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Biomedical subjects

H Kiesewetter

Publications and source records attributed to H Kiesewetter.

At least 19 recordsLinked to original sources

Replacement of intravenous administration of anti-D by subcutaneous administration in patients with autoimmune thrombocytopenia.

Intravenous (IV) administration of anti-D in patients with autoimmune thrombocytopenia (AITP) may result in severe hemolysis and even death. Over a 3-year period, we gave anti-D only subcutaneously (SC), and none of our patients have developed any acute adverse reaction. Most importantly, SC delivery of anti-D produces largely the same beneficial effect as obtained by IV anti-D. We recommend replacement of IV administration of anti-D by SC administration in AITP.

Adult↗

CD47 is expressed at normal levels in patients with autoimmune haemolytic anaemia and/or immune thrombocytopenia.

CD47 deficiency results in lethal autoimmune haemolytic anaemia (AIHA) and mild spontaneous thrombocytopenia in non-obese diabetic mice. It is unknown whether CD47 has an impact on AIHA of the warm type or autoimmune thrombocytopenia (ITP) in humans. Healthy blood donors (n= 11), patients with AIHA (n= 13), patients with ITP (n= 18) and one patient with Rh(null) phenotype were investigated. CD47 expression on red blood cells (RBC), platelets, granulocytes and lymphocytes and in plasma was determined by quantitative flow cytometry. All types of blood cells studied were found to carry CD47. Although CD47 expression on Rh(null) RBCs was decreased, there was no significant difference between CD47 expression on RBCs of healthy blood donors and on those of patients with AIHA or ITP. Similarly, CD47 was detectable in the plasma of the studied subjects. No evidence for a pathogenetic role of CD47 in autoimmune haemolysis or thrombocytopenia in humans could be demonstrated.

Anemia, Hemolytic, Autoimmune↗

A simple and practical agglutination assay for human leucocyte antigen-B27 typing.

BACKGROUND AND OBJECTIVES: The human leucocyte antigen (HLA) B27 is the most frequently typed single antigen that is associated with diseases. Here, we describe a simple and rapid particle agglutination assay (PaGIA) for HLA-B27 typing. MATERIALS AND METHODS: Superparamagnetic particles were coated with a monoclonal antibody to HLA-B27 and subsequently used for testing. Anticoagulated whole-blood samples were obtained from healthy blood donors (n = 194) with known HLA patterns and from patients (n = 51) who had been typed positive for HLA-B27 by flow cytometry. RESULTS: The particles agglutinated only after incubation with HLA-27-positive blood samples, using the ID-microtyping system. Positive reactions were clearly distinguishable from negative reactions in all samples tested. Flow cytometric HLA-B27 typing revealed an indeterminate result in one patient. CONCLUSIONS: The new HLA-B27 PaGIA is suitable for rapid typing of HLA-B27. The assay is simple and easy to perform, and can be implemented in any routine laboratory.

Agglutination Tests↗

A novel antigen-specific capture assay for the detection of platelet antibodies and HPA-1a phenotyping.

BACKGROUND AND OBJECTIVES: The antigen-specific assays currently used for the laboratory investigation of platelet antibodies and antigens are technically complex and cannot be used in most routine laboratories. Here, we describe a simple antigen-specific capture assay (ASCA) for the detection of serum platelet antibodies and for human platelet antigen-1a (HPA-1a) phenotyping. MATERIALS AND METHODS: For the detection of platelet antibodies, platelets from healthy blood donors were incubated with biotinylated monoclonal antibodies to platelet glycoprotein complexes (GP), then solubilized and mixed with superparamagnetic streptavidin particles. Serum samples from patients with autoimmune thrombocytopenia (n = 39), from patients with platelet alloantibodies (6 HPA-1a, 1 HPA-2b, 1 HPA-3a, 6 HPA-5b), and from healthy blood donors (n = 70), were tested. All serum samples from the patients were investigated in parallel by the indirect monoclonal antibody-specific immobilization of platelet antigen assay (MAIPA). For HPA-1a phenotyping, superparamagnetic particles were coated with a monoclonal antibody to HPA-1a and mixed with diluted whole blood samples from healthy blood donors (n = 139), who had previously been genotyped for platelet alloantigens. Results The indirect MAIPA detected autoantibodies in 18%, and the direct MAIPA in 50% of patients tested. In contrast, the new ASCA demonstrated positive results in 77% of patients. All tested alloantibodies reacted positive by the ASCA, and all serum samples from healthy blood donors were negative. The results of HPA-1a phenotyping were in concordance with those of genotyping in all cases. CONCLUSION: In our opinion, the ASCA is easy to perform and much more sensitive than the currently available antigen-specific assays for the detection of platelet antibodies.

Antigens, Human Platelet↗

Report of a patient with heparin-induced thrombocytopenia type II associated with IgA antibodies only.

Heparin-induced thrombocytopenia type II (HIT II) is usually mediated by immunoglobulin G (IgG) antibodies that lead to platelet activation via the FcgammaIIA-receptor. Here we describe a patient who developed HIT II after aortocoronary bypass surgery. His serum contained an antibody that was detectable by the heparin-induced platelet activation assay (HIPA) and by the ID-HPF4 particle agglutination assay. The flow cytometric analysis showed that the antibody was of the IgA class.

Autoantibodies↗

Observation of serum erythropoietin concentrations in female athletes for up to eight days after a marathon run.

Erythropoietin (EPO) was studied in 13 female marathon runners before and up to 8 days after a competition marathon run. The median baseline control value was 13.7 U/l. No change in EPO concentration was found immediately (15 min.) and one day after the run. However, a median increase in EPO concentration (18.1 U/l) was found on day three post-exercise (p< 0.05). On day 8 no change was found compared to pre-exercise values. This late increase in EPO concentration would seem to be responsible for the well known increase of red blood cell mass in long distance runners.

Adult↗

[Quality assurance of haemotherapy in anaesthesia. Auswertung eines 3-jahrigen Erfahrungszeitraums].

BACKGROUND: The measures for quality assurance in haemotherapy developed at the Charite Campus Mitte according to the transfusion law of 1998 are introduced and discussed. METHODS: In a 3-year study period the submission of transfusion documentation and the expiration of blood products were compared to the year 2001. The potentials for improvement for 2001-2003 were compared based on evaluation of the self-inspection documentation of the transfusion clinics. RESULTS: The submission rate for the transfusion documentation increased from 90% in 2001 to 98.3% in 2002 and 2003. The expiration of blood products decreased from 5.1% in 2001 to 1.7% in 2003 resulting in a cost reduction from 140,000 Euro to 50,000 Euro. Continuous need for improvement was found for the prescribed annual further education of staff on quality management. CONCLUSIONS: The self-inspection procedure in transfusion clinics still revealed potential for improvement, although documentation of transfusions were improved and the expiration of blood products and the resulting costs were substantially reduced.

Anesthesia↗

Antibodies against protein Z and fetal loss: current perspectives.

Protein Z (PZ) is a vitamin K-dependent plasma protein that serves as a cofactor for the inactivation of factor Xa. A number of investigators found low PZ levels in patients with haemorrhagic as well as thromboembolic diseases, although there is no clear evidence of a pathogenic link between PZ deficiency and these clinical disorders. Nevertheless, low PZ levels have been found in association with early fetal losses, especially those occurring before the 15th week of gestation and in patients with detectable antiphospholipid and anti-PZ antibodies. The current diagnostic relevance and therapeutic consequences of these parameters will be discussed.

Antibodies↗

Immunoadsorption patients with multiple sclerosis: an open-label pilot study.

BACKGROUND: Immunoadsorption (IA) is occasionally applied in patients with acute relapses of multiple sclerosis (MS). This pilot study was undertaken to determine whether IA might help in secondary progressive and relapsing-remitting multiple sclerosis. DESIGN: IA was performed at 1-week intervals in 12 patients with secondary progressive or relapsing-remitting MS. These patients had an extended disability status scale (EDSS) score of 4.5-7 and an EDSS increase of 0.5 within 6 months before inclusion in the study despite conventional drug therapy. The change in the EDSS and that in the MS functional composite (MSFC) score, which consisted of quantitative tests of arm function, ambulation, visual acuity and cognition, served as the primary outcome variables, which were measured at baseline and at 3, 6 and 12 months. Changes in quality of life and cerebral lesions by magnetic resonance imaging (MRI) were also assessed at baseline and after the last immunoadsorption (month 3). RESULTS: A significant reduction of the median EDSS change was observed after the treatment period, which reversed 3 months after the immunoadsorptions had been stopped. Ten of 12 patients remained stable during the first year of follow-up with no significant changes of the MSFC scores. No significant changes in magnetic resonance imaging T2-hyperintense brain lesions or in the number of gadolinium-positive lesions and in the patients' quality of life were observed. Western blot analyses demonstrated a reduction of serum myelin-specific antibodies, which were collected in the adsorber eluates. CONCLUSIONS: Removal of immunoglobulins, including myelin-specific antibodies by immunoadsorption, seems to delay disease progression as defined by EDSS, MSFC and MRI, while the patients' quality of life did not deteriorate.

Adult↗

B*4440: a novel HLA-B allele identified by sequence-specific oligonucleotide hybridization and sequence-specific amplification.

A novel human leukocyte antigen B (HLA-B) allele, B*4440, is described. The allele was identified in an adult stem cell donor of Caucasian origin. HLA-B*4440 most closely matches to B*4403 differing by a substitution of three nucleotides at codon 44, 45, and 50. Thus, low-resolution HLA typing using sequence-specific oligonucleotide hybridization or amplification using sequence-specific primers gave inconclusive results. DNA sequencing confirmed a variation of codons 44 and 45 (AGG AAG-->AGA GAG) and codon 50 (CCA-->CCG), resulting in an amino acid substitution Lys-->Glu at codon 45.

Adult↗

Identification and characterization of a novel HLA-DRB1 allele, DRB1*0830*.

Here, we report on the identification and characterization of a novel human leukocyte antigen (HLA) DRB1 allele, DRB1*0830. This allele was identified in a candidate for hematopoietic stem cell donation. While the DNA sequence of HLA-DRB1*0830 most closely matches to DRB1*080202, its amino acid sequence resembles DRB1*080202 and DRB1*0813. Due to a substitution at nucleotide position 286 HLA-typing using sequence-specific oligonucleotide hybridization or amplification using sequence-specific primers gave inconclusive results. Allele-specific DNA sequencing confirmed a 286 T-->A substitution resulting in Phe67Ile.

Alleles↗

Human platelet antigen genotyping by using sequence-specific primers and the particle gel agglutination assay.

BACKGROUND AND OBJECTIVES: Polymerase chain reaction using sequence-specific primers (PCR-SSP) is currently the most widely used technique for human platelet antigen (HPA) genotyping. Here, we describe a novel particle gel-agglutination technique for simplified visualization of the amplified products. MATERIALS AND METHODS: Biotinylated primers were used to amplify HPA-1, -2, -3, -4, -5, -6, and -15, and the PCR products were incubated with streptavidin particles. Fluorescein isothiocyanate (FITC)-labelled primers [amplifying a fragment of the human growth hormone (HGH) gene] and anti-FITC-coated particles were used as internal controls. Agglutination of the particles in or on top of the gel indicated specific amplification. A total of 100 samples from blood donors was tested by using this new technique and a standard PCR-SSP protocol. RESULTS: The use of biotinylated sequence-specific primers resulted in PCR products that agglutinated streptavidin particles, and the FITC-labelled HGH primers led to agglutination of anti-FITC-coated particles. Negative reactions were clearly distinguishable from positive reactions. The results of the particle gel agglutination method were in concordance with those of the electrophoretic visualization in all cases tested. CONCLUSIONS: The new particle agglutination method is reliable and easy to use.

Agglutination Tests↗

Amlodipine-induced immune thrombocytopenia.

BACKGROUND AND OBJECTIVES: Drug-induced thrombocytopenia is a serious, but rare, side-effect of treatment with a number of drugs. In this report, we investigate the suspicion that amlodipine, a calcium-channel blocker, was responsible for immune thrombocytopenia in a 79-year-old patient. PATIENT AND METHODS: Our patient experienced two attacks of thrombocytopenic purpura after 10 years of treatment with amlodipine. Antibodies to platelets were tested by standard methods. RESULTS: Initially, the platelet count increased owing to treatment with prednisolone and intravenous immunoglobulin G, but decreased shortly after discontinuation of this treatment. The patient's serum was found to contain amlodipine-dependent antibodies to platelets, and he recovered after stopping the drug. CONCLUSIONS: Amlodipine can induce immune thrombocytopenia, which may strongly resemble autoimmune thrombocytopenia.

Aged↗

Massive haemolysis after intramuscular diclofenac in a patient who apparently tolerated oral medication.

BACKGROUND AND OBJECTIVES: Administration of diclofenac may lead to immune haemolytic anaemia (IHA) owing to the presence of drug-dependent antibodies and/or autoantibodies. A relationship with oral or intramuscular drug administration is unknown. Here, we describe a patient who apparently tolerated oral diclofenac but developed severe IHA following intramuscular injection of the drug. PATIENTS AND METHODS: A 66-year-old-female was admitted to hospital because of jaundice and nausea, which were initially presumed to be manifestations of a postcholecystectomy syndrome. The patient soon developed haemolysis and renal failure. Although the symptoms and signs were suggestive of autoimmune haemolytic anaemia (AIHA), the patient had diclofenac-induced IHA. RESULTS: Serological testing, including detection of drug-dependent antibodies, was performed using standard techniques. The patient's serum was found to contain a highly reactive diclofenac-dependent red cell antibody of the immune complex type (titre 256 000). She recovered after 7 weeks of treatment with prednisolone, blood transfusions, haemodialysis and plasma exchange. CONCLUSIONS: Diclofenac-induced IHA should always be considered when a patient on diclofenac develops haemolysis.

Administration, Oral↗

Description of a novel HLA-B allele, B*5613, identified during HLA-typing using sequence-specific oligonucleotide hybridization and sequence-specific amplification.

Here, we report on the characterization of a novel human leukocyte antigen (HLA)-B allele, B*5613. The allele was identified in an adult male from North Africa who was suffering from sickle cell anemia. HLA-B*5613 most closely matches to B*5601 differing only by a substitution of three nucleotides of codon 180. Due to this substitution, low-resolution HLA-typing using sequence-specific oligonucleotide hybridization or amplification using sequence-specific primers gave inconclusive results. DNA sequencing confirmed a variation of codon 180 (CTG-->GAC) resulting in an amino acid substitution Leu156Asp.

Base Sequence↗