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J Bux

Publications and source records attributed to J Bux.

At least 55 records · Page 3Linked to original sources

[Immunoassays for determination of glycoprotein specificity, serum quantitation and determination of binding sites of granulocyte antibodies].

For the characterization of granulocyte antibodies an antigen capture assay and a method based upon quantitative determination of eluted IgG were developed. These assays allowed the quantitation of granulocyte antibodies in sera, the determination of their binding sites and their glycoprotein specificity, and the identification of NA-specific antibodies in sera containing mixtures of granulocyte antibodies.

Antibodies, Monoclonal↗

[Transfusion-associated acute pulmonary insufficiency: a rare, but life-threatening transfusion reaction].

Two patients developed within several hours after blood transfusion severe shortness of breath which required temporary artificial ventilation. X-ray pictures of the chest showed pulmonary edema in both patients. No cardiac causes for edema were found. Detection of granulocyte antibodies in the sera of one patient and one blood donor confirmed the suspected diagnosis of transfusion-related acute lung injury.

Acute Disease↗

Analysis of granulocyte-reactive antibodies using an immunoassay based upon monoclonal-antibody-specific immobilization of granulocyte antigens.

To detect human granulocyte-reactive antibodies, a glycoprotein-specific enzyme immunoassay for platelet antibodies was adapted for the use of granulocytes as target cells. Peripheral blood granulocytes were simultaneously incubated with a monoclonal antibody (mAb) and the serum to be investigated. After solubilization, aliquots of the cell lysate were transferred to plastic tubes coated with goat anti-mouse antibodies. Following immobilization of the trimolecular (mAb-glycoprotein-human antibody) complex it was detected by addition of enzyme-labelled goat anti-human antibodies using a luminescence technique. This assay allowed identification of different granulocyte-reactive antibodies present in the same sample without the need for complicated absorption studies. Alloantibodies against HLA and the granulocyte-specific NA antigens as well as isoantibodies against the Fc-gamma-receptor III (FcRIII) were detectable using mAb-specific immobilization of granulocyte antigens (MAIGA). Binding of autoantibodies to the FcRIII and to the CD 11b/CD18 complex could be shown.

Animals↗

Influence of granulocyte antibodies on granulocyte function.

The following substances were tested for their influence on granulocyte function: 8 sera that contained human granulocyte-specific alloantibodies against the antigens NA1, NA2 and NB1, two HLA antisera, and the monoclonal antibodies W6/32 and CLB-FcR-gran 1. The effects examined included spontaneous and directed migration, immune phagocytosis inhibition and the generation of oxygen radicals. Using the under-agarose technique, spontaneous migration of sensitized granulocytes was normal. For all antibodies tested, the chemotactic index for N-fMLP, LTB4 and opsonized zymosan was greater than 1. Granulocyte immune phagocytosis of sensitized sheep red blood cells was strongly inhibited by all alloantisera and monoclonal antibodies. The generation of oxygen radicals after triggering the respiratory burst with sensitized sheep red blood cells was also strongly inhibited in the chemiluminescence assay. Immune phagocytosis and chemiluminescent response of granulocytes lacking the corresponding antigen of the tested alloantibodies were not affected. Since sensitization of neutrophils with F(ab')2 fragments of the monoclonal antibodies W6/32 and CLB-FcR-gran 1 showed lower inhibition of generation of oxygen radicals after triggering, Fc-dependent interaction with the target cells seems to be necessary for inhibition. Our results suggest that binding of NA1-, NA2- or NB1-specific alloantibodies to granulocytes not only causes neutropenia, but also impairs granulocyte function.

Chemotaxis, Leukocyte↗

Quantitation of HLA class II antigens on B-cell lines.

By quantitation of elutable immunoglobulin G in a recently described ELISA, the number of HLA class I and II molecules on B-cell lines was determined using monomorphic monoclonal antibodies. An average number of 183,000 binding sites per cell for HLA class I-, 99,000 for HLA-DR-, 63,000 for DQ- and 42,000 for DP-specific monoclonal antibodies were determined. The small amount of HLA class II molecules can in part explain the difficulties observed in HLA class II typing or in detection of class II antibodies using the lymphocytotoxicity text.

Antibodies, Monoclonal↗

May-Hegglin anomaly: a rare cause of thrombocytopenia.

A family with four and an unrelated family with three individuals affected by the May-Hegglin anomaly are described. Platelet counts were markedly reduced and were correctly determined only in the counting chamber. Bleeding time and platelet aggregation were always normal, but platelet nucleotide concentrations (ATP and ADP) were elevated. The platelet glycoprotein complexes Ib/IX, IIb/IIIa and Ia/IIa were quantitatively normal. Platelet-associated IgG was slightly elevated, although thrombocytopenia was presumably not caused by an immunological mechanism. Morphological investigations showed giant platelets and spindle-shaped inclusion bodies in the granulocytes, while their function (phagocytic capacity, radical production) was normal. To exclude hereditary types of thrombocytopenia, morphological and family investigations are required to avoid misdiagnosis with far-reaching diagnostic and therapeutic consequences.

Blood Platelet Disorders↗

Alloimmune neonatal neutropenia is a potential side effect of immunization with leukocytes in women with recurrent spontaneous abortions.

Alloimmunization of a mother against granulocytes causing alloimmune neonatal neutropenia (ANN) in her newborn was found likely to be attributed to previous intradermal injections of paternal lymphocytes. Immunotherapy with leukocytes which was performed for recurrent spontaneous abortions hence provides the possibility of granulocyte alloimmunization and increases the risk of the occurrence of ANN.

Abortion, Habitual↗

Quantitation of granulocyte antibodies in sera and determination of their binding sites.

An enzyme immunoassay using eluates was developed for the quantitation of granulocyte antibodies in sera. Incubation of donor granulocytes with sera containing granulocyte alloantibodies (NA1, NA2, NB1, 5b, HLA) or autoantibodies resulted in a significantly higher amount of immunoglobulin G (IgG) per cell than did incubation with control sera. Ultracentrifugation of the sera prior to testing reduced unspecific binding of IgG to granulocytes. In eight of 10 assessed sera from patients with Felty's syndrome eluted IgG decreased from elevated to normal levels. Ultracentrifugation further allowed determination of the binding sites of granulocyte-reactive alloantibodies. Using sera containing NA1- and NA2-specific alloantibodies 173,000-188,000 binding sites on homozygous and 84,000-110,000 on heterozygous cells were determined. A similar number was found using a Fc gamma receptor III (FcRIII)-specific monoclonal antibody. Granulocytes from patients with paroxysmal nocturnal haemoglobinuria showed greatly reduced binding sites for NA-specific alloantibodies. The binding sites for the human alloantibody NB1 ranged from 36,000 to 318,000 and for the 5b alloantibody from 44,000 to 65,000. Mean values of 139,000 binding sites for HLA antigens and 27,000 binding sites for the FcRII were determined using monoclonal antibodies.

Autoantibodies↗

Serological and clinical aspects of granulocyte antibodies leading to alloimmune neonatal neutropenia.

Eighteen cases of alloimmune neonatal neutropenia (ANN) were analysed for their clinical and serological properties. Pregnancy was normal in all cases, but a 50% incidence of abortion is recorded. With the exception of two premature babies, all newborns were delivered at term. Omphalitis and mild infections of the skin were predominantly present. None of the new-borns died by overwhelming sepsis. The average duration of neutropenia was 11 weeks (range 3-28 weeks). Intravenous IgG therapy was followed by transient remission in 2 of 4 affected newborns. Antibody differentiation revealed in five sera NA1-, in four sera NA2- and in two sera NB1-specific antibodies. In two sera only HLA antibodies were detectable. Complement activating antibodies were determined in 72% of the sera. Screening for granulocyte-specific antibodies in 1016 postpartum sera of unselected women revealed a total of 11 sera (1.1%) reacting selectively with granulocytes, but only four (0.4%) were directed against a known granulocyte-specific antigen. None of the new-born of mothers alloimmunized to granulocyte antigens developed neutropenia, which suggests an incidence of ANN below 0.1%.

Abortion, Spontaneous↗

[Clinical importance of granulocyte-specific antibodies].

Clinical and laboratory data of 184 patients with immune neutropenia were evaluated. They suffered from autoimmune neutropenia (n = 165), alloimmune neonatal neutropenia (n = 18) and from transfusion-associated lung injury (n = 1). Autoimmune neutropenia was predominantly found in patients below 3 years. Patients were usually affected by benign bacterial infections. The peripheral blood count showed normal or diminished leukocyte counts with median absolute neutrophil counts of 285 cells/microliters. Bone marrow examination revealed in 60% of the cases a hypercellular marrow with a shift to the left. In 36% the bone marrow was normal and in 4% a hypocellular marrow was found. Spontaneous remission occurred in all newborns and, so far, in 4 patients with autoimmune neutropenia. Symptomatic treatment of the infections was sufficient in most of the patients.

Adolescent↗

[Granulocyte-specific and HLA antibodies in pregnancy: incidence and clinical value].

Postpartum sera of 1,016 unselected women were examined for granulocyte-specific and HLA antibodies. A total of 11 out of 1,016 sera (1.1%) were only reactive with neutrophils. Cytotoxic HLA antibodies were detected in 24%, noncytotoxic HLA antibodies in 4.8% of the sera. All antibodies belonged to the IgG 1 and IgG 3 subclasses. NA1 and NB1 specificity were each determined in one serum, two sera contained NA2-specific antibodies. After 1 year all antibodies were no more detectable. As none of the newborns from immunized mothers developed neutropenia, the incidence of alloimmune neonatal neutropenia seems to be lower than 0.1%.

Autoimmune Diseases↗

Autoimmune neutropenia: clinical and laboratory studies in 143 patients.

Clinical and laboratory data of 143 patients with primary or secondary autoimmune neutropenia (AIN) were evaluated. Primary AIN was found predominantly in children below 3 years, whereas secondary AIN was more frequent in patients 40-60 years of age. Female patients with primary AIN were slightly more prevalent (54%) than male patients (46%). The peripheral blood count showed normal or diminished leukocyte counts with median absolute neutrophil counts of 250 cells/microliters. In 38% of the patients neutropenia was accompanied by monocytosis. Bone marrow examination revealed in 95% a normo- or hypercellular marrow with a marked reduction of mature neutrophils in 56% of the specimens. Twenty-three percent of the sera showed specificity for the NA1 antigen. Patients were usually affected by benign bacterial infections of the skin and of the upper respiratory tract, as well as by recurrent otitis media. Infections were treated symptomatically, and only six patients required continuous administration of antibiotics. Remission of neutropenia during treatment occurred in three of six patients treated with intravenous immunoglobulin G and in three of four patients who received steroid therapy. Except for one patient neutropenia relapsed after discontinuation of therapy. During a follow-up of 6-36 months, spontaneous remission has been observed in four patients.

Antibodies, Anti-Idiotypic↗

Autoimmunization against the neutrophil-specific NA1 antigen is associated with HLA-DR2.

A significant association of autoimmune neutropenia (AIN) in infants due to NA1-specific autoantibodies and HLA-DR2 is reported. Nineteen of 26 infants presenting with AIN and NA1 autoantibodies possessed the DR2 antigen, while only one out of six children with AIN, but non-NA1-specific autoantibodies, was DR2 positive. Furthermore, one adult patient with primary biliary cirrhosis and periods of neutropenia due to NA1 autoantibodies also carried the DR2 antigen. All DR2-negative patients were positive for DRw6. These findings indicate that a close relationship exists between autoimmunization against the NA1 antigen and HLA-DR2 and possibly also DRw6.

Aged↗

Preparation of leukocyte-poor platelet concentrates from buffy coats by a new type of quadruple bag.

A new quadruple blood bag system for collection of platelet concentrates is described. The principle modification consists of a smaller (150 ml) buffy coat bag in conjunction with SAG-mannitol for red cell preservation. Platelet yield and function were comparable to conventional techniques, leukocyte contamination was lower (1.7 +/- 1.3 x 10(7) per unit). The system is recommended for specialized institutions with a high demand of blood components.

Blood Platelets↗