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

Restoring the haemolytic activity of heat-treated chicken serum was used to monitor an initial purification of chicken complement component factor B of the alternative complement pathway. The partially purified factor B was used to prepare a monospecific rabbit antiserum against factor B, and this antiserum was now used to monitor and establish a purification scheme for chicken factor B which could be used for large-scale purifications. The final purification involved precipitation with polyethyleneglycol, ion exchange chromatography on CM 52-cellulose, gel-filtration on Sephadex G 200, elution from a Con A-Sepharose column, and finally ion exchange chromatography on a DE 52-cellulose column. 2.8 mg of factor B was purified from 100 ml of chicken plasma with a yield of 28%. Purity was high as judged from crossed immunoelectrophoresis, SDS-PAGE, and from immunizations of rabbits with the factor B preparation. The purification gave preliminary indications that chicken factor B is a glycoprotein with beta-mobility and with a molecular weight around 95 Kd.

Blood mononuclear cells (MNC) from 9 randomly selected patients with autoimmune thyroiditis were stimulated in vitro with pokeweed mitogen (PWM), a polyclonal B lymphocyte activator. The secretion of immunoglobulins (Ig) and anti-thyroglobulin antibodies (TgAb) was assayed by means of haemolytic plaque-forming cell (PFC) assays, radio-immune assay (RIA) and enzyme-linked immunosorbent assays (ELISA). Total Ig and TgAb production was maximal using MNC cultured at 1.0 X 10(6)/ml as estimated by PFC, RIA and ELISA. The Ig and TgAb production as measured by RIA and ELISA was 1.5 - 3 times higher after 12 days' culture compared to 6 days' culture. Ig and TgAb production measured by PFC-assays at day 6 correlated positively to the results obtained by RIA and ELISA at day 12. PWM-induced TgAb secretion correlated positively to TgAb titres in serum. As judged by PFC, TgAb production was found in 8/9 patients; about 5% (range 0 - 7.9%) of the total PWM-stimulated IgG-secreting cells were involved in TgAb secretion. TgAb production as measured by ELISA and RIA was found in 6/9 patients. By reference to an affinity-purified human TgAb preparation, the TgAb secretion was about 0.7% (range 0 - 21.3%) of the total PWM-induced IgG secretion.

The aim of the study was to test the influence of thyroglobulin in vitro and in vivo on two different principles for quantification of serum antithyroglobulin antibody content. The methods employed were a previously described homogeneous phase radioassay and a newly developed enzyme-linked immunosorbent assay. Thyroglobulin inhibited the measurement of antithyroglobulin antibody content by both methods, not only by addition in vitro but also when present in vivo as a result of release from thyroid surgery. Both methods proved sufficiently sensitive as screening for presence of antithyroglobulin antibodies before measurement of thyroglobulin in serum. The study shows the importance of assessing the assay systems in each laboratory for interference before use, especially if intended for screening before quantifying serum thyroglobulin or for measurement of antithyroglobulin antibodies in other media (e.g. supernatants from lymphocyte cultures).

After administration of 0.2 mg adrenaline subcutaneously to 8 healthy individuals, natural killer (NK) cell activity of peripheral mononuclear cells increased within 15-30 min (p less than 0.005) and returned to baseline values within two hours. The activity was correlated to plasma adrenaline concentration (r = 0.52, p less than 0.001). No changes in NK cell activity were observed in 3 control subjects receiving an injection of 0.2 ml saline.

The concentration of factor D (D) was determined in the sera of 144 healthy adults and 29 healthy children by enzyme amplified electroimmunoassay (EAE), and by a hemolysis in gel (HIG) assay for D. The 95% geometric confidence areas for D in adults were 75-143% as determined by EAE, and 65-171% by the functional assay. The concentrations were given in per cent of a reference serum pool containing 1.6 mg/l of D as measured by EAE. The correlation coefficient between D values obtained by the two methods was r = 0.52 (p less than 0.001). D levels were lower in children than in adults. In the EAE purified D, and pathological sera with very high D Concentrations, gave lower precipitation peaks with dilutions performed in buffer than with dilutions in D depleted serum or human serum albumin.

Chronic granulomatous disease (CGD) in utero has recently been detected by a new qualitative nitro blue tetrazolium (NBT) reduction slide test using phorbol-myristate-acetate (PMA) as a stimulus. The technique is simple and inexpensive and requires only a few microlitres of blood. Reported here is an evaluation of this method as applied to routine clinical diagnosis. The blood granulocytes from 300 normal individuals and 49 CGD patients and their relatives were tested and the results compared to the conventional in vitro function tests. In normal individuals the number of abnormal cells was very low never exceeding 2% of granulocytes. CGD patients (14 out of 15) diagnosed by conventional functional tests showed no positive cells in the NBT-PMA test. One patient diagnosed by functional tests had 4% positive cells. Thirty-four relatives of these patients were tested and eleven were found to have fewer positive cells than normal in the NBT-PMA slide test ranging from 16-88% of all granulocytes. These are presumably carriers, a finding supported by granulocyte function tests. All these individuals were female, mothers, sisters or maternal aunts of male CGD patients, thus presumably X-linked heterozygote carriers of CGD. An example of successful prenatal diagnosis using the PMA-NBT test is described. The results show that the PMA-NBT test provides a simple and reproducible method for routine diagnosis of CGD and CGD X-linked heterozygote carriers.

The effect of temperature on the function of polymorphonuclear leukocytes (PMN) has been investigated in vitro. Increases in temperature from 37 degrees C to 40 degrees C progressively increased chemiluminescence (CL) responses by PMN after stimulation by Staphylococcus aureus, zymosan or phorbol myristate acetate (PMA) while increases above 40 degrees C decreased these functions. Temperature increases from 37 degrees C to 40 degrees C also produced increased PMN bactericidal activity against S. aureus. In contrast, similar increases in temperature did not change superoxide production by PMN stimulated by PMA. Incubation of PMN at the various temperatures did not cause release of LDH indicating that damage to PMN was not the cause of reduced PMN chemiluminescence and bactericidal activity seen within the temperature range studied. The discrepancy between the influence of temperature on PMN chemiluminescence and bactericidal activity of PMN compared to superoxide anion production by PMN suggests that superoxide anion production may not be solely, or at least directly, responsible for killing of bacteria. Careful temperature control is needed when assaying PMN function. Febrile responses up to 40 degrees C may play a beneficial role in host defense.

The degranulation and release of lysosomal (myeloperoxidase, beta-glucuronidase, lysozyme) and cytoplasmic (lactate dehydrogenase-LDH) enzymes from polymorphonuclear neutrophil granulocytes (PMG) during phagocytosis of inert latex particles or bacteria were studied. Degranulation was much faster and more pronounced by phagocytosis of bacteria than of inert particles. A high frequency of lysosome-lysosome as well as lysosome-phagosome fusions suggested that granular material was transported by lysosome- lysosome- phagosome fusions. During bacterial phagocytosis there was evidence of release of granular material into cytoplasm causing enzymatic disintegration. After 60 minutes cell lysis occurred in about 5 per cent of the cells during bacterial phagocytosis. There was non-specific release of LDH during phagocytosis of inert particles, probably due to erythro-phagocytosis. After 60 minutes the release during bacterial phagocytosis amounted to 20-30 per cent of the enzyme content of the cells. A nearly equal release of lysosomal and cytoplasmic enzymes gave support for the idea that cell lysis was the main mechanism of enzyme release.