Complement inhibition significantly decreases red blood cell lysis in a rat model of acute intravascular hemolysis

Tushar A. Shah, Clifford T. Mauriello, Pamela S. Hair, Julia A. Sharp, Parvathi S. Kumar, Frank A. Lattanzio, Alice L. Werner, Pamela H. Whitley, Lou Ann Maes, Kenji M. Cunnion, Neel K. Krishna

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Abstract

Background Prevention of acute hemolytic transfusion reactions is a worldwide concern. The objective of this study was to develop a simple rat model of complement-mediated acute intravascular hemolysis.

Study Design and Methods Human AB red blood cells (RBCs) were incubated with complement-sufficient or complement-deficient Wistar rat serum (WRS) in the presence and absence of human RBC antibody in vitro to elucidate the mechanism of hemolysis. To study the role of complement in acute intravascular hemolysis in vivo, Wistar rats were treated either with or without cobra venom factor (CVF) to deplete complement activity. Human AB RBCs were then injected into both groups of rats, followed by serial blood draws up to 2 hours. Venous blood clearance and lysis of transfused RBCs at each time point were measured by flow cytometry and spectrophotometry. RBC sequestration was determined in the liver, spleen, and kidney by immunohistochemistry.

Results In vitro incubation of human RBCs with WRS demonstrated that RBC lysis was mediated via the classical complement pathway and that hemolysis was antibody dependent. Transfusion of human RBCs into rats showed significantly less hemolysis in the CVF group versus untreated group. RBC sequestration in the spleen and liver 2 hours posttransfusion were not quantitatively different between the two groups.

Conclusions Given the much higher degree of similarity for rat and human complement compared to mice, this simple rat model is ideal for testing novel inhibitors of classical pathway activation for the prevention and treatment of acute intravascular hemolysis.

Original languageEnglish (US)
Pages (from-to)2892-2900
Number of pages9
JournalTransfusion
Volume54
Issue number11
DOIs
StatePublished - Nov 1 2014

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Hemolysis
Erythrocytes
Wistar Rats
Spleen
Classical Complement Pathway
Erythrocyte Transfusion
Antibodies
Spectrophotometry
Liver
Serum
Flow Cytometry
Immunohistochemistry
Kidney

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Hematology

Cite this

Shah, T. A., Mauriello, C. T., Hair, P. S., Sharp, J. A., Kumar, P. S., Lattanzio, F. A., ... Krishna, N. K. (2014). Complement inhibition significantly decreases red blood cell lysis in a rat model of acute intravascular hemolysis. Transfusion, 54(11), 2892-2900. https://doi.org/10.1111/trf.12695
Shah, Tushar A. ; Mauriello, Clifford T. ; Hair, Pamela S. ; Sharp, Julia A. ; Kumar, Parvathi S. ; Lattanzio, Frank A. ; Werner, Alice L. ; Whitley, Pamela H. ; Maes, Lou Ann ; Cunnion, Kenji M. ; Krishna, Neel K. / Complement inhibition significantly decreases red blood cell lysis in a rat model of acute intravascular hemolysis. In: Transfusion. 2014 ; Vol. 54, No. 11. pp. 2892-2900.
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abstract = "Background Prevention of acute hemolytic transfusion reactions is a worldwide concern. The objective of this study was to develop a simple rat model of complement-mediated acute intravascular hemolysis.Study Design and Methods Human AB red blood cells (RBCs) were incubated with complement-sufficient or complement-deficient Wistar rat serum (WRS) in the presence and absence of human RBC antibody in vitro to elucidate the mechanism of hemolysis. To study the role of complement in acute intravascular hemolysis in vivo, Wistar rats were treated either with or without cobra venom factor (CVF) to deplete complement activity. Human AB RBCs were then injected into both groups of rats, followed by serial blood draws up to 2 hours. Venous blood clearance and lysis of transfused RBCs at each time point were measured by flow cytometry and spectrophotometry. RBC sequestration was determined in the liver, spleen, and kidney by immunohistochemistry.Results In vitro incubation of human RBCs with WRS demonstrated that RBC lysis was mediated via the classical complement pathway and that hemolysis was antibody dependent. Transfusion of human RBCs into rats showed significantly less hemolysis in the CVF group versus untreated group. RBC sequestration in the spleen and liver 2 hours posttransfusion were not quantitatively different between the two groups.Conclusions Given the much higher degree of similarity for rat and human complement compared to mice, this simple rat model is ideal for testing novel inhibitors of classical pathway activation for the prevention and treatment of acute intravascular hemolysis.",
author = "Shah, {Tushar A.} and Mauriello, {Clifford T.} and Hair, {Pamela S.} and Sharp, {Julia A.} and Kumar, {Parvathi S.} and Lattanzio, {Frank A.} and Werner, {Alice L.} and Whitley, {Pamela H.} and Maes, {Lou Ann} and Cunnion, {Kenji M.} and Krishna, {Neel K.}",
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Shah, TA, Mauriello, CT, Hair, PS, Sharp, JA, Kumar, PS, Lattanzio, FA, Werner, AL, Whitley, PH, Maes, LA, Cunnion, KM & Krishna, NK 2014, 'Complement inhibition significantly decreases red blood cell lysis in a rat model of acute intravascular hemolysis', Transfusion, vol. 54, no. 11, pp. 2892-2900. https://doi.org/10.1111/trf.12695

Complement inhibition significantly decreases red blood cell lysis in a rat model of acute intravascular hemolysis. / Shah, Tushar A.; Mauriello, Clifford T.; Hair, Pamela S.; Sharp, Julia A.; Kumar, Parvathi S.; Lattanzio, Frank A.; Werner, Alice L.; Whitley, Pamela H.; Maes, Lou Ann; Cunnion, Kenji M.; Krishna, Neel K.

In: Transfusion, Vol. 54, No. 11, 01.11.2014, p. 2892-2900.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Complement inhibition significantly decreases red blood cell lysis in a rat model of acute intravascular hemolysis

AU - Shah, Tushar A.

AU - Mauriello, Clifford T.

AU - Hair, Pamela S.

AU - Sharp, Julia A.

AU - Kumar, Parvathi S.

AU - Lattanzio, Frank A.

AU - Werner, Alice L.

AU - Whitley, Pamela H.

AU - Maes, Lou Ann

AU - Cunnion, Kenji M.

AU - Krishna, Neel K.

PY - 2014/11/1

Y1 - 2014/11/1

N2 - Background Prevention of acute hemolytic transfusion reactions is a worldwide concern. The objective of this study was to develop a simple rat model of complement-mediated acute intravascular hemolysis.Study Design and Methods Human AB red blood cells (RBCs) were incubated with complement-sufficient or complement-deficient Wistar rat serum (WRS) in the presence and absence of human RBC antibody in vitro to elucidate the mechanism of hemolysis. To study the role of complement in acute intravascular hemolysis in vivo, Wistar rats were treated either with or without cobra venom factor (CVF) to deplete complement activity. Human AB RBCs were then injected into both groups of rats, followed by serial blood draws up to 2 hours. Venous blood clearance and lysis of transfused RBCs at each time point were measured by flow cytometry and spectrophotometry. RBC sequestration was determined in the liver, spleen, and kidney by immunohistochemistry.Results In vitro incubation of human RBCs with WRS demonstrated that RBC lysis was mediated via the classical complement pathway and that hemolysis was antibody dependent. Transfusion of human RBCs into rats showed significantly less hemolysis in the CVF group versus untreated group. RBC sequestration in the spleen and liver 2 hours posttransfusion were not quantitatively different between the two groups.Conclusions Given the much higher degree of similarity for rat and human complement compared to mice, this simple rat model is ideal for testing novel inhibitors of classical pathway activation for the prevention and treatment of acute intravascular hemolysis.

AB - Background Prevention of acute hemolytic transfusion reactions is a worldwide concern. The objective of this study was to develop a simple rat model of complement-mediated acute intravascular hemolysis.Study Design and Methods Human AB red blood cells (RBCs) were incubated with complement-sufficient or complement-deficient Wistar rat serum (WRS) in the presence and absence of human RBC antibody in vitro to elucidate the mechanism of hemolysis. To study the role of complement in acute intravascular hemolysis in vivo, Wistar rats were treated either with or without cobra venom factor (CVF) to deplete complement activity. Human AB RBCs were then injected into both groups of rats, followed by serial blood draws up to 2 hours. Venous blood clearance and lysis of transfused RBCs at each time point were measured by flow cytometry and spectrophotometry. RBC sequestration was determined in the liver, spleen, and kidney by immunohistochemistry.Results In vitro incubation of human RBCs with WRS demonstrated that RBC lysis was mediated via the classical complement pathway and that hemolysis was antibody dependent. Transfusion of human RBCs into rats showed significantly less hemolysis in the CVF group versus untreated group. RBC sequestration in the spleen and liver 2 hours posttransfusion were not quantitatively different between the two groups.Conclusions Given the much higher degree of similarity for rat and human complement compared to mice, this simple rat model is ideal for testing novel inhibitors of classical pathway activation for the prevention and treatment of acute intravascular hemolysis.

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