Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock

Severe injury and hemorrhagic shock (HS) result in multiple changes to hematopoietic differentiation, which contribute to the development of immunosuppression and multiple organ failure (MOF). Understanding the changes that take place during the acute injury phase may help predict which patients will develop MOF and provide potential targets for therapy. Obtaining bone marrow from humans during the acute injury phase is difficult so published data are largely derived from peripheral blood samples, which infer bone marrow changes that reflect the sustained inflammatory response. This preliminary and opportunistic study investigated leucopoietic changes in rat bone marrow 6 h following traumatic injury and HS. Terminally anesthetized male Porton Wistar rats were allocated randomly to receive a sham operation (cannulation with no injury) or femoral fracture and HS. Bone marrow cells were flushed from rat femurs and immunophenotypically stained with specific antibody panels for lymphoid (CD45R, CD127, CD90, and IgM) or myeloid (CD11b, CD45, and RP‐1) lineages. Subsequently, cell populations were fluorescence‐activated cell sorted for morphological assessment. Stage‐specific cell populations were identified using a limited number of antibodies, and leucopoietic changes were determined 6 h following trauma and HS. Myeloid subpopulations could be identified by varying levels CD11b expression, CD45, and RP‐1. Trauma and HS resulted in a significant reduction in total CD11b + myeloid cells including both immature (RP‐1(−)) and mature (RP‐1+) granulocytes. Multiple B‐cell lymphoid subsets were identified. The total percentage of CD90+ subsets remained unchanged following trauma and HS, but there was a reduction in the numbers of maturing CD90(−) cells suggesting movement into the periphery. © 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

[1]  Paul J. Harrison,et al.  Prehospital immune responses and development of multiple organ dysfunction syndrome following traumatic injury : A prospective cohort , 2019 .

[2]  T. Rasmussen,et al.  Cellular therapies in trauma and critical care medicine: Looking towards the future , 2017, PLoS medicine.

[3]  P. Efron,et al.  Persistent injury-associated anemia: the role of the bone marrow microenvironment. , 2017, The Journal of surgical research.

[4]  B. Brumback,et al.  Human Myeloid-derived Suppressor Cells are Associated With Chronic Immune Suppression After Severe Sepsis/Septic Shock , 2017, Annals of surgery.

[5]  P. Efron,et al.  Clonidine reduces norepinephrine and improves bone marrow function in a rodent model of lung contusion, hemorrhagic shock, and chronic stress , 2017, Surgery.

[6]  M. Kumar,et al.  Impaired hematopoietic progenitor cells in trauma hemorrhagic shock. , 2016, Journal of clinical orthopaedics and trauma.

[7]  S. Gardner,et al.  The Significance and Challenges of Monocyte Impairment: For the Ill Patient and the Surgeon. , 2016, Surgical infections.

[8]  M. Kumar,et al.  Hematopoietic stem cells: Can it be a therapeutic option for the hematopoietic failure in patients with trauma-hemorrhagic shock? , 2016, Journal of emergencies, trauma, and shock.

[9]  L. Koenderman,et al.  The role of neutrophils in immune dysfunction during severe inflammation , 2016, Critical Care.

[10]  Y. Chu,et al.  B cells expressing CD11b effectively inhibit CD4+ T‐cell responses and ameliorate experimental autoimmune hepatitis in mice , 2015, Hepatology.

[11]  G. Esendagli,et al.  Granulocytic subset of myeloid derived suppressor cells in rats with mammary carcinoma. , 2015, Cellular immunology.

[12]  J. Lord,et al.  The impact of trauma on neutrophil function. , 2014, Injury.

[13]  J. Lord,et al.  The systemic immune response to trauma: an overview of pathophysiology and treatment , 2014, The Lancet.

[14]  G. Crooks,et al.  Critical Differences in Hematopoiesis and Lymphoid Development between Humans and Mice , 2013, Journal of Clinical Immunology.

[15]  L. Harsányi,et al.  Experimental Models of Hemorrhagic Shock: A Review , 2013, European Surgical Research.

[16]  J. Pugin,et al.  Innate Immune Functions of Immature Neutrophils in Patients With Sepsis and Severe Systemic Inflammatory Response Syndrome* , 2013, Critical care medicine.

[17]  L. Koenderman,et al.  Immune suppression by neutrophils and granulocytic myeloid-derived suppressor cells: similarities and differences , 2013, Cellular and Molecular Life Sciences.

[18]  I. Pallister,et al.  Altered leucocyte progenitor profile in human bone marrow from patients with major trauma during the recovery phase , 2012, The British journal of surgery.

[19]  T. Billiar,et al.  Role of Macrophages in Mobilization of Hematopoietic Progenitor Cells From Bone Marrow After Hemorrhagic Shock , 2012, Shock.

[20]  M. Huber-Lang,et al.  A New Experimental Polytrauma Model in Rats: Molecular Characterization of the Early Inflammatory Response , 2012, Mediators of inflammation.

[21]  Huanbin Xu,et al.  Bone marrow and the control of immunity , 2011, Cellular and Molecular Immunology.

[22]  S. Lendemans,et al.  A new model of severe hemorrhagic shock in rats. , 2011, Comparative medicine.

[23]  P. Rameshwar,et al.  Beta-blockade prevents hematopoietic progenitor cell suppression after hemorrhagic shock. , 2011, Surgical infections.

[24]  Jun Seita,et al.  Hematopoietic stem cell: self‐renewal versus differentiation , 2010, Wiley interdisciplinary reviews. Systems biology and medicine.

[25]  P. Rameshwar,et al.  Hematopoietic progenitor cell mobilization is mediated through beta-2 and beta-3 receptors after injury. , 2010, The Journal of trauma.

[26]  Bing Li,et al.  Mobilized Human Hematopoietic Stem/Progenitor Cells Promote Kidney Repair After Ischemia/Reperfusion Injury , 2010, Circulation.

[27]  D. Cooper,et al.  IL-7 receptor is expressed on adult pre-B-cell acute lymphoblastic leukemia and other B-cell derived neoplasms and correlates with expression of proliferation and survival markers. , 2010, Cytokine.

[28]  D. Livingston,et al.  Mobilization of bone marrow cells to the site of injury is necessary for wound healing. , 2009, The Journal of trauma.

[29]  Srinivas Nagaraj,et al.  Myeloid-derived suppressor cells as regulators of the immune system , 2009, Nature Reviews Immunology.

[30]  K. Anderson,et al.  B-lineage commitment prior to surface expression of B220 and CD19 on hematopoietic progenitor cells. , 2008, Blood.

[31]  T. Fabian,et al.  Efficacy and safety of epoetin alfa in critically ill patients. , 2007, The New England journal of medicine.

[32]  E. Deitch,et al.  Hematopoietic progenitor cells mobilize to the site of injury after trauma and hemorrhagic shock in rats. , 2007, The Journal of trauma.

[33]  C. Thiemermann.  Beneficial effects of erythropoietin in preclinical models of shock and organ failure , 2007, Critical care.

[34]  A. McGuinness,et al.  Mobilization of endothelial precursor cells: Systemic vascular response to musculoskeletal trauma , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[35]  KA Brown,et al.  Neutrophils in development of multiple organ failure in sepsis , 2006, The Lancet.

[36]  N. Milićević,et al.  T cells are required for the peripheral phase of B‐cell maturation , 2005, Immunology.

[37]  P. Rameshwar,et al.  The impact of a hypercatecholamine state on erythropoiesis following severe injury and the role of IL-6. , 2005, The Journal of trauma.

[38]  Benjamin Geiger,et al.  Resolving and classifying haematopoietic bone‐marrow cell populations by multi‐dimensional analysis of flow‐cytometry data , 2005, British journal of haematology.

[39]  V. V. D. van der Velden,et al.  Immunophenotypic differentiation patterns of normal hematopoiesis in human bone marrow: Reference patterns for age‐related changes and disease‐induced shifts , 2004, Cytometry. Part B, Clinical cytometry.

[40]  Takashi Nagasawa,et al.  Cellular niches controlling B lymphocyte behavior within bone marrow during development. , 2004, Immunity.

[41]  D. Miura,et al.  Flow cytometric analyses on lineage-specific cell surface antigens of rat bone marrow to seek potential myelotoxic biomarkers: status after repeated dose of 5-fluorouracil. , 2004, The Journal of toxicological sciences.

[42]  Pranela Rameshwar,et al.  Bone Marrow Failure Following Severe Injury in Humans , 2003, Annals of surgery.

[43]  M. Heineman,et al.  Total white blood cell counts and LPS-induced TNF alpha production by monocytes of pregnant, pseudopregnant and cyclic rats. , 2003, Journal of reproductive immunology.

[44]  S. Minguet,et al.  The first 3 days of B-cell development in the mouse embryo. , 2002, Blood.

[45]  T. Scalea,et al.  Persistent systemic inflammatory response syndrome is predictive of nosocomial infection in trauma. , 2002, The Journal of trauma.

[46]  T. Scalea,et al.  Back to basics: validation of the admission systemic inflammatory response syndrome score in predicting outcome in trauma. , 2001, Journal of Trauma.

[47]  M. D. de Broe,et al.  Identification and kinetics of leukocytes after severe ischaemia/reperfusion renal injury. , 2000, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[48]  F. Kroese,et al.  The origin of marginal zone B cells in the rat , 1999, European journal of immunology.

[49]  E. Kreuzfelder,et al.  HLA-DR expression and soluble HLA-DR levels in septic patients after trauma. , 1999, Annals of surgery.

[50]  M. Durlik,et al.  Surgical Trauma Evokes a Rise in the Frequency of Hematopoietic Progenitor Cells and Cytokine Levels in Blood Circulation , 1998, European Surgical Research.

[51]  J. Liesveld,et al.  Expression of interleukin-7 receptor by lineage-negative human bone marrow progenitors with enhanced lymphoid proliferative potential and B-lineage differentiation capacity. , 1997, Blood.

[52]  M. Ogawa,et al.  Differentiation and proliferation of hematopoietic stem cells. , 1993, Blood.

[53]  T. Ulich,et al.  The hematopoietic and mature blood cells of the rat: their morphology and the kinetics of circulating leukocytes in control rats. , 1991, Experimental hematology.

[54]  J D Kemp,et al.  Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow , 1991, The Journal of experimental medicine.

[55]  E. Moore,et al.  Inadequate granulopoiesis after major torso trauma: a hematopoietic regulatory paradox. , 1990, Surgery.

[56]  B. Gupta,et al.  Vasopressors: Do they have any role in hemorrhagic shock? , 2017, Journal of anaesthesiology, clinical pharmacology.