Analysis of the cellular immune response induced by Bacillus thuringiensis Cry1A toxins in mice: effect of the hydrophobic motif from diphtheria toxin.
Insecticidal Cry1A toxins from Bacillus thuringiensis elicit strong humoral immune response in mice. Previously, we showed that an eight hydrophobic motif amino acid substitution in Domain I did not affect the antibody inducing capacity of the Cry1A toxins, on the contrary, it was enhanced after intranasal immunization. In addition, Cry1A mutants (carrying a substitution of a motif from fragment B of diphtheria toxin into the structurally similar hydrophobic alpha-helix 7 motif of Cry1A toxins) were able to modulate the ratio of IgG subclasses, IgG1/IgG2a. However, the capacity of these toxins to induce cellular immune response has not been studied. Thus, in this work, we investigated the cytokine profile induced after in vitro stimulation with the toxins, in spleen cell cultures from unprimed mice, and intranasally primed mice, with either wild-type Cry1Aa or with mutant toxin Cry1Aa8. Spleen cells from unprimed mice stimulated with Cry1Aa produced very low levels of Th1 (IFN-gamma, IL-12p70) and Th2 type cytokines (IL-10, IL-4), whereas immunization with Cry1Aa8 toxin led to higher production of these cytokines. Restimulation of spleen cells from primed mice with the Cry1Aa induced the production of significant levels of IL-12p70 whereas with Cry1Aa8, IFN-gamma production was stimulated. Interestingly, we found that the capacity of Cry1A toxins to induce cytokine production by lymphocytes was inhibited by N-acetylgalactosamine. Altogether these data demonstrate the immunogenic properties of Cry1A toxins and show that amino acid substitution in Domain I principally affects its ability to induce Th1 cytokines in lymphocytes.
Guerrero
G G
GG
Unidad de Biomedicina Immunidad en Mucosas, FES-Iztacala, Universidad Nacional Autónoma de México. Av de los Barrios No 1 Los Reyes Iztacala CP 54090,Tlanepantla, Estado de México, México. gloriaguillermina@correo.unam.mx
Russell
W M
WM
Moreno-Fierros
L
L
D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology; D019943:Amino Acid Substitution / Q000235:genetics / Q000276:immunology; D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics / Q000276:immunology*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D004167:Diphtheria Toxin / Q000235:genetics / Q000276:immunology*; D004731:Endotoxins / Q000235:genetics / Q000276:immunology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000276:immunology*; D057927:Hydrophobic and Hydrophilic Interactions; D007111:Immunity, Cellular; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004167:Diphtheria Toxin; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis