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Biomedical subjects

E Maggi

Publications and source records attributed to E Maggi.

At least 181 records · Page 10Linked to original sources

Amniotic fluid insulin, C peptide concentrations, and fetal morbidity in infants of diabetic mothers.

Glucose, insulin, C peptide, and insulin antibody concentrations were measured in amniotic fluid collected under basal conditions and 2 hours after an arginine challenge from 61 insulin-treated diabetic women (12 basal and 49 after arginine challenge) and 31 nondiabetic pregnant women in late gestation (23 basal and eight after arginine challenge). The insulin, C peptide, and glucose concentrations were significantly higher in diabetic pregnant women than in nondiabetic pregnant women in each case. In the amniotic fluid obtained after arginine challenge in diabetic pregnant women, C peptide concentration was correlated with both insulin concentration (r = 0.61) and birth weight (r = 0.53). The insulin and C peptide concentrations were significantly higher (p less than 0.025) in samples from diabetic pregnancies associated with fetal morbidity than from diabetic pregnancies without fetal morbidity. Basal amniotic fluid insulin and C peptide concentrations were slightly greater in overweight infants of diabetic mothers compared to those of normal weight, whereas the differences for insulin and C peptide concentrations in the amniotic fluid obtained after arginine challenge were highly significant (p less than 0.0125 and p less than 0.0005, respectively). Finally insulin and C peptide concentrations in the amniotic fluid obtained after arginine challenge in diabetic pregnant women showed a correlation with maternal metabolic control but not with the degree (White classification) of maternal diabetes. No or negligible interference of insulin antibody in the radioimmunoassay of insulin in amniotic fluid was observed.

Amniocentesis↗

Different reactivity of activated human B cells to B-cell growth factor and interleukin 2 in the costimulation assay with anti-IgM antibody and in the preactivation assay with Staphylococcus bacteria.

The two main assay systems which have been developed for the study of lymphokine-mediated human B-cell proliferation, i.e., the costimulation assay with anti-mu antibody and the preactivation assay with Staphylococcus aureus Cowan I (SAC) bacteria, were compared. Purified interleukin 2 (IL-2), obtained by the recombinant DNA technology (r-IL-2), enhanced the proliferative response of anti-mu-stimulated human B cells in the costimulation assay with anti-mu antibody and maintained the B-cell proliferation induced by preactivation with SAC bacteria. Although the majority of T-cell clones, established from normal peripheral blood T lymphocytes, showed production of both IL-2 and B-cell growth factor (BCGF) following phytohemagglutinin (PHA)-stimulation, some T-cell clones were found whose supernatants (PHA-SN), apparently free of IL-2, manifested strong BCGF activity in the costimulation assay with anti-mu antibody. However, the same clonal, IL-2-free, T-cell SN displayed no BCGF activity in the preactivation assay with SAC bacteria. When B cells were activated for 3 days with anti-mu antibody, followed by the addition of r-IL-2 or clonal T-cell SN containing BCGF for an additional 3 days, r-IL-2 showed the ability to maintain B-cell proliferation, whereas clonal SN containing BCGF had virtually no effect. These data indicate that the costimulation assay with anti-mu antibody explores the reactivity of normal human B cells to both BCGF and IL-2, whereas the preactivation assay with SAC bacteria, due to a shorter reactivity to BCGF of activated human B cells, essentially represents a probe for the study of IL-2-promoted B-cell proliferation.

Antibodies, Anti-Idiotypic↗

Synergy of B cell growth factor and interleukin 2 in the proliferation of activated human B cells.

The activity of purified interleukin 2 (IL2), obtained by the recombinant DNA technology, on the proliferative response of human B cells stimulated with low concentrations of anti-mu antibody was investigated. Recombinant IL2 was capable of augmenting the proliferative response of anti-mu-activated B cells and the T cell activation (Tac) antigen was expressed on a substantial proportion of normal B cells stimulated with anti-mu antibody. However, crude supernatants from protein A-stimulated peripheral blood mononuclear cells, which were found to possess both IL2 and B cell growth factor (BCGF) activities, maintained the ability to promote proliferation of anti-mu-activated B cells after depletion of IL2. In addition, supernatants from some T cell clones, apparently free of IL2 activity, displayed strong BCGF activity in the co-stimulation assay with anti-mu antibody. This BCGF activity was found in 25 kDa fractions by gel filtration and it was unaffected by addition to the cultures of anti-Tac antibody, which consistently inhibited the B cell proliferative response promoted by recombinant IL2. The proliferative response of anti-mu-activated B cells to clonal, IL2-free supernatants containing BCGF and recombinant IL2 present together from the beginning of culture was close to the sum of responses to the two stimulants, separately. In addition, the presence of clonal supernatant containing BCGF from the beginning of culture had a synergistic effect in the response of activated B cells to the subsequent addition of IL2, whereas the initial presence of IL2 had no such an effect on the reactivity of anti-mu-stimulated B cells to the late addition of clonal supernatant containing BCGF. The synergistic effect of BCGF in the IL2-promoted B cell proliferation was probably the result of the recruitment of a greater number of IL2-reactive B cells. In fact, the number of Tac-positive cells was significantly higher in 36-h cultures established in the presence of anti-mu antibody plus clonal supernatant containing BCGF than in cultures stimulated with anti-mu antibody alone. Taken together, these data indicate that anti-mu antibody promotes the expression by normal human B cells of distinct receptors for IL2 and a BCGF distinct from IL2. They also suggest that BCGF can exert a synergistic effect in the IL2-promoted proliferation of activated B cells.

Antibodies, Anti-Idiotypic↗

Phenotypic and functional heterogeneity of human T lymphocytes producing B-cell growth factor(s): a clonal analysis.

Human T cells active on B-cell proliferation are phenotypically and functionally heterogeneous. A series of 43 human T-cell clones, selected according to their ability to release factors active on B-cell proliferation, were analyzed. B-Cell proliferation was evaluated by two different assays, namely, a costimulation assay with anti-mu antibody or by an assay based on B-cell preactivation with Staphylococcus aureus. Eight of these clones expressed the T4-/T8+ phenotype while the remaining were T4+/T8-. The large majority had T-cell growth factor activity as well. However, some clones appeared to have B-cell growth factor activity only. Fourteen clones (eight T8+ and six T4+) also displayed cytolytic activity in a phytohemagglutinin-dependent cytolytic assay.

Antibodies, Anti-Idiotypic↗

In vitro synthesis of human IgE: reappraisal of a 5-year study.

In the last 5 years some models of human IgE production in vitro have been investigated in our laboratory. Spontaneous IgE synthesis was found in cultures of B cells from most patients with atopic dermatitis or atopic patients with multiple sensitivities and from some patients with pollenosis, but only during the pollination period. A small and variable increase of the spontaneous IgE synthesis was induced by soluble factor(s) produced by T cells from patients with severe atopy. Selected helper T cell clones were also able to induce IgE synthesis in vitro by both atopic and normal B cells.

Antibody Formation↗

A pilot study of the pharmacokinetics and triglyceride lowering activity of acipimox in dialyzed uremic patients.

The pharmacokinetics of acipimox was studied in 6 dialyzed uremic patients given single oral doses of 50 mg. Acipimox was not significantly eliminated outside dialysis, whereas during dialysis it was efficiently cleared with plasma, t 1/2 is about 2.6 hours. Accordingly, a dosage schedule of 50 mg or 100 mg of acipimox after each dialysis session was selected for a second, 4-week study in 14 uremic patients with hypertriglyceridemia. Acipimox plasma levels, monitored during the study, proved in agreement with those expected on a theoretical pharmacokinetic basis. A clear-cut reduction of serum triglyceride levels was also achieved.

Adult↗

Frequent coexpression of cytolytic activity and lymphokine production among human T lymphocytes. Production of B cell growth factor and interleukin 2 by T8+ and T4+ cytolytic clones.

These studies were designed to determine the degree of overlapping between cytolytic function and lymphokine production among peripheral blood human T lymphocytes. T8+ and T4+ cells were obtained by sorting purified T cells using the fluorescence-activated cell sorter (FACS) and cloned at 0.25 cells/well in a microculture system that allows clonal growth of every T cell. 49/49 T8+ clones but only 5/102 T4+ clones displayed cytolytic activity, as assessed by a lectin-dependent assay. Cytolytic clones were further analyzed for their ability to release interleukin 2 (IL2) and B cell growth factor(s) (BCGF) upon 24 h stimulation with phytohemagglutinin. Fifteen percent and 18% of T8+ cytolytic clones were producing IL2 and BCGF, respectively; in addition 3 out of 5 T4+ cytolytic clones released both lymphokines. Although the majority of cytolytic clones producing IL2 were also producing BCGF, 5 clones released either BCGF or IL2. The present studies indicate that the coexpression of cytolytic and helper/inducer functions is relatively common among peripheral blood T lymphocytes, irrespective of their T8+ or T4+ phenotype.

B-Lymphocytes↗

Radioimmunoassay of acipimox in human plasma and urine.

A radioimmunoassay was developed for the determination of acipimox (5-methylpyrazinecarboxylic acid 4-oxide) in human plasma and urine. Acipimox was conjugated to bovine serum albumin through a spacer with 4 carbon atoms, and repeatedly injected into rabbits. Antisera raised in these animals were highly specific and virtually no cross-reaction was observed with 5-methylpyrazinecarboxylic acid and nicotinic acid. Despite the low specific activity of the labelled antigen used, concentrations of acipimox in human plasma and urine as low as 40 ng/ml could be determined. The intra- and inter-assay coefficients of variation ranged between 4.32-6.25% and 6.19-11.55% respectively, and mean recovery of the compound spiked to plasma was 100-103%. The method was applied to determine plasma levels and urinary excretion of acipimox after oral doses of 150 mg and 250 mg to four volunteers.

Animals↗

Surface markers and function of circulating thyroid autoantibody-producing cells.

The in vitro synthesis of antithyroglobulin (anti-Tg) and antithyroid microsomal (anti-M) autoantibodies by peripheral blood mononuclear cells (MNC) from patients with autoimmune thyroid diseases was investigated using sensitive immunoradiometric assays. Cultures were carried out in the presence or in the absence of pokeweed mitogen (PWM). Thyroid autoantibodies were undetectable in supernatants of MNC cultures from 9 normal subjects. Supernatants of MNC cultured without PWM had detectable levels of anti-Tg and anti-M in 5 (19.3%) and in 2 (7.7%) of 26 patients with autoimmune thyroid diseases, respectively. In the presence of PWM, a marked increment in the antibody concentrations occurred in all but 1 of these cultures, and the number of positive cultures increased to 13 (50.1%) for anti-Tg and to 15 (57.7%) for anti-M. Studies of MNC fractions depleted of T lymphocytes (non-T cells) were carried out on selected patients showing antibody synthesis only after PWM stimulation. Autoantibody production was not found with non-T cells, but the effect of the mitogen was restored by readdition of T cells. Irradiation (1000 rad) of T cells before coculturing significantly enhanced autoantibody production. With this model no significant functional difference was found between autologous and allogenic T cells from thyroid autoimmune disease patients or from normal subjects. The cells involved in PWM-driven thyroid autoantibody synthesis, as defined by depletion studies, were lymphocytes bearing DR antigens and surface immunoglobulin G (IgG) without detectable surface immunoglobulin M (IgM). Depletion from MNC suspensions of Tg-binding cells abolished PWM-stimulated anti-Tg production, but did not alter the synthesis of anti-M. Further studies were carried out on MNC from a single patient with Hashimoto's thyroiditis, whose non-T cells consistently produced large amounts of anti-M and total IgG in the absence of PWM. The addition of PWM to these unfractionated MNC slightly increased the production of anti-M, but inhibited antibody synthesis after depletion of T lymphocytes. Interestingly, the addition of autologous T lymphocytes to non-T cells inhibited the spontaneous synthesis of anti-M. These data indicate that in vitro synthesis of anti-Tg and anti-M by MNC may be frequently induced by stimulation with PWM in patients with thyroid autoimmune disorders. PWM-stimulated synthesis of thyroid autoantibodies appears to be T-cell dependent and modulated by radiosensitive T lymphocytes. The cells responsible for PWM-dependent thyroid autoantibody synthesis are B lymphocytes with surface membrane IgG and have receptors specific for the autoantigen.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

Displacement of T lymphocytes with the 'Helper/Inducer' phenotype from peripheral blood to lymphoid organs in untreated patients with Hodgkin's disease.

A panel of previously characterized monoclonal antibodies: B67.6, OKT3, OKT4, B53.4, Leu3a, OKT8, Leu2a, OKM1, M12 and B52.1 were used as a probe to assess mononuclear cells in peripheral blood (PB), lymph nodes (LN) and spleens of untreated patients with Hodgkin's disease (HD). The mean % and absolute number of T lymphocytes were significantly decreased in PB of HD patients when compared with control values. Reduction of circulating T lymphocytes reflected the selective loss of cells showing the 'helper/inducer' ('H/I') phenotype. In fact, a lower number of these cells was demonstrated in HD patients with advanced disease and, even though to a lesser extent, in those with localized disease. In contrast, decreased values of T cells with the 'cytotoxic/suppressor' ('C/S') phenotype were only found in patients with advanced disease, showing pan-lymphocytopenia. Unlike PB, LN and spleens involved by HD usually showed increased %s of T lymphocytes, especially of those possessing the 'H/I' phenotype. The displacement of T lymphocytes with 'H/I' phenotype from PB to lymphoid organs further supports the possibility of a chronic immune response against abnormal cells or unknown antigens in the affected organs of patients with HD.

Adolescent↗

Abnormalities of in vitro immunoglobulin synthesis by peripheral blood lymphocytes from untreated patients with Hodgkin's disease.

The immunoglobulin-synthesizing activities of peripheral blood mononuclear cells from 57 untreated patients with Hodgkin's disease and 47 normal subjects were compared. Cumulative amounts of IgM and IgG synthesized and secreted by unstimulated and pokeweed mitogen-stimulated cells over a 7-d period were determined in a solid-phase radioimmunoassay. Synthesis of IgM in unstimulated cultures and of both IgM and IgG in cultures stimulated with pokeweed mitogen was markedly reduced in patients with Hodgkin's disease, whereas the mean level of the spontaneous IgG synthesis was enhanced. The degree and frequency of in vitro abnormalities were not influenced by disease stage or histology. Depression of pokeweed mitogen-induced immunoglobulin synthesis did not correlate with excessive number of monocytes and it was unaffected by removal of phagocytic cells or addition to the cultures of monocytes from normal individuals. On the other hand, monocytes isolated from blood of patients with Hodgkin's disease were even more effective than normal monocytes in supporting pokeweed mitogen-induced immunoglobulin synthesis by normal phagocyte-depleted mononuclear cells. Synthesis of both IgM and IgG induced by pokeweed mitogen remained subnormal after addition to patient B cell cultures of autologous irradiated T cells or allogeneic normal T lymphocytes. T cells from patients with Hodgkin's disease appeared at least as effective as normal T cells in helping pokeweed mitogen-induced immunoglobulin production by normal B cells. However, when normal T cells were co-cultured with B cells from patients with Hodgkin's disease, spontaneous IgG synthesis declined, whereas the addition of patient T cells to normal B cells resulted in an increase of spontaneous IgG synthesis. In patients showing depression of pokeweed mitogen-induced immunoglobulin synthesis the lymphoproliferative response and immunoglobulin synthesis stimulated by Staphylococcus aureus bacteria of the Cowan first strain, a T cell independent B cell mitogen, were also markedly reduced. These studies demonstrate impairment of immunoglobulin synthesis by cultured lymphocytes from untreated patients with Hodgkin's disease after stimulation with polyclonal B cell activators and suggest that the in vitro abnormalities may be, at least in part, the result of a preexisting in vivo activation of lymphocytes in Hodgkin's disease patients.

Adolescent↗

In vitro selective expansion of allergen specific T cells from atopic patients.

Peripheral blood mononuclear cells from atopic donors were stimulated in vitro with allergens (Rye group I or Dermatophagoides pteronyssinus). T cell lines were originated and maintained in long term culture using IL-2 and periodical restimulations with allergen. The lines were antigen specific (i.e. responded to the allergen used to raise them and not to other antigens) and required that the antigen was presented by autologous cells (i.e. they were restricted). The restriction elements were probably at the level of HLA-DR antigens since the proliferative response was specifically blocked by anti-HLA-DR antibodies. Surface marker analysis revealed that the lines comprised mainly cells with an helper/inducer phenotype, although cells with markers of the suppressor/cytotoxic T cells were also present. The lines could be cloned by limiting dilution and clones with the same restriction and specificity as the parental line were isolated. These studies demonstrate the possibility of obtaining a large number of allergen specific human T cells that can be used for further in vitro studies on the regulation of the IgE response.

Allergens↗