Interleukin-2 receptor expression and interleukin-2 production in bullous pemphigoid.

Expression of interleukin-2 (IL2) receptors was studied on peripheral blood lymphocytes (PBLs) in 25 patients with bullous pemphigoid. Analysis was carried out by flow cytometry. Without immunosuppressive therapy expression of IL2 receptors only on T cells (CD5) was significantly increased as shown by double staining (11.9% +/- 7.8% vs 2.1% +/- 1% in controls). After 2-3 weeks under immunosuppressive treatment with prednisolone and azathioprine, however, BP lymphocytes did not exhibit any IL2 receptors. PBLs of 18 BP patients showed an increase in IL2 production (1027.4 +/- 670.5 U/ml vs 270 +/- 100 U/ml in controls) during the acute stage of the disease after stimulation with phytohaemagglutinin (PHA)/phorbol myristate acetate (PMA). On the contrary, IL2 production of the same cells in five patients was only in the lowest range of control values after PHA stimulation without PMA (12.5 +/- 29.6 U/ml vs 40 +/- 20 U/ml in controls). Under treatment with immunosuppressants the IL2 production normalized after PHA/PMA stimulation and slightly decreased following PHA stimulation. From these results we conclude that a T-cell activation via activation antigens, as IL2 receptors, and the production of the specific ligand, IL2, may play a role in the pathogenesis of bullous pemphigoid, especially in the earliest stages, and serve as a marker of disease activity.

[Characterization of a monoclonal antibody against a differentiation antigen of human natural killer cells (NK cells)]. / Charakterisierung eines monoklonalen Antikörpers gegen ein Differenzierungsantigen von humanen natürlichen Killerzellen (NK-Zellen).

The new murine monoclonal antibody BL-(H5) reacts with a novel surface molecule which is mainly expressed on human NK and B cells. The antigen is not expressed on peripheral T lymphocytes, thymocytes and different human T-cell lines. BL-(H5) does not bind to erythrocytes and platelets. The monoclonal antibody reacts in western blotting experiments with an antigen of 78kDa.

[Simple preparation of R-phycoerythrin of the red alga Ceramium rubrum (Huds. 1762)]. / Einfache Präparation von R-Phycoerythrin aus der Rotalge Ceramium rubrum (Huds. 1762).

Phycoerythrin is a highly fluorescent phycobiliprotein. Because of its special fluorescence characteristics R-phycoerythrin is very useful in flow cytometry. The preparation of R-phycoerythrin of the red alga Ceramium rubrum is described.

[A simple method for separating fresh and cryopreserved human sperm using dextran-visotrast density gradient centrifugation]. / Eine einfache Methode zur Separierung von frischen und kryokonservierten Humanspermien mittels einer Dextran-Visotrast-Dichtegradientenzentrifugation.

A procedure for the separation of fresh and cryopreserved human spermatozoa is presented, compared with the Ficoll method according to Harrison [7] and clinically tested. The principle includes a filtration through glass wool and a gradient centrifugation. The gradient medium consists of Ca2(+)-free buffer, Dextran "70" and Visotrast, a radio-opaque medium. The sperm concentration, the sperm-migration-index and the sperm morphology checked by FACS 440 corresponds to the results of the Ficoll-method described by Harrison [7]. Furthermore, our method is usable for semen samples after adding of a cryoprotective medium with egg yolk and glycerol. An improvement of the chance of conception after artificial insemination by husband (AIH) by our method could not be found up to now.

Lack of HLA-molecules on human spermatozoa and in seminal plasma.

The expression of human leukocyte antigens (HLA) on ejaculated spermatozoa and on lymphocytes was compared by flow cytometry using monoclonal antibodies towards HLA class I (pan-HLA-A, -B, -C) and class II (DR) antigens. Soluble antigens of HLA class I (s HLA-A, -B, -C) in seminal plasma and in blood plasma were monitored with an ELISA technique. Lymphocytes showed specific fluorescence after incubation with the antibodies against HLA class I and class II (DR), whereas, on spermatozoa no positive immunofluorescence could be detected. No antibodies were bound to any significant extent either after modifications of sperm preparation (density gradient centrifugation, swim up-technique, addition of azide, foetal calf serum or benzamidine chloride) or after treatment of spermatozoa with detergents. Furthermore, different concentrations of soluble HLA-A, -B, -C in seminal plasma and in blood plasma were detected. The latter one showed soluble HLA about four-fold more concentrated than the seminal plasma (means +/- SD: 262.5 +/- 144.4 nmol l-1 vs. 62.5 +/- 27.1 nmol l-1). These results suggest, that the HLA-expression differs between human spermatozoa and somatic cells.

[Expression of leukocyte antigens in human sperm]. / Expression von Leukozytenantigenen auf Humanspermien.

The expression of leukocyte antigens on human ejaculated spermatozoa and lymphocytes was compared by means of the immunofluorescence technique and monoclonal antibodies against anti-HLA class I (pan-HLA-A,B,C) and class II (DR, DR/DP). Lymphocytes showed specific fluorescence phenomena after incubation with the antibodies. However, in the case of spermatozoa no immunofluorescent labeling was detected. Modifications of the method of sperm preparation (Ficoll centrifugation, swim-up technique, addition of azide, fetal calf serum or benzamidinchloride) did not lead to different results. We were not able to find HLA-antigen expression on fresh human spermatozoa using immunofluorescence.

[Pre- and postoperative reactivity of breast cancer patients to tumor associated antigens and HEP in the macrophage electrophoresis mobility (MEM) test]. / Prä- und postoperative Reaktivität von Mammakarzinom-Patienten gegen tumorassoziierte Antigene und HEP im Makrophagen-Elektrophorese-Mobilitäts(MEM)-Test.

27 female patients with breast cancer and 23 with benign breast diseases have been investigated with the MEM-test. Antigens used in these experiments were human encephalitogenic protein (HEP) and 3M KC1 tissue extracts from carcinomas of breast and kidney. Preoperative examination of the patients showed an accuracy of the test of 89% (breast cancer extract) and 86% (HEP) respectively. The control persons gave less than 9% false positive tests. The results obtained with the extract from kidney cancer demonstrate the organ specificity (3% positive responses). Follow-up studies were carried out with cancer patients. We found, beginning with a delay, a strong decrease of reactivity 5 to 8 weeks after surgical treatment and later again increased reactivity in some patients. We suppose a relation between this restoration of MEM-reactivity and development of recurrent cancer whereby follow-up studies can get some clinical importance. The investigation of frequency distribution of MEM values obtained shows a division into low values of about 0% and high values of about 10% mobility inhibition without overlapping. The reasons are discussed regarding the levels of the patient and the test principle.

[Immunologic studies in patients with dilated cardiomyopathy]. / Immunologische Untersuchungen bei Patienten mit dilatativer Kardiomyopathie.

In 10 cases of dilated cardiomyopathy the HLA-loci (A, B, C, DR) and marker expression on lymphocytes were studied. In this group the T4/T8-ratio was decreased and interleukin-2-expression was increased in relation to normals or patients with chronic coronary heart disease, respectively. 9 patients had the HLA-A1 or HLA-A2-locus in their siblings. In correlation analysis between immunological and hemodynamic parameters a good correlation of the number of NK-cells to the ejection fraction (r = 0.79) and negative correlation of helpercells (CD4) to the cardiac output were found (r = -0.80). The results of this attempt suggest an immunological pathogenesis of the dilated cardiomyopathy.