Su.55. Cytotoxin Neutralization and Enzyme-Linked Immunosorbent Assays for Escherichia Coli Toxins

Ferreira, Tatiane; Scola, Monica C G; Cruz, Ana M.; Gaspari, Elizabeth De

doi:10.1016/j.clim.2006.04.482

Cited by 2 publications

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“…We have used a quantitative colorimetric method based on the uptake of NR dye, which accumulates in the lysosomes of uninjured cells. After detoxification with formaldehyde, neither stx1 nor stx2 was cytotoxic to Vero or HeLa cells (data not shown) [ 28 , 29 ].…”

Section: Resultsmentioning

confidence: 99%

“…For the evaluation of cytotoxicity, the average absorbance of the media control well, which contained no chemical, was regarded as 100%, and the percentage of viable cells in each well was calculated according to (O.D. × 100)/control media, which is equivalent to the percentage of viable cells per well [ 28 , 29 ]. The concentrations of cytokines were determined with reference to a standard curve for serial twofold dilutions of murine recombinant cytokines.…”

Section: Methodsmentioning

confidence: 99%

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The Design of New Adjuvants for Mucosal Immunity toNeisseria meningitidisB in Nasally Primed Neonatal Mice for Adult Immune Response

Ferreira

Gaspari

2012

The Scientific World Journal

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The aim of this study was to determine the value of detoxified Shiga toxins Stx1 and Stx2 (toxoids ofEscherichia coli) as mucosal adjuvants in neonatal mice for immunogenicity against the outer membrane proteins (OMPs) ofNeisseria meningitidisB. Mucosal immunization has been shown to be effective for the induction of antigen-specific immune responses in both the systemic and mucosal compartments. Systemic antibody levels (IgG, IgG1, IgG2a, IgG2b, IgM, and IgA) and mucosal IgM and IgA were measured by ELISA using anN. meningitidisas an antigen. In addition, IFN-γand IL-6 production were measured after stimulated proliferation of immune cells. Intranasal administration elicited a higher anti-OMP IgA response in both saliva and vaginal fluids. Our results suggest that both Stx1 and Stx2 toxoids are effective mucosal adjuvants for the induction of Ag-specific IgG, IgM, and IgA antibodies. The toxoids significantly enhanced the IgG and IgM response against OMPs with a potency equivalent to CT, with the response being characterized by both IgG1 and IgG2a isotypes, and increased IFN-gamma production. Additionally, bactericidal activity was induced with IgG and IgM antibodies of high avidity. These results support the use of the new toxoids as potent inducing adjuvants that are particularly suitable for mucosal immunization.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The Design of New Adjuvants for Mucosal Immunity toNeisseria meningitidisB in Nasally Primed Neonatal Mice for Adult Immune Response

Ferreira

Gaspari

2012

The Scientific World Journal

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Rapid and Efficient Preparation of Monoclonal Antibodies against 35 kDa Lipoprotein ofMycoplasma penetrans

et al. 2007

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To develop a rapid and efficient method for preparing monoclonal antibodies (MAb) against 35 kDa lipoprotein of Mycoplasma penetrans, BALB/c mice were injected into the footpads for immunization, and the popliteal lymph nodes were isolated 19 days later for MAb-producing hybridomas, from which the mAbs against the 35 kDa lipoprotein were screened. The identification of the mAb against the 35 kDa lipoprotein was performed using indirect enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Using popliteal lymph node procedures, we generated several positive clones, one of which we characterized by ELISA and immunoblot. The clone 1D41B8 was identified as the immunoglobulin G1 (IgG1) isotype, kappa chain with affinity constants (Ka) of 2.95 x 10(9) M(-1). The MAbs did not cross-react with a number of control bacteria, which included Mycoplasma fermentans, Mycoplasma hominis, and Mycoplasma genitalium. This is the first report on the preparation of mAbs against M. penetrans that is specific to 35 kDa lipoprotein using popliteal lymph nodes. The high-specificity and high-affinity MAbs produced by two methodologies used of hybridomas provide a basis for further research on the pathogenesis and early diagnosis of M. penetrans. This simple approach may become a method of choice for the generation and production of MAbs in a short period of time.

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Su.55. Cytotoxin Neutralization and Enzyme-Linked Immunosorbent Assays for Escherichia Coli Toxins

Cited by 2 publications

References 0 publications

The Design of New Adjuvants for Mucosal Immunity toNeisseria meningitidisB in Nasally Primed Neonatal Mice for Adult Immune Response

The Design of New Adjuvants for Mucosal Immunity toNeisseria meningitidisB in Nasally Primed Neonatal Mice for Adult Immune Response

Rapid and Efficient Preparation of Monoclonal Antibodies against 35 kDa Lipoprotein ofMycoplasma penetrans

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