Staphylococcus aureus-Cure-Associated Antigens Elicit Type 3 Immune Memory T Cells.

BACKGROUND: Staphylococcus aureus is one of the most frequently major mastitis pathogens that cause clinical and subclinical mastitis worldwide. Current antimicrobial treatments are usually ineffective, and the commercially available vaccines lack proven effectiveness. The immunological response elicited by the recombinant S. aureus-cure-associated proteins phosphoglycerate kinase (PGK), enolase (ENO), and elongation factor-G (EF-G) in combination with the granulocyte-macrophage colony-stimulating factor (GM-CSF) DNA vaccination was studied in this work. METHODS: Here, twenty-three C57BL/6 mice were divided into four groups and vaccinated with: G1: none (control); G2: GM-CSF DNA plasmid DNA vaccine; G3: the combination of EF-G+ENO+PGK; and G4: the combinations of EF-G+ENO+PGK proteins plus GM-CSF plasmid DNA vaccine. After 44 days, spleen cells were collected for immunophenotyping and lymphocyte proliferation evaluation by flow cytometry upon S. aureus stimulus. RESULTS: Immunization with the three S. aureus recombinant proteins alone resulted in a higher percentage of IL-17A+ cells among CD8+ T central memory cells, as well as the highest intensity of IL-17A production by overall lymphocytes indicating that the contribution of the combined lymphocyte populations is crucial to sustaining a type 3 cell immunity environment. CONCLUSION: The immunization with three S. aureus-cure-associated recombinant proteins triggered type 3 immunity, which is a highly interesting path to pursue an effective bovine S. aureus mastitis vaccine.

Alveolar macrophage functions during the transition phase to active immunity in calves1.

The first 3 to 6 mo of the life of calves is the period during which active immunity is established. During this period, greater morbidity and mortality is caused by bronchopneumonia because of the immaturity of the pulmonary immune system or the exaggerated cytotoxic response at subsequent infection. The aim of this study was to examine the maturity of the immune system during this phase of activation of acquired immunity in calves. For this purpose, the functions of phagocytosis and the reactive oxygen species (ROS) of alveolar macrophages CD14+ were evaluated. Further, the classes of immunoglobulins and the cytokines implicated in lymphocyte response patterns Th1 and Th2 in 10 healthy Holstein calves were quantified. Samples were taken from calves every 15 d, from the third to the sixth month of life. The alveolar macrophage CD14+ functions increased progressively until 150 d of age (phagocytosis, P = 0.02, ROS, P = 0.05), IgG1 and IgG2 isotype secretion reached an equilibrium, and the cytokine profiles were compatible with the Th1 response. At 165 d of age, there was a decrease in cellular function (phagocytosis P = 0.02, ROS P = 0.04) and an increase in IgG1 titers (P = 0.005) and IL-10 mRNA expression (P = 0.09). At 180 d of life, we observed an IgG1 and IgG2 secretion balance, a decrease in IL-10 mRNA expression, and an increase in IL-12 mRNA (P = 0.04) and tumor necrosis factor (TNF)-α mRNA expressions (P = 0.0003) and alveolar macrophage oxidative metabolism were observed. These results indicate that the calves had an active immune response that was distinctive for the age group. The CD14+ response is more reactive at 150 d. A regulatory and/or humoral response begins at 165 d of life as the equilibrium of Th1 and Th2 profiles is reached at 180 d of life. This may be clinically relevant for the development of specific therapies and prophylactic measures for bronchopneumonia in calves at 135 to 180 d of life.

Parenteral administration of vitamins A, D and E on the oxidative metabolism and function of polymorphonuclear leukocytes in swine / Administração parenteral das vitaminas A, D e E no metabolismo oxidativo e sobre a função de leucócitos polimorfonucleares em suínos

The weaning period of piglets is characterized by physiological alterations, such as decreased weight gain, increased reactive oxygen species (ROS) and increased serum cortisol levels with possible effects on the immune response. The effect of parenteral administration of vitamins A, D and E on production performance, oxidative metabolism, and the function of polymorphonuclear leukocytes (PMNLs) was assessed in piglets during the weaning period. The sample was comprised of 20 male piglets that were given an injectable ADE vitamin combination (135,000 IU vitamin A, 40,000 IU vitamin D and 40mg vitamin E/ animal) at 20 and 40 days of age. Weight gain, concentration of reduced glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD) and the microbicidal and phagocytic activity of PMNLs were assessed. No difference was observed in the average piglet weight during the study; however, a greater percentage of weight gain was observed after weaning in the treated group. The concentrations of GSH and SOD did not differ between groups, although lipid peroxidation was greater in the control group at 60 days of age. The investigated variables of oxidative metabolism were correlated as follows: -0.41 for GSH and MDA, -0.54 for GSH and SOD and 0.34 for MDA and SOD. The intensity of intracellular ROS production, the percentage of ROS-producing PMNLs and the intensity of phagocytosis by PMNLs did not differ between treatment groups. Administration of the injectable ADE combination improved the percentage of weight gain between 20 and 40 days of age, decreased oxidative stress at 60 days of age and did not influence the function of PMNLs in piglets.

O período de desmame nos leitões é caracterizado por alterações fisiológicas como menor ganho de peso, aumento na produção de espécies reativas de oxigênio (EROs) e aumento na concentração plasmática de cortisol com possíveis implicações para a resposta imune. Foi avaliado o efeito da administração parenteral das vitaminas A, D e E sobre o desempenho produtivo, o metabolismo oxidativo e a função de leucócitos polimorfonucleares (PMNLs) em suínos durante esta fase de crescimento. Foram utilizados 20 leitões, machos, com 20 dias de idade que receberam ADE injetável (135.000 UI vitamina A, 40.000 UI vitamina D e 40mg vitamina E/animal), aos 20 e 40 dias de idade. Foi determinado o ganho de peso e as concentrações de glutationa reduzida (GSH), malondialdeído (MDA) e superóxido dismutase (SOD) e a capacidade microbicida e fagocítica dos PMNLs. Não houve diferença entre o peso vivo médio durante o experimento, porém maior ganho de peso percentual foi observado 20 dias após o desmame para o grupo tratado. As concentrações de GSH e SOD não diferiram entre os grupos, porém a lipoperoxidação foi maior no grupo controle aos 60 dias de idade. As correlações entre as variáveis do metabolismo oxidativo foram -0,41 para GSH e o MDA, -0,54 para GSH e SOD e 0,34 para MDA e SOD. A intensidade da produção intracelular de EROs, a porcentagem de PMNLs que produziram EROs e a intensidade de fagocitose dos PMNLs não diferiram entre os tratamentos. A administração de ADE injetável melhorou o ganho de peso percentual no período de 20 a 40 dias de idade, diminuiu o estresse oxidativo aos 60 dias de idade e não influenciou função dos PMNLs dos leitões.