Sensitive Methods for the Detection and Characterization of Platelet Isoantibodies

Summary

Two sensitive methods for the detection and characterization of platelet isoantibodies are presented in this paper. The methods involve incubating platelets with either radioactive serotonin or adenine, interaction of the platelets with serum antibody and finally detection of supernatant radioactivity. Both tests can be used for HL-A typing of platelets and both tests detect antibodies undetectable by complement fixation. The requirement of a heat labile serum component is variable in the serotonin test but absolute in the adenine test. Antibody activity was found to be in the 7S gamma globulin fraction on DEAE cellulose column chromatography.

^* Current address: Children's Cancer R esearch Foundation, Boston, Massachusetts.

• References

• 1 Aster R, Enright S.E. 1969; A platelet and granulocyte membrane defect in paroxysmal nocturnal hemoglobinuria: usefulness for the detection of platelet antibodies. Journal of Clinical Investigation 48: 1199.
• 2 Aster R. H, Levin R. H, Cooper H. 1964; Complement fixing platelet iso-antibodies in serum of transfused persons. Correlation of antibodies with platelet survival in thrombocytopenic patients. Transfusion 04: 428.
• 3 Bridges J. M, Baldini M, Fechera C, Dameshek W. 1963; Effect of anti-platelet antibodies on the in vitro uptake of 5-hydroxytryptamine by blood platelets. Nature 197: 364.
• 4 Colombani M, Colombani J, Dehay C, Dausset J. 1970. A microtechnique of platelet complement fixation. Results obtained with sera and eluates as the source of antibody. In: Histocompatibility Testing 1970. The Williams and Wilkins Company; Baltimore: 553.
• 5 Freireich E. J. 1966; Effectiveness of platelet transfusion in leukemia and aplastic anemia. Transfusion 06: 50.
• 6 Gockermann J. R, Bowman R. P, Conrad M. E. 1972; A simple method for platelet typing for use in platelet transfusion in alloimmunized patients. Blood 40: 972.
• 7 Grumet CarlF, Yankee R. A. 1970; Long term platelet support of patients with aplastic anemia. Effect of splenectomy and steroid therapy. Annals of Internal Medicine 73: 1.
• 8 Hirschman R. J, Shulman N. R. 1972 Utilization of the platelet release reaction to measure ITP factor and platelet antibodies. Transactions of the Association of American Physicians. (In press)
• 9 Hirschman R. J, Shulman N. R. 1972 Utilization of the platelet release reaction to measure ITP factor and platelet antibodies. Transactions of the Association of American Physicians. (In press)
• 10 Holmsen H, Rozenberg M. C. 1968; Adenine nucleotide metabolism of blood platelets. Ill Adenine phosphoribosyl transferase and nucleotide formation from exogenous adenine. Biochimica et Biophysica Acta 157: 266.
• 11 Holmsen H, Day H. J, Setkowsky C. A. 1972; Behavior of adenine nucleotides during the platelet release reaction induced by adenosine diphosphate and adrenaline. Biochemical Journal 129: 67.
• 12 Holmsen H, Weiss H. J. 1972; Behaviour of adenine nucleotides during the platelet release reaction induced by adenosine diphosphate and adrenaline. Biochemical Journal 129: 67.
• 13 Holmsen H, Weiss H. J. 1972; Further evidence for a deficient storage pool of adenine nucleotides in platelets from some patients with Thrombocytopathia — “Storage Pool Disease”. Blood 39: 197.
• 14 Karpatkin S, Siskind G. W. 1969; In vitro detection of platelet antibody in patients with idiopathic thrombocytopenic purpura and systemic lupus erythematosus. Blood 33: 795.
• 15 Mittal K. M, Mickey M. R, Sengal D. P, Terasaki P. I. 1968; Serotyping for homotransplantation XVIII. Refinement of microdroplet lymphocyte cytotoxicity test. Transplantation 06: 913.
• 16 Mittal K. M, Mickey M. R, Terasaki P. I. 1971; Heterogeneity of HL-A specificities shown by platelet complement fixation. Tissue Antigens 01: 279.
• 17 Mittal K. M, Terasaki P. I. 1972; Cross-reactivity in the HL-A system. Tissue Antigens 02: 94.
• 18 Mtteller-Eckhardt C, Ltischer E.F. 1968; Immune reactions of human blood platelets. Thrombosis et Diatheses Haemorrhagica 20: 155.
• 19 Peterson E. A, Sober H. A. 1966. Chromatography of the plasma proteins. In: The Plasma Proteins I, F. W. Putman. Ed. Academic Press; New York, N. Y: 105.
• 20 Rossi E. C. 1972; The effect of albumin upon the loss of enzymes from washed platelets. Journal of Laboratory and Clinical Medicine 79: 240.
• 21 Shulman N. R. 1966; Immunological considerations attending platelet transfusion. Transfusion 06: 39.
• 22 Shulman N. R, Marder V. J, Hiller M. C, Collier E. M. 1964. Platelet and leukocyte isoantigens and their antibodies ; serologic, physiologic and clinical studies. In: Brown E. G, Moore C. V. (Eds.) Progress in Hematology. Vol 4 Grune and Stratton; New York.:
• 23 Thorsby E, Helgesen A, Gjemdal T. 1972; Repeated platelet transfusions from HL-A compatible unrelated and sibling donors. Tissue Antigens 02: 397.
• 24 Yankee R. A, Grumet F. C, Rogentine G. N. 1969; Platelet transfusion therapy. The selection of compatible platelet donors for refractory patients by lymphocyte HL-A typing. New England Journal of Medicine 281: 1208.