To evaluate the role of CCR2 in allergic asthma, mutant mice deficient in CCR2 (CCR2−/−) and intact mice were sensitized with i.p. OVA with alum on days 0 and 7, and challenged by inhalation with nebulization of either OVA or saline. Airway hyperreactivity, measured by the methacholine-provoked increase in enhanced pause, was significantly increased (p < 0.05) in OVA-challenged CCR2−/− mutant mice, compared with comparably challenged CCR2+/+ mice. OVA-challenged CCR2−/− mutants also were also found to have enhanced bronchoalveolar lavage fluid eosinophilia, peribronchiolar cellular cuffing, and Ig subclass switching, with increase in OVA-specific IgG1 and IgE. In addition, RNase protection assay revealed increased whole lung expression of IL-13 in OVA-challenged CCR2−/− mutants. Unexpectedly, serum monocyte chemotactic protein-1 levels were 8-fold higher in CCR2−/− mutants than in CCR2+/+ mice sensitized to OVA, but OVA challenge had no additional effect on circulating monocyte chemotactic protein-1 in either genotype. Ag stimulation of lymphocytes isolated from OVA-sensitized CCR2 mutants revealed a significant increase (p < 0.05) in IL-5 production, which differed from OVA-stimulated lymphocytes from sensitized CCR2+/+ mice. These experiments demonstrate an enhanced response in airway reactivity and in lung inflammation in CCR2−/− mutant mice compared with comparably sensitized and challenged CCR2+/+ mice. These observations suggest that CC chemokines and their receptors are involved in immunomodulation of atopic asthma.

Background:Lolium multiflorum (Lm) pollen allergens are the major causative agents for rhinoconjunctivitis in Southern Brazil. There have been no studies about the sensitization and allergenic cross-reactivity between Lm and other grass pollens. We evaluated the sensitization of Brazilian pollinosis patients to Lm pollen allergens through skin prick test (SPT) and immunoassays (ELISA and immunoblot). Methods: Serum samples from 60 patients with pollinosis and positive SPT to grass pollen extracts (Lm+ group), 30 patients with negative SPT to grass pollen, but positive SPT to mite extracts (Lm– group), and 30 nonatopic subjects (NA group) were tested by SPT, ELISA, and immunoblot using Lm extract. Inhibition immunoassays with Lolium perenne (Lp), mixed grass (Gmix) and Lm extracts were also performed. Results: A high concordance was found between the Gmix and Lm extracts in SPT. Positivity rates in SPT were also highly concordant with IgE-ELISA results. The assay was able to detect Lm-specific IgE in >95% of Lm+ patients. A significant self- and cross-inhibition was observed in IgE-ELISA, reflecting a high cross-reactivity between the grass pollen allergens. Immunoblot revealed 13 IgE-binding Lm fractions, from which the bands 28–30 kDa and 31–34 kDa were recognized by >90% of Lm+ patients. Conclusion: Lm-specific IgE antibodies are highly cross-reactive with pollen proteins from other grass species. The results indicate that Lm extracts could be used in both SPT and ELISA for a more specific evaluation of IgE responses to Lm grass pollen in Brazilian pollinosis patients.