ABSTRACT
RATIONALE: Nasal allergen provocations may be useful in investigating the pathophysiology of allergic rhinitis and effects of treatments. OBJECTIVE: To use grass pollen nasal allergen challenge (NAC) to investigate the effects of allergen immunotherapy in a cross-sectional study. METHODS: We studied nasal and cutaneous responses in untreated subjects with seasonal grass-pollen allergic rhinitis (n = 14) compared with immunotherapy-treated allergics (n = 14), plus a nonatopic control group (n = 14). Volunteers underwent a standardized NAC with 2000 biological units of timothy grass allergen (equivalent to 1.3 µg major allergen, Phl p5). Nasal fluid was collected and analysed by ImmunoCAP and multiplex assays. Clinical response was assessed by symptom scores and peak nasal inspiratory flow (PNIF). Cutaneous response was measured by intradermal allergen injection. Retrospective seasonal symptom questionnaires were also completed. RESULTS: Immunotherapy-treated patients had lower symptom scores (P = 0.04) and higher PNIF (P = 0.02) after challenge than untreated allergics. They had reduced early (P = 0.0007) and late (P < 0.0001) skin responses, and lower retrospective seasonal symptom scores (P < 0.0001). Compared to untreated allergics, immunotherapy-treated patients had reduced nasal fluid concentrations of IL-4, IL-9 and eotaxin (all P < 0.05, 8 h level and/or area under the curve comparison), and trends for reduced IL-13 (P = 0.07, area under the curve) and early-phase tryptase levels (P = 0.06). CONCLUSIONS: Nasal allergen challenge is sensitive in the detection of clinical and biological effects of allergen immunotherapy and may be a useful surrogate marker of treatment efficacy in future studies.
Subject(s)
Cytokines/immunology , Nasal Mucosa/immunology , Phleum/immunology , Plant Extracts/therapeutic use , Pollen/immunology , Rhinitis, Allergic, Seasonal/drug therapy , Rhinitis, Allergic/drug therapy , Administration, Intranasal , Adult , Bodily Secretions/immunology , Case-Control Studies , Cross-Sectional Studies , Desensitization, Immunologic , Female , Humans , Male , Middle Aged , Nasal Mucosa/metabolism , Rhinitis, Allergic/immunology , Rhinitis, Allergic, Seasonal/immunology , Sublingual Immunotherapy , Treatment Outcome , Young AdultABSTRACT
Erythropoietin (Epo) and thyroid hormone (T(3)) are key molecules in the development of red blood cells. We have shown previously that the tyrosine kinase Lyn is involved in differentiation signals emanating from an activated erythropoietin receptor. Here we demonstrate that Lyn interacts with thyroid hormone receptor-interacting protein 1 (Trip-1), a transcriptional regulator associated with the T(3) receptor, providing a link between the Epo and T(3) signaling pathways. Trip-1 co-localized with Lyn and the T(3) receptor alpha in the cytoplasm/plasma membrane of erythroid cells but translocated to discrete nuclear foci shortly after Epo-induced differentiation. Our data reveal that T(3) stimulated the proliferation of immature erythroid cells, and inhibited maturation promoted by erythropoietin. Removal of T(3) reduced cell division and enhanced terminal differentiation. This was accompanied by large increases in the cell cycle inhibitor p27(Kip1) and by increasing expression of erythroid transcription factors GATA-1, EKLF, and NF-E2. Strikingly, a truncated Trip-1 inhibited both erythropoietin-induced maturation and T(3)-initiated cell division. This mutant Trip-1 acted in a dominant negative fashion by eliminating endogenous Lyn, elevating p27(Kip1), and blocking T(3) response elements. These data demonstrate that Trip-1 can simultaneously modulate responses involving both cytokine and nuclear receptors.
Subject(s)
Adaptor Proteins, Signal Transducing , Transcription Factors/metabolism , Transcription Factors/physiology , ATPases Associated with Diverse Cellular Activities , Animals , Cell Cycle Proteins/metabolism , Cell Differentiation , Cell Division , Cell Nucleus/metabolism , Cells, Cultured , Cyclin-Dependent Kinase Inhibitor p27 , Cytokines/metabolism , DNA-Binding Proteins/biosynthesis , Erythrocytes/metabolism , Erythroid-Specific DNA-Binding Factors , Fluorescent Antibody Technique, Indirect , GATA1 Transcription Factor , Immunoblotting , Kruppel-Like Transcription Factors , LIM Domain Proteins , Mice , NF-E2 Transcription Factor , NF-E2 Transcription Factor, p45 Subunit , Precipitin Tests , Proteasome Endopeptidase Complex , Protein Binding , Retroviridae/metabolism , Signal Transduction , Transcription Factors/biosynthesis , Transcription, Genetic , Transfection , Tumor Suppressor Proteins/metabolism , Two-Hybrid System Techniques , src-Family Kinases/biosynthesisABSTRACT
Results of this study indicated that a protein-negative, blood-negative dipstick result may be used to rule out the necessity for performing a microscopic examination in "routine urinalysis" only if one is willing to accept 13% false-negative results. On the other hand, a protein-, blood-, and leukocyte esterase-negative dipstick result was associated with 1.4% to 3.3% false-negative results. The high sensitivity of the leukocyte esterase-measuring dipstick for microscopically abnormal urine samples was dependent on its ability to detect small numbers of leukocytes only when interpreted five minutes after immersing it in the sample. Various approaches may be used by which these findings could be applied to produce cost savings and also protect the small number of patients who may have dipstick-negative, microscopically positive urine.
