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CONTENTS
Immunohematology
Volume 11, Number 4, 1995

ABSTRACT

Review: antibodies and antigens in immune neutropenias
G.F. Lucas

Flow cytometric phenotyping of platelet HPA-1a antigen: donor screening for a case of neonatal alloimmune thrombocytopenia due to anti-HPA-1a antibodies
J.J.M.L. Hoffman, W.C.M. Janssen, and H.Y. von Hegedus

Trimeresurus venom inhibition of anti-HPA-1a and anti-HPA-1b antibody binding to human platelets
S.J. Wlodar, D.L. Stone, and L.T. Sinor

Review: comparing platelet compatibility to red cell compatibility protocols
S. Rolih

Application of the proteolytic enzyme papain in routine platelet serology
J.A.G. Lown and B.J. Dale

Detection of drug-dependent platelet antibodies by use of solid-phase red cell adherence techniques
M.F. Leach, L.K. Cooper, and J.P. AuBuchon

Case report: solid-phase platelet crossmatching to support the alloimmunized patient
B.A. O'Connell

Case report: a pregnant woman with immune thrombocytopenic purpura and unusual red cell antibodies
C.R. Nanton, S.M. Martin, L.O. Cook, and P.J. Larison


Review: antibodies and antigens in immune neutropenias

G.F. LUCAS

    In the 1960s and 1970s, the role of granulocyte-specific allo-, auto-, and drug-dependent antibodies in certain neutropenic conditions became established. In some of these conditions, neutropenia is associated with an increased risk of life-threatening infections or unexpected mortality during routine blood transfusion but, in the majority of conditions involving granulocyte antibodies, the clinical course tends to be benign. Consequently, because the detection and identification of granulocyte antibodies is time consuming and labor intensive, there has been little impetus for further development in this area. However, in the last few years, immunochemistry and molecular biology have provided advances in granulocyte immunology. The purpose of this review is to describe current knowledge about the antibodies and antigens implicated in immune neutropenias.

Geoff F. Lucas, PhD, Immunohaematology Section Manager, International Blood Group Reference Laboratory, Southmead Road, Bristol, BS10 5ND, UK

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Flow cytometric phenotyping of platelet HPA-1a antigen: donor screening for a case of neonatal alloimmune thrombocytopenia due to anti-HPA-1a antibodies

J.J.M.L. HOFFMANN, W.C.M. JANSSEN, AND H.Y. von HEGEDUS

Neonatal alloimmune thrombocytopenia can effectively be treated by transfusing compatible platelets to the affected newborn, but typed, compatible platelets are not generally available. For a case of probable neonatal alloimmune thrombocytopenia due to anti-HPA-1a, part of the donor population of the regional blood bank was phenotyped to find HPA-1a-negative platelets. A flow cytometric technique was used, which is reliable, rapid, and relatively simple and therefore well-suited for large-scale screening. Using this method, several HPA-1a-negative donors were identified, and one of them donated platelets that were successfully transfused to the thrombocytopenic newborn. Immunohematology 1995;11:4

Johannes J.M.L. Hoffmann, PhD, Director, Department of Clinical Laboratories, Catharina Hospital, P.O. Box 1350, 5602 ZA Eindhoven, The Netherlands; Willy C.M. Janssen, MT, Research Assistant, Department of Clinical Laboratories, Catharina Hospital, The Netherlands; Harry Y. von Hegedus, MT, Head of Laboratory, Red Cross Blood Bank Eindhoven, Eindhoven.

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Trimeresurus venom inhibition of anti-HPA-1a and anti-HPA-1b antibody binding to human platelets

S.J. WLODAR, D.L. STONE, AND L.T. SINOR

A solid-phase red cell adherence assay was used to demonstrate the specific inhibitory effect of seven species of Trimeresurus snake venom on the binding of HPA-1a- and HPA-1b-specific platelet antibodies. Trimeresurus venom did not inhibit the binding of HLA, HPA-3a, HPA-3b, HPA-4a, HPA-5a, and HPA-5b specific platelet antibodies. Venom from other genera of snakes, including representatives from Agkistrodon, Ancistrodon, Bitis, Bothrops, Bungarus, Causus, Crotalus, Dendroaspis, Ecis, Micrurus, Naja, Notechis, Ophiophagus, Pseudechis, Sepedon (Hemachatus), and Vipera, all failed to specifically inhibit anti-HPA-1a and HPA-1b binding. These results may indicate that the component in Trimeresurus snake venom previously reported to bind to the platelet GPIIb-IIIa complex, inhibiting fibrinogen binding, binds close to the HPA-1a and HPA-1b epitopes. Immunohematology 1995;11:4

Steve J. Wlodar, Immucor, Inc., 3130 Gateway Drive, Norcross, GA 30071; Darryl L. Stone, PhD, Immucor, Inc., Norcross, GA; Lyle T. Sinor, PhD, MAIC (to whom reprint requests should be sent), Immucor, Inc., Norcross, GA.

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Review: comparing platelet compatibility to
red cell compatibility protocols

S. ROLIH

    Between 1971 and 1980, the number of platelet concentrates transfused in the United States increased from 0.4 to 2.8 million units.1 That number increased to nearly 5 million in 1987 and continues to increase in the 1990s. The increase in platelet transfusions has led to a concomitant rise in the incidence of platelet alloimmunization. As a consequence, many transfusion services or blood products providers have developed platelet compatibility protocols to select platelet components that will have acceptable survival when transfused to alloimmunized patients. Most serologists are familiar with compatibility tests used to select red blood cells (RBCs) for transfusion. These tests are performed before transfusion, whether or not alloimmunization has occurred. Platelet compatibility tests, in contrast, are implemented only after alloimmunization and development of the refractory state.

Susan Rolih, MS, MT(ASCP)SBB, Vice President, Technical Services, Immucor, Inc., 3130 Gateway Drive, Norcross, GA 30091-5625.

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Application of the proteolytic enzyme papain
in routine platelet serology

J.A.G. LOWN AND B.J. DALE

The use of proteolytic enzymes is well established in red cell serology. These enzymes modify some antigen structures and remove sialic acid from the red cell membrane. Enzyme-sensitive structures have also been identified on the platelet membrane. The effect of papain, a proteolytic enzyme, used widely in red cell serology, on the detection of various platelet alloantibodies was examined to determine its usefulness in platelet serology. Antisera with the specificities anti-HPA-la, -2b, -3a, -4a, -5a, -5b, and -Naka were examined. HLA antibodies were also included. All sera were tested by a solid-phase red cell adherence technique in parallel with untreated platelets (UP) and papain-treated platelets (PP) for 15 minutes at 37oC. The reactivity of anti-HPA-2b was eliminated and that of anti- HPA-3a was either eliminated or almost eliminated with PP. Antisera specific for the other alloantigens tested reacted similarly or more strongly with PP compared with UP. These findings were confirmed by flow cytometry. The reactivity of HLA antibodies with PP was generally enhanced. Inactivation by papain of platelet alloantigens in the HPA-2 and HPA-3 systems, but not in other systems, may assist in resolving mixtures of platelet alloantibodies. Also, detection of weak antibodies of other specificities may be enhanced. The use of PP may be a simple and useful serologic tool for investigating platelet alloantibodies. Immunohematology 1995;11:4

John A.G. Lown, FAIMS, Transfusion Medicine Unit, Royal Perth Hospital, Wellington St., Perth, Western Australia, 6000; Brian J. Dale, BAppSci, Department of Haematology, Royal Perth Hospital, Perth, Western Australia.

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Detection of drug-dependent platelet antibodies by
use of solid-phase red cell adherence techniques

M.F. LEACH, L.K. COOPER, AND J.P. AUBUCHON

Many drugs have been reported to cause drug-dependent thrombocytopenia, either by the immune complex or by hapten mechanisms. Testing for the presence of these platelet antibodies has not been considered feasible for transfusion services because their presence was thought to be rare, and their detection involved complex and costly methods. We have developed a new technique for detection of these antibodies which can be performed without the need for specialized and expensive instrumentation. A solid-phase red cell adherence assay was used to detect drug-dependent platelet antibodies active by either the immune complex or the hapten mechanism. Three cases were evaluated for the presence of drug-dependent platelet antibodies. Two patients presented with thrombocytopenia that could not be attributed to other causes. The third case was evaluated for the presence of drug-dependent antibodies after poor responses to platelet transfusions. In these three cases, discontinuation of the implicated drugs, i.e., porcine heparin, quinine sulfate, amoxicillin, Bactrim, and albuterol, was followed by a correction of thrombocytopenia or improved platelet transfusion response within 72 hours. This test methodology and protocol has proven very useful in avoiding transfusions with little likelihood of benefit, and in identifying drugs interfering with platelet recovery or survival. Further investigations with this technique may expand our knowledge of the capability of this technique and of the observed frequency of drug-related immunologic platelet destruction. Immunohematology 1995;11:4

Miriam F. Leach, MT(ASCP)SBB, Transfusion Service, Dartmouth- Hitchcock Medical Center, One Medical Center Drive, Lebanon, NH 03756; Linda K. Cooper, MT(ASCP), Transfusion Service, and James P. AuBuchon, MD, Professor of Pathology, Dartmouth-Hitchock Medical Center, Lebanon, NH.

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Case report: solid-phase platelet crossmatching
to support the alloimmunized patient

B.A. O'CONNELL

Platelet crossmatching by a solid-phase red cell adherence assay was used to provide compatible platelets for two alloimmunized patients with leukemia. In this study, a successful platelet transfusion was defined as giving a corrected count increment (CCI) of >7,500 in a posttransfusion sample. For patient A, a total of 205 random platelet concentrates (PCs) were crossmatched. Eleven were considered compatible. These 11 PCs were transfused during five transfusion episodes. Four of the five transfusions produced CCIs of >7,500 and were considered successful. Individually, eight of the eleven units were considered in vivo compatible, and five of the eight donors of these units agreed to become apheresis donors. Platelets from three of these five apheresis donors gave CCIs of >7,500. For patient B, 1,074 random PCs were crossmatched, and 332 were considered compatible. These units were administered during 78 different transfusions. Seventy-one of these transfusion episodes resulted in CCIs of >7,500. In addition, 19 apheresis donors were identified by platelet crossmatching, and they provided platelets for 38 of 39 successful transfusions for Patient B. Platelet crossmatching should therefore be considered when a blood bank is called upon to support a refractory thrombocytopenic patient. Immunohematology 1995;11:4

Bernadette A. O'Connell, MT(ASCP)SBB, Cell Component Specialist, University of Maryland Cancer Center, 22 South Greene Street, Baltimore, MD 21201.

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Case report: a pregnant woman with immune thrombocytopenia purpura and unusual red cell antibodies

C.R. NANTON, S.M. MARTIN, L.O. COOK, AND P.J. LARISON

Maternal immune thrombocytopenia purpura (ITP) may lead to fetal platelet destruction. This process is mediated by IgG platelet autoantibodies that cross the placenta. In this case, not only were platelet autoantibodies present, but red cell alloantibodies anti-E, anti-M, and anti-He were also present. Anti-E, present as an IgG antibody, crossed the placenta, but did not cause clinical problems in the E+ newborn, other than possible hyperbilirubinemia that was treated by phototherapy.

Carmela R. Nanton, SBB (ASCP), Blood Bank, West Palm Beach VA Medical Center, 7305 N. Military Trail, Palm Beach Gardens, FL 33410; Sharon M. Martin, Ed.D., MT(ASCP), Lloyd O. Cook, MD, and Patricia J. Larison, MT(ASCP)SBB, Medical College of Georgia, Augusta, GA.

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