Identification of naturally processed T cell epitopes from glutamic acid decarboxylase presented in the context of HLA-DR alleles by T lymphocytes of recent onset IDDM patients.

Glutamic acid decarboxylase (GAD) has been defined as a major target antigen in insulin-dependent diabetes mellitus (IDDM). To identify the molecular ligands triggering a T cell response to GAD, a panel of human GAD65-specific T lymphocyte lines was generated from peripheral blood of three recent onset IDDM patients. All lines derived from a patient expressing the high-risk-conferring HLA-DR*0301/ *0401 haplotypes recognized a single epitope localized between amino acid positions 270 and 283 of GAD65, a stretch that is located in close proximity to the homology region shared with Coxsackie virus P2-C protein. All lines with this specificity were restricted to the DRA, B1*0401 product of the DR4 haplotype. Analysis of the GAD-specific T cell response in a second patient homozygous for DR4 haplotypes demonstrated that the same DRA, B1*0401 allele selected T cells specific for a different determinant. The T cell response profile in a third patient showed that DR*1501/ *1601-encoding haplotypes could present at least three different epitopes to GAD65-specific T lymphocytes. One of these epitopes was presented by a DR allele associated with the resistance-conferring DRB1*1501 haplotype. GAD-specific T cell lines could not be isolated from HLA class II-matched normal individuals. Our data reveal that (a) the T cell response to GAD65 is quite heterogenous in recent onset IDDM patients; (b) HLA-DR, not DQ, seems to be the principal restriction element used by T cells present at the onset of the disease; and (c) T cells responding to epitopes containing identical sequences to Coxsackie virus P2-C protein were not detected.

Autoantibody epitopes to the smaller isoform of glutamate decarboxylase do not differ in Swedish and Japanese type 1 diabetes patients and may be associated with high-risk human leucocyte antigen class II alleles.

Type 1 diabetes (T1D) is an autoimmune disease with a strong human leucocyte antigen (HLA) class II association. Depending on geographic locations, the disease-associated HLA class II alleles vary. We evaluated the beta cell-specific autoimmunity reflected in autoantibodies directed to the smaller isoform of glutamate decarboxylase (GAD65) in Japanese and Swedish T1D patients. GAD65Ab epitope specificities were assessed using GAD65-specific recombinant Fab. GAD65Ab epitope specificities did not differ between Swedish and Japanese patients. Only recognition of the MICA-4-defined middle epitope was significantly stronger in the Japanese T1D patient group compared to the Swedish T1D patients (P = 0.001). Binding to the b96.11-defined middle epitope was substantial in both groups and showed significant associations with high-risk HLA class II haplotypes. In the Japanese T1D group the association was with haplotype DRB1*0802-DQB1*0302 (P = 0.0008), while in the Swedish T1D patients binding to the b96.11-defined epitope as associated with the presence of high-risk HLA genotypes DR3-DQB1*0201 and/or DR4-DQB1*0302 (P = 0.02). A significant association between reduction in binding in the presence of recombinant Fab (rFab) DPD and high-risk allele DQB1*0201 was found (P = 0.008) in the Swedish T1D patients only. We hypothesize that epitope-specific autoantibodies effect the peptide presentation on HLA class II molecules by modulating antigen uptake and processing. Molecular modelling of the high-risk HLA class II molecules will be necessary to test whether these different molecules present similar peptide-binding specificities.

Characterization of preparations of GAD65, proinsulin, and the islet tyrosine phosphatase IA-2 for use in detection of autoreactive T-cells in type 1 diabetes: report of phase II of the Second International Immunology of Diabetes Society Workshop for Standardization of T-cell assays in type 1 diabetes.

The identification, quantification, and characterization of T-cells reactive with the islet autoantigens GAD65, proinsulin (PI), and tyrosine phosphatase-like molecules IA-2 and phogrin are major research goals in type 1 diabetes. In the Immunology of Diabetes Society First Workshop on Autoreactive T-Cells, the quality of recombinant preparations of these autoantigens was identified as a significant weakness, a finding that may account for much of the inconsistency in published studies of peripheral blood T-cell reactivity to islet autoantigens. Poor antigen quality has also hampered the development of novel technologies for the detection of islet-reactive T-cells. For these reasons, in the present study, several preparations of GAD65, PI, and IA-2 were collected and evaluated for endotoxin content, ability to stimulate a panel of relevant T-cell clones, and inhibitory effects on proliferation to unrelated third-party antigens. Through this process, we have been able to identify preparations of GAD65 and IA-2, generated in insect cells using the baculovirus expression system, that stimulate relevant clones and display low inhibitory effects on third-party antigens. In addition, we characterized a PI preparation generated in bacteria as being free of effects on proliferation to third-party antigens and low in endotoxin content. These preparations are important to promote the development of robust and sensitive assays of islet-reactive T-cells in patients with type 1 diabetes or patients at high risk for developing the disease.

A new ELISA-based assay for quantitation of human T-lymphocyte subpopulations.

In order to circumvent the problems associated with the available methods, we have developed a simple, reliable ELISA for quantitation of T-cells and their subpopulations, T-helper/inducer and T-suppressor/cytotoxic cells. Standard curves with three concentrations of sheep anti-mouse IgG-coupled beads for each T-cell population were used for the determination of the T-cell content in blood. Enzyme-labelled monoclonal antibodies against T-Pan, T-S and T-H cell surface markers were readily able to bind to such beads and the test system was calibrated with T-lymphocytes by comparing cytofluorographic and enzyme immunometric results. Purified preparations of monocytes and granulocytes were negative in the test. Lymphocytes from 50 healthy blood donors gave results which correlated closely with cytofluorograph determinations.

Intradermal administration of GAD & evaluation of diabetes incidence in mice: possible relevance for skin tests in humans.

In vitro cell mediated reactivity to Glutamic Acid Decarboxylase (GAD) has been reported in man and in the non-obese diabetic (NOD) mouse. The demonstration of such reactivity in vivo using GAD in a simple intradermal skin test would be useful for mass screening of subjects at risk of Type 1 diabetes. Such a skin test could be simply applied to the forearm, then signs of local reaction would indicate patients at risk. However, in order to safely apply a skin test of this type it must be certain that administration of the antigen does not itself provoke or start the process leading to diabetes in susceptible individuals. In the present study the NOD mouse model was used. GAD and two peptides of GAD, which may have relevance to the disease process, were applied intradermally to these mice to determine whether a local reaction could be seen and to see if the diabetes rate was altered. Moreover, Balb/c mice, which can be considered to be at zero risk of developing the disease, were also injected with the same GAD and GAD peptides. No significant differences were seen in the diabetes incidence of the treatment groups compared to the control groups in either the NOD or Balb/c mice although a local swelling was seen in female NOD mice susceptible to diabetes after GAD administration in the footpad. We conclude that the administration of GAD and/or GAD peptides does not provoke or accelerate diabetes incidence in the NOD mouse and that an intradermal skin-test with GAD may be suitable for preliminary trials aimed at large scale screening of humans for their potential to develop type 1 diabetes.

[Graphic representation of normal and pathological pregnancies by means of an ultrasonic-growth curve of the amniotic sac and of the biparietal diameter (author's transl)]. / Die graphische Darstellung normaler und pathologischer Schwangerschaftsverläufe mittels Ultraschallwachstumskurve des Fruchtsackes und des biparietalen Durchmessers.

A diagram is presented which enables the continuous recording of biometric values from the earliest possible time right up to the end of pregnancy. It has been tested in practice. The duration of pregnancy and fetal development and growth can be evaluated from the diagram.

[Early detection of postoperative deep-vein thrombosis in gynaecological patients by the 125I-Fibrinogen test (author's transl)]. / Frühdiagnose der postoperativen tiefen Beinvenenthrombose mit dem 125Jod-Fibrinogentest an einem gynäkologisch operierten Patientengut

Deep-vein thrombosis was detected by means of the 125I-fibrinogen test in 37.2% of patients undergoing gynaecological surgical procedures within the first six days postoperatively; 50% of the cases occurred within the first 48 hours. There was no difference in the incidence of thrombosis according to whether the abdominal or vaginal approach had been used. Varicose veins proved to be a high-risk factor in the development of deep-vein thrombosis. Prophylactic administration of horse-chestnut extract has to begin preoperatively in order to achieve a beneficial result.

[Results of a questionnaire issued to expectant and delivered mothers (author's transl)]. / Ergebnisse einer Schwangeren- und Wöchnerinnenbefragung.

At the Department of Obstetrics and Gynaecology, Wilhelminenspital der Stadt Wien, Vienna, 98% of all deliveries are continuously monitored. 30% (1979) received epidural anaesthesia. The presence of the husband in the delivery room and partial rooming-in is available to all mothers. To find out if the service to expectant and delivered mothers is according to the requirement of our patients, questionnaires were distributed indiscriminately over a two-month period to 350 pregnant women and to 240 women in the puerperium. Fetal monitoring was valued positively in the majority of cases, the presence of the husband during delivery is requested in a minority of cases only, but would be welcomed by a higher proportion of puerperal women for the next delivery. Expectant mothers wished epidural anaesthesia in 50% of cases. Not only pregnant, but also delivered women demanded rooming-in, the latter group agreeing in the main with our partial form of rooming-in. Nearly 90% of mothers with rooming-in felt well prepared for baby care on leaving the hospital. We believe in the possibility of a synthesis of continuous monitoring to achieve optimum safety of delivery and family-orientated obstetrics in the hospital management of labour and the puerperium.

[Intravaginal treatment of colpitis maculosa with an oestriol-containing vaginal cream (author's transl)]. / Intravaginale Behandlung der Colpitis maculosa mit einer östriolhaltigen Vaginalcreme.

43 patients with colpitis maculosa (average age 60.2 years) were selected for an open control therapeutic study with Ortho-Gynest vaginal cream (Ortho-Cilag). The cream contains 0.5 mg of oestriol per single applicator filling. The treatment lasted from 3 to 4 weeks, success being evaluated by clinical documentation and cytological evaluation of vaginal smears before and after treatment. 10 patients (23.3%) were treated successfully, 29 (67.4%) showed a distinct improvement both clinically and cytologically, whilst the remaining 4 (9.3%) showed only moderate improvement. Hence, 39 patients (90.7%) were classified as having been successfully or partly successfully treated. Severe symptoms disappeared completely or were greatly alleviated in 91.4% cases. Moderate symptoms vanished in 59.7%. 58.1% showed a complete normalisation of the former atrophic vaginal skin. Blood spotting and reddening of the vaginal wall vanished completely. A change from dry to moist vagina occurred in 77.3% patients. Discharge vanished completely in 80.6% cases. No untoward side effects were recorded.

[Problems and results of skull x-ray following vacuum extraction (author's transl)]. / Röntgenuntersuchungen am kindlichen Schädel nach Vakuumextraktion, Probleme und Ergebnisse

The vacuum extraction supplies the forceps nearly complete on the IInd Department of obstetrics and gynecology Vienna in the last years. Skull X-ray of 104 newborns following vacuum extraction and of 101 spontaneous delivered newborns are examined on the incidence of fractures, fracture-fissures, elevations and widening of skull sutures. Early pathological findings are rigid criteria for the possible danger due to a method of operative delivery but we have no typical late damage after vacuum extraction or forceps delivery, we don't find in spontaneous born children too.

[Spontaneous rupture of uterus during delivery. Interpretation of cardiotocogram (author's transl)]. / Falbericht -- Spontane Uterusruptur Interpretation des Kardiotokogramms.

The authors report a case of silent rupture of uterus during delivery. The whole delivery was monitored by intrauterine pressure recording and fetal scalp electrode. Severe fetal bradycardia induced cesarian section. The retrospectively study of the papercover of fetal monitoring indicates the rupture of uterus.

Chemical composition and structure of cell wall teichoic acids of staphylococci.

The cell wall teichoic acid structures of 22 staphylococci including 13 type strains were determined. Most of the strains contain a poly(polyolphosphate) teichoic acid with glycerol and/or ribitol as polyol component. The polyolphosphate backbone is partially substituted with various combinations of sugars and/or amino sugars. Most of the substituents occur in a monomeric form but some strains also contain dimers of N-acetylglucosamine as substituents. Staphylococcus hyicus subsp. hyicus NCTC 10350 and S. sciuri DSM 20352 revealed rather complex cell wall teichoic acids. They consist of repeating sequences of phosphate-glycerol-phosphate-N-acetylglucosamine. The amino sugar component is present in this case as a monomer or an oligomer (n less than or equal to 3). Moreover, the glycerol residues are partially substituted with N-acetylglucosamine. The cell wall teichoic acid of S. auricularis is a poly(N-acetylglucosaminyl-phosphate) polymer similar to that found in S. caseolyticus ATCC29750. The cell wall teichoic acid structures for type strains of S. auricularis, S. capitis, S. cohnii, S. haemolyticus, S. hominis, S. hyicus subsp. hyicus, S. sciuri, S. xylosus and S. warneri were determined for the first time in detail. The structures of some of the previously described teichoic acids had to be revised (S. epidermidis, S. simulans, S. aureus phage type 187).

Determination of cell wall teichoic acid structure of staphylococci by rapid chemical and serological screening methods.

Investigations of cell wall teichoic acid structures of various staphylococci were carried out by a rapid method based on the gas-liquid chromatographic separation of products obtained after treatment of phenol-extracted cells with 70% hydrofluoric acid. In most of the strains teichoic acids of the poly(glycerolphosphate) and/or poly(ribitol-phosphate) type were found. Teichoic acids of the poly(glycerolphosphate-N-acetylglucosaminephosphate) type and polymers consisting of N-acetylglucosaminephosphate were present in few strains. The results obtained by the rapid chemical screening method were compared with data obtained by serological analysis of teichoic acid structures using specific antisera and the lectin wheat germ agglutinin. Teichoic acid components occurring in low concentrations could only be detected with the chemical and not with the serological method. A number of strains of species of the genus Staphylococcus have been studied using these rapid methods. With a few exceptions, the teichoic acid structure proved to be a constant marker within a given species.