Evaluation of the immune responses against reduced doses of Brucella abortus S19 (calfhood) vaccine in water buffaloes (Bubalus bubalis), India
VACCINE
Authors: Shome, Rajeswari; Kilari, Sreenivasulu; Sahare, Amol; Kalleshamurthy, Triveni; Niranjanamurthy, Harish Heballi; Shome, Bibek Ranjan; Hiremath, Jagadish; Misri, Jyoti; Rahman, Habibar
Abstract
Background: Brucella abortus S19 is the most widely used vaccine for the prevention of bovine brucellosis which remains the reference vaccine to which many other vaccine/s are compared. Considering the larger vaccination coverage by reduced dose of vaccine, the study aimed to compare reduced graded doses (1/10th, 1/20th and 1/100th) with standard dose of S19 vaccine (40 x 10(9)CFU/dose) to determine the effective immunizing dose in water buffaloes. Methods: A total of 25 female buffalo calves (Bubalus bubalis) in the age group of 4-5 months were equally grouped into five animals each in four test and one control groups and given with specified vaccine dose. The blood samples were collected on post vaccination days 14, 28, 45, 60, 90 and 120 for assessing innate (TNF-alpha and IL-12), humoral (IgG antibodies against Brucella LPS) and cell mediated immune responses (IFN-gamma, CD4 + and CD8 + counts). Results: The full dose, 1/10th and 1/20th reduced doses of S19 vaccine was capable of eliciting pathogen-specific antibody response, vaccine induced secretion of IL-12, TNF-alpha and IFN-gamma with CD4 + and CD8 + effector T cell responses. Persistence of antibody and magnitude of immune responses were found dose dependent. Conclusion: Comparable immune responses were noticed with 1/10th reduced dose similar to standard dose. With this observation, decline of antibody titre will reduce the number of false positives and reduced dose of vaccine will facilitate larger vaccination coverage in the country. (C) 2020 Elsevier Ltd. All rights reserved.
Efficient Plant Production of Recombinant NS1 Protein for Diagnosis of Dengue
FRONTIERS IN PLANT SCIENCE
Authors: Xisto, Mariana Fonseca; Dias, Roberto Sousa; Feitosa-Araujo, Elias; Prates, John Willians Oliveira; da Silva, Cynthia Canedo; de Paula, Sergio Oliveira
Abstract
Dengue fever is endemic in more than 120 countries, which account for 3.9 billion people at risk of infection worldwide. The absence of a vaccine with effective protection against the four serotypes of this virus makes differential molecular diagnosis the key step for the correct treatment of the disease. Rapid and efficient diagnosis prevents progression to a more severe stage of this disease. Currently, the limiting factor in the manufacture of dengue (DENV) diagnostic kits is the lack of large-scale production of the non-structural 1 (NS1) protein (antigen) to be used in the capture of antibodies from the blood serum of infected patients. In this work, we use plant biotechnology and genetic engineering as tools for the study of protein production for research and commercial purposes. Gene transfer, integration and expression in plants is a valid strategy for obtaining large-scale and low-cost heterologous protein production. The authors produced NS1 protein of the dengue virus serotype 2 (NS1DENV2) in the Arabidopsis thaliana plant. Transgenic plants obtained by genetic transformation expressed the recombinant protein that was purified and characterized for diagnostic use. The yield was 203 mu g of the recombinant protein per gram of fresh leaf. By in situ immunolocalization, transgenic protein was observed within the plant tissue, located in aggregates bodies. These antigens showed high sensitivity and specificity to both IgM (84.29% and 91.43%, respectively) and IgG (83.08% and 87.69%, respectively). The study goes a step further to validate the use of plants as a strategy for obtaining large-scale and efficient protein production to be used in dengue virus diagnostic tests.