Zeitschrift fur Immunitatsforschung, experimentelle und klinische Immunologie最新文献

By means of free-flow electrophoresis a preparative separation of cells according to their surface charge is achieved. The method has been applied to human lymphocytes of blood and of tonsils. Technical details are described and discussed with respect to optimal separation conditions, reproducibility of results and viability of the separated cells. Unseparated cells, pooled separated cells and cells corresponding to different electrophoretic mobilities are compared for stimulation in response to PHA, determination of cells bearing membrane immunoglobulins, colony growth of hemopoietic stem cells and DNA syntheses in vitro. It is concluded that reproducible results can be obtained, that the biological activities of the cells after separation are unaltered and that a separation or at least an enrichment of cells with certain properties can be achieved.

Transferrin phenotypes were determined in 3380 sera of unrelated persons of the western region of Germany with 97.60 percent for TfC and 2.40 percent for Tf variants. Identification was achieved by immunochemical means or through autoradiography. Relative mobilities in some variants were measured using Tf B2C (0.7) as reference. Application of Tf variants is demonstrated in paternity cases.

The antibody response of rabbits to Micrococcus lysodeikticus is characterized by the production of a high concentration of antibodies which manifest markedly reduced heterogenicity. The specificity of these antibodies was studied and it revealed that M. lysodeikticus contains 2 major antigens: both the glucose-N-acetyl-aminomannuronic acid polymer obtained by formamide extraction of the cell walls and peptidoglycan solubilized by ultrasonic treatment gave precipitin reactions with hyperimmune antisera. By means of inhibition studies of the glucose-mannose polymer specificity, glucose appeared as the immunodominant sugar in the majority of antibodies studied. Inhibitions studies also confirmed that both the glycan and peptide moieties constitute antigenic determinants of M. lysodeikticus peptidoglycan. Antibodies to the glucose-mannose-polymer and the peptidoglycan were specifically fractionated by use of immunoadsorbents formed from lysozyme solubilized cell walls and activated Sepharose. Both antibody specificities showed a limited heterogeneity by isoelectric focusing. Finally, because antisera to M. lysodeikticus are a rich source of antibodies to peptidoglycan, emphasis is placed on the possible usefulness of this system for studies of clonal dominance.

The frequencies of 36 HL-A antigens of the SD-1, SD-2 and SD-3 locus have been determined in 20 patients with idiopathic Addison's disease and compared with those of 450 controls. No significantly different HL-A antigen frequencies could be observed in both groups. These findings are in contradiction to data reported by a Copenhagen group. Possible mechanisms which might have led to this discrepancy are discussed.

Migration inhibition factor (MIF) was produced by peritoneal exudate (PE) cells from guinea pigs sensitized with heat-killed cells of Staphylococcus aureus cultured in vitro with staphylococcal cell walls or defined subunits of the cell walls. MIF activity was assayed by inhibition of migration of alveolar macrophages from lungs of normal guinea pigs. Specificity of inhibition was established using PE cells from tuberculin sensitive guinea pigs and tuberculin as appropriate controls. Staphylococcal cell walls, their peptidoglycan complex, and teichoic acid-peptidoglycan fragments stimulated MIF production by staphylococcus sensitive PE cells. Peptidoglycan fragments and teichoic acid were ineffective antigens.

A transfer factor (TF) was prepared and characterized from the leucocytes of Mantoux-positiv donors using a variety of techniques. The results allowed the following conclusions: Whilst extraction by dialysis and phenol (without precipitation) led to nearly the same positiv immunological activities, when assayed in the MEM-test, phenol extracts subjected to RNA-precipitation lost considerable activity. The residual activity was associated with the lower molecular fractions, and RNA itself has shown no activity. Residues after leucocyte dialysation have also shown considerable TF-activity when extracted by phenol. It was found that phenol extraction followed by gel filtration allowed recovery of considerable quantities of active TF, confined to several specific filtration fractions. Combination of active and inactive gel filtration fractions suggested that there existed an inhibitory factor within the extracted leucocyte preparations. This modified preparation and purification procedure may allow the preparation of TF for therapeutic purposes.

The peptidoglycan of group A Streptococcus prepared by hot formamide or TCA extraction has an expressive ability to lyse rabbit blood platelets in vitro. Initial changes in the ultrastructure of platelets can be observed after 10 minutes incubation of rabbit platelets with 0.1 mug of Streptococcus peptidoglycan per ml. The submicroscopic alterations are characterized by changes of the shape and damage of the limiting membrane of the platelets. Larger amounts of peptidoglycan (10 mug/ml and more) cause total destruction of platelets; only remainders of the limiting membrane and free granulomers can be seen. Peptidoglycans of Streptococcus pneumoniae and Staphylococcus aureus exhibit a comparable effect on rabbit platelets. There are substantial differences in the sensitivity of blood platelets of different animal species to the streptococcal peptidoglycan. Rat platelets exhibit a similar thrombocytolytic effect after the Streptococcus peptidoglycan treatment as rabbit ones; however, the development of comparable changes in their submicroscopic structure needs a dose of peptidoglycan 10(2)-10(3) times higher. Platelets of guinea-pig, dog, cat, calf and human appear to be quite resistant under analogous conditions to as such a high dose of Streptococcus peptidoglycan as 500 mug/ml.

Lipopolysaccharides (endotoxins) of gram-negative bacteria consist of 2 components with distinct physico-chemical character: a heteropolysaccharide and a covalently linked lipid, termed lipid A. The chemical structure of lipid A, which represents the toxic center of lipopolysaccharides, is discussed. Evidence is presented that lipid A antiserum suppresses the pyrogenic effect of lipid A and lipopolysaccharides in rabbits. The protective power of lipid A antiserum, however, is only expressed in animals which have been pretreated with lipid A or lipopolysaccharide indicating that other than humoral factors, perhaps cellular, also participate in endotoxin fever (cross) immunity. The fever resistance mediated by lipid A antiserum seems to be endotoxin-specific with regard to both the preparative and the challenging injection. Lipid A antiserum therefore may serve as a tool to discriminate between fever caused by endotoxins and that induced by other pyrogens.

In 250 paternity actions (one-man one-child affairs 1965-1970) without exclusion by blood group opinion, which include 48 cases involving persons from abroad, 41 men acknowledged the paternity or withdrew the suit during the process. 32 cases were judged by default. 174 men were judged of child support according to the former 1717 BGB, 13 of these after an anthropological expertise. In 2 cases the anthropological opinion gave an indication of nonpaternity; accordingly, the court dismissed the action. In 157 cases the plausibility of paternity according to Essen-Möller's formula was calculated by the expert and dealt with in the blood group opinion. It appears, however, that the W values had no remarkable influence on the decision of the court.

Staphylococcus epidermidis peptidoglycans solubilized by sonication or lysozyme digestion, and synthetic peptidoglycan analogs such as HSA-carboxymethyl-Gly-L-Ala-L-Ala-D-Ala-D-Ala (HSA-pentapeptide) or L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala (pentapeptide) have been labeled with 125I and tested for their applicability in the radioactive antigen binding assay. Use of radioiodinated Staph. epidermidis peptidoglycans was found to be considerably impeded by the presence of at least 2 different antigenic sites on such molecules, the pentapeptide and the glycan determinant. Application of labeled HSA-pentapeptide was limited by the necessity to use PEG for precipitation of Ag-Ab-complexes and by short linear potions of binding curves. However, the synthetic pentapeptide hapten, radioiodinated by the active ester method of BOLTON and HUNTER, proved to be a most useful regent for the selective measurement of pentapeptide antibody. Inhibition studies indicated that the immunological specificity of the labeled hapten was retained. Pentapeptide binding curves were linear from 15-500 g/ml of antibody. Generally, there was good agreement between pentapeptide antibody concentrations measured by radioimmunoassay and quantitative precipitation.