Lolium multiflorum grass is the major pollen allergen source in the southern region of Brazil, but most of its allergens remain poorly characterized. The aim of this study was to investigate antibody reactivity to L. multiflorum crude and carboxymethyl-ligand extracts in allergic patients and healthy individuals. Ion exchange carboxymethyl (CM) chromatography (CM-Sepharose) was used to isolate proteins (S2) from L. multiflorum crude extract (S1), which were assessed by SDS-PAGE. S1- and S2-specific IgE and IgG4 levels were measured by ELISA using sera from 55 atopic and 16 non-atopic subjects. Reactive polypeptide bands in S1 and S2 were detected by immunoblotting, and the most prominent bands in S2 were analyzed by mass spectrometry (MS-MS). Similar IgE and IgG4 levels were observed to both S1 (IgE median absorbance: 1.22; IgG4 median absorbance: 0.68) and S2 (IgE median absorbance: 1.26; IgG4 median absorbance: 0.85) in atopic subjects. S1 and S2 had positive correlations for IgE and IgG4 (IgE: r=0.9567; IgG4: r=0.9229; P<0.0001) levels. Homology between S1 and S2 was confirmed by IgE (84%) and IgG4 (83%) inhibition. Immunoblotting revealed that the 29-32 kDa band was recognized by 100% of atopic subjects in both S1 and S2. MS-MS analysis identified similarity profile to groups 1 and 5 grass allergens. This study revealed that carboxymethyl-ligand fraction played an important role for pollen allergy diagnosis by containing clinically relevant allergens and constituted a promising candidate for allergen-specific immunotherapy.

Blomia tropicalis (Bt) and Dermatophagoides pteronyssinus (Dp) are the prevalent house dust mites in tropical countries and are associated with allergic diseases. Glycosylated antigens are highly immunogenic and involved in different pathologies. We evaluated the presence of IgE, IgG1, and IgG4 to concanavalin A-binding antigens (Bt-Con-A) isolated from Bt-total extract in sera of allergic and non-allergic subjects. Bt-total and Bt-Con-A extracts were evaluated by SDS-PAGE and ELISA for reacting with IgE, IgG1, and IgG4 in sera of 121 patients with allergic rhinitis and 36 non-allergic individuals. All subjects were skin prick tested with Bt-total extract and inhibition tests were performed for IgE, IgG1, and IgG4 using both extracts (Bt-total and Bt-Con-A). Skin prick test showed that 58% of the patients were sensitized to Bt (Bt+), with 52% reactive to both mites (Bt and Dp) and 6% to Bt only. A broad spectrum of proteins (14-152 kDa) was visualized in Bt-total and components >27 kDa for the Bt-Con-A extract. ELISA showed a similar profile of IgE, IgG1 and IgG4 levels in response to Bt-total and Bt-Con-A extracts in different groups, although Bt+ patients showed a lower IgG4 reactivity to Bt-Con-A extract. Specific IgG1 levels were higher in Bt+ patients than in control subjects, and IgG4 levels showed no significant difference among groups. ELISA inhibition showed a partial IgE and total IgG1 and IgG4 cross-reactivity with Dp extract for Bt-total and Bt-Con-A extracts. We conclude that Con-A-binding components isolated from Bt constitute major allergens and are involved in both allergen sensitization (IgE response) and homeostasis maintenance (IgG1 and IgG4 responses).

Couepia grandiflora Benth. é conhecida popularmente como fruta-de-ema, pertence à família Chrysobalanaceae, e ocorre em regiões tropicais e subtropicais. Plantas dessa família são utilizadas pelas populações, principalmente, para o tratamento da diarréia, disenteria e malária. Neste trabalho, avaliou-se as atividades citotóxica, antibacteriana, antifúngica e antioxidante dos extratos hexânico e etanólico de C. grandiflora. O teste com Artemia salina usado para avaliar a citotoxicidade dos extratos hexânico e etanálico apresentou valores de CL50 de 0,1082 mg/mL e 0,1408 mg/mL, respectivamente. O extrato etanálico de C. grandiflora mostrou atividade antibacteriana contra Staphylococcus aureus e Pseudomonas aeruginosa, enquanto que o extrato hexânico mostrou atividade somente contra P aeruginosa. Os dois extratos não mostraram atividade contra Escherichia coli. As Concentrações Inibitórias Mínimas (CIM) do extrato etanólico foram de 188, 125, 188 μL/mL, para Staphylococcus aureus, Escherichia coli e Pseudomonas aeruginosa, respectivamente. Quanto à avaliação da atividade antioxidante, os extratos hexânico e etanólico mostraram-se ativos com valores de CE50= 3,1 e 5,6 μg/mL, respectivamente, quando comparados com os padrões. Na avaliação da atividade antifúngica, os extratos etanálico e hexânico de C. grandiflora não apresentaram tal atividade. Conclui-se que os extratos etanólico e hexânico de C. grandiflora demostraram atividades antibacteriana, antioxidante e citotoxicidade frente à Artemia salina e ausência de atividade antifúngica.

Couepia grandiflora Benth., known popularly as ema fruit, belongs to the family Chrysobalanaceae, and is distributed in tropical and subtropical areas. Plants of this family are used mainly for the treatrnent of the diarrhea. dysentery and malaria. In this work, it was evaluated the cytotoxic, antibacterial, antifungal and antioxidant activíties of the hexane and ethanolic extracts from C. grandiflora. The cytotoxic activíty of the hexane and ethanolic extracts showed a LC50 of 0.1082 mg/mL and 0.1408 mg/mL, respectively. The ethanolic extract of C. grandiflora showed antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa. while the hexane extract showed activity just against P. aeruginosa. The two extracts did not show activity against Escherichia coli. The Minímum Inhibitory Concentrations (MIC) of the ethanolic extract were 188, 125 and 188 μL/mL for Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, respectively. The evaluation of the antioxidant activity of the hexane and ethanolic extracts showed good results when compared to the reference drugs(EC50=3.1 and 5.6 mg/mL, respectively). It was not detected antifungal activity for hexane and ethanolic extracts from C. grandiflora.