Abstract
Aim
Extracorporeal photochemotherapy (ECP) is emphasized chiefly as it has a high safety
profile. However, the genotoxic effects of ECP are not known. This experimental study
aimed to assess the potential genotoxic impact of ECP treatment by the AKLIDES system,
a new generation standardized and automated evaluation method.
Materials and Methods
Buffy coats were obtained from the blood of 26 healthy volunteers, and ECP was applied
to 2 j/cm2 UV-A for two hours. After the DNA isolation procedure, all slides were
stained with DAPI to visualize lymphocytes, FITC for visualization of damage foci
marker (γH2AX), and APC for visualization of repair foci marker (53BP1). With the
AKLIDES imaging system, all parameters were evaluated.
Results
Median damage marker Foci γ-H2AX before and after ECP were 11.42 and 18.65 arbitrary
units, respectively (p = 0.153). Median repair marker foci 53BP1 (repair biomarker)
before and after ECP were measured as 4.17 and 6.7 arbitrary units. The difference
was also not statistically significant (p = 0.088). Although 58 % of cells were affected
by ECP irradiation, as shown by FITC fluorescent staining, no statistical difference
was found in any genotoxicity parameters.
Conclusion
We found an increase in the foci γ-H2AX parameter, one of the objective indicators
of DNA breaks, and an increase in the foci 53BP1 parameter, which indicates the post-damage
repair mechanisms after ECP. However, further in vitro, and in vivo studies are needed
with large sample volumes to demonstrate the significance.
Abbreviations:
ECP (Extracorporeal photochemotherapy), UV-A (Ultraviolet-A), DSBs (Double-strand breaks), γH2AX (Phosphorylated histone H2AX), 51BP1 (p53-binding protein), 8-MOP (8-Methoxsalen), FTIC (Fluorescein isothiocyanate), DAPI (4′,6-diamidino-2-phenylindole), JACIE (The Joint Accreditation Committee ISCT-Europe & EBMT)Keywords
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Article info
Publication history
Published online: May 25, 2022
Accepted:
May 24,
2022
Received in revised form:
May 17,
2022
Received:
April 4,
2022
Identification
Copyright
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