Acta pathologica, microbiologica, et immunologica Scandinavica. Section C, Immunology最新文献

Human leukocyte phagocytosis of fluorescein-isothiocyanate (FITC)-labelled zymosan particles was studied by a flow cytometric (FCM) assay allowing discrimination of adhered and ingested zymosan particles. Free zymosan particles, non-phagocytes and phagocytes could be discriminated and quantified by simultaneous registration of fluorescence and light scatter. All leukocytes capable of phagocytosis were phagocytosing, and within 15 min 80% of the zymosan particles were adhered or ingested. Compared to the FITC-fluorescence of free zymosan particles, the mean fluorescence of phagocyte-associated zymosan particles was reduced by about 35%, indicating ingestion and processing of zymosan particles. Abolishing the FITC-fluorescence of extracellular zymosan particles by crystal violet, the number of zymosan particles adhered and ingested could be calculated from FCM measurements of phagocyte fluorescence. This showed that in 15 min 83% of the phagocyte-associated zymosan particles were actually ingested.

As shown previously, the suppressor activity measured by the Con A mixed lymphocyte culture assay exhibited a pronounced dependence on the technical parameters. In the present study it is shown that the length of the suppressor generation period, between 24 and 96 hours, had only a modest influence on the suppressor activity. However, when the activity in the responder system was high, there seemed to be a trend towards a slight increase of the suppressor level using a long generation period, i.e. 72 to 96 hours. An increase of the suppressor cell/responder cell ratio from 1/10 to 1/2 was followed by a rising suppressor level; a further increase of the ratio had no significant influence on the suppressor activity. A decrease of the ratio below 1/10 showed variable results: stimulation, no effect and suppressor activity. This variability might be due to technical reasons. DNA-synthesis was not necessary for the mechanisms involved in the Con A-elicited suppressor activity. No suppressor activity was obtained by Con A treatment of cells other than peripheral blood mononuclear cells, viz. autologous erythrocytes, allogeneic fibroblasts and allogeneic osteogenic sarcoma cells.

Epithelioid cells, giant cells and mononuclear cells in cryostat sections of lesional skin from 5 patients with chronic cutaneous sarcoidosis were characterized using an indirect immunofluorescence test with monoclonal antibodies. OKM1 (monocyte) antibody stained both epithelioid cells and giant cells as well as some cells in the periphery of the granulomas. OKM1 which reacts with most monocytes in peripheral blood, did not react with tissue macrophages in sections of normal skin, liver, spleen and kidney. The results indicate that the epithelioid and giant cells in the sarcoid granulomas are derived from blood monocytes. OKT3 (pan T cell) antibody stained the majority of cells surrounding the epithelioid cell islands. Examination of adjacent sections showed that more OKT4+ (helper/inducer) than OKT8+ (suppressor/cytotoxic) cells were present in the sarcoid lesions examined.

Antibodies against pneumococcal capsular polysaccharide types 1, 3, 6A, 7F, 8 and 9N were determined before and 4 weeks after pneumococcal vaccination by both enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) in 49 children, 42 of whom had been splenectomized. Using linear regression analysis a significant correlation between total antibody concentrations determined by ELISA and RIA was found for all 6 antigens (p less than 0.001 for each). The degree of correlation varied between types, antibodies against type 3 showing the best concordance, those against type 6A the poorest. In 84% of the samples assayed both the RIA and the ELISA determined antibody concentrations were either above (55%) or below (29%) the hypothetical protective antibody concentration.

A newly developed experimental model in which poststreptococcal glomerulonephritis (PSGN) was established in rabbits utilizing viable group A streptococci, was used to study the possibility of hindering the development of renal disease by early penicillin treatment. The development of renal injury was followed by proteinuria, creatinine clearance, histological and immunological examinations of the kidneys. Therapy within the first 3 days of infection with i.m. pc-G prevented the nephritic process.

The in vitro immunoglobulin (Ig) secretion of pokeweed mitogen (PWM)-activated peripheral blood lymphocytes (PBL) from individuals splenectomized post-trauma was monitored with a protein A plaque-forming cell (PFC) assay. Cultures of unfractionated as well as reconstituted cultures of isolated erythrocyte rosette-forming (E-RFC)-positive (T lymphocytes) and E-RFC-negative (B lymphocytes) cells were established. Using unfractionated cells, the response was substantially reduced or absent, whereas cultures of autologous untreated B and 2000 rads irradiated T cells restored the response to normal levels. Normal T cells were not able to stimulate patients' B cells to Ig-secretion and patients' untreated T cells did not induce plaque formation in normal B cells, whereas irradiated patients' T cells induced development of approximately 50% of the response induced by normal irradiated T cells. These results indicate that the immunological defect in splenectomized individuals is not merely restricted to a high level of radiosensitive T cell suppression but also involves an impaired B cell function and T/B cell cooperation.

Viable group A, M56 streptococci were inoculated into subcutaneously implanted steel net cages in rabbits, thereby establishing a local infection. The infectious process was monitored by quantitating the number of bacteria in the cage fluids. Functional signs of renal lesion were followed by urine analysis and measurement of serum creatinine. All animals were sacrificed 2-8 weeks after the inoculation and showed histological and immunological signs of kidney lesions. The morphological and functional abnormalities observed closely mimic those of post-streptococcal glomerulonephritis in man.

The IgG-, IgM- and IgA- anti-pneumococcal antibody response to pneumococcal vaccination in 29 splenectomized adults and adolescents with hereditary spherocytosis or previous traumatic splenic rupture was determined by an enzyme-linked immunosorbent assay. It was not significantly different from that of 12 healthy controls except for a lower IgM class antibody increase in the splenectomized against one of four antigens studied. The antibody response was predominantly of IgG class, but significant increases in IgM and IgA class antibodies against all four antigens (polysaccharide types 2, 6A, 12F and 14) studied were observed. In 1/29 splenectomized and 2/12 healthy individuals (7%) the IgG antibody class did not predominate. In 36 adults and adolescents splenectomized due to traumatic rupture or during surgery for gastric ulcer, 77% of the peak geometric mean total antibody concentration four weeks after vaccination was still present after 21 months (16-26 months).

Peritoneal cells from C57/BL/6J mice untreated or injected with saline, proteose-peptone or Corynebacterium parvum intraperitoneally or Sauton medium or BCG intravenously were fractionated on discontinuous Percoll gradients. Six subsets of macrophages were obtained in all the groups. Injection of BCG and proteose-peptone increased the percentage of heavy density macrophages whereas C. parvum caused an increase in macrophages in the lighter density fractions. The macrophages in all the subsets were tested for their ability to kill L929 target cells and cytochemically for expression of beta-galactosidase. The highest degree of cytotoxicity was found in the higher density subsets, although the other subsets in the C. parvum group were also cytotoxic. There was a marked reduction in beta-galactosidase in macrophages obtained after injection of C. parvum and to a varying degree after injection of BCG, proteose-peptone or saline. Cells which were devoid of enzyme were evenly distributed throughout the subsets. These studies indicate that a certain degree of separation of cytotoxic macrophages can be achieved by density centrifugation, but not of cells containing different amounts of beta-galactosidase.

Crossed immunoelectrophoresis was used to study the antigens of chronic lymphocytic leukemia lymphocytes. A reference pattern was obtained and 20 samples from 18 patients were compared with this pattern. Extensive variation in the expression of individual antigens was observed. The antigens in the reference pattern were further characterized by modifications of crossed immunoelectrophoresis, viz. labelling of intact lymphocytes by lactoperoxidase-catalyzed iodination, affinity crossed immunoelectrophoresis with phenyl-Sepharose and Lentil lectin-Sepharose, charge-shift crossed immunoelectrophoresis and postelectrophoretic incubation in (125I) Lentil lectin. Six antigens were identified as surface membrane glycoproteins, 5 as cytoplasmic proteins while 5 could not be classified. Two antigens were finally identified as HLA-ABC and HLA-DR by application of small amounts of monospecific rabbit antiserum and monoclonal antibody, respectively.