TY - JOUR
T1 - Instant intra-operative neutropenia despite the emergence of banded (CD16dim/CD62Lbright) neutrophils in peripheral blood - An observational study during extensive trauma-surgery in pigs
AU - Teuben, Michel
AU - Heeres, Marjolein
AU - Blokhuis, Taco
AU - Hollman, Arne
AU - Vrisekoop, Nienke
AU - Tan, Edward
AU - Pfeifer, Roman
AU - Pape, Hans-Christoph
AU - Koenderman, Leo
AU - Leenen, Luke P H
N1 - Funding Information:
We would like to thank both the local and international faculty-members of the Definitive Surgical Trauma Care?-Course in Nijmegen (The Netherlands) as well as the members of the Animal Research Facility Nijmegen (The Netherlands) and the participating members of the mobile laboratory of the Royal Dutch Army for their support during the preparation and execution of the experiments. The current study was partly funded by the Alexandre Suermanfund (University Medical Centre Utrecht, MT). The funding source had not role in the design/execution of this study.
Funding Information:
We would like to thank both the local and international faculty-members of the Definitive Surgical Trauma Care™-Course in Nijmegen (The Netherlands) as well as the members of the Animal Research Facility Nijmegen (The Netherlands) and the participating members of the mobile laboratory of the Royal Dutch Army for their support during the preparation and execution of the experiments. The current study was partly funded by the Alexandre Suermanfund (University Medical Centre Utrecht, MT). The funding source had not role in the design/execution of this study.
Publisher Copyright:
© 2020
PY - 2021/3
Y1 - 2021/3
N2 - Introduction: Deregulation of polymorphonuclear neutrophils (PMNs) is an essential step in the development of inflammatory complications upon trauma. Different neutrophil subtypes have been identified recently, however, the role of neutrophil subtypes in immunoregulation upon trauma is unclear. We hypothesize that extensive trauma surgery causes instant progressive heterogeneity of the blood neutrophil pool, and increased appearance of young (CD16
dim/CD62L
bright) neutrophils in peripheral blood. Material and methods: A standardized extensive thoraco-abdominal porcine trauma surgery model was utilized, and 12 animals were included. Blood was collected at defined timepoints and neutrophil numbers and subtypes were studied by flowcytometry. Neutrophil subtypes were identified by differences in cell surface expression levels of CD16 (FcγRIII) and CD62L (L-selectin). Porcine neutrophil subtypes were further characterized after flow sorting. RESULTS: Eleven animals survived the 3-hour surgical protocol. Neutrophil numbers dropped significantly from a mean of 8,6 ± 3,5 × 10
6 to 2,4 ± 1,8 × 10
6 cells/ml during 180 min, (p<0.001). Simultaneously, the blood PMN population became increasingly heterogeneous due to the appearance of new neutrophil subtypes. Cell sorting experiments and cytological analysis revealed that these porcine subtypes had specific morphological characteristics, mimicking their human counterparts. At baseline, 88% ± 1 percent of circulatory PMNs comprised of mature (CD16
bright/CD62L
bright) PMNs, while at 3 h the blood PMN pool consisted of 59% ± 2 percent of mature subtypes (p<0.001). Despite a marked drop in neutrophil levels during surgery, absolute and relative numbers of banded (CD16
dim/CD62L
bright) neutrophils continued to rise throughout surgery. Conclusion: Standardized extensive trauma surgery was associated with instant progressive neutropenia and increased heterogeneity of the blood neutrophil pool. Furthermore, three different neutrophil subsets in peripheral porcine blood were identified over the course of surgery. Further studies should clarify their precise role in the development of early organ failure upon extensive trauma surgery. This for the first time exemplifies experimentally the time constraints and impact of damage control surgery after severe trauma.
AB - Introduction: Deregulation of polymorphonuclear neutrophils (PMNs) is an essential step in the development of inflammatory complications upon trauma. Different neutrophil subtypes have been identified recently, however, the role of neutrophil subtypes in immunoregulation upon trauma is unclear. We hypothesize that extensive trauma surgery causes instant progressive heterogeneity of the blood neutrophil pool, and increased appearance of young (CD16
dim/CD62L
bright) neutrophils in peripheral blood. Material and methods: A standardized extensive thoraco-abdominal porcine trauma surgery model was utilized, and 12 animals were included. Blood was collected at defined timepoints and neutrophil numbers and subtypes were studied by flowcytometry. Neutrophil subtypes were identified by differences in cell surface expression levels of CD16 (FcγRIII) and CD62L (L-selectin). Porcine neutrophil subtypes were further characterized after flow sorting. RESULTS: Eleven animals survived the 3-hour surgical protocol. Neutrophil numbers dropped significantly from a mean of 8,6 ± 3,5 × 10
6 to 2,4 ± 1,8 × 10
6 cells/ml during 180 min, (p<0.001). Simultaneously, the blood PMN population became increasingly heterogeneous due to the appearance of new neutrophil subtypes. Cell sorting experiments and cytological analysis revealed that these porcine subtypes had specific morphological characteristics, mimicking their human counterparts. At baseline, 88% ± 1 percent of circulatory PMNs comprised of mature (CD16
bright/CD62L
bright) PMNs, while at 3 h the blood PMN pool consisted of 59% ± 2 percent of mature subtypes (p<0.001). Despite a marked drop in neutrophil levels during surgery, absolute and relative numbers of banded (CD16
dim/CD62L
bright) neutrophils continued to rise throughout surgery. Conclusion: Standardized extensive trauma surgery was associated with instant progressive neutropenia and increased heterogeneity of the blood neutrophil pool. Furthermore, three different neutrophil subsets in peripheral porcine blood were identified over the course of surgery. Further studies should clarify their precise role in the development of early organ failure upon extensive trauma surgery. This for the first time exemplifies experimentally the time constraints and impact of damage control surgery after severe trauma.
KW - CD16
KW - CD62L
KW - Cell mobilization
KW - Neutrophil subsets
KW - Porcine
KW - Trauma
UR - http://www.scopus.com/inward/record.url?scp=85096085117&partnerID=8YFLogxK
U2 - 10.1016/j.injury.2020.11.018
DO - 10.1016/j.injury.2020.11.018
M3 - Article
C2 - 33208273
SN - 0020-1383
VL - 52
SP - 426
EP - 433
JO - Injury
JF - Injury
IS - 3
ER -