Reconstitution of bactericidal activity in chronic granulomatous disease cells by glucose-oxidase-containing liposomes.

Chronic granulomatous disease (CGD) is an inherited primary immunodeficiency characterized by phagocytes devoid of a functioning nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. The failure of CGD phagocytes to produce reactive oxygen species (ROS) results in a marked increase in the susceptibility of affected patients to life-threatening bacterial and fungal infections. This study investigated whether loading of CGD phagocytes with glucose oxidase (GO)-containing liposomes (GOLs) could restore cellular production of bactericidal ROS (eg, H2O2 and HOCl) in vitro. Results indicate that GO encapsulated in liposomes enabled NADPH oxidase-deficient phagocytes to use H2O2 for the production of highly bactericidal HOCl. The intracellular colocalization of bacteria and liposomes (or liposome-derived ferritin) was demonstrated by confocal laser microscopy and electron microscopy. After uptake of GOLs (approximately 0.2 U/mL at 1 mM total lipid concentration, size approximately 180 nm), CGD granulocytes produced HOCl levels comparable to those of normal phagocytes. Remarkably, after treatment with GOLs, CGD phagocytes killed Staphylococcus aureus as efficiently as normal granulocytes. Moreover, treated cells retained sufficient motility toward chemotactic stimuli as measured by chemotaxis assay. Side effects were evaluated by measuring the H2O2 concentrations and the production of methemoglobin in whole blood. These studies revealed that H2O2 produced by GOLs was degraded immediately by the antioxidative capacity of whole blood. Elevated methemoglobin levels were observed only after application of extremely high amounts of GOLs (2 U/mL). In summary, the application of negatively charged GOLs might provide a novel effective approach in the treatment of patients with CGD at high risk for life-threatening infections.

Influence of anticoagulants on the level of soluble HLA class I and class II antigens measured in blood samples.

The existence of soluble forms of HLA class I and class II antigens in human serum is well established and altered concentrations of these serum proteins have been described to be associated with various diseases. Since soluble HLA antigens (sHLA) can be measured both in serum and plasma samples, we investigated whether anticoagulant treatment influences the determined levels of soluble HLA class I (sHLA-I) or soluble HLA-DR (sHLA-DR). Analyzing paired samples of serum and plasma of 40 healthy individuals we found significantly lower serum levels of sHLA-DR (0.31 +/- 0.15 ng/ml) compared to EDTA plasma levels (0.58 +/- 0.20 ng/ml). By contrast, serum levels of sHLA-I (0.89 +/- 0.74 micrograms/ml) were only slightly lower than EDTA plasma values (0.95 +/- 0.86 micrograms/ml), a situation similar to that of sIL-2R and sCD4 levels. Further experiments intended to clarify the reasons of the reduced sHLA-DR serum levels revealed that (i) the blood storage time before centrifugation did not influence the sHLA-DR level, (ii) treatment of serum with anticoagulant did not augment the measured sHLA-DR concentration, and (iii) the recovery of spiked sHLA-DR was significantly lower when added to native blood than to serum or anticoagulant-treated blood. These results suggest that sHLA-DR is partly removed by the process of blood clotting thus resulting in diminished sHLA-DR serum levels.

Quantitation of soluble HLA-DR antigens in human serum and other body fluids.

The existence of soluble forms of MHC class II molecules is well established. To quantify soluble HLA-DR antigens (sHLA-DR) in human serum and other body fluids, we developed an enzyme immunoassay using two non-overlapping HLA-DR-specific monoclonal antibodies (RoDR, BL-la/5) and an immunoaffinity chromatography-purified sHLA-DR standard. In serum of healthy individuals, sHLA-DR levels were found in the range between 0.6 and 3 ng/ml (median 0.85 ng/ml) whereas EDTA plasma samples showed concentrations about 20 times higher (median 21 ng/ml). In tears, saliva, sweat, urine, amniotic fluid, cerebrospinal fluid, and bronchoalveolar lavage, sHLA-DR could also be detected. No association was found between sHLA-DR serum levels and distinct HLA specificities. In the sera of patients with autoimmune diseases, slightly enhanced sHLA-DR values were found (juvenile rheumatoid arthritis: median 2.0 ng/ml, lupus erythematosus: 1.5 ng/ml, diabetes mellitus: 2.1 ng/ml).

[11th International Histocompatibility Workshop 1991: Personal data for the Multiple Sclerosis Study]. / 11. Internationaler Histokompatibilitäts-Workshop (IHWS) 1991: Eigene Daten zur Multiplen-Sklerose (MS)-Studie.

In our multiple sclerosis (MS) study as a part of the 11th IHWS we HLA-typed 6 MS families with 9 patients and defined the complement polymorphisms (BF, C2, C4) of these families. The aims of the study were the definition of the MS susceptibility gene and the investigation of the involvement of other factors in the etiopathogenesis of MS. The MS study of the IHWS demonstrated a strong association with HLA-DRw15 and -DWw6 in a Caucasian population. The heterozygous C2 deficiency in our family PD1 linked with the haplotype A25 B18 DR2 BFS C4A4 C4B2 confirmed by complement titration may express the participation of complement factors in the etiopathogenesis of MS resulting in immunogenetic heterogeneity of MS. Analysis of the 3 MS pairs of sisters shows the linkage of HLA with the assumed MS susceptibility gene. This could not be confirmed in the whole MS family study of the 11th IHWS.

Acquisition and reversal of taste/tactile discrimination after forebrain noradrenaline depletion.

Two experiments were performed to assess the role of noradrenaline (NA) on the acquisition of an aversively motivated discrimination task and its reversal. A conditioned taste aversion procedure was used. The NA depletions were achieved through two different pharmacological means: systemic N-2-chloroethyl-N-ethyl-2-bromo-benzylamine (DSP4) and destruction of the dorsal noradrenergic bundle (DNAB) with 6-hydroxydopamine. Both procedures caused marked reductions of NA in the frontal cortex and hippocampus. In neither of the studies (Experiment 1, DSP4, and in Experiment 2, DNAB) were there any significant changes between controls and NA-depleted rats in either the rate of acquisition of the original discrimination (Phase 1) or the subsequent reversal (Phase 2). This occurred irrespective of which of the two stimuli (a taste cue, i.e., saccharin presented in bottles with nozzles that do not have ball bearings, "silent bottles," or a tongue-tactile cue, i.e., water in bottles with nozzles that had ball bearings "noisy bottles") initially was used as the conditioned stimulus (CS1, i.e., the stimulus first followed by contingent administration of lithium chloride, and later, in Phase 2, followed by saline injections). Thus NA does not appear to be critically involved in the acquisition and reversal of a taste/tactile discrimination task. The significance of forebrain NA for other discrimination tasks is discussed.

[Immunologic reactivity in the rabbit after the application of Xenoderm]. / Studie zur Immunreaktivität beim Kaninchen nach Applikation von Xenoderm.

UNLABELLED: The immunological compatibility of the pig skin preparation, Xenoderm was tested in an animal model. Firstly an experimental conception was selected which seized the immune degree of Xenoderm on the humoral and cellular level in animal test. Specific immune reactions should be provoked in rabbit after a repeated artificial immunisation by soluble protein extracts respectively by means of recured implantation of Xenoderm tissue pieces and native pig skin. Test period 3-6 months. RESULTS: Native pig skin of the offered configuration induced an intensive humoral and cellular immune reaction in experimental animal. Xenoderm was immunologically inert after application as a soluble protein extract in different concentrations and also as an implant. Xenoderm (desantigenic splitted pig skin preserve) is suitable as a temporary skin replacement for the therapy of skin defects.

Radioimmunoassay for serum thyroglobulin designed for early detection of metastases and recurrencies in the follow-up of patients with differentiated thyroid carcinoma.

A radioimmunoassay (RIA) for the measurement of thyroglobulin in human serum was developed and factors that influence sensitivity were investigated. In a comparison of 3 different labeling procedures (chloramine T, iodogen, lactoperoxidase) iodogen-prepared tracer proved to be slightly superior with respect to sensitivity and stability. The shelf life of the tracer was improved by a protein-enriched buffer, which serves as a radical scavenger. The binding kinetics of tracer to antibody at different temperature ranges were examined, and the most rapid and complete binding was found at room temperature. For the preparation of standard curves, several artificial media were compared with thyroglobulin-free serum. Second antibody separation was investigated and optimized. By employing sequential saturation, sensitivity of 0.75 microgram/1 (B0-3 SD) and 50% intercept of less than 5 micrograms/l were achieved. The results of RIA measurements of thyroglobulin in 142 patients with papillary and follicular thyroid carcinoma after thyroidectomy and 131I treatment were compared with 131I whole-body scans. The results confirmed that serum thyroglobulin is an early indicator of recurrency.

Monozygotic vs. dizygotic twin behavior in artificial mouse twins.

Adult inbred mice of an isogenic strain (AKR/NHan or C57BL/6J Han) differ in social (sexual and agonistic), emotional and psychomotoric behavior, depending on the kind of manipulation to which they were subjected at an early ontogenetic stage. Monozygotic twins (MZT) from eight-cell stages halved and transferred to uterine foster mothers were compared with dizygotic twins (DZT) from nonreduced but transferred eight-cell stages and with naturally born animals (NBA). Generally, early embryonic conditions predict the behavioral characteristics of the adult animals to a high degree. The MZT are motorially less active, less emotional, less aggressive and less socially interested than DZT and NBA. In tests of spontaneous social behavior (allogrooming, anogenital licking, mounting, fighting), as well as in tests for emotionality (open field: crossed fields and defecation), these behavioral patterns occurred less frequently in MZT than in DZT; the NBA were mostly intermediate. The copulatory pattern of male MZT differs from that of male DZT by a shortage of intromission latency and duration; furthermore, MZT pairs do not build up a steady rank order in competitive copulation tests, as opposed to DZT and NBA pairs. In a test for psychomotoric behavior (swimming), the MZT prefer "floating" as a survival strategy, whereas the DZT and NBA prefer "adult swimming." Therefore, it can be concluded that these behavioral differences may be caused by the particular psychosocial environment in which the twins grow up or may be due to early prenatal peculiarities, such as inadequate synchronization of the developmental status of uterus and embryo.

Vaccinations in the presence of skin diseases.

The authors collected data from the literature and from experts on the subject of vaccination of patients with dermatologic disorders. They assembled these data into categories of specific skin conditions and vaccinations. These data may be used in deciding whether or not to vaccinate, and at which stage of the dermatologic condition.