Yoshizawa I

Induction of allergen-specific IL-2 responsiveness of lymphocytes after respiratory syncytial virus infection and prediction of onset of recurrent wheezing and bronchial asthma.

BACKGROUND In pediatric patients with bronchial asthma and/or atopic dermatitis, peripheral lymphocytes are activated if they are stimulated with the responsible antigen, resulting in induction of responsiveness to IL-2. Because some nursing infants experience recurrent wheezing after respiratory syncytial virus (RSV) infection, attention is being directed to progression of the disease to bronchial asthma. OBJECTIVE The study was designed to elucidate the mechanism of the onset of allergic diseases after RSV infection. METHODS We examined allergen-specific IL-2 responsiveness induced in lymphocytes in the peripheral blood of infants after infection by RSV. The relationship between the onset of recurrent wheezing and antigen-specific IL-2 responsiveness was analyzed in 25 pediatric patients who could be followed up for 3 years after RSV infection. RESULTS Stimulation of lymphocytes with ovalbumin, alpha-casein, and mite (Dermatophagoides farinae) antigens induced significantly higher responsiveness to IL-2 in the RSV-infected infant group than in the healthy infant and disease control groups of the same age. There was no clear correlation between the IgE RAST scores for D. farinae, ovalbumin, and alpha-casein and IL-2 responsiveness. The families of RSV-infected infants had a high incidence of history of allergy (67%), but there was no significant difference in the incidence of patients with positive test results for IL-2 responsiveness between the groups with and without a familial history of allergy. The D. farinae-specific IL-2 responsiveness was significantly increased in the group with the symptom (16 patients) for a value of 1.64 +/- 0.13 (mean +/- SEM) compared with the value of 1.31 +/- 0.21 in the asymptomatic group (9 patients). The incidence of patients with positive test results for IL-2 responsiveness was 68.8% in the symptomatic group and 44.4% in the asymptomatic group. Similarly, the ovalbumin-specific IL-2 responsiveness was significantly increased in the symptomatic group (1.63 +/- 0.17) compared with the asymptomatic group (1.12 +/- 0.26). The incidence of patients with positive test results was 62.5% and 22.2%, respectively. alpha-Casein-specific IL-2 responsiveness was also higher in the symptomatic group than in the asymptomatic group, but the difference was not statistically significant. In the patient groups without RSV infection, on the other hand, the D. farinae-, ovalbumin-, and alpha-casein-specific IL-2 responsiveness in the symptomatic group were all similar to that in the asymptomatic group; no significant increases were detected. CONCLUSION The results indicated that after RSV infection, lymphocytes acquire specific susceptibility to D. farinae, a mite antigen, and food antigens, particularly ovalbumin. Hence, it is thought that positive IL-2 responsiveness specific for D. farinae and/or ovalbumin, detected several months after RSV infection, can be a prediction factor for the onset of allergic diseases, such as recurrent wheezing and bronchial asthma.

Cytokine production in children outgrowing hen egg allergy

Background Approximately 40 to 80% of egg‐allergic children outgrow egg allergy after 2 to 5 years.

Monoclonal proliferation of Epstein-Barr virus-infected T-cells in a patient with virus-associated haemophagocytic syndrome

Virus-associated haemophagocytic syndrome (VAHS) is a non-neoplastic, generalized histiocytic proliferation disorder showing marked haemophagocytosis associated with systemic viral infection. We describe the case of a 1-year-old girl with Epstein-Barr virus (EBV)-related VAHS, in whom Southern blot analysis showed monoclonal proliferation of bone marrow cells with the EBV genome; detected with the Xho-1 fragment of the latent infection membrane protein genome. EBV serology showed anti-Epstein-Barr virus nuclear associated antigen (EBNA), anti-viral capsid antigen (VCA)-IgG, anti-VCA-IgA elevation and positive EBNA of Sheep red blood cells (SRBC)-rosette-forming bone marrow cells in the late period of her clinical course, indicative of EBV infection. DNA analysis of her bone marrow cells showed monoclonal rearrangement of the T-cell receptor-β and-ψ chain genes but not of the immunoglobulin heavy chain genes. Those results suggest that EBV may infect T-cells, after which the cells proliferate monoclonally. Repeated administration of epipodophyllotoxin VP-16-213 induced remission, but adrenocortical steroid, vincristine, and cyclophosphamide had no effect on the patients condition. Ours is a first case report of VAHS showing monoclonal proliferation of EBV-infected T-cells.

Pattern of cytokine production by T cells from adolescents with asthma in remission after stimulation with dermatophagoides farinae antigen

ABSTRACT: Children with asthma usually become asymptomatic by the time they reach age 20 y. To clarify the immunologic mechanisms responsible for this phenomenon, we studied patients in remission and others who still had frequent asthma attacks. Patients were grouped by clinical status, and three variables were measured: serum levels of lgE, production of 11.4 and interferon (IFN)-γ, and the activation of T cells induced by Dermaophagoides farinae (DF) antigen. Df-induced activation of T cells (as measured by antigen-induced 11.2 responsiveness) or 11.2 synthesis itself was induced in patients with active asthma but not in normal subjects. These responses were much weaker in patients in remission. When stimulated by Df antigen in vitro, lymphocytes from patients with active asthma produced much more 11.4 than did the cells from normal subjects, and cells from patients in remission produced only a small amount. In contrast, under similar conditions lymphocytes from patients with active asthma produced less IFN-γ than did the cells from normal subjects. Production of IFN-γ stimulated by Df antigen was high in patients in remission but not in normal subjects. Thus, up-regulated IFN-γ production after exposure to Df antigen might reduce 11.4 secretion, which would suppress IgE production and would improve clinical status. Df antigen may suppress Df-induced allergic responses in patients with asthma in remission.

Effect of ketotifen on antigen-induced interleukin 2 (IL-2) responsiveness in lymphocytes from patients with atopic dermatitis and/or bronchial asthma

We tested the effect of Ketotifen (4-(1-methyl-4-piperidylidene)-4H- benzo[4,5] cyclohepta[1,2-b]thiophen-10(9H)-one hydrogen (fumarate) on the induction of allergen-induced IL-2 responsiveness in lymphocytes from patients with atopic dermatitis and/or bronchial asthma. Ovalbumin (OVA)- and/or Dermatophagoides farinae(Df)-induced IL-2 responsiveness was increased in almost all patients (1-15 years old) before Ketotifen treatment. Two to 12 months administration of Ketotifen (0.06 mg/kg/day) decreased activity of the response in 7 out of 9 cases corresponding to improvement of clinical symptoms. In in-vitro studies, antigen presenting cells (adherent cells) from the patient pretreated with 5, 50 and 500 ng/ml doses of Ketotifen for 12 h failed to present OVA or Df antigen to T-cells for induction of IL-2 responsiveness. Antigen-pulsed adherent cells also failed to induce the response of the T-cells pretreated with 50 and 500 ng/ml doses of Ketotifen but not with a 5 ng/ml dose. A 50 ng/ml dose of Ketotifen did not affect T-cells for induction of the response. In contrast, the treated adherent cells are capable of presenting PPD antigen or Con A for the induced response. The combined data indicate that induction of IL-2 responsiveness of peripheral blood lymphocytes on stimulation with nominal antigen may reflect an immune response to allergen in patients with allergy and a weak immunosuppressive effect of Ketotifen seems to block the response in the pathogenic process of allergic diseases.

A comparative investigation of the restorative effects of roxithromycin on neutrophil activities

The effects of roxithromycin, a macrolide antibiotic, on neutrophil activities were investigated in six seriously handicapped patients with severe mental retardation. Neutrophil activities were evaluated by flow cytometry using a heparinized blood analysis method. All six patients showed decreased levels of neutrophil phagocytosis, intracellular killing, and CD11b expression. Treatment with roxithromycin in vitro selectively restored the decreased phagocytic and bactericidal activities of neutrophils in these patients. There was no significant restorative effect with cefaclor, ofloxacin, or aztreonam. These results suggest the need to consider therapeutic effects of antibiotics on neutrophil functions in patients at increased risk for bacterial infections due to decreased neutrophil activities.

Correlation between antigen‐specific IL‐2 response test and provocation test for egg allergy in atopic dermatitis

The antigen‐specific interleukin‐2 response (AIR) test using lymphocytes is effective in searching for the antigen which causes allergic diseases and understanding their disease activity .

Effect of oxatomide on T-cell activation and the production of interferon-γ in mite sensitive asthma

Interleukin-2 responsiveness of lymphocytes induced by Dermatophagoides farinae antigen was suppressed upon exposure to 20 to 2000 ng/ml of oxatomide for 24 h in a dose-related manner in children with mite-sensitive bronchial asthma. Suppression was greater in the plastic-adherent antigen-presenting cells than in the T-cells. Oxatomide suppressed the production of interleukin-1alpha induced by Dermatophagoides farinae antigen in plastic-adherent cells. These results indicate that the target cells of oxatomide are antigen-presenting cells and not T-cells. Oxatomide also suppressed interleukin-2 responsiveness in lymphocytes exposed to purified protein derivative, but not in those exposed to concanavalin A. Unlike its effect on cell proliferation, oxatomide potentiated the Dermatophagoides farinae-induced production of interferon-gamma, which was suppressed by stimulation with Dermatophagoides farinae antigen in lymphocytes from the patients. In contrast, production of interferon-gamma induced by concanavalin A was not affected by this drug. These results indicate that oxatomide suppresses interleukin-2 responsiveness of allergen-activated helper T-cells and increases the production of interferon-gamma induced by Dermatophagoides farinae antigen, without causing cell proliferation.

Association of increased numbers of peripheral blood double-negative T-lymphocytes with elevated serum IgG levels in severely handicapped children.

CD3+4−8− double negative cells in peripheral blood lymphocytes were examined in 21 severely handicapped children divided into two groups according to serum IgG level. All children were bedridden and were taking multiple anticonvulsants and there were no apparent clinical differences between these two groups. Serum levels of IgG correlated well with percentages of CD3+4−8− double negative lymphocytes in patients of both groups. In comparisons between the two groups, the high IgG group had higher counts of CD3+4−8− double negative lymphocytes in peripheral blood than the normal IgG group. Two distinct types of double negative cells were identified in the patients with high IgG: one had T-cell antigen receptors of γδ heterodimers, the other had receptors of αβ chains on their surface. As double negative T-cells are reported to have an important role in defence against bacterial infections, the increased numbers of CD3+4−8− T-cells of both phenotypes in the high IgG patients may reflect exposure to repetitive bacterial stimuli or persistent subclinical infection which in many cases, may be undetectable clinically. Moreover, the hyperimmune states shown by the high serum IgG of these patients may result from the appearance of these unique lymphocytes because they are reported to have a helper function for IgG synthesis in vitro. Taken together, the increased numbers of double negative cells in patients with high IgG may reflect activated defence mechanisms and the development of hyperimmune status.

Reduced IL-1 production in adolescents with mite antigen asthma in remission

To determine the immunological mechanisms associated with outgrowing mite antigen‐induced bronchial asthma during adolescence, we studied the relationship between clinical status and Dermatophagoides farinae (Df) antigen‐induced peripheral cell activation by measuring IL‐1α and IL‐lβ production in patients with bronchial asthma. After antigen‐driven restimulation in vitro, there was increased IL‐1α, IL‐1β production by peripheral blood mononuclear cells (PBMC) from patients with active bronchial asthma, while cellular IL‐1α, IL‐1β production was reduced in patients with asthma in remission. IL‐1α and IL‐1β production by PBMC (possibly reflecting airway inflammation) after exposure to Df antigen might be down‐regulated in patients outgrowing mite antigen‐induced asthma, because lipopolysaccharide‐induced IL‐1α, IL‐1β production (seen in both normal individuals and patients with active asthma) was also reduced when patients were in remission.