CROSS-REACTION BETWEEN PROTEINS OF Larrea divaricata Cav. (JARILLA) AND BACTERIAL PROTEINS

M.A. MATTAR; DAVICINO R; MARTINO, RENZO; CASALI Y; MICALIZZI B

Mendoza

Congreso; XXVI Reunión Anual Científica de la Sociedad de Biología de Cuyo.; 2008

Sociedad de Biología de Cuyo

Larrea divaricata is employed to treat different pathologies. We aimed to characterize the cross-reaction of proteins from a partially purified crude aqueous extract (PPCE) of jarilla and whole cell-bacterial proteins (W-CBP) of Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris to characterize the cross-reaction of proteins from a partially purified crude aqueous extract (PPCE) of jarilla and whole cell-bacterial proteins (W-CBP) of Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris is employed to treat different pathologies. We aimed to characterize the cross-reaction of proteins from a partially purified crude aqueous extract (PPCE) of jarilla and whole cell-bacterial proteins (W-CBP) of Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgaris and Klebsiella pneumoniae using a mouse anti-PPCE serum. Proteins of jarilla were concentrated and partially purified by using membrane concentrators (Centriplus Amicon) with a 10 kDa cut off. Animals were immunized subcutaneously with PPCE. Levels of IgG against PPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-PPCE serum. Crossreactivity between proteins PPCE and W-CBP was studied by inhibition ELISA, using a poliyclonal antiserum specific for PPCE. Different concentrations of inhibitory proteins (PPCE and W-CBP) and antiserum were preincubated separately from the coated antigen. The antiserum dilution was previously determined by titration assays for each antigen: 1:1600 for PPCE, 1:800 for P. aeruginosa, P. vulgarisP. aeruginosa, P. vulgaris and K. pneumoniae and 1:1600 for E. coli. The binding of antibodies to PPCE was inhibited (>70%) by preincubation of the antibodies with 3.9, 15.6, 62.5, 250 and 1000 ìg/ml of PPCE. When the polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. to PPCE was inhibited (>70%) by preincubation of the antibodies with 3.9, 15.6, 62.5, 250 and 1000 ìg/ml of PPCE. When the polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. K. pneumoniae and 1:1600 for E. coli. The binding of antibodies to PPCE was inhibited (>70%) by preincubation of the antibodies with 3.9, 15.6, 62.5, 250 and 1000 ìg/ml of PPCE. When the polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. ìg/ml of PPCE. When the polyclonal antiserum was preincubated with E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. E. coli, P. vulgaris, K. pneumoniae and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. and P. aeruginosa W-CBP antigens, an inhibition of approximately 65% was obtained. The stronger inhibitory activity was observed with P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP. P. aeruginosa whole cell antigens. These results demonstrates that the PPCE-specific antiserum cross reacts with components present in the W-CBP.