Frequency of allergen-specific T lymphocytes in blood and bronchial response to allergen in asthma☆
Abstract
Background: This study was designed to investigate whether the bronchial response to the sensitizing allergen in asthma is correlated with the frequency of allergen-specific T lymphocytes.
Methods: Twenty-three asthmatic patients sensitized to Dermatophagoides pteryonyssinus who had never received hyposensitizing therapy and 11 healthy control subjects were studied. Allergen-specific T lymphocytes were enumerated in peripheral blood with limiting dilution cultures. Bronchial challenge with methacholine was performed in all subjects; patients with asthma also underwent an allergen bronchial challenge. Correlations between allergen-specific T cell frequencies and nonspecific bronchial hyperresponsiveness to methacholine as independent variables and early and late bronchial responsiveness to allergen challenge as dependent variables were investigated by means of stepwise-multiple regression analysis.
Results: We found that the frequency of allergen-specific T lymphocytes was higher than in control subjects in both patients with asthma with (p < 0.001) and those without (p < 0.05) late phase asthmatic response to allergen. Moreover, the provocative does of allergen necessary to produce an early 15% fall of forced expiratory volume in 1 second could be predicted in part (59%) by an equation that incorporates methacholine sensitivity and allergen-specific T cell frequency.
Conclusions: We conclude that allergen-specific T lymphocytes, which have an established influence on immunoglobulin E production, play an additional role in the induction of the bronchospastic response to inhaled allergen.
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Cited by (41)
Human basophils and the immune response
2004, Revue Francaise d'Allergologie et d'Immunologie CliniqueL’inflammation allergique répond à des mécanismes immunologiques complexes, contrôlés par les cytokines de type 2, d’où un rôle pivot de la cellule T. Après activation antigénique, le basophile exprime aussi les IL-4 et IL-13, cytokines clés de la réponse inflammatoire allergique. Ainsi, après stimulation antigénique spécifique, 5–20 % des basophiles de patients allergiques uniquement expriment l’IL-4 et l’IL-13, (80 % des cellules productrices d’IL-4). La stimulation par la ionomycine est responsable d’une production des deux cytokines par la majorité des basophiles, et intervient quel que soit le statut clinique. L’expression de l’IL-4 par le basophile est rapidement maximale, celle de l’IL-13 est retardée, permettant au basophile d’initier, et d’amplifier la réponse allergique. De plus l’augmentation précoce de l’expression de CD40 ligand par le basophile, suggère son intervention dans la production initiale des IgE. Les CC chimiokines sont responsables d’une augmentation de la production d’IL-4 par le basophile, et des concentrations d’antigène 40 fois inférieures sont alors requises pour assurer une production équivalente d’IL-4. Les particules de diesel induisent une production précoce d’IL-4, par la majorité des basophiles, observée chez des sujets allergiques ou non, dépendante du stress oxydant. L’ensemble de ces résultats suggère un rôle déterminant du basophile dans la physiopathologie de la maladie allergique. À côté de cette action déterminante dans la réaction inflammatoire allergique, le basophile pourrait aussi être impliqué dans la lutte anti-infectieuse, en utilisant notamment des lignes de défenses non spécifiques, dépendantes de l’immunité innée.
Allergic inflammation is the result of complex immune mechanisms that depends on cytokines produced by Th2 lymphocytes, which gives these cells a pivotal role in the allergic reaction. When activated by allergen, basophils produce IL-4 and IL-13, cytokines key in the allergic inflammatory response. Antigen activation induces expression of these two cytokines by basophils of only about 5–20% of allergic patients; basophils represent 80% of the IL-4 producing cells. Ionomycine can induce the production of these cytokines by basophils from most individuals, regardless of their clinical status. IL-4 production by basophils peaks at 2 h, whereas IL-13 expression is delayed, suggesting that basophils may be involved both in initiation and amplification of the allergic response. Furthermore, the rapid increase in CD40L expression on peripheral blood basophils suggests that they may be involved in IgE production. The CC chemokines increase the production of IL-4 by basophils, and stimulation with a 40-fold lower concentration of antigen then results in an equivalent level of IL-4 production. Diesel exhaust particles induce early IL-4 production by the majority of basophils, regardless of the individual’s allergic status; this effect is dependent on the generation of reactive oxygen species. Taken as a whole, these data add weight to the conclusion that basophils are involved in the pathophysiology of allergic diseases. In addition to their critical role in allergic inflammation, basophils may also be involved in anti-infectious immunity through the nonspecific mechanisms involved in innate immunity
Specific immune response to Parietaria judaica plant profilin: A low T cell proliferative response supports high IgE and skin prick test
2002, Allergologia et immunopathologiaallergic disease caused by Parietaria judaica (Pj) has been widely documented in Mediterranean area. Profilins have been identified as widely distributed allergenic proteins. The role of Pj profilin in specific immune response in Pj-sensitized patients is unknown
skin prick test and determination of specific and total IgE levels in serum were performed in all patients (n = 28) and non-allergic controls (n = 18). Peripheral blood mononuclear cells (PBMC) were isolated from both groups and stimulated with crude extract or highly purified Pj profilin. The production of type I and type II cytokines was determined by specific and polyclonal stimuli in patients and controls. T-cell lines specific to Pj profilin were established and cross-reactivity with another highly purified profilin from Phleum pratense (Phl p) was evaluated
Pj profilin-sensitized patients showed a small but significantly increased in T-cell proliferative response to this profilin compared with non-atopic controls. The production of interleukin (IL)-4 and interferon (IFN)-γ in response to the specific stimulus was undetectable. However, the production of IL-4 in response to a polyclonal stimulus [phytohemagglutinin (PHA)] was significantly higher in atopic patients than in controls. The T-cell response did not correlate with the magnitude of response to skin prick tests with Pj profilin or with Pj-specific serum IgE levels. In addition, the production of IL-4 in response to a polyclonal stimulus (PHA) did not correlate with the individual skin prick tests to Pj profilin or with Pj-specific IgE levels in serum. The T-cell lines tested showed no cross-reactivity with Phl p profilin
our results suggest that Pj profilin is partly responsible for the T-cell-mediated response in patients allergic to Pj. The high skin reactivity to Pj profilin is these patients was accompanied by a small increase in the T-cell response to this profilin. The response was highly specific since Pj profilin specific T-cell lines showed no cross-reactivity with a highly homologous profilin from Phl p. The lack of correlation between the proliferative T-cell response and polyclonal IL-4 production with allergen-specific serum IgE and skin reactivity probably indicates that some of the responding T-cells may be involved in immune reactions other than those supporting IgE production
La enfermedad alérgica originada por Parietaria judaica (Pj) ha sido ampliamente documentada en la zona Mediterránea. Las profilinas han sido identificadas como proteínas alergénicas de amplia distribución. El papel de la profilina de Pj en la respuesta inmunológica específica en pacientes sensibilizados a la Pj es desconocida
Se determinaron en todos los pacientes (n = 28) y controles normales no atópicos (n = 18) el test de reactividad cutánea y los niveles tanto de IgE específicas como totales en suero. Se aislaron células mononucleares de sangre periféica (PBMC) en ambos grupos y fueron estimuladas con el extracto crudo o con la profilina altamente purificada de Pj. Se determinaron también la producción de citocinas de tipo I y de tipo II, con estimulaciones específicas o policlonales en pacientes y controles. Establecimos líneas de células T específicas por la profilina de Pj y evaluamos la reactividad cruzada con otra profilina altamente purificada de Phleum pratense (Phl p)
Los pacientes sensibilizados a la profilina de Pj muestran un ligero pero significativo incremento en la respuesta proliferativa T frente la profilina respecto con el grupo de controles no alérgicos. La producción de IL-4 y IFN-γ en respuesta a estímulos específicos fue indetectable. Sin embargo, la producción de IL-4 en respuesta a un estímulo policlonal (PHA), fue significativamente mayor en pacientes atópicos que en los controles no alérgicos. La respuesta celular T no se corresponde ni con la magnitud de la repuesta en la reactividad cutánea a la profilina de Pj ni con los niveles de IgE en suero específicos para la Pj. La producción de IL-4 frente a un estímulo policlonals (PHA), no se correlaciona con la reactividade cutánea a la profilina ni con los niveles de IgE en suero. Las líneas específicas T testadas no mostraron ninguna reactividad cruzada con la profilina de Phl p
Nuestros resultados sugieren que la profilina de Pj es responsable en parte de la respuesta mediada por células T en pacientes alérgicos a la Pj. La elevada reactividad cutánea de estos pacientes, se acompaña de un pequeño incremento en la respuesta de células T a esta profilina. La respuesta es muy específica porque las líneas de células T no presentaron respuesta cruzada con la profilina altamente homóloga de Phl p. La falta de correlación entre la respuesta proliferativa de las células T y la producción policlonal de IL-4 con los niveles de IgE específicos en suero y la reactividad cutánea probablemente indica que algunas de las células T respondedoras estarían involucradas en una reacción inmunitaria diferente a la que promueve la producción de IgE
Specific immune response to Phleum pratense plant profilin in atopic patients and control subjects
2001, Allergologia et ImmunopathologiaPhleum pratense (Phl p) pollen is a known cause of allergic disease worldwide. Profilins have been identified as functional plant pan-allergens. The role of Phl p profilin in the specific immune response in sensitized Phl patients is unknown
skin prick test and specific serum IgE levels were performed in 26 patients allergic to Phl p and in 18 onallergic control donors. Peripheral blood mononuclear cells were isolated from both groups and stimulated with crude extract or highly purified Phl p profilin, and the production of type I and type ll cytokines was determined in patients and controls stimulated with specific and polyclonal stimulus. T-cell lines specific to Phl p profilin were established from PBMCs and cross-reactivity with another highly purified profilin from Parietaria judaica (Pj) was evaluated
patients allergic to Phl p profilin showed increased T-cell-proliferative responses to this profilin compared with control subjects. The production of IL-4 and IFN-g in response to the specific stimulus was undetectable. However, the production of IL-4 and IFN-g in response to a polyclonal stimulus (PHA) was measurable and different for atopic patients and control subjects: IL-4 was higher (p < 0.001) in allergic patients and IFN-g lower (although not significant) in controls. Neither the T-cell responses nor the production of IL-4 in response to a polyclonal stimulus (PHA) correlated with the individual degree of cutaneous response to Phl p profilin or to the levels of specific Phl p IgE. The T-cell lines tested did not show any cross-reactivity with Pj profilin
Phl p profilin is in part responsible for the T-cell mediated immunological response in patients allergic to Phl p. The response is very specific since Phl p profilin specific T-cell lines did not show cross-reactivity with a highly homologous profilin from Parietaria judaica (Pj). The lack of correlation between the proliferative T-cell response and polyclonal IL-4 production with allergen-specific serum IgE and SPT probably indicates that some of the responding T-cells may be involved in immune reactions other than the support of IgE production
el polen de Phleum pratense (Phl p) es una de las causas más frecuentes de alergia respiratoria en el mundo occidental. Las profilinas han sido identificadas como proteínas pan-alergénicas con una amplia distribución en el reino vegetal. El papel de la profilina de Phl p en la respuesta inmunológica específica en pacientes sensibilizados al Phl p es desconocida
se practicaron a 26 pacientes alérgicos a Phl p y a 18 controles no atópicos pruebas cutáneas cuantificadas a profilina de Phl p, así como determinación de IgE específica a polen de Phl p. Se aislaron células mononucleares de sangre periférica (PBMC) en ambos grupos y tras estímulo con el extracto crudo o con la profilina altamente purificada de Phl p, se determinó la producción de citocinas de tipo I y de tipo II, por medio de estimulaciones específicas o policlonales tanto en pacientes como en controles. Se establecieron líneas de células T específicas por la profilina de Phl p y se evaluó la posible reactividad cruzada con otra profilina altamente purificada de Parietaria judaica (Pj)
los pacientes sensibilizados a la profilina de Phl p mostraron un incremento en la respuesta respuesta proliferativa T frente la profilina respecto al grupo de controles no alérgicos. La producción de IL-4 e IFN-g en respuesta al estímulo específico fue indetectable. Sin embargo, la producción de IL-4 en respuesta a un estímulo policlonal (PHA) fue significativamente mayor en pacientes alérgicos que en los controles y la producción de IFN-g fue menor en el grupo de alérgicos que en los controles. Ni la respuesta celular T ni la producción de IL-4 frente a un estímulo policlonal se corresponden con la magnitud de la respuesta en la reactividad cutánea a la profilina de Phl p, o con los niveles de IgE específica a Phl p en suero. Las líneas específicas T testadas no mostraron ninguna reactividad cruzada con la profilina de Pj
nuestros resultados sugieren que la profilina de Phl p es responsable en parte de la respuesta mediada por células T en pacientes alérgicos a Phl p. La respuesta es muy específica porque las líneas de células T no presentaron respuesta cruzada con la profilina altamente homóloga de Pj. La falta de correlación entre la respuesta proliferativa de las células T y la producción policlonal de IL-4 con los niveles de IgE específicos en suero y la reactividad cutánea probablemente indica que algunas de las células T respondedoras podrían estar involucradas en reacciones inmunitarias distintas de las que promueven la producción de IgE
PBMCs from both atopic asthmatic and nonatopic children show a T(H)2 cytokine response to house dust mite allergen
2000, Journal of Allergy and Clinical ImmunologyBackground: The hypothesis that in atopic diseases the T-helper response is skewed toward a TH2-type cytokine response was based on studies with mitogen stimulation, T-cell clones, or both. Objective: Using primary cultures, we investigated (1) whether atopic asthmatic patients have a TH2 response and nonatopic subjects have a TH1 response to allergen and (2) whether atopic patients have a decreased ability to mount TH1 immune responses to mycobacterial antigens. Methods: The responses of PBMCs to allergen (house dust mite [HDM]) or purified protein derivative of Mycobacterium tuberculosis (PPD) stimulation from 10 severely and 14 moderately asthmatic patients (all allergic to HDM) were compared with those of 17 nonatopic healthy black (Xhosa) children. Results: HDM-stimulated proliferation, IL-5 release, and the IL-5/IFN-γ ratio were significantly increased in subjects with atopic asthma, whereas IFN-γ release was not significantly different. IL-4 levels were below the level of detection. PPD-stimulated proliferation, IL-5 release, IFN-γ release, and the IL-5/IFN-γ ratio were not significantly different among the groups. Each group had a significantly higher IL-5/IFN-γ ratio in response to HDM than to PPD (a TH1 stimulus). Conclusion: Our study, which used primary cultures to investigate the hypothesis that nonatopic subjects have a TH1 response to allergens, indicates that HDM stimulates a TH2 cytokine response in both atopic and nonatopic subjects but that the response is enhanced in atopic patients. Our results with PPD suggest that normal and atopic asthmatic subjects can have a TH1 cytokine response to mycobacteria, but there is a subgroup of atopic subjects that have a TH2 response. (J Allergy Clin Immunol 2000;106:84-91.)
In childhood asthma the degree of allergen-induced T-lymphocyte proliferation is related to serum IgE levels and to blood eosinophilia
2000, Annals of Allergy, Asthma and ImmunologyTo investigate whether the state of activation of circulating T-cells in childhood asthma could be related to serum IgE levels and/or to blood eosinophilia.
Seventeen atopic asthmatic children, sensitized to Dermatophagoides pteronyssinus (Der p), in stable condition at the time of the study and 15 sex-matched and age-matched controls were studied. The expression of activation surface markers (HLA-DR and CD25) on peripheral blood mononuclear cells (PBMCs) was tested by monoclonal antibodies and FACS analysis, while the PBMC proliferative response to Der p antigens was measured by tritiated thymidine (3HTdR) incorporation.
As compared to controls, atopic children showed higher eosinophil counts (P < .01), similar lymphocyte counts (P > .1, each comparison) but higher proportion of HLA-DR+ and CD25+ T-lymphocytes (P < .05, each comparison). A significant Der p allergen-induced PBMC proliferation was observed in atopic children (P < .01) but not in controls (P > .1). Both in controls and in atopic children, no correlations were found between lymphocyte counts and eosinophil counts or total or allergen-specific IgE levels (P > .1, each comparison). In contrast, weak correlations were detected between the degree of allergen-induced PBMC proliferation and: a) allergen-specific IgE levels in serum (P < .05) and b) eosinophil counts (P < .05).
These data support the concept that the degree of activation of allergen-specific T-lymphocytes in blood may reflect the intensity of allergic sensitization in childhood asthma.
Activation of telomerase is induced by a natural antigen in allergen-specific memory T lymphocytes in bronchial asthma
1999, Biochemical and Biophysical Research CommunicationsThe function of the immune system is known to be dependent on the cellular differentiation and clonal expansion of allergen-specific lymphocytes. Telomerase, a ribonucleoprotein enzyme, is believed to be essential for the indefinite proliferation of human cells. To clarify whether telomerase is involved in the pathogenesis of immune diseases as well as of malignancies, we investigated the upregulation of telomerase activity in allergen-specific T lymphocytes. Upregulation of telomerase in allergen-sensitized lymphocytes was induced not only by artificial mitogenic stimulations but also by the natural antigen, house dust mite, which causes allergic diseases. Moreover, the upregulation of telomerase activity in memory T cells activated during allergen-specific immune responses might be associated with the enduring allergen-specific atopic propensity in asthmatics.
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Supported in part by a grant from the Italian Ministry of University and Technological and Scientific Research.