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Updated: Aug 8, 2026
Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets
Published on: March 28, 2014
[Lymphocyte interactions in platelet concentrates from buffy coats. Determination of proliferating cells using Ki-67
M Müller-Steinhardt1, H Klüter, H Kirchner
1Institute für Immunologie und Transfusionsmedizin, Medizinische Universität zu Lübeck, Deutschland.
Insights
Platelet concentrates (PC) from pooled buffy coats do not show lymphocyte activation during storage. This research indicates that transfusing activated lymphocytes is highly unlikely, ensuring patient safety.
Area of Science:
- Immunology
- Transfusion Medicine
- Cell Biology
Context:
- Platelet concentrates (PC) are prepared from pooled buffy coats.
- Concerns exist regarding potential lymphocyte activation during PC storage.
- Activated lymphocytes, particularly HLA-DR-positive cells, can trigger mixed lymphocyte reactions (MLR).
Purpose:
- To investigate leukocyte contamination in PCs.
- To quantify proliferating leukocytes using the KI-67 monoclonal antibody.
- To assess allogeneic lymphocyte activation during PC storage.
Summary:
- Leukocyte content in PCs was determined to be (0.07 +/- 0.05) x 10(9).
- Immunostaining with KI-67 showed only single activated leukocytes immediately after preparation and on days 1, 3, and 5.
- No significant increase in proliferating cells indicative of MLR was observed.
Impact:
- The study concludes that allogeneic lymphocyte activation does not occur in PCs during storage.
- Transfusion of activated lymphocytes from these PCs is considered extremely unlikely.
- This finding has implications for transfusion safety and patient outcomes.
Abstract:
The preparation of platelet concentrates (PC) from pooled buffy coats has raised concern as to whether activation of lymphocytes may take place during the period of storage. Lymphocytes and HLA-DR-positive cells are of particular relevance, as they may induce a mixed lymphocyte reaction (MLR). Therefore we investigated the degree of leukocyte contamination and determined the quantity of proliferating leukocytes using the monoclonal antibody KI-67. The content of total leukocytes was (0.07 +/- 0.05) x 10(9). Immunostaining of KI-67-positive cells proved that, directly after preparation, only single leukocytes were activated. On days 1, 3 and 5 after preparation the results were identical. Isolated cells were found to be KI-67-positive but there was no significant increase in proliferating cells as would have been expected in an MLR. We conclude that there is no allogenic activation of lymphocytes taking place in these PC during storage. Therefore transfusion of activated lymphocytes seems to be extremely unlikely.
