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Biomedical subjects

V Andersen

Publications and source records attributed to V Andersen.

At least 109 records · Page 6Linked to original sources

Induction of immunoglobulin secretion in cultured human lymphocytes by 4 Staphylococcus aureus strains and their extracts.

Human blood lymphocytes were stimulated in vitro by four Staphylococcus aureus strains. Activation of immunoglobulin-secreting cells was determined by a reverse plaque forming cell (PFC) assay, and proliferation by quantitation of thymidine incorporation. Whole killed S. aureus were slightly more efficient than water-soluble preparations in the form of sonicated extracts and culture supernatants. Two S. aureus strains rich in protein A (Cowan I and E 2371) and one S. aureus strain deficient in protein A (E 1369) were potent B-lymphocyte stimulators inducing maximal activity on day 6 of culture. Another S. aureus strain deficient in protein A (Wood 46) did not possess++ human lymphocyte stimulating capacity.

Hemolytic Plaque Technique↗

Interaction of human IgG preparations with polymorphonuclear leukocytes in vitro.

The effect of purified human immunoglobulin G, prepared for intravenous administration by polyethylene glycol precipitation and ion exchange chromatography, on polymorphonuclear leukocytes (PMN) in vitro was studied. IgG induced a dose-dependent stimulation of the oxidative metabolism of the cells, as measured by release of superoxide anion and by chemiluminescence. The lowest concentration giving a detectable effect was 0.001 mg/ml. Two commercially available preparations of IgG for clinical intravenous use caused similar stimulation of the PMN although they differ in methods of isolation and purification. The addition of 10% maltose markedly reduced the stimulation by all three preparations of IgG. We suggest that i.v. infusion of purified IgG may lead to activation in vivo of host phagocytic cells without participation of complement, and that this interaction may be responsible for the hitherto unexplained side-effects caused by i.v. infusion of such preparations.

Energy Metabolism↗

Spontaneous in vitro immunoglobulin secretion at the diagnosis of insulin-dependent diabetes.

Blood mononuclear cells obtained from 17 newly diagnosed insulin-dependent diabetic (IDDM) patients treated with insulin for 5-7 days were assessed for the number of spontaneous and pokeweed mitogen (PWM)-stimulated immunoglobulin-secreting cells in a reverse haemolytic plaque assay. The spontaneous in vitro immunoglobulin secretion was evanescent and decreased in individual patients within 1-4 months of insulin treatment. Compared to matched controls, 53% (9/17) of the IDDM patients had an elevated spontaneous secretion of immunoglobulin, 41% (7/17) for IgG, 35% (6/17) for IgM, and 35% (6/17) for IgA. The quantities of PWM-stimulated IgG, IgM, or IgA secreting cells in IDDM were comparable to the controls. The IDDM patients with spontaneous immunoglobulin secreting cells had higher fasting C-peptide levels compared to the patients with immunoglobulin-producing cells within the normal range (P less than 0.05). The average titre of islet cell cytoplasmic antibodies was 1:26 in (9 out of 9 were positive) patients with, compared to 1:1 in patients (4 out of 8 were positive) without spontaneous secretion (P = 0.025). These results suggest that the clinical onset of IDDM is associated with a polyclonal B lymphocyte activation and that higher levels of fasting C-peptide islet cell antibodies are associated with this immunoregulatory abnormality.

Adolescent↗

Storage of protein A-conjugated sheep erythrocytes for use in plaque-forming cell (PFC) assays.

Sheep red blood cells conjugated with protein A from Staphylococcus aureus were frozen in liquid nitrogen. After thawing these cells were used as indicator cells in a plaque-forming cell assay visualizing human B lymphocytes secreting IgG, IgM or IgA. The thawed erythrocytes performed as well in the assay as erythrocytes freshly conjugated with protein A. Recovery of conjugated erythrocytes after freezing and thawing averaged 66%. By the method described indicator cells for a number of PFC assays may be prepared in bulk, frozen in suitable portions and thawed when needed. This prevents possible batch-to-batch variations and saves time.

Animals↗

Lymphocyte function and chromosome aberrations in patients with early mycosis fungoides and parapsoriasis en plaques.

Thirteen patients with stage I or II mycosis fungoides (MF) and 10 patients with large-plaque parapsoriasis en plaques (PEP) were examined for immunologic and cytogenetic disturbances. Total lymphocyte counts and immunoglobulin concentrations in the blood were normal. In vitro lymphocyte responses to polyclonal activators and various antigens in standard concentrations were normal. However, titration of phytohemagglutinin and concanavalin A (ConA) disclosed significantly lowered responses to suboptimal concentrations in the patient group, most pronounced in patients with MF II. ConA-induced leukocyte migration inhibitory factor (LIF) production, tested in an indirect leukocyte migration inhibitory assay, was low in the patient group. Furthermore spontaneous LIF production in vitro and small amounts of serum LIF were demonstrated in a few patients. The chromosomal banding pattern, sister chromatid exchange, and break frequency were within normal limits except for 3 translocations in the MF group. It is concluded that even in early-stage MF a pathologic function of blood lymphocytes can be demonstrated, when sensitive methods are applied. The findings might be important for monitoring disease activity and effect of treatment.

Adult↗

Immunological studies in two children with recurrent parotitis.

In two children with recurrent parotitis, labial salivary gland biopsies showed chronic sialoadenitis. Immunofluorescence studies disclosed deposits of immunoglobulins and complement in juxta-acinar small vessels. Case 1 had gluten enteropathy, IgA deficiency and high titres of antinuclear antibodies (ANA), and in vivo fixation of ANA to nuclei of different cells in lip, skin and jejunum was present. Case 2 showed deposition of IgM in the dermo-epidermal junction of the skin. These findings suggest that autoimmune reactivity and immune complexes may play a role in the pathogenesis of this disorder.

Adolescent↗

Suppression of B lymphocytes in mature newborn infants.

The ability of cord blood lymphocytes to secrete immunoglobulins during in vitro culture was investigated by means of a reverse hemolytic plaque forming cell (PFC) assay. Mononuclear cord blood cells did not differentiate into immunoglobulin-secreting cells after stimulation with the polyclonal B lymphocyte activator pokeweed mitogen (PWM), contrary to the findings in normal adults. Mononuclear cord blood cells were then separated into T-enriched and T-depleted blood lymphocyte subsets. When these were co-cultured, the PWM-induced immunoglobulin secretion was still low; following irradiation of the T-enriched cells, the numbers of IgM-PFC but not of IgG- or IgA-PFC increased considerably. The effect of irradiation of the T-enriched cells on the PWM-induced IgM response was dose-dependent, with maximal effect at 2500 rad. It is concluded that the low PWM responses obtained using cord blood lymphocytes are in part due to suppression by radiosensitive T suppressor cells. Following removal of this suppression by means of irradiation, B lymphocytes can be induced to secrete IgM, but not IgG or IgA.

Adult↗

Synovial fluid and blood monocyte influence on lymphocyte proliferation in rheumatoid arthritis and traumatic synovitis.

The influence of synovial fluid and blood monocytes on autologous lymphocyte proliferation in vitro was investigated in patients with rheumatoid arthritis (RA) and traumatic synovitis (TS). In cultures stimulated with PHA, PWM and ConA, thymidine incorporation by RA blood mononuclear cells (BMC) increased considerably in most cases after addition of 10% or 30% synovial fluid monocytes and a similar effect was seen after stimulation with Candida extract or PPD. A comparable helper activity was observed after addition of synovial fluid monocytes to monocyte-depleted blood cells in RA. The augmenting effect of synovial fluid monocytes was likewise demonstrated in TS patients. Synovial fluid mononuclear cells (SMC) from RA and TS patients displayed low proliferative responses to PHA and ConA in comparison with BMC, while the responses to microbial antigens were similar or greater. The proliferative activity of SMC or of monocyte-depleted synovial fluid cells after polyclonal and antigenic stimulation was not affected by addition of blood monocytes. This was observed in both RA and TS patients. It is concluded that the regulatory properties of synovial fluid monocytes in RA and TS do not explain the low proliferative responses of SMC to polyclonal activators.

Adult↗

Secretion of IgM-rheumatoid factor and IgM by blood lymphocytes in rheumatoid arthritis.

A direct plaque forming cell (PFC) assay for the detection of complement fixing IgM-rheumatoid factor (RF) secreting lymphocytes was evaluated. The specificity of the RF-PFC assay was demonstrated by the inhibitory action of exogenous human IgG. The sensitivity of the RF-PFC assay was similar to that of a reverse PFC assay detecting all cells secreting IgM (IgM-PFC). In blood from 75% of seropositive patients with rheumatoid arthritis (RA) RF-PFC were demonstrated, median value 6.8 (range: 0.4-1477) RF-PFC/10(6) mononuclear cells. Practically no RF-PFC were detected in seronegative RA patients and controls. Despite the fact that most IgM-RF undoubtedly is produced outside the blood, a positive correlation was found between the number of RF-PFC and the Waaler-Rose titer in serum. The number of circulating IgM secreting cells did not differ significantly between seropositive RA patients, seronegative RA patients and controls. Comparison of RF-PFC and IgM-PFC in seropositive patients revealed that in mean 7% of IgM-secreting cells in blood secrete IgM-RF.

Antibody Specificity↗

Production and secretion of immunoglobulins by in vitro-activated human B lymphocytes.

Activation induced by pokeweed mitogen in cultures of mononuclear cells from human blood was followed sequentially by simultaneous quantitation of live cells, thymidine incorporation, cells displaying cytoplasmic, IgM, IgG, IgA or IgD, cells secreting IgM, IgG or IgA and cumulated IgM secretion. Maximal cellular activity was found after 7 days of cultures, with means of 16000 IgM-, 20700 IgG- and 9900 IgA-secreting cells per 10(6) originally cultured cells. The cumulated IgM secretion after 21 days of culture averaged 10400 ng per 10(6) originally cultured cells. A close correlation was found between the number of IgM-secreting cells and the cumulated IgM secretion.

Antibody-Producing Cells↗

Influence of autologous monocytes on the pokeweed mitogen-induced generation of immunoglobulin-secreting cells in man.

The regulatory influence of autologous blood monocytes on the PWM-induced generation of Ig-secreting cells was assessed using a reverse haemolytic plaque forming cell (PFC) assay. The PWM-induced PFC responses of monocyte-depleted cells were low or absent in most cases. Addition of 12-42% freshly isolated monocytes fully reconstituted the IgM-, IgG- and IgA-PFC responses. With more monocytes added, the PFC responses declined. Monocytes precultured for 48 h supported the PFC responses of monocyte-depleted cells less well and addition of monocytes stimulated with phorbol myristate acetate (PMA) did not increase the response. The low responses of monocyte-depleted cells co-cultured with precultured monocytes were not increased by addition of supernatant from monocyte cultures. The PFC responses of mononuclear cells induced by PWM were significantly inhibited by unstimulated precultured monocytes, and to a larger degree by PMA-treated monocytes, indicating the presence of suppressor cells among the precultured monocytes. The PWM-induced thymidine incorporation by monocyte-depleted cells with precultured monocytes added was only slightly lower than that obtained with freshly isolated monocytes added, suggesting that the suppressive role of precultured monocytes was not due to cytotoxicity.

Antibody-Producing Cells↗