Analysis of Oxidative Burst Capacity of Neutrophils in Buffy-Coat Concentrates

Salih Haldun Bal; Diğdem Yöyen Ermiş; Hilal Vardar; İrem Özveri; Yasemin Heper; Haluk Barbaros Oral

Article Details

Analysis of Oxidative Burst Capacity of Neutrophils in Buffy-Coat Concentrates

Authors:

Salih Haldun Bal | Diğdem Yöyen Ermiş | Hilal Vardar | İrem Özveri | Yasemin Heper | Haluk Barbaros Oral

Publisher:

Bursa Uludağ University

Main Topics:

İmmünoloji (Diğer)

Journal:

Uludağ Üniversitesi Tıp Fakültesi Dergisi Vol.51 Issue 2, pp.175-180

Document Type:

Article

Publication Date:

8/2025

Document Language:

English

Document ID (Org No.):

2001350310

pISSN:

1300-414X

Keywords:

Abstract:

 Buffy-Coat concentrates BFCs can be transfused to provide neutrophil support in patients with febrile neutropenia. In this study, the oxidative burst capacity of neutrophils in BFC was investigated in 12 hours-stored BFCs n16. Samples taken from healthy volunteers n7 fresh EDTA; fEDTA and taken before donation from blood donors who donated BFCs n16 donor EDTA; dEDTA were included. fEDTA and dEDTA samples were used to compare the effect of time. Nitroblue tetrazolium NBT test was performed to evaluate the oxidative burst. Different titrations 1:16, 1:32, 1:64, 1:128 of phorbol myristate acetate PMA were used. BFC samples without PMA addition were used as negative control NC. While similar results were obtained in BFC samples with fEDTA samples in NC, oxidative activity was statistically high in the dEDTA samples. These results suggested that neutrophils in dEDTA may be spontaneously activated. In all PMA titrations except 1:64 PMA titers, the oxidative burst capacity in dEDTA and BFC samples was significantly lower than that in fEDTA samples. Also, oxidative burst significantly decreased in fEDTA-dEDTA-BFC in all PMA titrations. Spontaneous burst was detected in dEDTA samples under PMA-free conditions, but not in BFC samples, indicating that the additive solution in BFC protected neutrophils from spontaneous activation. Despite the statistically significant decrease, neutrophils in BFCs can remain functional due to their high oxidative activity at all titers above 75. This indicates that they can function when transfused. This result also shows that we can use BFCs as a source of neutrophils in experiments.

 Buffy-Coat concentrates BFCs can be transfused to provide neutrophil support in patients with febrile neutropenia. In this study, the oxidative burst capacity of neutrophils in BFC was investigated in 12 hours-stored BFCs n16. Samples taken from healthy volunteers n7 fresh EDTA; fEDTA and taken before donation from blood donors who donated BFCs n16 donor EDTA; dEDTA were included. fEDTA and dEDTA samples were used to compare the effect of time. Nitroblue tetrazolium NBT test was performed to evaluate the oxidative burst. Different titrations 1:16, 1:32, 1:64, 1:128 of phorbol myristate acetate PMA were used. BFC samples without PMA addition were used as negative control NC. While similar results were obtained in BFC samples with fEDTA samples in NC, oxidative activity was statistically high in the dEDTA samples. These results suggested that neutrophils in dEDTA may be spontaneously activated. In all PMA titrations except 1:64 PMA titers, the oxidative burst capacity in dEDTA and BFC samples was significantly lower than that in fEDTA samples. Also, oxidative burst significantly decreased in fEDTA-dEDTA-BFC in all PMA titrations. Spontaneous burst was detected in dEDTA samples under PMA-free conditions, but not in BFC samples, indicating that the additive solution in BFC protected neutrophils from spontaneous activation. Despite the statistically significant decrease, neutrophils in BFCs can remain functional due to their high oxidative activity at all titers above 75. This indicates that they can function when transfused. This result also shows that we can use BFCs as a source of neutrophils in experiments.

 Buffy-Coat concentrates BFCs can be transfused to provide neutrophil support in patients with febrile neutropenia. In this study, the oxidative burst capacity of neutrophils in BFC was investigated in 12 hours-stored BFCs n16. Samples taken from healthy volunteers n7 fresh EDTA; fEDTA and taken before donation from blood donors who donated BFCs n16 donor EDTA; dEDTA were included. fEDTA and dEDTA samples were used to compare the effect of time. Nitroblue tetrazolium NBT test was performed to evaluate the oxidative burst. Different titrations 1:16, 1:32, 1:64, 1:128 of phorbol myristate acetate PMA were used. BFC samples without PMA addition were used as negative control NC. While similar results were obtained in BFC samples with fEDTA samples in NC, oxidative activity was statistically high in the dEDTA samples. These results suggested that neutrophils in dEDTA may be spontaneously activated. In all PMA titrations except 1:64 PMA titers, the oxidative burst capacity in dEDTA and BFC samples was significantly lower than that in fEDTA samples. Also, oxidative burst significantly decreased in fEDTA-dEDTA-BFC in all PMA titrations. Spontaneous burst was detected in dEDTA samples under PMA-free conditions, but not in BFC samples, indicating that the additive solution in BFC protected neutrophils from spontaneous activation. Despite the statistically significant decrease, neutrophils in BFCs can remain functional due to their high oxidative activity at all titers above 75. This indicates that they can function when transfused. This result also shows that we can use BFCs as a source of neutrophils in experiments.

 Buffy-Coat konsantreleri BFCler febril nötropenisi olan hastalarda nötrofil desteği sağlamak için transfuze edilmektedir. Bu çalışmada, BFCdeki nötrofillerin oksidatif patlama kapasitesi araştırıldı. 12 saat depolanmış BFCler n16, sağlıklı gönullulerden alınan örnekler n7 taze EDTA; fEDTA ve BFC bağışlayan kan bağışçılarından bağış öncesi alınan örnekler n16 donör EDTA; dEDTA çalışmaya dahil edildi. fEDTA ve dEDTA örnekleri zamanın oksidatif patlama uzerindeki etkisini değerlendirmek amacıyla çalışmaya dahil edildi. Oksidatif patlamayı değerlendirmek için Nitroblue tetrazolium NBT testi farklı titrasyonlarda phorbol miristat asetat PMA ile gerçekleştirildi. PMAnın 1:1, 1:2, 1:4, 1:8 titrasyonlarında oksidatif patlama 95ten yuksek bulunduğundan, çalışmaya 1:16, 1:32, 1:64, 1:128 titrasyonları ile devam edildi. Ayrıca PMA eklenmemiş BFC örnekleri negatif kontrol olarak kullanıldı. Negatif kontroldeki fEDTA örnekleri ile BFC örneklerinde benzer oksidatif patlama duzeyleri belirlenirken, dEDTA örneklerinde oksidatif aktivite istatistiksel olarak yuksek bulundu. Bu sonuçlar dEDTAda bulunan nötrofillerin spontan olarak aktive olabileceğini duşundurdu. dEDTA ve BFC örneklerindeki oksidatif patlama kapasitesi 1:64 PMA titrasyonu hariç tum PMA titrasyonlarında fEDTA örneklerine göre anlamlı duzeyde duşuk bulundu. Ayrıca artan titrasyona bağlı olarak oksidatif kapasitenin tum gruplarda önemli ölçude azaldığı tespit edildi. PMAsız dEDTA örneklerinde spontan patlama tespit edilirken BFC örneklerinde tespit edilmemesi, BFCdeki ek solusyonun nötrofilleri spontan aktivasyondan koruduğunu göstermektedir. fEDTA örneklerine göre anlamlı azalmaya rağmen, BFClerdeki nötrofillerin tum titrelerde 75in uzerinde yuksek oksidatif aktivite göstermeleri işlevsel kalabildiklerini duşunulmektedir. Elde edilen sonuçlar BFC içindeki nötrofillerin transfuzyon yapıldığında işlev görebileceklerini duşundurmekte, ayrıca deneylerde BFCleri nötrofil kaynağı olarak kullanabileceğimizi göstermektedir.

Article Details

Analysis of Oxidative Burst Capacity of Neutrophils in Buffy-Coat Concentrates

Authors:

Salih Haldun Bal | Diğdem Yöyen Ermiş | Hilal Vardar | İrem Özveri | Yasemin Heper | Haluk Barbaros Oral

Publisher:

Bursa Uludağ University

Main Topics:

İmmünoloji (Diğer)

Journal:

Uludağ Üniversitesi Tıp Fakültesi Dergisi Vol.51 Issue 2, pp.175-180

Document Type:

Article

Publication Date:

8/2025

Document Language:

English

Document ID (Org No.):

2001350310

pISSN:

1300-414X

Buffy-Coat Concentrates

Nitroblue Tetrazolium Test

Oxidative Burst

NeutrophilBuffy-Coat Konsantreleri

Nitroblue Tetrazolium Testi

Oksidatif Patlama

Nötrofil

Issues

Article Details

Authors: Salih Haldun Bal | Diğdem Yöyen Ermiş | Hilal Vardar | İrem Özveri | Yasemin Heper | Haluk Barbaros Oral

Publisher: Bursa Uludağ University

Main Topics: İmmünoloji (Diğer)

Journal: Uludağ Üniversitesi Tıp Fakültesi Dergisi Vol.51 Issue 2, pp.175-180

Document Type: Article

Publication Date: 8/2025

Document Language: English

Document ID (Org No.): 2001350310

pISSN: 1300-414X

Issues

Authors:

Salih Haldun Bal | Diğdem Yöyen Ermiş | Hilal Vardar | İrem Özveri | Yasemin Heper | Haluk Barbaros Oral

Publisher:

Bursa Uludağ University

Main Topics:

İmmünoloji (Diğer)

Journal:

Uludağ Üniversitesi Tıp Fakültesi Dergisi Vol.51 Issue 2, pp.175-180

Document Type:

Article

Publication Date:

8/2025

Document Language:

English

Document ID (Org No.):

2001350310

pISSN:

1300-414X