bluuebunny/crossref_metadata_2025 huggingface dataset:
A Simple Method for the Standardization of Centrifugation Procedures in Blood Component Preparation
Authors: Högman Claes F. , Johansson Anne , Bergius Bengt
Published: March 2009

The total sedimentation effect in centrifugation procedures is described by including the phases of acceleration and retardation and expressing the effect by the product g.s. 91 CPD whole blood units were centrifuged at different speeds and times, between 373 g for 9 min 52s and 2,606 g for 22s. All procedures gave a sedimentation effect of 244,090 g.s. The platelet yield in the platelet‐rich plasma was 78–92% of whole blood. No significant difference was observed between low g force for a long time as compared to high g force for a short time.
Platelet Concentrates
Authors: Snyder Edward L. , Koerner Theodore A.W. , Hezzey Anne
Published: March 2009

Abstract. To determine what effect different preparative protocols and different platelet storage bags have on in vitro release, platelet concentrates were prepared according to two different protocols (group I: 2,200 g for 3.5 min followed by 4,900 g for 5 min; group II: 1,000 g for 9 min and 3,000 g for 20 min). The storage bags used were either a standard polyvinyl chloride (PL‐146) or a polyolefin bag (PL‐732). The results for platelet concentrates stored in PL‐146 showed that while group I had undergone more alpha granule release at 24 and 48 h of storage than had group II, by 72 h both groups showed a similar degree of release. Results for platelets stored in PL‐732, however, did not show any significant difference in release between the two protocol groups at any storage time up to 120 h. We conclude that although the preparative protocol used may induce different degrees of platelet release in vitro, the platelet storage bag also has a significant influence on the degree of in vitro damage which occurs during subsequent storage.
Proficiency in Testing for Hepatitis B Surface Antigen: Experience with Ausria II®1
Authors: Nath Nrapendra , Fang Chyang T. , Pielech Mitchell , Lawson William , Dodd Roger Y.
Published: March 2009

Abstract. Eight hepatitis B surface antigen (HBsAg) proficiency panels were tested using a radioimmunoassay (RIA) test kit during 1978 and 1979 by routine processing laboratories within the 57 American Red Cross Blood Services Regions. The test results were analyzed and it was found that lot‐to‐lot variability in the reagents may result in a measurable loss of sensitivity of the test leading to an increased error rate. On the basis of these studies, it is recommended that the positive to negative (P/N) ratio for validation of the test be raised to at least 25.
Analysis of the Immunogenetic Background of Japanese Leprosy Patients by the HLA System
Authors: Izumi Shinzo , Sugiyama Kazuko , Matsumoto Yoshiko , Ohkawa Susumu
Published: March 2009

Abstract. 295 lepromatous and 74 tuberculoid leprosy patients were typed for HLA‐A, ‐B and ‐C and compared to 110 healthy controls. It was found that frequency of HLA‐B7 was significantly high and that of Bw54 was significantly low in lepromatous leprosy. HLA‐DR and MT types were investigated in 84 lepromatous and 28 tuberculoid patients and compared to 55 controls. Both lepromatous and tuberculoid patients showed a marked increase in DR2 frequency. The relative risk is 8.7 and 5.9. Lepromatous leprosy showed an increase in frequency of MT1 and decreases in those of DRw9 and MT3 too.
A ‘New’ Allele Giving Further Insight into the LW Blood Group System
Authors: Sistonen Pertti , Tippett Patricia
Published: March 2009

Abstract. Nea, a red cell antigen present in 6% of Finnish people but in less than 1% of other Europeans tested, is shown to be part of the LW system. Tests on 10,025 Finnish donors show the antithetical relationship of anti‐Nea (proposed name anti‐LWb) and anti‐LW made by LW3 people (proposed name anti‐LWa). Tests on families of previously reported LW3 propositi and of LW(a‐b+) Finnish donors support the hypothesis that Nea (L Wb) is an allele at the L W locus which when homozygous produces the phenotype once called LW3.
Chronic Cold Haemagglutinin Disease Due to an Anti‐M‐Like Autoantibody
Authors: Chapman J. , Murphy M.F. , Waters A.H.
Published: March 2009

Abstract. A case of chronic cold haemagglutinin disease due to an IgM monoclonal (kappa) autoantibody with anti‐M‐like specificity is described in a patient with the MN phenotype. The autoantibody was present in very high titre and active at body temperature, but haemolysis was only mild. The direct antiglobulin test was positive due to C3d on the patient's red cells, and the autoantibody was able to bind complement to normal MM and MN cells with a marked dosage effect.
Immunochemical Studies of Infectious Mononucleosis IX. Heterophile Antigen Associated with a Glycoprotein from the Bovine Erythrocyte Membrane
Authors: Fletcher Mary Ann , Caldwell Karen E. , Latif Zuhair
Published: March 2009

A glycoprotein was isolated from the membrane of the bovine erythrocyte by refluxing the acetone‐ and ethanol‐extracted stroma residue with 75% ethanol. The glycoprotein was purified by phosphocellulose chromatography, ethanol precipitation, lipid‐solvent extraction and DEAE chromatography. The glycoprotein appeared to have two serological determinants, both reactive with antibodies present in the sera of patients with infectious mononucleosis. One of the determinants is similar to the Paul‐Bunnell heterophile antigen found on sheep erythrocytes. It is dependent on carbohydrate, including sialic acid residues. Another specificity, seemingly not shared by sheep erythrocytes to any great extent, is resistance to neuraminidase and to alkaline borohydride treatment and thus it may be located either on the polypeptide portion of the molecule or on an alkali‐stable oligosaccharide. The purified glycoprotein comprises 73% amino acids. Carbohydrate components and their molar ratios were sialic acid (1.0): galactose (1.5): N‐acetylglucosamine (1.1): N‐acetylgalactosamine (0.5): mannose (0.1).
G6PD Varadero A New Variant of Glucose‐6‐Phosphate Dehydrogenase Associated with Congenital Nonspherocytic Hemolytic Anemia
Authors: Estrada M. , García M. , Gutiérrez A. , Quintero I. , González R.
Published: March 2009

A glucose‐6‐phosphate dehydrogenase (G6PD) variant was studied in a mulatto patient with chronic nonspherocytic hemolytic anemia. This variant has reduced activity, increased thermolability, a reduced Michaelis constant for glucose‐6‐phosphate, slightly increased electrophoretic mobility, a biphasic pH activity profile, high 2‐deoxy‐glucose‐6‐phosphate utilization, normal diamino nicotinamide adenine dinucleotide phosphate utilization and a peak of elution profile after G6PD B. The electrophoretic, kinetic, and chromatographic properties of this erythrocyte G6PD variant allow the conclusion that G6PD Varadero is probably a new variant.
Hemostasis Testing during Massive Blood Replacement A Study of 172 Cases
Authors: Mannucci P.M. , Federici A.B. , Sirchia G.
Published: March 2009

Abstract. A 24‐hour service was organized to study changes in the hemostatic system in surgical patients undergoing massive transfusion for excessive bleeding during operation or in the early postoperative period. Hemostasis tests gave normal results in only 12 (7%) of the 172 patients, while in the remaining 160 (93%) one or more tests gave abnormal results. The platelet count was the most frequently abnormal, followed by the prothrombin time and plasma fibrinogen. Well‐defined hemostatic disorders (such as DIC, heparinization and liver disease) were ascertained in 82 patients (48%). 78 patients (45%) had less specific laboratory abnormalities, with a particularly high incidence of thrombocytopenia and less pronounced alterations in the coagulation tests. Unlike the patients with defined disorders, the strong inverse correlation in this group between platelet count, prothrombin time, plasma fibrinogen, and the number of transfusions suggests that the laboratory abnormalities were induced by massive blood replacement. Standard schemas involving the administration of platelet concentrates and/or fresh‐frozen plasma without evaluation of hemostasis did not help to reduce the incidence of abnormalities. These measures also failed to decrease the requirements for whole blood and/or packed red cells. Therefore, indiscriminate administration in the massively transfused postoperative patient of blood components based on preestablished schemes appears to be unjustified. An approach based on hemostasis screening, identification of the underlying disorder, directed therapeutic intervention and laboratory monitoring is likely to be more effective.
An Inherited RBC Characteristic, NOR, Resulting in Erythrocyte Polyagglutination
Authors: Harris P. A. , Roman G.K. , Moulds J.J. , Bird G. W. G. , Shah N. G.
Published: March 2009

Abstract. This report describes a new inheritable red blood cell characteristic, termed NOR, that is associated with polyagglutination. The NOR factor was shown to be inherited in an apparent dominant manner by 5 members of a two‐generation family. NOR was clearly demonstrated to be a unique form of polyagglutination distinguishable from previously reported forms by serological tests using seed lectins, snail agglutinins, enzymes and absorption techniques. The NOR factor was characterized by the lack of reactivity with various seed and snail lectins, enhanced serum agglutinability after RBC proteolytic enzyme treatment, and the ability of hydatid cyst fluid to inhibit the serum hemagglutination reaction.
Effectiveness of Red Blood Cells Filtered Through Cotton Wool to Prevent Antileukocyte Antibody Production in Multitransfused Patients
Authors: Sirchia G. , Parravicini A. , Rebulla P. , Greppi N. , Scalamogna M. , Morelati F.
Published: March 2009

Abstract. The effectiveness of red blood cells made leukocyte‐free by filtration through cotton wool to prevent the production of antileukocyte antibodies was evaluated in children suffering from Cooley's anemia. Two studies were performed: study I was carried out prospectively in two groups of non transfused patients, one group treated with leukocyte‐free filtered red cells, the other with buffy‐coat‐free packed red cell units. Different types of antileukocyte antibodies were looked for in both groups and the results were compared. In study II the behavior of pre‐existing lymphocytotoxic antibodies found in the serum of children previously transfused with standard or buffy‐coat‐free packed red cell units was followed after the patients had been passed to a program of transfusion with leukocyte‐free filtered red cells. Study I showed that none of the patients transfused with leukocyte‐free filtered red cell units have produced antileukocyte antibodies, while these could be found in 2/3 of the patients transfused with buffy‐coat‐free packed red cell units. Study II showed that the repeated transfusion of leukocyte‐free filtered red cells to patients who possessed in their serum preformed lymphocytotoxic antibodies did not cause any increase in the potency or spectrum of these antibodies, but was in fact accompanied in some cases by their decrease or disappearance. It is concluded that filtration through cotton wool is an easy and inexpensive means of preparing leukocyte‐free red blood cells for transfusion capable of preventing (or reducing) the production of antileukocyte antibodies in multitransfused patients.
Hepatitis‐Associated Markers in the American Red Cross Volunteer Blood Donor Population
Authors: Bastiaans M.J.S. , Nath N. , Dodd R. Y. , Barker L.F.
Published: March 2009

Abstract. The present study was undertaken to determine whether there were any differences in the distribution of hepatitis B virus‐(HBV) associated serologic markers among hepatitis B surface antigen‐(HBsAg) positive first‐time and repeat blood donors. The markers examined in samples from 412 newly identified HBsAg‐positive donors (254 first‐time and 158 repeat) included HBsAg titer and subtype, HBeAgIanti‐HBe, and anti‐HBc. Repeat donors were more frequently HBeAg‐positive (25.9%) than were first‐time donors (17.7%). Anti‐HBc and anti‐HBe were observed more often among first‐time (99.6 and 76%) than repeat (91.8 and 60.8%) donors. No differences were found in the mean HBsAg titer nor in the subtype distribution in the two populations. The frequency of HBeAg positivity and the mean HBsAg titer in blood were significantly lower among first‐time donors aged 30 or older as compared to those younger than 30. Such age‐related tendencies did not occur among the repeat donors. The profiles of HBV markers suggest that the HBsAg‐positive first‐time donor group consists predominantly of long‐term HBsAg carriers who may have acquired HBV at an early age, while the HBsAg‐positive repeat donors have newly acquired infections.
A Case Study
Authors: Pegels J. G. , Bruynes E.C.E. , Altes H.R. Korthals , Engelfriet C.P. , von dem Borne A. E. G. Kr.
Published: March 2009

Abstract. Both 51Cr‐survival studies with donor platelets and allogenic skin transplantations were performed in a patient with immune unresponsiveness to platelet antigens, i.e., HLA‐ and platelet‐specific antigens. The patient, who suffered from hypoplastic anaemia, was successfully transfused with random donor platelets during 13 months. The serum of this patient contained only granulocyte‐ and mononuclear‐cell‐reactive antibodies, but no platelet‐reactive antibodies. A nearly normal survival time of the donor platelets as well as a prolonged rejection time of a skin allograft of the same donor support the serological findings. The 51Cr‐platelet survival time was not influenced by a leucocyte concentrate from the same donor, which was administered at the same time. Thus, in our patient, no increased platelet destruction could be induced via the so‐called innocent bystander mechanism.
Removal of Leukocytes from Blood by Fibre Filtration: A Comparison Study on the Performance of Two Commercially Available Filters
Authors: Reesink H. W. , Veldman H. , Henrichs H.J. , Prins H.K. , Loos J.A.
Published: March 2009

Abstract: Two different kinds of filters suitable for the almost complete removal of leukocytes from blood‐cell concentrates were tested. The maximal retention of filter I was 1.9‐3.4×109 leukocytes per filter, whereas filter II could retain 3.6–7.8×109 leukocytes per filter before the leukocyte concentration in the filtrate passed the level of 500 leukocytes/μl. The leukocytes, once absorbed by the fibre material, could be released by washing the filters I, whereas the leukocytes were retained by the material of the filters II. No detectable particles were released after the first 100 ml of filtrate during the washing procedure of either kind of filters. From more than 20% of the filters I, more than 500 pg/ml of endotoxin could be released during the prewashing, whereas none of the filters II was contaminated with endotoxin. The filter II released acetic acid which could be completely removed during the prewashing with 250 ml of saline solution. Operation according to the prescribed conditions of 25 filters of both kinds revealed that the residual leukocyte content in the filtrate was more than 0.25 ×109 leukocytes in 8 out of 25 of the filtrates when filters I were used, whereas with all filters II, this content remained lower than 0.1×109 leukocytes per filtrate. It was concluded that only filter II has sufficient capacity to guarantee the removal of 97% of all leukocytes and 90% of the thrombocytes present in 500 ml of fresh human blood.
Ulex Seed Extracts: Lymphocyte Growth Inhibition and the Anti‐H Hemagglutinins1
Authors: Pirofsky Bernard , Ingebo Kristy R. , August Anne V. , Hefeneider Stephen H.
Published: March 2009

Abstract: Extracts produced from the seeds of Ulex europaeus are commonly used for their ability to react with the H basic substance present on erythrocytes and secreted in body fluids. Such extracts were found to also contain a potent inhibitor of human and murine lymphocyte growth. Inhibition of growth does not result from cytotoxicity and is easily reversible. Ulex seed extract (USE) solutions were modified in various ways to produce reagents in which the anti‐H hemagglutinins were either retained or removed. The fractionated solutions were then analyzed for hemagglutination and lymphocyte growth‐inhibiting activity. Such studies clearly indicated that these two biological functions resulted from the action of different materials. The lymphocyte growth inhibitor is not a glycoprotein lectin. It does not mediate its effect through the H basic substance and is a heat‐stable, small molecule. The data suggest that plant seed extracts employed for their lectin content may contain an additional class of biologically active agents potentially useful in man.
Immunological Monitoring of a Patient with Guillain‐Barré Syndrome Successfully Treated with Plasma Exchange
Authors: Fiorini G. , Bigi G. , Paracchini M. L. , Marinig C. , Gibelli A.
Published: March 2009

Abstract: A patient with Guillain‐Barré syndrome was successfully treated by plasma exchange (without any other therapy), and the relationship between changes in his peripheral blood lymphocyte subpopulations and the clinical improvement induced by plasma‐pheresis was studied. At the recovery, the most interesting findings were an increase in T suppressor lymphocytes, and in Fc receptor‐bearing lymphocytes (above normal values), together with a decrease in B lymphocytes. In this patient, plasma exchange not only removed toxic humoral factors, but also restored a normal balance among peripheral lymphocytes.
BgbExpression in Relation to the HLA‐B17 Antigen Splits Bw57 and Bw58 and the Cross‐Reactions of Anti‐BgbAntibodies1
Authors: Pollack Marilyn S. , Crawford Mary N. , Robinson Harriet M. , Berger Rachel , Sabo Bernice , O'Neill Geoffrey J.
Published: March 2009

A substantial number of individuals typed as HLA‐B17 do not have Bgb. To determine whether or not this discrepancy reflects genetic or ethnic group differences or the influence of B17 antigen subtypes, a large number of unrelated and related B17 individuals from the Caucasian, Hispanic, Black and Chinese ethnic groups were tested in parallel for Bgb and Bw57‐ or Bw58‐ B17 subtype. Higher Bgb expression was found in association with Bw57, but differences in expression of Bgb were also seen in different ethnic groups and in different related individuals carrying the same Bw57 or Bw58 haplotype. Genetic factors other than HLA thus influence the expression of HLA antigens on red cells. Standard and antiglobulin lymphocytotoxicity tests indicate that all Bgb typing sera contain anti‐Bw57 antilymphocyte antibodies and most also contain anti‐Bw58 and cross‐reacting anti‐B12, B15 and/or Bw49 antibodies.
The Significance of Blood Transfusion in Non‐A, Non‐B Chronic Liver Disease in Japan1
Authors: Kiyosawa Kendo , Akahane Yoshihiro , Nagata Atsuo , Koike Yuriko , Furuta Seiichi
Published: March 2009

We performed a follow‐up study on 70 patients with acute non‐A, non‐B (NANB) posttransfusion hepatitis and a retrospective study on 283 chronic hepatitis, 70 cirrhosis and 53 hepatocellular carcinoma patients of type NANB. In acute NANB posttransfusion hepatitis, as judged by the transaminase levels, the duration of the disease exceeded 6 months in 46/70 = 65.7% and 1 year in 32/70 = 45.7%. The histological diagnosis of the 32 cases persisting for more than 1 year was chronic active hepatitis in 5, chronic persistent hepatitis in 2 and unresolved hepatitis in 6. The frequency of previous transfusion in chronic hepatitis, cirrhosis and hepatocellular carcinoma of type NANB was 42.8, 37.1 and 15.1%, respectively, whereas the incidence of early posttransfusion hepatitis was 8.5, 8.6 and 7.5%, respectively. In chronic liver diseases with a history of jaundice and/or hepatitis, previous transfusions are more frequently associated with type NANB than with type B disease. The present study demonstrates that NANB posttransfusion hepatitis tends to develop to chronic liver disease when analyzed prospectively as well as retrospectively.
Individualizing the Dose of Intravenous Immune Serum Globulin for Therapy of Patients with Primary Humoral Immunodeficiency
Authors: Schiff Richard I.
Published: March 2009

The availability of safe and effective IgG preparations for replacement therapy for patients with primary disorders of humoral immunity allows almost unlimited doses of IgG to be used. Thus, knowledge of the optimal dose and serum concentration of IgG to be achieved is of practical importance. In a multicenter study of a reduced and alkylated IgG preparation (MISG), it was shown that the MISG was well tolerated and as effective as the intramuscular immune serum globulin (ISG) used for more than 30 years. During the course of this study, we observed that the standard dose of 100 mg/kg/month resulted in a wide range of IgG levels. We began a study using another intravenous IgG preparation to determine whether the variable IgG levels were due to variability in the half‐life of IgG in these patients and whether an individualized dose of IgG could be calculated based on the half‐life for each patient. The half‐life was greatly prolonged, ranging from 26 to 86 days compared with 21 days in normal individuals. However, there was no correlation between the half‐life and the serum IgG concentration over the range of concentrations measured. When patients were treated with higher doses of IgG, based on the amount calculated to raise the serum IgG concentration to 200 mg/dl, only 1 patient actually achieved that level and only 3 had a significant increase in the serum concentration despite the higher dose. Failure to achieve the predicted levels could not be explained by a shortened half‐life and seemed to be related to increased losses in the early, redistribution, phase. The half‐life was not reduced in patients who did achieve higher serum IgG concentrations, suggesting that the half‐life is characteristic for each patient and not directly dependent on the serum IgG concentration. However, only modest increases were observed and additional studies using much higher doses of IgG will be necessary before this issue can be settled. Similarly, we did not see an additional benefit in those patients receiving higher doses of IgG, and further studies will be necessary before it can be determined whether higher levels of IgG can reduce the incidence of chronic infections and justify the increased time and expense. The results of the completed study have been published elsewhere [7].
Intravenous Immune Serum Globulin in Immune Thrombocytopenia: Clinical Results and Biochemical Evaluation
Authors: Bussel James
Published: March 2009

Intravenous gamma‐globulin of the pH 4‐treated Swiss Red Cross (SRC) preparation is an effective therapy for acute and chronic immune thrombocytopenia (ITP) of childhood and chronic ITP in adults. Patients who may respond satisfactorily to treatment include those without detectable immune complexes and those with poor response to pre‐treatment. Results are especially good in children, half of whom with chronic conditions are able to discontinue therapy.
HTLV‐III Antibody Testing in Three Danish Blood Banks
Authors: Lindhardt B. Ørskov , Ulrich K. , Ryder L. , Dickmeiss E. , Sørensen H. , Jørgensen J. , Jersild C. , Grunnet N.
Published: March 2009

Abstract. The Organon Teknika Vironostika anti‐HTLV‐III/LAV test was evaluated in three Danish blood banks. The evaluation comprised in total 3,940 consecutive donors. In all three blood banks the tests were carried out exactly according to the manufacturer's instructions, using a low cut‐off value defined as (4N̄ + P̄)/5, where N̄ and P̄ are means of optical densities of known negative and positive samples. By this method the overall frequency of repeatably positive samples was 0.30%. When tested by Western blot, however, none of these samples were shown to contain specific antibodies against HTLV‐III/LAV proteins. When testing different categories of patients, only sera containing HLA antibodies gave rise to false‐positive reactions. Finally, important differences in the results were observed regarding sample preparation, single or dual wavelength optical density readings, and the experience of the technical staff.
Transfusion‐Transmitted Cytomegalovirus Infections: Significance and Control
Authors: Tegtmeier Gary E.
Published: March 2009

Abstract. Cytomegalovirus (CMV) is a herpes virus which can give rise to primary infections, reactivated infections, or reinfections in humans. Seroepidemiologic studies have shown CMV infection to be worldwide with the highest antibody prevalences detected in Third World countries; however, significant regional variations can be seen within a given country. Antibody prevalence varies directly with age and inversely according to socioeconomic status.

Numerous prospective studies of blood transfusion recipients carried out since 1966 have shown marked differences in infection rates but relatively little associated disease. Infection rates were highest in seronegative recipients given large amounts of fresh blood. Recently published reports have shown substantially lower infection rates than earlier studies, a change likely to be due to the current practice of transfusing fewer units of older blood. CMV has not been found to play a significant role in the etiology of posttransfusion hepatitis.

CMV infections have been found to be an important source of morbidity and mortality in immunocompromised patients. Several studies of transfused, premature infants have shown significant differences in infection rates and disease expression. Seronegative low‐birth‐weight infants receiving blood from seropositive donors are at greatest risk. Blood from CMV‐seronegative donors substantially lowers the risk of infection.

Receiving a kidney or heart from a CMV‐seropositive donor appears to be a more salient risk factor than blood transfusion in renal and cardiac transplant patients who are also more likely to have symptomatic CMV infections. Leukocyte transfusions have been found to be a significant source of CMV infection and disease in bone marrow transplant patients. Random donor blood and blood components are likely sources of CMV in marrow recipients although this has yet to be proved in a controlled study.

Methods for preventing or ameliorating CMV infection in immunocompromised patients include the use of CMV‐seronegative blood, frozen‐washed red cells, and CMV immune globulin.
Starting a Plasmapheresis Program with Hollow‐Fiber Filtration
Authors: van der Does Johan A. , van Antwerpen Carlize E.
Published: March 2009

Abstract. In a routine donor center, a program of hollow‐fiber filtration can be instituted. With the Organon Teknika PLASMAPUR system the collection of 600 ml plasma was completed within 50 min in 76% of the procedures. Changing the tubing set took 12–15 min. From the plasma a frozen cryoprecipitate is made with a recovery of 59%. After pooling, lyophilization and heat treatment, the recovery is 42%.
Hematologic Disorders
Authors: Harrington William
Published: March 2009

Abstract. A distinction is made between postinfection thrombocytopenia of childhood and idiopathic thrombocytopenia (ITP) of adulthood. Relative advantages of treatment modalities, including splenectomy, are weighed, with an emphasis on the administration of intravenous immunoglobulin (IVIG) as an emergency measure in the adult patient with ITP. Management of thrombotic thrombocytopenic purpura (TTP) is also discussed, as is the case of a patient who became alloimmunized after 2 years of multiple transfusions used to treat an arteriovenous malformation of the small bowel.
Recognition and Treatment of Immune Disorders at the Time of Birth
Authors: Stiehm E. Richard
Published: March 2009

Abstract. Neonatal immunology offers a fertile field for the investigator. Not only does it supply a great deal of information about immunology in general, but it also presents challenges peculiar to this critical period of life. The complement, phagocytic, antibody, and T cell systems in the neonate have characteristic defects and inadequacies. The genetic conditions that compromise the newborn's immune system are discussed in terms of clinical manifestations as well as laboratory abnormalities. In addition, patterns that may suggest the presence of immunodeficiency are described. Finally, methods of managing the immunodeficient infant are outlined, including clinical and laboratory modalities.
Immunoglobulin in Chronic Inflammatory Diseases
Authors: Cottier Hans , Hässig Alfred
Published: March 2009

Abstract. Several case studies are presented to illustrate the success of intravenous gammaglobulin (IVIG) therapy in ulcerative colitis, Crohn's disease, chronic anterior uveitis, herpes zoster, and relapsing genital herpes simplex. Hypothetical mechanisms explain the marked improvement of the chronic disorders, with particular emphasis on substitution of specific antibodies. The recently recognized heterogeneity of the major histocompatibility complex (MHC) is explored as one possible explanation for low and high responders to particular antigens. It is suggested that various chronic inflammatory disorders result from ineffective immune responses, and that the administration of IVIG may shift the delicate balance between the pathogen and the host to favor the latter.
IgG Subclass Deficiencies: Identifying and Treating Patients at Risk
Authors: Heiner Douglas C.
Published: March 2009

Abstract. IgG subclass deficiencies can increase the risk of pulmonary infections or other severe recurrent infections. Various groups of patients were studied in an attempt to learn more about (1) measuring the IgG subclass levels (particularly IgG4 levels) of possible high‐risk patients; (2) identifying those patients who are at risk of developing recurrent infections, and (3) treating these patients once they have been identified. Individual cases are presented, and associations of isolated subclass deficiencies are examined with respect to infection. Replacement therapy with plasma or intravenous immunoglobulin (IVIG) to increase titers of deficient or absent subclasses is suggested.
Therapy in Cytopenia
Authors: Bussel James
Published: March 2009

Abstract. Intravenous immunoglobulin (IVIG) may be considered first‐line maintenance therapy for idiopathic thrombocytopenic purpura (ITP) because it has been proven to be the least toxic. In a study of 25 children with acute ITP, treatment with IVIG maintained platelet counts above 40,000/mm3 in all of the children. After 1 year, none of these patients required further therapy. In another study group of 25 pediatric patients with chronic ITP, treatment with IVIG circumvented splenectomy in 60% of the cases. The therapeutic regimens for adults and children are described, as is a strategy to overcome IVIG resistance. Experiences with IVIG in hemolytic anemia and neutropenia are discussed. The mechanism of action is explored in some detail, specifically as it relates to reticuloendothelial system (RES) Fc receptor blockade and suppression of antiplatelet antibody synthesis.
Bone Marrow Transplantation as Treatment of Hematologic Disease
Authors: Sanders Jean E.
Published: March 2009

Abstract. The clinical achievements in bone marrow transplantation are the culmination of more than 20 years of study of the principles of transplant biology, tissue typing, supportive care, and pregrafting and postgrafting regimens. Bone marrow transplantation, which involves replacing defective or abnormal marrow with normal donor marrow, is explored. The marrow transplant preparative regimen must suppress the recipient's response and destroy any malignant cells. Indications for bone marrow transplantation are outlined, and studies of the use of transplantation in the treatment of various disorders are discussed. Research continues to focus on the problems associated with marrow transplantation, which include immune deficiency, opportunistic infections, graft failure, the recurrence of the original disease, and acute and chronic graft‐versus‐host disease (GVHD).
Progress in Broadening the Uses of Marrow Transplantation: Donor Availability
Authors: Filipovich A. H.
Published: March 2009

Abstract. The use of bone marrow transplantation as a curative treatment for congenital immunodeficiencies is limited by the lack of available histocompatible sibling donors. The problems associated with mismatched donor transplantation and efforts to overcome these difficulties are discussed, as are results obtained in transplantation with partially matched related and unrelated donors. The use of haploidentical relative donors, an attractive option due to the wider availability of such donors, is explored, along with the consequent need for T cell depletion. Methods of T cell depletion are compared, and efforts at improving depletion methods are discussed.
Management of Serious Alloimmunization in Pregnancy
Authors: Gravenhorst J. Bennebroek
Published: March 2009

Abstract. In the past 40 years, tremendous advances in the treatment of hemolytic disease have been made. The perinatal mortality has been reduced considerably. Rh immunoprophylaxis has decreased the incidence of Rh immunization to around 6 per 100,000 live births. Nevertheless, treatment of severe hemolytic disease is still essential. Increased reliability of diagnostic procedures have made early detection and optimal treatment of hemolytic disease possible. Treatment by plasmapheresis is still controversial. Although believed to be beneficial by some authors, conclusive evidence of its efficacy has not been obtained. With preterm induction of birth at 32 weeks of gestation, the chances for survival are 95%. Improvement of techniques and the use of modern real‐time ultrasound equipment have made intrauterine intraperitoneal transfusion, in experienced hands, a relatively safe and successful procedure. Intravascular transfusion appears beneficial in early pregnancy. Combination of both techniques will probably be the answer for the future. The development of neonatal intensive care facilities has made a major contribution to the survival of children with haemolytic disease. The relative low incidence of haemolytic disease has made it imperative that treatment should take place in centers where experience and special skill are available.
Fluorocarbon‐Based in vivo Oxygen Transport and Delivery Systems
Authors: Riess Jean G.
Published: March 2009

Abstract. The approval of Fluosol, a fluorocarbon emulsion for oxygenating the myocardium during the transluminal coronary angioplasty procedure, is a landmark in the field of injectable oxygen carriers, the so‐called blood substitutes. This review discusses the advances made since this first emulsion was initially developed about 12 years ago. Attention is focused on the progress achieved in the preparation and selection of new, better‐defined and faster‐excreted fluorocarbons, and better surfactants, improved emulsions, knowledge of structure/property relationships along with an improved understanding of the physiologic response to their administration. These advances have led to the development of a second generation of highly concentrated, fluid and stable injectable oxygen carriers suitable for a broad range of clinical applications. Prospects for further progress and future generations of emulsions are also outlined.
Has Hepatitis A Virus Been Transmitted by Clotting Factor Concentrates among Hemophiliacs in the United States?
Authors: Mosley J.W.
Published: March 2009

Hepatitis A virus (HAV) infection occurred after administration of factor VIII concentrate in Europe associated with one manufacturing process. We determined if there was an excess prevalence of anti‐HAV among hemophiliacs in the United States, and whether any infections here were potentially concentrate‐associated. We observed clotting disorder patients and household members for up to 7 years at 6‐month intervals. Selected specimens were titered for anti‐HAV‐immunoglobulin G content to distinguish whether they were passively or actively acquired. Our results show that anti‐HAV prevalence was 20–25%, but males with clotting deficiencies had a 44% rate (p < 0.001). Passive anti‐HAV (≥ 30 to < 950 mIU/ml) from intermediate‐purity factor VIII may have protected against community HAV exposure. Factor IX concentrates, which contain no anti‐HAV, did not protect and a few instances of past transmission are possible. Criteria for present‐day concentrate‐associated cases were developed; no subject conclusively met these criteria during the period between late 1985 and May 1993. In conclusion, the excess of anti‐HAV prevalence in male clotting disorders is attributable to both passive administration in concentrate, and past therapy prior to concentrates. No untoward HAV event related to concentrate was found in our population, but measures to prevent transmission of nonenveloped viruses should be instituted.
Ultracentrifugal Analysis of Factor VIII and von Willebrand Factor in Therapeutic Preparations
Authors: Aronson David L. , Chang Ping
Published: March 2009

Plasma and therapeutic preparations of factor VIII (1 recombinant factor VIII and two monoclonally purified plasma‐derived factor VIII preparations, Kogenate, and AHF‐M and Monoclate, respectively) were centrifuged in a sucrose density gradient, and the fractions were analyzed for factor VIII and von Willebrand factor (vWF). The residual vWF in the monoclonally purified factor VIII preparations sediments more slowly than the vWF of plasma. In the absence of added vWF, the factor VIII in all preparations sediments more slowly than plasma factor VIII. These same preparations of factor VIII added to hemophilic plasma as a source of vWF sediment differently. The addition of either recombinant factor VIII or AHF‐M results in sedimentation of the factor VIII with the plasma vFW and in a position indistinguishable from factor VIII in plasma. In contrast, when Monoclate is added to hemophilic plasma in vitro, the factor VIII sediments more slowly than the vWF of the hemophilic plasma. However, 5 min after the infusion of Monoclate into a patient with hemophilia A, the factor VIII sediments with the plasma vWF. These results indicate that the addition of recombinant factor VIII and AHF‐M results in random binding to all vWF multimers of plasma, while there is little exchange between the added factor VIII in Monoclate and the plasma vWF in vitro. In contrast, when the Monoclate is infused, there is rapid binding of factor VIII to the plasma vWF. This is presumably due to tight binding of the factor VIII to the vWF in Monoclate, whereas the factor VIII in Kogenate and AHF‐M, containing no or very low amounts of vWF, are free to bind to plasma vWF. In vivo there are other conditions, as yet uninvestigated, which allow for the binding of factor VIII to vWF.
Large‐Scale Production and Properties of Immunoaffinity‐Purified Human Activated Protein C Concentrate
Authors: Orthner Carolyn L. , Ralston Annemarie H. , Gee Dan , Kent Randy , Kolen Billy , McGriff J.D. , Drohan William N.
Published: March 2009

Activated protein C (APC) is a highly specific serine proteinase which functions as an important naturally occurring antithrombotic enzyme. APC also has antiinflammatory properties. We have developed a large‐scale process for the production of APC for therapeutic use starting with cryoprecipitate‐poor human plasma. This report describes the process, its performance at the pilot plant scale, and the characteristics of immunoaffinity‐purified human APC concentrate referred to as APC (human). The process consists of three chromatographic steps, an enzymatic conversion step, and incorporates a solvent/detergent treatment step for the inactivation of lipid‐enveloped viruses. Solvent/detergent was shown to rapidly inactivate spiked HIV‐1, as well as three marker viruses to nondetectable levels under process conditions. The immunoaffinity‐purified protein C (PC) intermediate was enriched 13,600‐fold over plasma and had a specific activity of 231 U/mg. The overall yield of the process following enzymatic conversion of the PC intermediate to APC and its processing by anion exchange chromatography was 36%. APC (human) was shown to be highly purified, functional and stable.
Monitoring Anti‐HPA‐1a Platelet Antibody Levels during Pregnancy Using the MAIPA Test
Authors: Dawkins B.
Published: March 2009

Anti‐HPA‐1a platelet antibody levels in pregnant women with a history of fetomaternal alloimmune thrombocytopenia (FMAIT) were monitored longitudinally using the monoclonal antibody immobilisation of platelet antigens (MAIPA) assay, in order to examine any variation in optical density (OD) readings obtained over the course of pregnancy and after delivery. Seven women were selected; 4 were studied retrospectively and 3 prospectively (the latter being treated with intravenous gammaglobulin; IVGG). Levels of anti‐HPA‐1a were measured at various intervals after delivery of the first affected infant, to post delivery of the following affected infant. A decrease in MAIPA OD was demonstrated in all patients during the course of these pregnancies. This assay is a useful tool for monitoring anti‐HPA‐1a in women with a history of infants affected with FMAIT. A maternal antibody ‘resting’ level prior to, or early in the first trimester, must be established for comparison.
Incidence and Diagnosis of EDTA‐Dependent Pseudothrombocytopenia in a Consecutive Outpatient Population Referred for Isolated Thrombocytopenia
Authors: Silvestri Federico , Virgolini Luigi , Savignano Chiara , Zaja Francesco , Velisig Marinella , Baccarani Michele
Published: March 2009

Among 111 patients referred to our outpatient clinic for isolated thrombocytopenia during a 24‐month period, 17 (15.3%) cases of EDTA‐dependent pseudothrombocytopenia (EDTA‐PTCP) were identified. EDTA‐PTCP represented the second most frequent cause of thrombocytopenia in this population. The diagnosis was confirmed by the following findings: (a) normal platelet numbers immediately after blood withdrawal; (b) progressive fall of platelet counts and evident platelet clumping over time, only in EDTA‐anticoagulated blood. A simple, inexpensive and quick diagnostic method was devised, that consists in evaluating the platelet number in a blood sample anticoagulated with EDTA immediately after blood withdrawl and 4 h later.
Reemphasis on Leukocyte Transfusions: Induction of Myeloid Marrow Recovery in Critically ill Neutropenic Children with Cancer
Authors: Saarinen U. M. , Hovi L. , Vilinikka L. , Juvonen E. , Myllylä G.
Published: March 2009

Neutropenia is the major factor predisposing to sepsis in cancer patients, and its duration is important for survival. We administered leukocyte transfusions (LT) to 10 children suffering from documented life‐threatening infections during profound, prolonged neutropenia. Nine had acute leukemia, and one had aplastic anemia; three were bone marrow transplant recipients. These 10 were the only patients in our unit who received LT during the past 7 years. The median leukocyte dose was 0.6×109/kg in total. Instead of leukapheresis products, we used pooled buffy coats from random donors, with a high relative content of lymphocytes and especially T lymphocytes. The leukocyte concentrates were irradiated with 15 Gy. In all 10 patients, we observed prompt and sustained myeloid marrow recovery following LT. Such an effect of LT has never been described before. We hypothesize that in the internal milieu of these aplastic patients the transfused leukocytes were stimulated to secrete cytokines, the result being a cascade‐like phenomenon and stimulation and proliferation of the patient's own bone marrow cells. The bone marrow‐stimulating effect of LT merits further study.
Standardization of the Flow Cytometric Determination of HLA Class I Antigens, ‘Platelet‐Specific’ Glycoproteins and Activation Markers
Authors: Neumüller J. , Tohidast‐Akrad M. , Jilch R. , Schwartz D.W. M. , Mayr W.R.
Published: March 2009

Flow cytometry (FC) provides a reproducible investigation of cell surface antigens on platelets. The aim of this study was to elaborate appropriate protocols and to compare them with other techniques that have already been published. (1) Venipuncture with tubes containing citrate was better for the preservation of the antigenicity than using ACD tubes. The isolated platelets could not be completely distinguished from detritus and protein aggregates. Therefore a platelet concentration between 107 and 108/ml measurement buffer was necessary to obtain a sufficient resolution by FC. (2) Isolation methods using either differential centrifugation or diluted Ficoll‐Hypaque as a flotation medium provided platelets of equal purity. The method with Ficoll‐Hypaque resulted in a higher number of isolated platelets than differential centrifugation. The demonstration of platelets and their antigens in whole blood without isolation gave good results provided the platelets were not activated. Activation of platelets with 1 NIH‐U thrombin/1 resulted in the loss of a part of the highly activated platelets because of their aggregation. (3) Comparing different concentrations of paraformaldehyde in PBS, fixation with 1% for 15 min provided the best antigen preservation for most of the antigens investigated. Isolation induced platelet activation. In order to avoid this effect, the whole anticoagulated blood was fixed with 1% paraformaldehyde for 15 min immediately after venipuncture. Then the platelets were isolated using diluted Ficoll‐Hypaque. In this way, systemic activation of platelets can be detected with antibodies against glycoproteins which are translocated from the α‐granules or lysosomes to the cell membrane. These activation markers can be determined on immediately fixed platelets (already in the whole blood) without any interference due to unspecific activation caused by the isolation procedure. (4) Platelet treatment with citric acid at pH 3, in order to remove the antigenicity of HLA‐class I molecules, was sensitive to immediate fixation with paraformaldehyde in the whole blood. Fixation after isolating the platelets made it possible to demonstrate antigen stripping, and the free heavy chain, devoid of the β2‐microglobulin, could be clearly demonstrated. (5) Using standardization beads, the average number of antigenic sites per platelet could be determined for the investigated specificities. It was shown that antibodies which have been directly conjugated or biotinylated and combined with streptavidin‐phycoerythrin yielded similar results in terms of the number of antigenic binding sites while unconjugated antibodies in combination with FITC‐conjugated anti‐mouse‐IgG led to overestimation of antigenic binding sites.
Significance of Anti‐LU8 in Pregnancy
Authors: Watt J. , Jones M.L. , Rose P. , Pepper R.
Published: March 2009

Anti‐LU8, an antibody defining a high‐frequency antigen in the Lutheran system, was identified in the serum of a gravida 4 para 0 pregnant woman who had never been transfused. The antibody consisted of both an IgM and an IgG1 component and was nonreactive in the antibody‐dependent cell‐mediated cytotoxicity (ADCC) assay. The red cells of the mother and 3 siblings of the patient typed as LU:8,14 and were incompatible with the patient's serum. The baby was born at term with a negative direct antiglobulin test and typed as LU:8,14. There was no clinical evidence of haemolytic disease of the newborn.