A bivalent recombinant mycobacterium bovis BCG expressing the S1 subunit of the pertussis toxin induces a polyfunctional CD4 ⁺ T cell immune response

© 2019 Alex I. Kanno et al. Background. A recombinant BCG strain expressing the genetically detoxified S1 subunit of pertussis toxin 9K/129G (rBCG-S1PT), previously constructed by our research group, demonstrated the ability to develop high protection in mouse models of pertussis challenge which correlated with the induction of a Th1 immune response pattern. The Th1 immune response induced by rBCG-S1PT treatment was also confirmed in the murine orthotopic bladder cancer model, in which the intravesical instillation of rBCG-S1PT resulted in an improved antitumor effect. Based on these observations, we hypothesize that the reengineering of the S1PT expression in BCG could increase the efficiency of the protective Th1 immune response in order to develop a new alternative of immunotherapy in bladder cancer treatment. Objectives. To construct rBCG strains expressing S1PT from extrachromosomal (rBCG-S1PT) and integrative vectors (rBCG-Sli), or their combination, generating the bivalent strain (rBCG-S1+S1i), and to evaluate the respective immunogenicity of rBCG strains in mice. Methods. Mycobacterial plasmids were constructed by cloning the s1pt gene under integrative and extrachromosomal vectors and used to transform BCG, individually or in combination. Antigen expression and localization were confirmed by Western blot. Mice were immunized with wild-type BCG or the rBCG strains, and cytokines quantification and flow cytometry analysis were performed in splenocytes culture stimulated with mycobacterial-specific proteins. Findings. S1PT expression was confirmed in all rBCG strains. The extrachromosomal vector directs S1PT to the cell wall-associated fraction, while the integrative vector directs its expression mainly to the intracellular fraction. Higher levels of IFN-γ were observed in the splenocytes culture from the group immunized with rBCG-S1i in comparison to BCG or rBCG-S1PT. rBCG-S1+S1i showed higher levels of CD4 + IFN-γ + and double-positive CD4 + IFN-γ + TNF-α + T cells. Conclusions. rBCG-S1+S1i was able to express the two forms of S1PT and elicited higher induction of polyfunctional CD4 + T cells, indicating enhanced immunogenicity and suggesting its use as immunotherapy for bladder cancer

Charcoal chronology of the Amazon forest: A record of biodiversity preserved by ancient fires

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this record.he Amazon region holds a wide variety of ethnic groups and microclimates, enabling different interactions between humans and environment. To better understand the evolution of this region, ancient remains need to be analysed by all possible means. In this context, the study of natural and/or anthropogenic fires through the analysis of carbonized remains can give information on past climate, species diversity, and human intervention in forests and landscapes. In the present work, we undertook an anthracological analysis along with the 14 C dating of charcoal fragments using accelerator mass spectrometry (AMS). Charcoal samples from forest soils collected from seven different locations in the Amazon Basin were taxonomically classified and dated. Out of the 16 groups of charcoal fragments identified, five contained more than one taxonomic type, with the Fabaceae, Combretaceae and Sapotaceae families having the highest frequencies. 14 C charcoal dates span ∼6000 years (from 6876 to 365 yr BP) among different families, with the most significant variation observed for two fragments from the same sampling location (spanning 4000 14 C yr). Some sample sets resulted in up to five different families. These findings demonstrate the importance of the association between anthracological identification and radiocarbon dating in the reconstruction of paleo-forest composition and fire history.The authors thank the Brazilian agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ). KDM thanks CNPq for fellowship 305079/2014–0. CL thanks FAPEAM/FAPESP (09/53369-6, led by Flávia Regina Capellotto Costa) for financial support and Thaise Emílio, José Luiz Purri da Veiga Pinto, Rosineide Machado and Francislaide da Silva Costa for help with charcoal collection. TRF, BSM, and BHM acknowledge financial support from NERC (NE/N011570/1), CAPES/CNPq Science without Borders (PVE 177/2012 and PVE 401279/2014-6), CNPq/PPBio (457602/2012-0), CNPq/PELD (403725/2012-7) and the University of Exeter - College of Life and Environmental Sciences

Brazil in the Era of Fascism: The “New State” of Getúlio Vargas

The New State established in Brazil by Getúlio Vargas (1937–1945) is the most important case of the institutionalisation of a dictatorship of the fascism era in Latin America. During this time, an impressive spectrum of authoritarian regimes was established, some of which were very instable and poorly institutionalised, while others were more consolidated. Roger Griffin coined the concept of para-fascism for some of them, and the “New State” of Getúlio Vargas in Brazil is a paradigmatic case. In this essay, we analyse the processes of institutional reform in 1930s Brazil paying particular attention to how domestic political actors look at institutional models of fascism and corporatism.info:eu-repo/semantics/publishedVersio