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Biomedical subjects

F Hiepe

Publications and source records attributed to F Hiepe.

122 records · Page 7Linked to original sources
PubMed1985

The use of glass as solid phase in enzyme immunoassay as exemplified by the detection of circulating immune complexes.

This article describes a solid-phase enzyme immunoassay for the quantitative determination of circulating immune complexes based on the covalent binding of proteins to glass. A quantitative comparison between adsorption to polystyrene and covalent binding to glass gave a protein higher concentration on the glass support. This leads, in connection with the strong attachment of the protein, to a higher sensitivity and precision in immunoassay. The binding of C1q to glass as solid phase offers the possibility to reuse the protein as well as the glass support after accomplishment of the immunoassay which can reduce the cost of the assay in routine work dramatically. Furthermore, the immobilization of C1q to glass gives an extreme stability of the protein which means that the c1q-coated glass supports can be used up to one year under routine conditions.

Adsorption↗
PubMed1983

[Demonstration of circulating immune complexes using a C1q-solid phase enzyme immunoassay].

An enzyme-immunoassay for the quantitative determination of circulating immune complexes in human serum is described. The technique uses alkaline phosphatase-conjugated anti-human IgG to detect immune complexes bound to solid phase C1q. This assay is sensitive detecting 4 ng/ml to 60 micrograms/ml aggregated IgG. First data from normal individuals and patients with systemic lupus erythematosus show that this method is specific and sensitive for detection of circulating immune complexes.

Antigen-Antibody Complex↗
PubMed1979

[Experimental infection in man and monkey (Cercopithecus callitrichus) by sarcosporidian cysts of cattle and swine].

Reported in this paper is experimental sarcosporidial infection of man and lower primates (grass monkey). The courses of infection were followed up by coprological and serological tests. Oral-alimentary administration of 100 Sarcocystis (S.)-fusiformis cysts obtained from cattle muscle caused patent infection in man. Gastro-intestinal symptoms developed in one of the probands, between five and seven days after infection. No sporocysts were detectable from the stools of seven probands who had consumed 200 g each of commercially available minced pork which contained small amounts of Sarcocystis miescherania villi. Rise in antibody to Sarcocystes following infection was detected however, by means of indirect fluorescence antibody reaction (IFAR). Gastrointestinal complaints were reported by one of the probands, one day after infection. No sporocysts were detectable from the stools of four grass monkeys, following oral-alimentary administration of 50 or 25 S. miescherania cysts, but rise in antibody to S. miescherania was recorded in these cases. Those grass monkeys which had received 50 cysts responded by exhibiting, after infection, severe clinical phenomena with intestinal signs and symptoms of the central nervous sytem.

Adult↗
PubMed1977

Some unusual properties of globin-mRNA isolated from rabbit reticulocytes.

Globin mRNA isolated from rabbit reticulocytes showed an unusually high molecular weight for the beta-chain mRNA (280000 daltons), a large difference between the molecular weight of the alpha-chain mRNA (230000 daltons) and the beta-chain mRNA and a dense and compact structure in the electron microscope (average length of the molecules 0.14 micron), which can be transformed to a linear structure (average length of the molecules 0.44 micron corresponding to a molecular weight of about 420000 daltons). The poly(A)-tail of the mRNA estimated by polyacrylamide gel electrophoresis with poly(A)-markers as reference substances has a size between 12 and 52 nucleotides. The mRNA works as a good template in the cDNA-synthesis with reverse transcriptase and full length copies of the mRNA were obtained by this reaction. The cDNA hybridizes to an amount of 70% with the globin mRNA at a 1:4 cDNA/mRNA ratio.

Animals↗
PubMedDate not supplied

[Cyclophosphamide therapy in systemic lupus erythematosus].

Cyclophosphamide is a potent immunosuppressive drug which is widely used to treat renal and central nervous system manifestation of Systemic Lupus Erythematosus (SLE). It is employed especially to prevent renal failure in lupus nephritis. Within the last decade intravenous cyclophosphamide bolus therapy has become the standard therapy in severe SLE due to its tendency towards a higher efficacy and fewer side effects as compared to oral application. Studies on the slightly more cost-effective oral cyclophosphamide bolus therapy have recently been published, however, there are no data on long-term results yet. Here, we review the effects and side effects of cyclophosphamide as well as recent clinical studies on cyclophosphamide bolus therapy in SLE. Because of the possible side effects, especially the high risk of malignancies, which sharply increases at a cumulative dose of approximately 60 g, cyclophosphamide should be used cautiously. Cyclophosphamide bolus therapy should only be performed in hospitals with special experience.

Cyclophosphamide↗
PubMedDate not supplied

Development of accessory phenotype and function during the differentiation of monocyte-derived dendritic cells.

Dendritic cells (DCs) are professional antigen-presenting cells (APCs), and classical DCs, such as Langerhans cells (LCs) or interdigitating DCs (IDCs) are known to be the most potent stimulators of T lymphocytes. Earlier, several groups described the generation of DCs from monocytes, starting with peripheral blood mononuclear cells (PBMCs), adherent cells or magnetic bead-purified CD14+ cells. Although modifications of the original protocols have already been described, some questions relevant to clinical application and basic studies have not yet been addressed. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL4) appear to be necessary, but are not sufficient for the differentiation of monocyte-derived dendritic cells (MoDCs), as indicated by the failure to generate such cells under serum-free conditions. Using adherence purified monocytes, we first investigated the amount of GM-CSF and IL4 required for the differentiation of DCs. Consecutive kinetic studies during the differentiation period were designed to demonstrate how monocytes acquire the phenotype and function of DCs. The results showed that small amounts of GM-CSF and IL4 were required to generate MoDC which acquired their phenotype and function within 4 days. IL13 may substitute for IL4, whereas IL10, TNF alpha or IFN gamma inhibited the generation of MoDCs.

Cell Culture Techniques↗
PubMedDate not supplied

Co-occurrence of spondyloarthropathy and connective tissue disease: development of Sjögren's syndrome and mixed connective tissue disease (MCTD) in a patient with ankylosing spondylitis.

Spondylarthropathies (SpA) and connective tissue diseases (CTD) are clinically distinct entities which, at first glance, seem to have little in common. However, a link between SpA and CTD has recently been suggested by a study in which a higher prevalence of Sjögren's syndrome (SS) and sicca symptoms was reported in patients with ankylosing spondylitis (AS) and undifferentiated SpA (1). Another link between SpA and CTD is a possible side effect of a DMARD widely used to treat SpA: sulfasalazine (SAS). SAS was reported to induce antinuclear antibodies (ANA) and systemic lupus erythematosus (SLE)-like syndromes such as drug-induced lupus. This report describes a 54-year-old white male, HLA B27-positive AS patient with some syndesmophytes who, after 15 years of disease, developed SS with salivary gland involvement, Raynaud's syndrome and anti-Ro antibodies. Then, 20 years after the onset of AS, he became acutely ill, suffering severe myositis and myocarditis along with swollen hands and highly elevated autoantibody titers recognizing UIRNP; his condition was interpreted as mixed connective tissue disease (MCTD). The patient had been treated with SAS and azathioprine (AZA) alone several times during the last years because he had not tolerated other DMARDs. A combination of both drugs had been prescribed 3 weeks before a severe flair because of progredient high disease activity with painful peripheral arthritis of the MCP and PIP joints which, however, had not shown radiographic erosions. We describe the rare development of MCTD in an AS patient and report, for the first time, the onset of MCTD potentially triggered by sulfasalazine.

Humans↗
PubMedDate not supplied

[Detection of autoantibodies to extractable nuclear antigens in autoimmune diseases of rheumatic origin using immunoblotting--comparison with counter-electrophoresis].

Sera from 82 patients with rheumatic autoimmune disease were tested for anti-ENA antibodies by immunoblotting and counterimmunoelectrophoresis (CIE), using HeLa cell extract and rabbit thymus extract, respectively, as antigens. Anti-ENA antibodies were more frequently detected and could be better differentiated by immunoblotting rather than by CIE. There was especially an increase in anti-Sm antibodies (23, in contrast to four positive results), which was only detectable in serum samples for the 59 SLE patients. The sera of the six patients with MCTD all reacted with the 68 U1-RNP antigen. The sera of the five patients with Sjögren's syndrome only recognized the 50k La antigen, while other anti-ENA antibodies were not observed. In SLE anti-La antibodies were often associated with other anti-ENA antibodies. Seven out of eight SLE patients showing a combined detection of antibodies against Sm, U1-RNP and La by immunoblotting demonstrated severe organ involvements, especially lupus nephritis. Therefore, the characterization of anti-ENA antibodies by immunoblotting may contribute to improve the differentiation of connective tissue diseases.

Antibodies, Antinuclear↗
PubMedDate not supplied

[New experimental findings on immunity and autoimmunity in old age].

The peripheral blood lymphocytes of subjects of old age can be characterized as follows: deteriorated immunoglobulin synthesis under PWM stimulation, improved immunoglobulin synthesis under autoantigen (DNA) addition, increase in the anti-DNA auto-antibody synthesis, 4. higher sensitivity of the PHA-induced lymphocyte proliferation to prednisolone. The in-vitro methods have a model character for further investigations on the regulation of the immunological system and its possible therapeutical influencing.

Aged↗
PubMedDate not supplied

Dimensions of homo- and heteropolymeric sections of DNA synthesized by reverse transcription from globin mRNA matrices.

The lengths of globin messenger RNAs, isolated from rabbit and pigeon reticulocytes, and single-stranded complementary DNAs, synthesized from mRNA in presence of RNA-directed DNA-polymerase and oligo(dt) have been investigated by polyacrylamide gel electrophoresis under denaturing conditions. The cDNA preparations contained a fraction which corresponded in length to the alpha and beta chains of the mRNA used as a matrix. The lengthwise distribution of poly(A) sequences, isolated from globin mRNAs, corresponded to a statistical distribution, with a maximum at 75-80 nucleotides and with a maximum length of approximately 150 nucleotides. The lengthwise distribution of poly(dt) sequences, isolated from cDNA, corresponded to the poly(A) sequence distribution. It was concluded that: 1) cDNA is heterogeneous with respect to the length of poly(dt) sequences located at the 5' end of the molecule; 2) there was no selective use as primer of only that oligo(dt) which was located within the complex formed with the 5' end of the mRNA poly(A) sequence; 3) the heterogeneity of poly(DT) corresponded qualitatively to two models of template-primer interaction: selective use as a true primer of the oligonucleotide located in the complex formed with the 3' end of mRNA, and a statistical distribution of primer along the poly(A) sequence; and 4) the heterogeneity of the homopolymeric section of the cDNA may affect the kinetics of hybridization with mRNA, which possibly explains the previously observed [1] anomalous dependence of the hybridization rate on time.

Animals↗
PubMedDate not supplied

[Circulating immune complexes and age].

The circulating immune complexes were quantitatively determined by PEG-precipitation in the serum samples of 413 from the clinical point of view healthy persons subdivided into several age groups. In comparison with the other age groups the age group of 0-15 years showed the lowest immune complex values (mean = 3.14 +/- 0.96 mg protein/ml) with a significant higher distribution (p = 5%). Significant differences between the persons of the age of 16-30, 31-45 and 46-60 years were not traceable. The average values of the circulating immune complexes amounted to 3.82 +/- 0.75, 3.9 +/- 0.67 respectively 3.91 +/- 0.38 mg protein/ml. In comparison with these three age groups the circulating immune complexes showed with an average value of 3.62 +/- 0.75 mg protein/ml a falling tendency for the persons more than 60 years old. There existed a statistical significant difference (p = 5%) for the persons of the age of 31-45 years. A sexual dependence of the immune complexes could not be found within the age groups. Autoantibodies (ANF, anti-dsDNA antibodies) were traceable in the serum of 23 (24.7%) of the persons more than 60 years old. The comparison of the sera with and without autoantibodies didn't show any significant difference with regard to the immune complexes.

Adolescent↗
PubMedDate not supplied

[An unusual constellation of findings in polymyositis--differential diagnosis and clinical course].

Clinical and biochemical findings are described in a 34-year-old female with atypical polymyositis. Uncommon clinical features in this patient included distally accented decreased muscle strength and myalgias, atypical electromyographic findings, a remarkable discrepancy between clinical findings and laboratory parameters of myolysis, unexplained episodes of somnolism, presence of increased serum lactate levels, and a unilateral mamma aplasia. For this combination of signs a polymyositis or an inclusion body myositis, but also a metabolic or heredodegenerative myopathy was considered. Finally, the idiopathic polymyositis was confirmed histologically and a marked improvement in the clinical and biochemical signs occurred after commencement of high-dose methylprednisolone.

Adult↗
PubMedDate not supplied

[Successful therapeutic management of risk pregnancy in primary Sjögren syndrome with plasmapheresis and dexamethasone].

We describe a 28-year-old woman with primary Sjögren's syndrome who had a miscarriage in the 10th week of gestation in December 1993. In her second pregnancy plasma hyperviscosity and high antibody reactivities to 52/60 kD Ro(SS-A) and La(SS-B) represented elevated risk factors for abortion and fetal congenital heart block. Therefore, we performed plasmapheresis and administered dexamethason. At the end of 37th week of gestation a healthy boy was born by caesarean operation. Interestingly, antibody levels to the anti-Ro(SS-A)/La(SS-B) complex showed an increase during both pregnancies with a decline after the abortion as well as the partus. The combined therapy led to a decrease in both the autoantibody reactivities to 52 kD Ro(SS-A) and La(SS-B) and the plasmaviscosity. Plasmapheresis and dexamethason are safe treatment modalities in identified high risk pregnancies for the birth of a child with CHB.

Abortion, Missed↗