Monoclonal Antibodies Against the 43,000 Da Glycoprotein from<i>Paracoccidioides brasiliensis</i>Modulate Laminin-Mediated Fungal Adhesion to Epithelial Cells and Pathogenesis

Gesztesi, J.-L.; Puccia, Rosana; Travassos, Luiz R.; Vicentini, Adriana Pardini; Moraes, Jane Zveiter de; Franco, Marcello; Lopes, José Daniel

doi:10.1089/hyb.1996.15.415

Cited by 67 publications

(53 citation statements)

References 29 publications

(5 reference statements)

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“…For gp43 purification, exoantigen of P. brasiliensis B-339 was prepared as previously described (34) and passed through an adsorbent column consisting of murine antigp43 monoclonal antibody (mAb) 17c (35) coupled to an Affi-Gel 10 column (Bio-Rad Laboratories, Hercules, CA, USA). Gp43 was eluted with 0.1 M citric acid buffer, pH 2.8, neutralized with 1 M Tris, pH 9.0, and further concentrated in a 10-K Amicon apparatus (Amicon Division, Beverly, MA, USA).…”

Section: Preparation Of Fungal Antigensmentioning

confidence: 99%

The low efficiency of dendritic cells and macrophages from mice susceptible to Paracoccidioides brasiliensis in inducing a Th1 response

Almeida

Lopes

2001

Braz J Med Biol Res

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In the present study we evaluated T cell proliferation and Th lymphokine patterns in response to gp43 from Paracoccidioides brasiliensis presented by isolated dendritic cells from susceptible and resistant mice. T cell proliferation assays showed that dendritic cells from susceptible mice were less efficient than those from resistant mice. The pattern of T cell lymphokines stimulated by dendritic cells was always Th1, although the levels of IL-2 and IFN-g were lower in T cell cultures from susceptible mice. To determie whether different antigen-presenting cells such as macrophages and dendritic cells stimulated different concentrations of Th1 lymphokines, the production of IFN-g and IL-2 was measured. It was observed that dendritic cells were more efficient than macrophages in stimulating lymphoproliferation in resistant mice. However, no significant difference was observed for IFN-g or IL-2 production. When cells from susceptible mice were used, macrophages were more efficient in stimulating lymphoproliferation than dendritic cells, but no difference was observed in the production of Th1 cytokine. Taken together, these results suggest the lower efficiency of dendritic cells and macrophages from B10.A mice in stimulating T cells that secrete Th1 lymphokines in vitro, an effect that may be involved in the progression of the disease in vivo.

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Section: Preparation Of Fungal Antigensmentioning

confidence: 99%

The low efficiency of dendritic cells and macrophages from mice susceptible to Paracoccidioides brasiliensis in inducing a Th1 response

Almeida

Lopes

2001

Braz J Med Biol Res

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“…For the first fusion, hybridomas were distributed on 48-well plates (Costar), and at 10 days postfusion the colonies were screened by immunoblotting, as described below. For the second fusion, hybridomas were distributed on 96-well plates (Costar), and screening was performed by an enzyme immunoassay, as described elsewhere (18). After the hybridomas were cloned by limiting dilution and expansion of the positive clones, large amounts of antibodies were obtained from the ascitic fluid induced in BALB/c mice injected with Pristane (Sigma) before inoculation of hybridoma cells.…”

Section: Methodsmentioning

confidence: 99%

Detection of Paracoccidioides brasiliensis gp70 Circulating Antigen and Follow-Up of Patients Undergoing Antimycotic Therapy

Silva¹,

Grosso²,

Lopes³

et al. 2004

J Clin Microbiol

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Paracoccidioidomycosis (PCM), one of the most important systemic mycoses in Central and South America, is caused by the dimorphic fungus Paracoccidioides brasiliensis and has a high prevalence in Brazil. Glycoproteins of 43 and 70 kDa are the main antigenic compounds of P. brasiliensis and are recognized by Western blotting by 100 and 96% of PCM patient sera, respectively. In the present study, an inhibition enzyme-linked immunosorbent assay (ELISA) was used to detect gp70 in different biological samples from patients with PCM. gp70 was detected in 98.76% of 81 serum samples, with an average concentration of 8.19 g/ml. The test was positive for 100% of the patients with the acute and chronic unifocal forms of PCM and 98.43% of the patients with the multifocal chronic form, with average concentrations of 11.86, 4.83, and 7.87 g/ml, respectively. Bronchoalveolar lavage fluid from 23 patients with pulmonary unifocal PCM and 14 samples of cerebrospinal fluid from patients with neurological PCM were also tested for gp70 detection, with the test showing 100% sensitivity and 100% specificity, with mean gp70 concentrations of 7.5 and 6.78 g/ml, respectively. To investigate the potential of gp70 detection by inhibition ELISA for the follow-up of PCM patients during antimycotic therapy with itraconazole (ITZ), the sera of 23 patients presenting with the chronic multifocal form of PCM were monitored at regular intervals of 1 month for 12 months. The results showed a decrease in circulating gp70 levels during treatment which paralleled the reduction in anti-P. brasiliensis antibody levels. The detection of P. brasiliensis gp70 from the biological fluids of patients suspected of having PCM proved to be a promising method for diagnosing infection and evaluating the efficacy of ITZ treatment.

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“…The higher antibody titer mouse had the spleen cells fused to the myeloma cell mouse line SP2/Ag14 (7,13). After fusion, cells were distributed over ten 96-well microtiter plates macrophage feeder layered and incubated in 5% CO 2 atmosphere at 37ºC (water jacked chamber, Forma Scientic, model 3158).…”

Section: Methodsmentioning

confidence: 99%

Monoclonal Antibodies to Identify Tomato Mosaic Tobamovirus (Tomv)

Duarte

Gomes

Gesztesi

et al. 2001

Braz. J. Microbiol.

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Monoclonal Antibodies Against the 43,000 Da Glycoprotein fromParacoccidioides brasiliensisModulate Laminin-Mediated Fungal Adhesion to Epithelial Cells and Pathogenesis

Abstract: ABSTRACT

Cited by 67 publications

References 29 publications

The low efficiency of dendritic cells and macrophages from mice susceptible to Paracoccidioides brasiliensis in inducing a Th1 response

The low efficiency of dendritic cells and macrophages from mice susceptible to Paracoccidioides brasiliensis in inducing a Th1 response

Detection of Paracoccidioides brasiliensis gp70 Circulating Antigen and Follow-Up of Patients Undergoing Antimycotic Therapy

Monoclonal Antibodies to Identify Tomato Mosaic Tobamovirus (Tomv)

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