Anti-D initially stimulates an Fc-dependent leukocyte oxidative burst and subsequently suppresses erythrophagocytosis via interleukin-1 receptor antagonist.
Previous results have demonstrated that anti-D therapy in children with chronic auto-immune thrombocytopenic purpura (AITP) induced a significant increase in several pro- and anti-inflammatory plasma cytokines within 2 hours of administration. To investigate the biologic basis of these early in vivo responses, we developed a flow cytometric assay to measure Fc-dependent responses of human peripheral leukocytes with fluorescently labeled and anti-D-opsonized red blood cells (RBCs). When anti-D-opsonized RBCs were incubated with peripheral blood leukocytes, the earliest detectible event observed was a significant oxidative burst in both monocytes (P <.05) and granulocytes (P <.0001), characterized by the production of hydrogen peroxide (H2O2), peroxynitrite (ONOO-), superoxide (O -2), and hydroxyl (OH) by 10 minutes which declined by 1 hour. By 2 hours, the opsonized RBCs were phagocytosed, particularly by granulocytes (P <.001), but the phagocytosis subsequently declined by 6 hours of incubation. The decline in phagocytosis was correlated with a significant production of interleukin-1 receptor antagonist (IL1ra) by both monocytes (P =.036) and granulocytes (P =.0002) within 4 hours. None of these events occurred if the RBCs were coated with anti-D F(ab)'2 fragments. When recombinant IL1ra was titrated into the assay, phagocytosis of the opsonized RBCs was significantly inhibited (P =.002). Taken together, these results suggest that at least one mechanism of action of anti-D is via the production of the anti-inflammatory cytokine IL1ra which can negatively regulate the ability of leukocytes to phagocytose opsonized cells.
Coopamah
Malini D
MD
Department of Laboratory Medicine and Pathobiology, St Michael's Hospital, 30 Bond St, Toronto, M5B 1W8 ON, Canada.
Freedman
John
J
Semple
John W
JW
D016207:Cytokines / Q000096:biosynthesis / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000378:metabolism*; D005434:Flow Cytometry; D006098:Granulocytes / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000378:metabolism*; D007136:Immunoglobulins / Q000378:metabolism; D053590:Interleukin 1 Receptor Antagonist Protein; D007962:Leukocytes / Q000187:drug effects* / Q000378:metabolism; D009000:Monocytes; D010587:Phagocytosis; D017382:Reactive Oxygen Species; D016897:Respiratory Burst; D018029:Rho(D) Immune Globulin / Q000008:administration & dosage / Q000627:therapeutic use*; D012795:Sialoglycoproteins / Q000378:metabolism*; D013997:Time Factors
D016207:Cytokines; C507763:IL1RN protein, human; D007128:Immunoglobulin Fragments; D007136:Immunoglobulins; D053590:Interleukin 1 Receptor Antagonist Protein; D017382:Reactive Oxygen Species; D018029:Rho(D) Immune Globulin; D012795:Sialoglycoproteins