Protective effect of rPb40 as an adjuvant for chemotherapy in experimental paracoccidioidomycosis.

Abstract

The conventional treatment for the most prevalent mycosis in Latin America, paracoccidioidomycosis (PCM), involves long periods of therapy that results in side effects and a high frequency of relapses. The search for a new, alternative treatment is necessary. Pb40 is an antigenic protein from P. brasiliensis fraction F0. This fraction has already been shown to have significant protective activity when used as a PCM vaccine in experimental models. The complete cDNA sequence corresponding to Pb40 was cloned into a pET-21a plasmid, expressed in E. coli with a his-tag and purified by affinity chromatography. The predicted protein sequence exhibited nearly 100% homology to a fragment of the hypothetical EF-hand domain containing protein of P. brasiliensis. Immunization with this recombinant protein was used together with chemotherapy in an attempt to improve PCM treatment. The combined drug/rPb40 treatment exhibited long-lasting control of PCM in the liver and spleen and largely preserved the tissue structures of these organs. Despite the lack of a reduction in CFUs in the group that received the combined treatment, there was a significant reduction in the size of the lesions in the lungs after 70 days of infection. At the same time, the IL-10 levels were higher in the treated mice than in the infected-only mice. Moreover, significant levels of rPb40-specific IgG antibodies were detected in the sera of immunized mice. Thus, the treatment protocol consisting of rPb40 immunization in addition to fluconazole chemotherapy showed an additive protective effect after intratracheal challenge, preventing fungal dissemination to other sites of infection and preventing relapses. These results provide new prospects for PCM immunotherapy.

• Authors: V C Fernandes, E M N Martins, J N Boeloni, R Serakides, A M Goes
• Journal: Mycopathologia (Mycopathologia) Vol. 174 Issue 2 Pg. 93-105 (Aug 2012) ISSN: 1573-0832 [Electronic] Netherlands
• PMID: 22391822 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Adjuvants, Immunologic
• Antibodies, Fungal
• Antifungal Agents
• Antigens, Fungal
• Immunoglobulin G
• Recombinant Proteins
• Interleukin-10
• Fluconazole

Topics

• Adjuvants, Immunologic (administration & dosage, genetics, isolation & purification)
• Animals
• Antibodies, Fungal (blood)
• Antifungal Agents (administration & dosage)
• Antigens, Fungal (administration & dosage, genetics, isolation & purification)
• Drug Therapy (methods)
• Fluconazole (administration & dosage)
• Immunoglobulin G (blood)
• Immunotherapy (methods)
• Interleukin-10 (blood)
• Liver (microbiology)
• Mice
• Mice, Inbred BALB C
• Paracoccidioidomycosis (drug therapy)
• Recombinant Proteins (administration & dosage, genetics, isolation & purification)
• Spleen (microbiology)
• Treatment Outcome