| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Allergy and Clinical Immunology, Imperial College School of Medicine at the National Heart and Lung Institute, London, United Kingdom
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
(10, 11, 12, 13, 14, 15, 16). Although in some circumstances the related molecule
CTLA-4, which has higher affinity for B7 molecules than CD28 and is
expressed on activation of T cells, can act in costimulation, recent
data suggest that it is predominantly a negative regulator of T cell
proliferation (9, 17, 18). CD86 and CD80 are also differentially
regulated on activation of APC (9).
Roles for both CD80 and CD86 costimulation pathways in airway
hyperresponsiveness and Ag-induced inflammatory airway eosinophilia in
mice have been suggested (19, 20, 21). In addition, some reports suggest
that CD86, but not CD80, costimulation can preferentially activate IL-4
expression and Th2 development (22, 23). However, recent data suggest
that although human PBMC memory responses, including those to allergen,
are dependent on CD86 costimulation, recently activated T cells and
human Th clones are not susceptible to inhibition by anti-CD86 Abs
(24, 25). We therefore examined the ability of CTLA4-Ig, a soluble
fusion protein of CTLA4 and human -1 constant region that binds B7
molecules with high affinity, or Abs to CD80 and CD86, to inhibit
allergen-induced peripheral blood T cell proliferation and IL-5
production and the dependence on CD80 and CD86 of airway APC and airway
T cell responses to allergen.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Eight atopic asthmatic subjects (mean age, 25.8; range, 22–36; four male) and seven nonatopic nonasthmatic control subjects (mean age, 25.3; range, 20–40; five male) were studied. Asthmatics had a history compatible with asthma and demonstrated reversible airflow obstruction of at least 15% and histamine PC203 of <4 mg/ml, and all had a positive skin prick test to house dust mite extract (Aluprick, ALK Horsholm, Denmark). Atopy was defined as previously (5) (at least one positive skin prick test to a common aeroallergen). Volunteers were assessed at an initial visit with a questionnaire on asthma symptoms, allergen skin prick tests, measurement of serum IgE concentration, and spirometry. Bronchial responsiveness to histamine was measured using a Wright nebulizer by the method of Cockcroft et al. (26). Asthmatic subjects completed a 2-week peak expiratory flow record to document variability in airflow obstruction and discontinued inhaled corticosteroids for at least 2 weeks before bronchoscopy. The asthmatic subjects had a mean serum IgE concentration of 324 IU/ml (range, 62–1009), mean FEV1 of 90.5% predicted (range, 63–102%), and geometric mean PC20 to histamine of 1.74 mg/ml (range, 0.4–3.6). None had received oral corticosteroids in the 2 months before the study, and permitted medication was inhaled ß2-agonists as required only. Control subjects all had no history of respiratory disease or allergy, negative skin prick tests, and normal IgE (mean, 27; range, 4–100) and had mean FEV1 of 100% predicted (range, 88–119). All had histamine PC20 of >16 mg/ml. None of the study volunteers had smoked in the preceding year, and none had more than 5 pack years in the past. All study volunteers had received Calmette-Guérin bacillus vaccination against tuberculosis. The study was approved by the ethics committee of the Royal Brompton Hospital, London, U.K.
Fiberoptic bronchoscopy was performed as described (5). In brief, all subjects were premedicated with nebulized albuterol, 2.5 mg, and sedated with i.v. midazolam. BAL was collected from the right middle lobe following instillation of 180–240 ml of sterile 0.9% saline in 60-ml aliquots.
Cell preparation
PBMC were isolated from heparinized blood samples by density gradient centrifugation over Ficoll-Paque (Pharmacia, Uppsala, Sweden). Cells were washed twice in HEPES-buffered RPMI 1640 (Sigma, Poole, U.K.) and resuspended in RPMI 1640 (Sigma) supplemented with 5% human AB serum (Sigma), 100 U/ml penicillin, 100 µg/ml streptomycin (Life Technologies, Paisley, U.K.), and 2 mM L-glutamine (Life Technologies) (complete RPMI). BAL cells were isolated by passing BAL through two layers of sterile gauze to remove mucus and washed twice in complete RPMI 1640 (Sigma). T lymphocytes were then purified by passage of BAL cells through a T cell enrichment column containing anti-human Ig-coated glass beads (R&D Systems, Abingdon, U.K.). This gave T cell purity of 85–90% for both blood and BAL, with <0.5% monocytes or B cells as judged by flow cytometric analysis with anti-CD3, anti-CD14, and anti-CD20 mAbs, respectively. To isolate CD4+ CD45RA+, and CD45RO+ cells from peripheral blood, depletion columns were used, with the same human Ig-coated glass bead separation columns after cells had been incubated with a mixture of Abs to remove CD8 and CD45RA/O cells, according to the manufacturer’s instructions (R&D Systems). Resulting cell populations were >97% pure, as judged by flow cytometry. Adherent cells (from BAL or PBMC) were isolated by culture on 90-mm plastic petri dishes for 2 h at 37°C with 5% CO2 in air. Nonadherent cells were removed by three vigorous washes with RPMI 1640 medium containing 1 U/ml preservative-free heparin (Evans Medical, Leatherhead, U.K.). Adherent cells were recovered by incubation at 37°C for 15 min in PBS containing 10 mM EDTA (British Drug Houses, Poole, U.K.) followed by harvesting using a rubber cell scraper. Cells were resuspended and counted before assay. BAL-adherent cells contained no detectable B cells by immunocytochemical staining of cytocentrifuge preparation with anti-CD20 Ab.
mAbs and cytokines
Anti-CD80 (BB1; IgM) was a kind gift of Dr Edward Clark, University of Washington, Seattle, WA. Anti-CD86 (IT2.2; IgG2b) was purchased from PharMingen (San Diego, CA). Anti-CD86 (BU63; IgG1) was a kind gift of Professor I. McClennan and Dr. D. Harding, Birmingham University, Birmingham, U.K. Abs directed against tumor-associated Ags (IgM and IgG1, respectively) were used as isotype controls and were a kind gift of Professor M. Ritter, Royal Postgraduate Medical School, London, U.K. An IgG2b control mAb (TNP) tri-nitrol phenol was purchased from PharMingen. CTLA-4-Ig fusion protein was a kind gift of Dr J. Ledbetter, Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, WA, and control fusion protein was a kind gift from Dr. M. Dallman, Imperial College, London, U.K. Phycoerythrin-conjugated anti-CD3, anti-CD14, anti-CD20, anti-CD45RA, and anti-CD45RO were from Becton Dickinson, Cowley, U.K. Phycoerythrin-labeled goat anti-mouse Ig and goat anti-mouse IgM were from Caltag (San Francisco, CA). IL-2 was from R&D Systems.
Proliferation studies and cytokine generation
T cells were incubated at 106 cells/ml in 200-µl volumes with 10 µg/ml Dermatophagoides pteronyssinus (ALK), or 10 µg/ml Mycobacterium tuberculosis PPD (Evans Medical, Leatherhead, U.K.). To measure allergen-specific proliferation of BAL T cells, 105 irradiated (3000 rads) autologous PBMC were added as APCs to 2 x 104 BAL lymphocytes, in the presence or absence of 10 µg/ml D. pternonyssinus extract (ALK). To assess BAL or peripheral blood-adherent cells as APC, these were added to isolated peripheral blood T cells in 96-well plates at a ratio of 2 x 104 APC to 1 x 105 T cells in quadruplicate cultures. Proliferation was measured on day 7 by adding 0.5 µCi of [methyl-3H]thymidine (Amersham, Amersham, U.K.) for the last 16 h of culture, and measuring label incorporation. Optimal cell ratios were determined for each of the different assays in preliminary experiments (data not shown). To assess dependence on CD80 or CD86, mAbs to CD80 (BB1) (1.0–10 µg/ml) or CD86 (BU63 or IT2), 0.01–0.5 µg/ml, or CTLA4Ig, 0.1–10 µg/ml, were added to quadruplicate wells. For cytokine generation, PBMC were stimulated with allergen at 5 x 106 cells/ml, and supernatants were collected on day 6. In some experiments, PBMC cultures were supplemented with 0.1, 1.0, or 10 ng/ml IL-2.
Cell lines
To generate allergen-specific T cell lines, PBMC were cultured with 10 µg/ml allergen at 106 cells/ml for 7 days. Cultures were then supplemented with fresh medium containing 20 U/ml recombinant human IL-2 (R&D Systems) and expanded for a further 7 days. Lymphoblasts were then washed and restimulated with irradiated autologous PBMC (3000 rads) and allergen and expanded in IL-2 (20 U/ml) for a further 7 days. After a further restimulation cycle, lymphoblasts were stimulated with APC and allergen, to measure allergen-induced proliferation or to generate cytokines. Briefly, 5 x 104 cells were incubated with an equal number of irradiated PBMC, with or without allergen in quadruplicate 200-µl cultures. After incubation for 48 h, 0.5 µCi of [methyl-3H]thymidine was added to each well, and label incorporation was assayed 16 h later. For cytokine generation, T cell lines were resuspended with irradiated PBMC (both at 0.5 x 106 cells/ml), and allergen and supernatants were collected 48 h later.
Mixed lymphocyte reaction
PBMC were isolated from two unrelated donors by density gradient centrifugation, as above, and resuspended in complete RPMI 1640 medium (containing 5% human AB serum). Cells were cultured alone or together at 5 x 104 cells/well (each donor), in the presence or absence of Abs, BB-1 (CD80; 5.0 and 10.0 µg/ml) or BU63 (CD86; 1.0 and 0.1 µg/ml), for 6 days before the addition of 1 µCi of [methyl-3H]thymidine for the last 8 h of culture. Proliferation was measured by thymidine uptake measured by scintillation spectroscopy.
Cytokine assays
IL-5 concentrations in culture supernatants were assayed by a
specific sandwich ELISA sensitive above 6.5 pg/ml, as previously
described (27). IL-4, IL-13, and IFN- were also measured by ELISA
(CMB, Amsterdam, The Netherlands), with sensitivities of 1, 0.5, and 2
pg/ml, respectively.
Flow cytometry
BAL cells were centrifuged at 500 x g for 7 min and then washed once in RPMI 1640 medium (Life Technologies). Cells were resuspended in medium at 5–10 x 106/ml, and an aliquot was taken for cytocentrifuge preparations. These were stained with May-Grünwald-Giemsa (Sigma) for differential cell counts. Aliquots of BAL cells and PBMC (5 x 105 cells in 100 µl, isolated as below) were incubated with control Abs, BU63, IT2, or BB1 to a final concentration of 5 µg/ml on ice for 15 min, washed twice in PBS with 1% BSA and 0.1% sodium azide (Sigma) (PAB), and then incubated with phycoerythrin-conjugated goat anti-mouse Ab. Following two further washes in PAB, cells were fixed in 2% formaldehyde in Isoton (Coulter, Luton, U.K.) and then analyzed on a EPICS Elite flow cytometer (Coulter). Alveolar macrophages and monocytes were identified on the basis of forward and side light scatter characteristics, and expression of CD80 or CD86 was analyzed in terms of specific mean fluorescence (relative to isotype control Ab).
Statistical analysis
Comparison of fluorescence intensity and T cell proliferation
(
cpm) between groups was by Student’s t test. Data were
analyzed using Minitab Release 9.2 (Minitab, State College, PA).
Differences were considered significant if p < 0.05.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Both atopic and nonatopic individuals showed proliferative
responses of PBMC to allergen and PPD, in accordance with previous
reports (28). To examine the costimulator dependence this proliferation
CTLA4-Ig or mAbs to CD80 or CD86 were titrated into cultures. As shown
in Fig. 1
, both CTLA4-Ig and
anti-CD86 inhibited allergen-induced proliferation in a
dose-dependent manner, whereas anti-CD80 Ab had no effect. Control
fusion protein did not inhibit proliferation (data not shown).
Allergen-stimulated proliferation of PBMC was also inhibited by
anti-CD86 in nonatopic donors, as was PBMC proliferation to PPD
(data not shown). To confirm biologic activity of the Abs at the
concentrations used, both anti-CD80 (BB-1) and anti-CD86 (BU63)
were shown to inhibit proliferation in a two-way mixed lymphocyte
reaction (Fig. 1).
|
As previously reported (28), culture of PBMC from atopic
individuals with allergen resulted in IL-5 production. This was
inhibited by both CTLA4-Ig and anti-CD86 mAbs to a similar extent,
whereas anti-CD80 or control fusion protein had no effect (Fig. 2). Titration of CTLA4-Ig showed that
maximal inhibition of allergen-induced PBMC IL-5 production was 89.5%
(±2.1 SEM) and 81.1 (±6.2)% for anti-CD86. In addition, allergen
induced IL-13 production (mean 651pg/ml, SEM 332 pg/ml), and IFN-
production (mean 870 ± 402pg/ml) from PBMC from atopic donors,
and this was also inhibited by anti-CD86 (mean, 71.3 ± 7.3%
for IL-13 and 94.4 ± 3.9% for IFN-), but not anti-CD80
mAbs. IL-4 production was not detected in these cultures.
|
When CD45RA+ and CD45RO+ CD4+
T cells were separated from peripheral blood and cultured separately
with irradiated PBMC and allergen, proliferation and IL-5 production
was seen from both subsets (Table I).
|
In some in vitro systems, T cell IL-5 production is IL-2 dependent
(29). Since blockade of CD86 costimulation inhibits IL-2 synthesis
(10), it was possible that the effects we observed on allergen-induced
IL-5 production were an indirect result of inhibition allergen-induced
IL-2 production. To test this, we attempted to restore proliferation
and IL-5 production by adding exogenous IL-2 to allergen-stimulated
PBMC cultured in the presence of anti-CD86 Abs. As shown in Fig. 3, although titration of exogenous IL-2
restored T cell proliferation to allergen, IL-2 did not overcome
anti-CD86 inhibition of allergen-induced IL-5 production.
|
To examine the CD86 dependence of recently activated allergen-specific
T cells, T cell lines prepared from atopic subjects were used 7 days
after their last Ag restimulation. Allergen-induced proliferation, IL-4
and IL-5 production from these CD4+ T cell lines were not
affected by mAbs to CD86 (Fig. 4) or CD80
(data not shown).
|
Resting monocytes express low levels of CD80 and CD86, and CD86 is
rapidly up-regulated following activation, with later induction of CD80
(8, 9). For this reason, we examined expression of CD80 and CD86 on
freshly isolated alveolar macrophages from atopic asthmatic subjects
and control individuals. Although alveolar macrophages from healthy
control subjects have been characterized as poor APC (30) and suppress
T cell responses to other APC (31), previous reports suggest that BAL
cells from asthmatic subjects have APC activity (32, 33). We therefore
examined the ability of BAL-adherent cells from asthmatic and control
subjects to stimulate allergen- and PPD-induced proliferation of
autologous peripheral blood T cells and the CD80 and CD86 dependence of
such proliferation. Alveolar macrophages from both asthmatic and
control subjects expressed CD86 and, to a lesser extent CD80 (Table II). BAL-adherent cells from asthmatics
induced proliferation of autologous T cells to both allergen and PPD,
to an extent comparable with peripheral blood-adherent cells (no
significant difference in counts per minute), whereas BAL cells from
nonasthmatic control subjects were poor APC (p
< 0.05 for proliferation with blood cells compared with BAL) (Fig. 5). T cell proliferation to allergen or
PPD, presented by airway APC, was inhibited by anti-CD86, but not
CD80 (Fig. 5). In addition, proliferation of T cells from nonasthmatic
subjects to allergen and PPD was inhibited by anti-CD86 (Fig. 5).
|
|
To examine the CD86 dependence of airway T cells, BAL T
lymphocytes were isolated from asthmatic subjects and cultured with
allergen presented by autologous irradiated PBMC. As shown in Fig. 6, house dust mite allergen-induced
proliferation of BAL T cells from asthmatic subjects was inhibited by
anti-CD86 mAbs.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The role of CD28/CTLA4-CD80/CD86 costimulatory signals in the development and maintenance of Th1 and Th2 appears to depend on the system studied (9, 22, 35, 36, 37). This may result from differential engagement of the different ligand pairs and the kinetics of expression of the molecules in different systems. Although studies examining repetitive stimulation of human peripheral blood naive T cells suggest a preferential role for CD86 in primary development of IL-4-producing cells (23, 38), other studies suggest that CD80 and CD86 provide similar costimulatory signals for T cell proliferation and cytokine production (35, 37). CTLA4-Ig treatment reduced pathology in a number of animal models of autoimmune disease (39, 40). CTLA4-Ig blocked aerosol Ag-induced bronchial hyperresponsiveness, IgE synthesis, and eosinophilia in at least three different mouse strains (19, 20, 21). Anti-CD86 Abs introduced into the airway in one mouse Ag challenge model inhibited eosinophil infiltration, IgE, and bronchial hyperresponsiveness, whereas anti-CD80 had little effect (21). In contrast, Harris et al. (41) suggested CD80 dependence of airway eosinophilia in a similar model. We studied allergen-induced T cell responses in both peripheral blood and airway T cells and airway APC from patients with chronic stable asthma, because this situation is most amenable to therapy. We show blockade of allergen and PPD responses by anti-CD86 but not anti-CD80, and the lack of effect of anti-CD80 was supported by the similar degree of inhibition of allergen-induced proliferation and cytokine production by CTLA4-Ig and CD86. This presumably reflects the predominant expression of CD86 by both PBMC and BAL cells. Thus, CD86 would seem most relevant to chronic asthma, although we cannot rule out a role of CD80 in T cell activation in acute exacerbations of disease. To confirm that the Ab to anti-CD80 was indeed biologicly active at the concentrations used, we showed inhibition of mixed lymphocyte reaction by both anti-CD80 and anti-CD86.
In contrast to PBMC, proliferation and cytokine production by recently activated allergen-specific T cell lines was not inhibited by anti-B7 Abs. These findings are in agreement with the recent demonstration by Bashian et al. (24) that human Th1 and Th2 clones were not susceptible to inhibition by CD80 or CD86, and the report by Yi-qun et al. (25) that T cell clones and in vitro preactivated T cells could be restimulated in the absence of CD80 or CD86 costimulation. However, both groups also found that peripheral blood memory cells were dependent on CD28 costimulation. It was therefore important to determine whether airway memory T cells, presumably frequently exposed to environmental allergen stimulation, are CD86 dependent or, like clones and T cell lines, have lost costimulator dependence. The current results suggest that in chronic asthma, allergen-specific airway cell proliferation is costimulator dependent. This is in agreement with animal models where administration of CTLA4-Ig or anti-CD86 after initial priming still lead to inhibition of subsequent Th2 activation after aerosol challenge (19, 20, 21). Because of the limitation of cell numbers, we did not test the effect of anti-CD80 on proliferation of BAL T cells; therefore, it remains possible that costimulation through this molecule plays a role in airway T cell activation. We were not able to detect cytokine production from the limited number of BAL T cells obtained, and it will require studies of the effect of CTLA-4Ig or CD86 blockade on allergen challenge or chronic asthma patients to confirm the importance of this potential therapeutic target.
The relatively low levels of allergen-induced proliferation compared with recall Ags such as PPD are in accordance with previous reports and may reflect frequency of responding cells (42, 43, 44). We saw equivalent allergen-induced T cell proliferation in cultures using isolated T cells and adherent peripheral blood APCs (predominantly monocytes) between atopic and nonatopic subjects. Previous reports have variously shown greater proliferative responses in atopic subjects, or equivalent T cell responses with different cytokine profiles, depending on the allergen and culture conditions (43, 44, 45, 46). We did not detect IL-4 in cultures of peripheral blood or BAL T cells. This may reflect the Ag dose used and the relative lack of B cells in the stimulating APC population (none were detected in the BAL-adherent cell population), since low allergen dose and B cell Ag presentation have been shown to favor IL-4 production (47).
Previous studies show that IL-5 production is dependent on IL-2 (29);
therefore, it was possible that inhibition of allergen-induced IL-5 by
anti-CD86 was an indirect result from diminished IL-2. We performed
cultures in the presence of allergen and anti-CD86 Ab, which
inhibits both proliferation and IL-5 production (Figs. 1
and 2).
Exogenous IL-2 did not restore allergen-induced IL-5 production in
cultures inhibited by anti-CD86. This raises the possibility that
inhibition of IL-5 production by anti-CD86 is a direct effect on
the T cell, rather than an indirect consequence of inhibition of
proliferation secondary to reduced IL-2 production.
The BAL-adherent cell population studied here was predominantly alveolar macrophages. However, APC activity likely also included airway dendritic cells, such as the CD1a+ popluation described by Van Haarst et al. (48). There was no increase in CD1a+ cell numbers in BAL from asthmatics when compared with control subjects (data not shown). However, we did not fully phenotype the adherent population studied and cannot exclude a difference in airway macrophage subpopulations between asthmatics and control subjects. B cells were not present in the BAL-adherent cell population studied here, although they may have potential as APCs in the airway mucosa. Chelen et al. (49) reported a relative deficiency in B7 expression by alveolar macrophages and showed that these cells did not increase B7 expression on activation. We studied BAL cells freshly isolated from asthmatics and control subjects, but did not examine the effects of in vitro activation. Since alveolar macrophages lack CD14, we were not able to use a specific phenotypic marker for these cells, but there was a clear separation of this population from other BAL cells by forward and side scatter at flow cytometry (50). It is possible that in vitro regulation of CD86 expression differed between the two groups studied, or that the relative signals from CD28 and CTLA4 differ between T cells from asthmatic subjects and controls, and that the balance between activating and inhibitory T cell molecules leads to differing responses to CD86 costimulation. Both CD45RA+ and CD45RO+ T cells from peripheral blood showed proliferation and IL-5 production to allergen, and, in agreement with previous reports (50, 51), >90% of the T cells in BAL were of CD45RO phenotype (data not shown), so that the inhibition we observed likely includes interruption of costimulation of a memory T cell response.
Cells other than T cells are thought to contribute to airway inflammation in asthma (52). In this regard, in studies of mice infected with Heligmosomooides polygyrus (40) or Leishmania major (53) or treated with anti-IgD (54), CTLA4-Ig reduced IL-4 production, but not IL-5 and IL-10. Non-T cell cytokine production would not be directly affected by therapy directed at CD86, although T cell cytokines are thought to have a pivotal role in IgE switching and eosinophil infiltration. Corticosteroid action in asthma are likely mediated through inhibition of T cell cytokine production (55), and we have previously shown therapeutic responses to T cell-directed therapy with cyclosporin A (56) in chronic asthma. Tsuyuki et al. (21) showed diminished bronchial hyperresponsiveness and local Th2, IgE, and eosinophil responses after local airway instillation of CTLA4-Ig or anti-CD86, without generalized systemic effects of this treatment. Taken together with our current findings, this suggests that T cell costimulation-targeted therapy will be of considerable interest in atopic asthma.
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Douglas S. Robinson, Allergy and Clinical Immunology, Imperial College School of Medicine at the National Heart and Lung Institute, London, SW3 6LY, U.K. E-mail address:
3 Abbreviations used in this paper: PC20, provocative concentration causing a 20% reduction in FEV1 from baseline; FEV1, forced expiratory volume in 1 s; BAL, bronchoalveolar lavage; PPD, purified protein derivative of Mycobacterium tuberculosis.
Received for publication January 22, 1998. Accepted for publication July 21, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
production. J. Exp. Med. 179:299.This article has been cited by other articles:
|
|
 |
|
  S.-E. Lamhamedi-Cherradi, R. E. Martin, T. Ito, F. Kheradmand, D. B. Corry, Y.-J. Liu, and M. Moyle Fungal Proteases Induce Th2 Polarization through Limited Dendritic Cell Maturation and Reduced Production of IL-12 J. Immunol., May 1, 2008; 180(9): 6000 - 6009. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Kallinich, K. C. Beier, U. Wahn, P. Stock, and E. Hamelmann T-cell co-stimulatory molecules: their role in allergic immune reactions Eur. Respir. J., June 1, 2007; 29(6): 1246 - 1255. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Crosby, M. Guha, D. Tung, D. A. Miller, B. Bender, T. P. Condon, C. York-DeFalco, R. S. Geary, B. P. Monia, J. G. Karras, et al. Inhaled CD86 Antisense Oligonucleotide Suppresses Pulmonary Inflammation and Airway Hyper-Responsiveness in Allergic Mice J. Pharmacol. Exp. Ther., June 1, 2007; 321(3): 938 - 946. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Linhart, S. Bigenzahn, A. Hartl, C. Lupinek, J. Thalhamer, R. Valenta, and T. Wekerle Costimulation Blockade Inhibits Allergic Sensitization but Does Not Affect Established Allergy in a Murine Model of Grass Pollen Allergy J. Immunol., March 15, 2007; 178(6): 3924 - 3931. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. T. Nouri-Aria, P. A. Wachholz, J. N. Francis, M. R. Jacobson, S. M. Walker, L. K. Wilcock, S. Q. Staple, R. C. Aalberse, S. J. Till, and S. R. Durham Grass Pollen Immunotherapy Induces Mucosal and Peripheral IL-10 Responses and Blocking IgG Activity J. Immunol., March 1, 2004; 172(5): 3252 - 3259. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Gosset, F. Bureau, V. Angeli, M. Pichavant, C. Faveeuw, A.-B. Tonnel, and F. Trottein Prostaglandin D2 Affects the Maturation of Human Monocyte-Derived Dendritic Cells: Consequence on the Polarization of Naive Th Cells J. Immunol., May 15, 2003; 170(10): 4943 - 4952. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Fahy, S. Senechal, J. Pene, A. Scherpereel, P. Lassalle, A.-B. Tonnel, H. Yssel, B. Wallaert, and A. Tsicopoulos Diesel Exposure Favors Th2 Cell Recruitment by Mononuclear Cells and Alveolar Macrophages from Allergic Patients by Differentially Regulating Macrophage-Derived Chemokine and IFN-{gamma}-Induced Protein-10 Production J. Immunol., June 1, 2002; 168(11): 5912 - 5919. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Ganea and M. Delgado VASOACTIVE INTESTINAL PEPTIDE (VIP) AND PITUITARY ADENYLATE CYCLASE-ACTIVATING POLYPEPTIDE (PACAP) AS MODULATORS OF BOTH INNATE AND ADAPTIVE IMMUNITY Critical Reviews in Oral Biology & Medicine, May 1, 2002; 13(3): 229 - 237. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. TIKKANEN, K. B. LEMSTROM, and P. K. KOSKINEN Blockade of CD28/B7-2 Costimulation Inhibits Experimental Obliterative Bronchiolitis in Rat Tracheal Allografts . Suppression of Helper T Cell Type1-dominated Immune Response Am. J. Respir. Crit. Care Med., March 1, 2002; 165(5): 724 - 729. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. Ford, D. Rennick, D. D. Donaldson, R. Venkayya, C. McArthur, E. Hansell, V. P. Kurup, M. Warnock, and G. Grunig IL-13 and IFN-{gamma}: Interactions in Lung Inflammation J. Immunol., August 1, 2001; 167(3): 1769 - 1777. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A.B. Kay Allergy and Allergic Diseases- First of Two Parts N. Engl. J. Med., January 4, 2001; 344(1): 30 - 37. [Full Text] [PDF]
|
|
|
|
|
|
Y. Yi, M. McNerney, and S. K. Datta Regulatory Defects in Cbl and Mitogen-Activated Protein Kinase (Extracellular Signal-Related Kinase) Pathways Cause Persistent Hyperexpression of CD40 Ligand in Human Lupus T Cells J. Immunol., December 1, 2000; 165(11): 6627 - 6634. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. M. Varga, P. Wachholz, K. T. Nouri-Aria, A. Verhoef, C. J. Corrigan, S. J. Till, and S. R. Durham T Cells from Human Allergen-Induced Late Asthmatic Responses Express IL-12 Receptor {beta}2 Subunit mRNA and Respond to IL-12 In Vitro J. Immunol., September 1, 2000; 165(5): 2877 - 2885. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Morel, B. P. Croker, K. R. Blenman, C. Mohan, G. Huang, G. Gilkeson, and E. K. Wakeland Genetic reconstitution of systemic lupus erythematosus immunopathology with polycongenic murine strains PNAS, June 6, 2000; 97(12): 6670 - 6675. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. N. Vallejo, L. O. Mugge, P. A. Klimiuk, C. M. Weyand, and J. J. Goronzy Central Role of Thrombospondin-1 in the Activation and Clonal Expansion of Inflammatory T Cells J. Immunol., March 15, 2000; 164(6): 2947 - 2954. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. H. Jaffar, L. Stanciu, A. Pandit, J. Lordan, S. T. Holgate, and K. Roberts Essential Role for Both CD80 and CD86 Costimulation, But Not CD40 Interactions, in Allergen-Induced Th2 Cytokine Production from Asthmatic Bronchial Tissue: Role for {alpha}{beta}, But Not {gamma}{delta}, T Cells J. Immunol., December 1, 1999; 163(11): 6283 - 6291. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Delgado, J. Leceta, R. P. Gomariz, and D. Ganea Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide Stimulate the Induction of Th2 Responses by Up-Regulating B7.2 Expression J. Immunol., October 1, 1999; 163(7): 3629 - 3635. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. E Donovan and P. W Finn Immune mechanisms of childhood asthma Thorax, October 1, 1999; 54(10): 938 - 946. [Full Text]
|
|
|
|
|
|
D. C. Tsitoura, R. H. DeKruyff, J. R. Lamb, and D. T. Umetsu Intranasal Exposure to Protein Antigen Induces Immunological Tolerance Mediated by Functionally Disabled CD4+ T Cells J. Immunol., September 1, 1999; 163(5): 2592 - 2600. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. E Burastero and G. A Rossi Immunomodulation by interference with co-stimulatory molecules: therapeutic perspectives in asthma Thorax, June 1, 1999; 54(6): 554 - 557. [Full Text]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
cpm) and are means from seven donors with standard error bars.
Culture without Ag gave background counts with a mean of 934 ±
736 cpm. In all cases, proliferation with allergen gave a stimulation
index (with allergen divided by no allergen background) of >3. The
activity of both anti-CD80 and anti-CD86 Abs was confirmed in a
mixed lymphocyte reaction in which both Abs inhibited proliferation.
Control fusion protein had no effect on proliferation (not shown). *,
p < 0.05 compared with isotype control.
). Results are plotted as percentage of
allergen-induced proliferation and IL-5 production in the presence of
isotype control Ab and are means with SEs for six separate
experiments.
