The lack of immunogenicity of most malaria antigens and the complex immune responses required for achieving protective immunity against this infectious disease have traditionally hampered the development of an efficient human malaria vaccine. The current boom in development of recombinant viral vectors and their use in prime-boost protocols that result in enhanced immune outcomes have increased the number of malaria vaccine candidates that access pre-clinical and clinical trials. In the frontline, adenoviruses and poxviruses seem to be giving the best immunization results in experimental animals and their mutual combination, or their combination with recombinant proteins (formulated in adjuvants and given in sequence or being given as protein/virus admixtures), has been shown to reach unprecedented levels of anti-malaria immunity that predictably will be somehow reproduced in the human setting. However, all this optimism was previously seen in the malaria vaccine development field without many real applicable results to date. We describe here the current state-of-the-art in the field of recombinant adenovirus research for malaria vaccine development, in particular referring to their use in combination with other immunogens in heterologous prime-boost protocols, while trying to simultaneously show our contributions and point of view on this subject.
Production of protoplasts is an important tool for genetic transformation of fungi. A protocol for protoplasts production in Aspergillus ochraceus was developed, evaluating culture aging of mycelium, different commercial enzymes and osmotic stabilizers. The best results were obtained with NH4Cl 0.8mol L-1 as osmotic stabilizer, mycelial age of 24 hours and Lysing Enzymes (20mg mL-1) plus Meicelase (20mg mL-1) as lytic enzymes. Good results were also obtained with Lysing Enzymes alone.
A obtenção de protoplastos é uma ferramenta importante para a transformação genética de fungos. Neste trabalho foi estudada a influência de fatores como idade micelial, tipo e concentração de enzimas e estabilizadores osmóticos na produção de protoplastos de Aspergillus ochraceus. Os melhores resultados de produção de protoplastos foram obtidos utilizando-se NH4Cl 0,8mol L-1 como estabilizador osmótico, micélio com 24h de crescimento e a combinação de Lysing Enzymes e Meicelase ambas, a 20mg mL-1. Entretanto, bons resultados foram também obtidos com a utilização apenas de Lysing Enzymes.