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Immunization of Experimental Dogs With Salivary Proteins From Lutzomyia longipalpis, Using DNA and Recombinant Canarypox Virus Induces Immune Responses Consistent With Protection Against Leishmania infantum
(Frontiers Media S A, 2018-11-16) Abbehusen MMC; Cunha J; Suarez MS; Teixeira C; Almeida VDA; Pereira LDS; Bordoni M; Gil-Santana L; Solcà MDS; Fraga DBM; Fischer L; Bozza PT; Veras PST; Valenzuela JG; Kamhawi S; Andrade BB; Brodskyn CI; Ali N
Metacyclic Leishmania promastigotes are transmitted by sand flies that inject parasites and saliva into the host's skin. Previous studies have demonstrated that DNA plasmids encoding Lutzomyia longipalpis salivary proteins LJM17 and LJL143, when used to immunize dogs, resulted in a systemic and local Th1 cell-mediated immunity that interfered in parasite survival in vitro. Here we evaluated the ability of these same salivary antigens to induce anti-Leishmania immunity and to confer protection by immunizing dogs using a novel vaccination strategy more suitable for use in the field. The strategy consisted of a single dose of plasmid followed by two doses of recombinant Canarypoxvirus (rCanarypoxvirus) expressing L. longipalpis salivary proteins (LJM17 or LJL143). Thirty days after the final immunization, dogs were intradermally challenged with 107Leishmania infantum promastigotes in the presence of L. longipalpis saliva. We followed the experimentally infected dogs for 10 months to characterize clinical, parasitological, and immunological parameters. Upon vaccination, all immunized dogs presented strong and specific humoral responses with increased serum concentrations of IFN-γ, TNF, IL-7, and IL-15. The serum of dogs immunized with LJM17 also exhibited high levels of IL-2, IL-6, and IL-18. L. infantum infection was established in all experimental groups as evidenced by the presence of anti-Leishmania IgG, and by parasite detection in the spleen and skin. Dogs immunized with LJM17-based vaccines presented higher circulating levels of IFN-γ, IL-2, IL-6, IL-7, IL-15, IL-18, TNF, CXCL10, and GM-CSF post-infection when compared with controls. Results demonstrated that relevant Leishmania-specific immune responses were induced following vaccination of dogs with L. longipalpis salivary antigen LJM17 administered in a single priming dose of plasmid DNA, followed by two booster doses of recombinant Canarypox vector. Importantly, a significant increase in pro-inflammatory cytokines and chemokines known to be relevant for protection against leishmaniasis was evidenced after challenging LJM17-vaccinated dogs as compared to controls. Although similar results were observed following immunization with LJL143, the pro-inflammatory response observed after immunization was attenuated following infection. Collectively, these data suggest that the LJM17-based vaccine induced an immune profile consistent with the expected protective immunity against canine leishmaniosis. These results clearly support the need for further evaluation of the LJM17 antigen, using a heterologous prime-boost vaccination strategy against canine visceral leishmaniosis (CVL).
Antigenicity of Leishmania-Activated C-Kinase Antigen (LACK) in Human Peripheral Blood Mononuclear Cells, and Protective Effect of Prime-Boost Vaccination With pCI-neo-LACK Plus Attenuated LACK-Expressing Vaccinia Viruses in Hamsters
(Frontiers Media S A, 2018-04-23) Fernández L; Carrillo E; Sánchez-Sampedro L; Sánchez C; Ibarra-Meneses AV; Jimenez MA; Almeida VDA; Esteban M; Moreno J; Ritter U
Leishmania-activated C-kinase antigen (LACK) is a highly conserved protein among Leishmania species and is considered a viable vaccine candidate for human leishmaniasis. In animal models, prime-boost vaccination with LACK-expressing plasmids plus attenuated vaccinia viruses (modified vaccinia Ankara [MVA] and mutant M65) expressing LACK, has been shown to protect against cutaneous leishmaniasis (CL). Further, LACK demonstrated to induce the production of protective cytokines in patients with active CL or cured visceral leishmaniasis, as well as in asymptomatic individuals from endemic areas. However, whether LACK is capable to trigger cytokine release by peripheral blood mononuclear cells from patients cured of CL due to Leishmania infantum (L. infantum) or induce protection in L. infantum-infected hamsters [visceral leishmaniasis (VL) model], has not yet been analyzed. The present work examines the ex vivo immunogenicity of LACK in cured VL and CL patients, and asymptomatic subjects from an L. infantum area. It also evaluates the vaccine potential of LACK against L. infantum infection in hamsters, in a protocol of priming with plasmid pCI-neo-LACK (DNA-LACK) followed by a booster with the poxvirus vectors MVA-LACK or M65-LACK. LACK-stimulated PBMC from both asymptomatic and cured subjects responded by producing IFN-γ, TNF-α, and granzyme B (Th1-type response). Further, 78% of PBMC samples that responded to soluble Leishmania antigen showed IFN-γ secretion following stimulation with LACK. In hamsters, the protocol of DNA-LACK prime/MVA-LACK or M65-LACK virus boost vaccination significantly reduced the amount of Leishmania DNA in the liver and bone marrow, with no differences recorded between the use of MVA or M65 virus vector options. In summary, the Th1-type and cytotoxic responses elicited by LACK in PBMC from human subjects infected with L. infantum, and the parasite protective effect of prime/boost vaccination in hamsters with DNA-LACK/MVA-LACK and DNA-LACK/M65-LACK, revealed the significance of LACK in activating human and hamster immune responses and support LACK to be a valuable candidate for inclusion in a vaccine against human VL.
Developing digital learning to help reduce laboratory inequalities: a viral haemorrhagic fever case study
(BioMed Central Ltd, 2025-12-01) Knox MA; Bromhead C; Anthony W; Kamara V; Wendland C; Connor-Douglas F; Hayman DTS
Countries with the highest potential exposure to viral haemorrhagic fever viruses are also those with low expenditure on health services, limiting the capacity for surveillance and detection of these viruses, and effective treatment and outbreak containment. The COVID-19 pandemic further limited travel and in-person collaborative training opportunities for researchers, laboratory and public health professionals. Digital learning offers the prospect of addressing some of the shortfall in training needs. In this short report, we describe our experiences in the development of effective laboratory training tools using digital learning methods. We describe the teaching methodology, list barriers to successful implementation and offer some potential solutions.
Dental aging offers new insights to the first epigenetic clock for common dolphins (Delphinus delphis)
(2025-10-13) Hanninger E-MF; Peters KJ; Gerber L; Barratclough A; Betty EL; Palmer EI; Horvath S; Stockin KA
Bioprocessing of Mulberry Leaf Juice With High-GABA Producing Lactobacillus plantarum: A Strategy for Flavor, GABA Enrichment, and Antioxidant Enhancement
(John Wiley and Sons Ltd on behalf of the Institute of Food Technologists, 2026-03-02) Xie J; Yang S; Matia-Merino L; Chen J; Tian HS
The purpose of this work is to isolate γ-aminobutyric acid (GABA)-producing lactic acid bacteria (LAB) from traditional Chinese pickles and to evaluate their fermentation characteristics and functional effects in a mulberry leaf juice system. Six GABA-producing strains were isolated, comprising five Lactobacillus plantarum strains and one Enterococcus faecium strain. These strains demonstrated significant probiotic properties, exhibiting traits such as acid tolerance, bile salt resistance, in vitro gastrointestinal resistance, cell surface hydrophobicity, auto-aggregation, γ-hemolysis, and antibiotic sensitivity. The GABA content in fermented mulberry leaf juice ranged from 0.663 ± 0.007 to 0.879 ± 0.023 mg/mL, which was 9.61 to 12.74 times that of the initial content. Mulberry leaf juice's flavor, α-glucosidase inhibitory activity, DPPH and hydroxyl radical scavenging capacity, and ferric-reducing antioxidant power (FRAP) were all greatly improved by the LAB fermentation. A quality evaluation model for fermented mulberry leaf juice was developed using principal component analysis (PCA), identifying L. plantarum L10 as the optimal probiotic strain for fermentation.