Basis for defective proliferation of peripheral blood T cells to anti-CD2 antibodies in primary Sjögren's syndrome.

Anti-CD2-induced T cell proliferation was analyzed in the peripheral blood samples of 31 primary and 8 secondary untreated Sjögren's syndrome patients. Anti-CD2-stimulated PBMC proliferation was very low in about one-third of primary Sjögren's syndrome samples, despite the number of CD2+ cells being similar in primary and secondary Sjögren's syndrome and normal PBMC samples. The depressed response to anti-CD2 was mainly found in anti-Ro+/La+ patients. Experiments on purified T cells demonstrated that a defect at the T cell level was responsible for the anti-CD2 unresponsiveness. Cell proliferation failure was associated with poor IL-2 and IL-2 receptor mRNA expression and, consequently, IL-2 and IL-2 receptor synthesis. Since defective anti-CD2-induced mitogenesis could be reversed by phorbol myristate acetate, but not calcium ionophore A23187, it is probably correlated with impaired protein kinase C activation. Comparison of anti-CD2-triggered PBMC proliferation in treated and untreated patients and a long-term study of nine patients showed that the defect is a stable characteristic in primary Sjögren's syndrome patients, but that it can be reversed by pharmacological immunosuppression.

Intracellular calcium levels are differentially regulated in T lymphocytes triggered by anti-CD2 and anti-CD3 monoclonal antibodies.

Antigen and/or mitogen-driven T-cell activation is mediated by a rise in intracellular free Ca2+, as second messenger. A regulatory key role for this process is represented by membrane-associated [Ca2+/Mg2+] ATP-ase that is mainly devoted to extrusion of intracellular ion excess. In the present study we have investigated the kinetics of CA2+ fluxes in both resting and already activated (Jurkat T-cell line) T lymphocytes after CD3 and CD2 (T11(2) and T11(3)) triggering and focused our attention on plasma membrane [Ca2+/Mg2+] ATP-ase activity. In both resting T cells and Jurkat cell line, the CD2 stimulation was able to determine a rise in intracellular free Ca2+ higher than that observed after CD3 triggering. In addition, this calcium signal was independent of negative feedback control exerted by [Ca2+/Mg2+] ATP-ase, as well as of IP3 generation. Thus the CD2 molecular system may, together with cell-adhesion properties, act as an amplifier of Ca2+ signals that, if delivered in the context of other molecular systems, such as CD3 or MHC class II antigens, are essentially devoted to the polyclonal co-stimulatory recruitment of a larger cellular repertoire.

IgG subclass deficiency and sinopulmonary bacterial infections in patients with alcoholic liver disease.

Abnormalities in IgG subclass distribution were sought in serum samples and bronchoalveolar lavage fluid from 15 patients with alcoholic liver disease to explain their increased susceptibility to bacterial respiratory infections. Serum IgG4 deficiency alone or in association with low IgG2 levels was revealed in approximately 30% of patients with alcoholic liver disease. This fact prompted us to further investigate the immunoglobulin concentrations in broncho-alveolar lavage fluid, paying special attention to the distribution of IgA and IgG subclasses. IgA levels were found to be normal or slightly elevated. However, there were substantial defects in total IgG and IgG1 concentrations, often associated with reduced IgG2 and IgG4 levels, in approximately 70% of patients with alcoholic liver disease, which proved to be closely correlated with the number and type (pneumonia) of bacterial respiratory infections. A prospective study of intravenous immunoglobulin substitutive therapy involving two patients with recurrent pneumonia and very low serum IgG2 values demonstrated a reduction in the number of respiratory infectious episodes as well as an increase in both serum and, to a lesser extent, bronchoalveolar lavage fluid IgG1 and IgG2 levels. We identified immune defects that may represent an important pathogenetic mechanism that, when considered together with the alcohol-related suppression of alveolar macrophage and ciliary functions and the inhibition of leukocyte migration into the lungs, should help clarify the complex relationships between alcohol and immune defense.

Anti-CD3 and anti-CD2-induced T-cell activation in primary Sjögren's syndrome.

Because T-cell dysfunctions have been reported in patients with primary Sjögren's syndrome (SS), peripheral blood mononuclear cell (PBMC) proliferation obtained with anti-CD3 and anti-CD2 monoclonal antibodies was evaluated in these patients. Anti-CD3-induced mitogenesis, which varied widely among the patients, was lower in subjects with evidence of anti-SSA and anti-SSB antibodies than in controls. Moreover, the anti-CD2-induced response was depressed in about half the patients and the nonresponders were mainly those with anti-SSA and anti-SSB antibodies. Phorbol myristate acetate, a protein kinase C activator, used alone or added to anti-CD3, induced greater proliferation in patients than in control PBMC. In contrast, exogenous recombinant interleukin 2 (rIL-2) did not significantly enhance the anti-CD2-induced response of patients' PBMC, as it did in normal PBMC. Peripheral blood and parotid T cells from a patient with well-defined primary SS and parotid enlargement also responded poorly to anti-CD2 stimulation. Exogenous rIL-2 restored T-cell proliferation only in the salivary gland cultures of this patient. The present findings suggest that there is a T-cell activation defect in subjects with primary SS, particularly in those with circulating anti-SSA and anti-SSB antibodies. In addition, the difference in the response to IL-2 of peripheral blood and parotid-infiltrating T cells would seem to indicate that T-cell subsets are differently distributed in the blood and inflammation site.

Functional characterization of T cells bearing the gamma/delta T-cell receptor in patients with primary Sjögren's syndrome.

High percentages of gamma/delta+ T cells in the peripheral blood of a subgroup of patients with primary Sjögren's syndrome (SS) were found. This allowed us to purify and analyze them without their being previously expanded in vitro, and to investigate, therefore, the role of these cells in the pathological immune response which characterizes such systemic autoimmune disorders. The results showed poor proliferation of patient gamma/delta+ T cells in response to anti-CD3, due not to macrophage-dependent suppression but to defective interleukin 2 (IL-2) synthesis. Despite the defective proliferation patient gamma/delta+ cells, unlike those of the normal controls, provided a helper effect in inducing B cells to secrete immunoglobulins (Ig), particularly when they were preincubated with IL-2. The relative increase in a gamma/delta+ T cell subset which, although it secretes low levels of IL-2, is able to provide help for B-cell Ig synthesis, suggests that this T-cell subpopulation may be functional in vivo and may be involved in the pathological immune response encountered in pSS.

Heretical thoughts about food hypersensitivity: small bowel manometry as an objective way to document gut reactions.

BACKGROUND: Food hypersensitivity is a frequent complaint by both pediatric and adult subjects. However, notwithstanding patient' belief about 'allergies' related to food stuffs, only a minority of them have actually such a diagnosis substantiated. Moreover, the diagnostic approach to these problems is cumbersome and unsatisfactory, and the objectivation of a food hypersensitivity is often difficult. PATIENTS AND METHODS: For these reasons we studied by means of small bowel manometry a small group of patients with food hypersensitivity, and showed abnormal fasting and postprandial findings in those with the gut as a target organ on clinical grounds. RESULTS: Manometric abnormalities were somewhat similar to those previously described in celiac disease, a well recognized food allergy disease. The possible usefulness of this technique in the investigative approach of food hypersensitivity is discussed.

Interferon-gamma (r-IFN-gamma) induced activation of alveolar macrophages (AM) from anergic patients with chronic obstructive pulmonary disease (COPD).

Forty-six anergic patients (37 males and 9 females, age range 55-79 yr) were selected from ninety-one patients suffering from COPD due to frequent exacerbations and impaired delayed cutaneous reactivity (43.9%). The phenotype of circulating lymphocytes, their proliferative response to a panel of polyclonal T-cell activators and the candidacidal activity (CA) of circulating PMNs (polymorphonuclear cells) were measured. In 13 patients presenting a defective CA of circulating PMNs, the in vitro response of alveolar macrophage CA to r-IFN-gamma was also determined. We found: 1) a significant reduction in the CL response to PHA in COPD patients vs controls; 2) a low PMN-CA in 23 (57%) COPD patients; 3) a non-significant difference in phenotype analysis in patients and controls; 4) lower CA of AMs in COPD patients than in controls; 5) restoration in vitro of CA by r-IFN-gamma in the group of anergic COPD patients presenting depressed CA. We conclude that a defective cell-mediated immunity could be the basis of the enhanced susceptibility to infectious exacerbations in many COPD patients and that, in vitro, it could be reversed by r-IFN-gamma treatment.

CD40 ligand expression on the surface of colostral T cells.

The proportion of T lymphocytes, mainly CD4 positive, co-expressing the CD40 ligand (CD40-L) was significantly greater (P = 0.001) in the colostrum of 10 breast-feeding mothers than in either autologous or heterologous blood. This surface glycoprotein is a T cell molecule involved in B cell isotype switching and immunoglobulin production with its natural counter-receptor, CD40, expressed by both adult and infant B lymphocytes. As the T cells of newborn infants fail to express the CD40-L when stimulated in vitro, the in vivo upregulation on milk T lymphocytes may be one of the mechanisms through which the mother transfers immune protection to the suckling infant.

Protein tyrosine kinase inhibition and cell proliferation: is the [3H]-thymidine uptake assay representative of the T-lymphocyte proliferation rate?

T-cell growth is controlled to a large degree by extracellular signals that bind to specific receptors on the surface of cells. A number of these receptors have intrinsic protein tyrosine kinase (PTK) activity. Their action on second messenger generation, and thus on cell proliferation, has been indirectly demonstrated by the decrease in [3H]-thymidine (TdR) uptake that follows co-stimulation of T-cells with mitogens and PTK inhibitors such as genistein (GEN). In this paper we report that the [3H]-TdR uptake assay is not a valid and reliable tool for investigating the proliferative activity of certain T-cell lines. In fact, a concomitant assessment of both [3H]-TdR uptake and cell cycle progression demonstrated that GEN is able to block G2/M progression of Jurkat T-lymphocytes even at doses (5 micrograms/ml) that do not influence [3H]-TdR uptake. Pretreatment with sodium o-vanadate (100 nM) could not reverse the GEN-related cell cycle perturbation, but was able to restore optimal [3H]-TdR uptake. Finally, GEN treatment was able to induce concentration-dependent apoptotic cell death of Jurkat T-cells. The control of cell activation, proliferation and programmed cell death is undoubtedly influenced by receptor-associated PTKs. The final effect on cell survival is almost entirely dependent on the activation state of the cell. The [3H]-TdR uptake assay seems to be inadequate for a correct interpretation of the expected results.

Lymphocyte surface phenotypes in Down's syndrome.

The lymphocyte surface phenotypes in the blood of 7 non-institutionalized adults with trisomy-21, and in 10 karyotypically normal control subjects, were analysed by fluorescent microscopy with an OKT series of monoclonal antibodies. Whereas the proportions of OKT3+, OKT4+ and OKT6+ cells were similar in the two groups, the percentages of both OKT8+ and OKT10+ trisomic lymphocytes were significantly higher. The biological implications of these findings with respect to the underlying immunopathology of Down's syndrome are discussed.

Lymphocyte surface antigens: a longitudinal study during the first month of human life.

A longitudinal study on human T lymphoid surface antigens was performed on blood samples from 6 infants during their first month of life. Although the results indicate a sudden increase in the percentage of OKT3+ and E-RF+ cell types a few hours after birth and a less rapid decrease of OKT6+ and OKT9+ circulating cells, high levels of OKT10+ cells persisted. This finding suggests that infant blood contains immature phenotypic T lymphocytes after birth.

Phenotypic analysis of T cell subsets in the blood of chronically aborting women.

A relative decrease in helper and a parallel increase in suppressor-cytotoxic T lymphocytes was found in the blood of six habitually aborting women, as compared with the T-cell subset distribution in ten normal multiparae and eight multigravid post-partum women. It is suggested that an imbalance between the immunoregulatory T-cell subpopulations may contribute to the rejection of the semiallogenic fetus.

[The determination of specific IgE in allergy diagnosis. The clinical significance of specific IgE in drug allergies]. / Il dosaggio delle IgE-specifiche nella diagnostica allergica. Significato clinico delle IgE-specifiche nelle allergie a farmaci.

IgE-mediated allergic reactions to drugs may be diagnosed on the basis of anamnestic criteria, clinico-pathological manifestations as well as by the measurement of allergen-specific IgE. The concordance of these diagnostic procedures was investigated in 50 patients with a history of sensitivity to penicillin (12), sulphamethoxazole (9) or both (14), aspirin (2) and pyrazolones (13). All subjects displayed chronic urticaria/angioedema syndrome. Optimal concordance values were observed for penicillin and sulphamethoxazole, while no specific IgE were detected in the ASA-sensitive group. False positive results were noted in pyrazolone-sensitive patients with high total IgE levels. Based on these results, serological methods that detect drug-specific IgE may be carefully used as complementary diagnostic procedure only in those patients in whom an adverse reaction to antibiotics is suspected.

[Recent findings on the phenotype and function of T-lymphocytes in the human colostrum]. / Recenti acquisizioni sul fenotipo e la funzione dei T linfociti presenti nel colostro umano.

Recent studies in our laboratory have demonstrated that the great majority of human colostral T cells display the phenotypic and functional characteristics of memory T lymphocytes, e.g. were able to proliferate in response to anti-CD3 and anti-CD2 monoclonal antibodies, and to a lesser extent, to the lectin mitogen phytohaemagglutinin. In addition, their production of interferon-gamma after anti-CD3 and anti-CD2 stimuli was similar to that calculated in autologous blood lymphocyte cultures. More interestingly, the proportion of T lymphocytes bearing the gamma/delta T-cell receptor was found to be significantly higher in the mammary secretion than in autologous and heterologous blood samples. Furthermore, these cells were mostly delta-TCS-1+, thereby suggesting that they are actively motile cells capable of migrating from lymphoid to extra-lymphoid body tissues. The fact that the phenotypic pattern of colostral gamma/delta T cells is similar, if not identical, to that of the intestinal intraepithelial counterpart suggests that these cells might originate in the gut-associated lymphoid system and home selectively to the mammary gland late in pregnancy and throughout lactation. However, additional studies are needed to confirm whether milk T lymphocytes are actively involved in the adoptive lactation transmission of cellular immunity to the suckling infant.

[Initial experience in the treatment of bronchiolitis using heparin]. / Prime esperienze sul trattamento della bronchiolite con eparina.

The aim of this paper was to evaluate the efficacy of treatment with heparin in children with bronchiolitis. We studied 30 children (average age 11.3 months) with bronchiolitis; 15 subjects were submitted to a supportive therapy with an adjunct of sodium heparin (50 U/Kg/24 h i.v.) for a time lasting 24-36 hours. The control group was managed with supportive therapy only. We considered the days of the disease and the persistence of symptoms since the admission as peculiar parameters between the two groups. The results appeared to be highly significant in the heparin treated group, with a mean duration of the diseases of 3.06 days compared to 5.53 days of the controls (p less than 0.001). These data are supportive for a possible utilization of the heparin in bronchiolitis but much more experiments are needed to confirm these results.