Cytokine levels as performance indicators for white blood cell reduction of platelet concentrates.
OBJECTIVE
With the implementation of universal white blood cell (WBC) reduction in the UK, in-process WBC-reduction filters for pooled buffy coat (BC)-derived platelet concentrates (PCs) and apheresis methods are used routinely for the production of WBC-reduced PCs. While these strategies meet the specification for WBC reduction (< 5 x 10(6) WBCs/unit), the products from these processes may differ depending on the process employed and its performance. The aim of this study was therefore to investigate whether PCs prepared using various WBC-reduction processes are sufficiently depleted of WBCs to limit cytokine accumulation during storage and to assess if cytokine levels detected in platelet products can serve as indicators of acceptable platelet activation as a result of the WBC-reduction process.


METHODS
We measured the levels of cytokines predominantly derived from WBCs [e.g. interleukin-8 (IL-8)] and platelets [e.g. regulated on activation, normal, T-cell expressed, and secreted (RANTES) and transforming growth factor-beta(1) (TGF-beta(1))] under the present experimental conditions in different WBC-reduced PCs, i.e. PCs prepared from three different WBC-reduction filters and control non-filtered PCs using pooled BCs from the same donors and three apheresis types. Supernatant plasma was collected at the beginning (day 1) and end (day 5) of the shelf life of each PC, and the cytokine content was determined using appropriate enzyme-linked immunosorbent assays (ELISAs). Process efficiency was assessed by platelet yield and residual WBC count.


RESULTS
We found that products from the apheresis process involving a filtration step (Haemonetics MCS+) showed a lower cytokine content on both day 1 and day 5 in comparison with the fluidized bed (COBE Spectra) or elutriation (Amicus) processes. WBC reduction of BC-PCs of the same origin using three different filters showed comparable levels of cytokines on day 1 in all units. After storage for 5 days, the levels of IL-8 remained essentially unchanged in filtered BC-PCs but increased by more than threefold in control non-filtered BC-PCs, suggesting IL-8 release by residual WBCs present in the control PCs. The concentration of platelet-derived cytokines such as RANTES and TGF-beta(1), however, increased significantly in all filtered and control non-filtered PCs during the storage period.


CONCLUSIONS
These results show that markers of cytokine release from both WBCs and platelets are useful indicators of the performance and efficacy of the WBC-reduction process and of platelet quality.
Wadhwa
Meenu
M
Division of Immunobiology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Herts EN6 3QG, UK. mwadhwa@nibsc.ac.uk
Krailadsiri
P
P
Dilger
P
P
Gaines Das
R
R
Seghatchian
M J
MJ
Thorpe
R
R
D015415:Biomarkers / Q000032:analysis; D001792:Blood Platelets / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D016207:Cytokines / Q000032:analysis*; D005374:Filtration; D006801:Humans; D007958:Leukocyte Count; D007962:Leukocytes / Q000378:metabolism; D015539:Platelet Activation; D017713:Platelet Transfusion / Q000592:standards*; D010983:Plateletpheresis; D011786:Quality Control; D013997:Time Factors
D015415:Biomarkers; D016207:Cytokines