BACKGROUND: Phthalocyanines are useful sensitizers for photodynamic sterilization of red cell concentrates. Various lipid-enveloped viruses can be inactivated with only limited red cell damage. Because white cells are involved in the immunomodulatory effects of blood transfusions, the study of the effect of photodynamic treatment on these cells is imperative. STUDY DESIGN AND METHODS: White cell-enriched red cell suspensions were photodynamically treated with either the hydrophobic Pc4 (HOSiPcOSi-(CH 3 ) 2 (CH 2 ) 3 N(CH 3 ) 2 ) or water-soluble aluminum phthalocyanine tetrasulfonate (AlPCS 4 ) under virucidal conditions. Viability of white cell subpopulations on Days 0, 1, and 4 after treatment was determined by fluorescenceactivated cell sorting by flow cytometric analysis of propidium iodide uptake. Apoptosis induction was studied by DNA ladder formation and staining for an early marker of apoptosis (annexin V). RESULTS: Treatment with Pc4 causes a significant decrease in cell viability of all white cells, as shown by prodidium iodide uptake. Monocytes and granulocytes are the most sensitive, and lymphocytes are relatively more resistant. Some of the cells die by apoptosis, which is induced within 30 minutes after treatment. Treatment with AlPCS 4 damages only monocytes; other cell populations are not affected. CONCLUSIONS: Physicochemical properties of the photosensitizers partly determine their effect on white cells. Differences in intracellular localization are likely to be responsible for the effects observed. ABBREVIATIONS: AlPcS 4 = aluminum phthalocyanine tetrasulfonate; FCS = fetal calf serum; FITC = fluoresceinisothiocyanate; HSA = human serum albumin; PBMNC(s) = peripheral blood mononuclear cell(s); PBS = phosphate-buffered saline; Pc4 = HOSiPcOSi-(CH 3 ) 2 (CH 2 ) 3 N(CH 3 ) 2 ; PDT = photodynamic treatment; PI = propidium iodide; RBC(s) = red cell(s); TA-GVHD = transfusion-associated graft-versus-host disease; VSV = vesicular stomatitis virus; WBC(s) = white cell(s).
Photochemistry and Photobiology, 1997
Abstract— Photodynamic treatment (PDT) using phthalocyanines and red light appears to be a promising procedure for decontamination of red blood cell (RBC) concentrates for transfusion. A possible complication of this treatment may be induced aggregation of RBC. The production of RBC aggregates was measured with a novel computerized cell flow properties analyzer (CFA). The PDT of RBC concentrates with sulfonated aluminum phthalocy-anine (AIPcS4) and the silicon phthalocyanine Pc 4 under virucidal conditions markedly enhanced RBC aggregation and higher shear stress was required to disperse these aggregates. The clusters of cells were huge and abnormally shaped, unlike the rouleaux formed by untreated RBC. This aggregation was prevented when a mixture of antioxidants was included during PDT. Addition of the antioxidants after PDT reduced aggregation only partially. It is concluded that inclusion of antioxidants during PDT of RBC concentrates prior to transfusion may reduce or eliminate the hemodynamic risk that the virucidal treatment may present to the recipient.
Photodynamic Inactivation of Viruses and Its Application for Blood Banking
Baylor University Medical Center Proceedings, 1988
A better understanding ofimmunological donor-recipient incompatibility has encouraged the use of blood and its components (1). In 1985, an estimated 10 million units of whole blood were processed by over 800 blood banks in the United States and 14 million units of blood components were transfused (2). This increase reflects major advances in managing trauma, hemorrhagic and neoplastic disorders, and recipients of bone marrow or solid organ transplants. Use of blood products still involves significant risk to the recipient because of the potential for transmitting infectious agents. Hepatitis B virus (HBV); cytomegalovirus (CMV); Epstein-Barr virus (EBV); herpes simplex virus (HSV); human T cell Iymphotropic virus, type 1 (HTLV-1); non-A, non-B (NANB) hepatitis virus; human immunodeficiency virus (HlV); and malaria can be transmitted by blood transfusion (3-10). Screening of donors and serologic testing reduce the risk, but these precautions still provide insufficient protection since detectable HlV antibody may not be present during the early stage of infection (11). Because a requisite to killing infectious agents in banked blood is the maintenance ofintegrity and full function ofblood elements after prophylactic measures are taken, many antiviral agents are unsuitable for this use. One approach to prophylactic treatment-photodynamic inactivation-uses no potent cytotoxic agents but does offer potential target selectivity (12). The photosensitizer used for selective destruction of tumor cells and viruses is hematoporphyrin derivative (Hpd), a complex mixture of ringed tetrapyrroles.
Differential sensitivities of pathogens in red cell concentrates to Tri-P(4)-photoinactivation
Vox Sanguinis, 2006
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.
The application of photosensitisers to tropical pathogens in the blood supply
Photodiagnosis and Photodynamic Therapy, 2011
The onset of the HIV pandemic led both to significant alterations in blood collection and screening practice and to the development of more sophisticated methods of inactivation of infectious agents from the blood supply. Photodynamic (i.e. light activated) pathogen inactivation is one such method currently in limited use in various European states. The approach is based on the generation of a burst of reactive oxygen and nitrogen species, resulting in the activation of several cell death mechanisms. However, its application to tropical pathogens is perhaps less appreciated, despite the fact that the efficacies of photoantimicrobial agents such as methylene blue were originally reported following screening against organisms such as Trypanosoma cruzi and viruses such as those responsible for dengue and yellow fever.
Use of laser-UV for inactivation of virus in blood products
Blood, 1987
Inactivation of virus by UV radiation was examined as a potential method for sterilization of blood products. Samples of attenuated poliovirus, platelets and plasma were uniformly irradiated with a XeCl excimer laser that delivered 40 nsec pulses of UV at 308 nm (UVB308). Intensities and exposure does were varied from 0.11 to 1.40 MW/cm2 and 0.51 to 56.0 J/cm2, respectively. In studies conducted with low intensity UVB308 (less than or equal to 0.17 MW/cm2), using exposure doses greater than or equal to 10.8 J/cm2, it was possible to inactivate poliovirus by 4 to 6 log10. Platelets irradiated with doses less than or equal to 21.5 J/cm2 exhibited minimal damage as assessed by aggregation activity and spontaneous release of serotonin. Examination of the coagulation activity of irradiated plasma indicated that exposure doses less than or equal to 21.5 J/cm2 resulted in less than 20% increase in prothrombin and partial thromboplastin times. The use of UVB308 at a higher intensity (1.4 MW...
Iranian Journal of Virology, 2013
Background and Aims: Fresh Frozen Plasma (FFP) is one of blood components. The risk of transmission of viruses from blood components regardless selection of blood donors and screening donated blood still remains. There are several methods for viral inactivation. In this study methylene blue(MB) photo inactivation process was used for inactivating viruses Materials and Methods: In this study Methylene Blue(MB) was used in final concentration of 1µM. Infected Fresh Frozen Plasma (FFP) illuminated by 143Pieces (PCs) of 1 w red Light Emitting Diodes (LEDs) from two side for 10,15 and 30 minutes and shacked 30 cycle in minutes. the central wavelength of these LED is 627 nm with 20 nm Full Width at Half Maximum (FWHM).Herpes simplex virus-1(HSV-1) and vesicular stomatitis virus(VSV) were used as model viruses to evaluated illumination effects on viral inactivation. level of fresh frozen plasma (FFP) coagulation factors such as fibrinogen, FV, FVIII, protein C, antitrombin measured pre and post illumination. Results: Initial HSV-1 and VSV titer were calculated to be 107 and 106.5 TCID50/ml, respectively .The level of viral inactivation was expressed as log-reduction. Titer reduction of HSV for 10, 15 and 30 minutes irradiation with shaking was > 6, ≥ 7 and ≥ 7 log, respectively. the ratio of coagulation factors activity remaining unchanged after pathogen inactivation with MB calculated. illumination had a major effects on the mean levels of fibrinogen and FVIII. Significant differences between level of factors before and after illumination were evaluated with a t test for paired samples. No significant differences were seen in the FFP coagulation factors before and after illumination.(P˃0.05) Conclusions: As results show the optimum time for viral inactivation were adjusted to be 15 minutes. Due to the reduction of virus titer at various times, agitation with illumination is effective.
Transfusion, 2005
BACKGROUND: Viral contamination of platelet (PLT) concentrates can result in transfusion-transmitted diseases. A photochemical treatment (PCT) process with amotosalen-HCl and long-wavelength ultraviolet light (UVA), which cross-links nucleic acids, was developed to inactivate viruses and other pathogens in PLT concentrates. STUDY DESIGN AND METHODS: High titers of pathogenic or blood-borne viruses, representing 10 different families, were added to single-donor PLT concentrates containing 3.0 ¥ 10 11 to 6.0 ¥ 10 11 PLTs in approximately 300 mL of 35 percent plasma and 65 percent PLT additive solution (InterSol). After PCT with 150 m mol per L amotosalen and 3 J per cm 2 UVA, residual viral infectivity was assayed by sensitive cell culture or animal systems. RESULTS: Enveloped viruses were uniformly sensitive to inactivation by PCT whereas nonenveloped viruses demonstrated variable inactivation. Log reduction of enveloped viruses for cell-free HIV-1 was > 6.
Avicenna Journal of Medical Biotechnology, 2015
Background: This study investigated the effects of Riboflavin (RB) combined with different doses of UV on Platelet Concentrate (PC) which was infected by three models of virus. Platelet quality after treatment was also assessed. Methods: Three models of virus used in this study were Vesicular Stomatitis Virus (VSV), Herpes Simplex Virus (HSV), and Polio virus, which were added to PC. After photochemical treatment with RB and UV light, residual viral infectivity was titrated using 50% Tissue Culture Infective Dose (TCID 50)/ml. This treatment was done with concentration of 50 μM of RB and different doses of UV light (0.24, 0.48, 0.97, 1.29 J/cm 2). Platelet quality was assessed by measuring pH, Lactate Dehydrogenase (LDH), MTT assay and cell count after treatments and during 4 days of storage against control groups. Results: Concentration of 50 μM RB with combination of 1.29 J/cm 2 dose of UV resulted in the highest titer reduction of VSV (4 log 10) and HSV (4.26 log 10) and lowest titer reduction of Polio virus (2.6 log 10). No significant difference was observed between different doses in comparison with control groups. In all treatment groups, the storage stability of platelets in PC was in the acceptable range in comparison with control group. Conclusion: This study indicated that RB/UV treatment was a promising pathogen reduction technique in PC and had limited effects on platelet quality. However, further optimization of this method is necessary to deal with blood-borne viruses like nonenveloped viruses.
Transfusion, 2007
BACKGROUND: Pathogen contamination, causing transfusion-transmitted diseases, is an ongoing concern in transfusion of cellular blood products. In this explorative study, the pathogen-inactivating capacity of UVC irradiation in platelet (PLT) concentrates was investigated. The dose dependencies of inactivation of several viruses and bacteria were compared with the effect on PLT quality. STUDY DESIGN AND METHODS: The potential of UVC irradiation was studied with a range of lipidenveloped (LE) and non-lipid-enveloped viruses (NLE) and bacteria. LE viruses were bovine viral diarrhea virus (BVDV), human immunodeficiency virus (HIV), pseudorabies virus (PRV), transmissible gastroenteritis virus (TGEV), and vesicular stomatitis virus (VSV). NLE viruses were canine parvovirus (CPV) and simian virus 40 (SV40). Bacteria were Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, and Bacillus cereus. After spiking and irradiation, samples were tested for residual infectivity and reduction factors (RFs) were calculated. Furthermore, the effect of UVC irradiation on PLT quality was determined by measuring in vitro quality variables. RESULTS: A UVC dose of 500 J per m 2 resulted in acceptable PLT quality (as measured by pH, lactate production, CD62P expression, and exposure of phosphatidylserine) and high RFs (>4 log) for CPV, TGEV, VSV, S. epidermidis, S. aureus, and E. coli. Intermediate RFs (approx. 3 log) were observed for BVDV, PRV, and B. cereus. Low RFs (approx. 1 log) were found for HIV and SV40. No differences in virus reduction were observed between cell-free and cell-associated virus. CONCLUSION: UVC irradiation is a promising pathogen-reducing technique in PLT concentrates, inactivating bacteria, and a broad range of viruses (with the exception of HIV) under conditions that have limited effects on PLT quality. Further optimization of the UVC procedure, however, is necessary to deal with bloodborne viruses like HIV. ABBREVIATIONS: BVDV = bovine viral diarrhea virus; CA virus = cell-associated virus; CPV = canine parvovirus; LE virus = lipid-enveloped virus; NLE virus = non-lipidenveloped virus; PC(s) = platelet concentrate(s); PRV = pseudorabies virus; PS = phosphatidylserine; RF(s) = reduction factor(s); SV40 = simian virus 40; TGEV = transmissible gastroenteritis virus; VSV = vesicular stomatitis virus.