Staphylococcus aureus Protection-Related Type 3 Cell-Mediated Immune Response Elicited by Recombinant Proteins and GM-CSF DNA Vaccine

Abstract

Staphylococcus aureus mastitis remains a major challenge for dairy farming. Here, 24 mice were immunized and divided into four groups: G1: control; G2: Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) DNA vaccine; G3: F0F1 ATP synthase subunit α (SAS), succinyl-diaminopimelate (SDD), and cysteinyl-tRNA synthetase (CTS) recombinant proteins; and G4: SAS+SDD+CTS plus GM-CSF DNA vaccine. The lymphocyte subpopulations, and the intracellular interleukin-17A (IL-17A) and interferon-γ production in the draining lymph node cells were immunophenotyped by flow cytometry. The immunophenotyping and lymphocyte proliferation was determined in spleen cells cultured with and without S. aureus stimulus. Immunization with S. aureus recombinant proteins generated memory cells in draining lymph nodes. Immunization with the three recombinant proteins plus GM-CSF DNA led to an increase in the percentage of IL-17A+ cells among overall CD44+ (memory), T CD4+, CD4+ T CD44+ CD27−, γδ TCR, γδ TCR+ CD44+ CD27+, and TCRVγ4+ cells. Vaccination with S. aureus recombinant proteins associated with GM-CSF DNA vaccine downregulated TH2 immunity. Immunization with the three recombinant proteins plus the GM-CSF DNA led to a proliferation of overall memory T, CD4+, and CD4+ TEM cells upon S. aureus stimulus. This approach fostered type 3 immunity, suggesting the development of a protective immune response against S. aureus.

Expression of L-selectin (CD62L) and CD44 on Bovine Lymphocytes and WC1.1+ γδ T Cells under Stimulation with Staphylococcus aureus

“…Similar to our findings, [6] reported that the expression of CD44 (i.e., GMFI) was higher in proliferative cells than in nonproliferative cells upon stimulation by PPD in all T cell subsets evaluated in M. bovis-infected cattle. In this regard, it should be noted that dairy cows are constantly exposed to S. aureus, as they are part of the skin microbiota [3,4].…”

“…Bovine mastitis is the costliest disease affecting dairy farming and afflicts a high proportion of dairy cows. One of the most common bovine mastitis pathogens is Staphylococcus aureus, which is a notable issue for dairy farming due to its pathogenicity, contagiousness, the likelihood of persistence in the form of intramammary infections, and poor cure rates associated with current antimicrobial therapies; this pathogen has unquestionable importance in veterinary and human medicine [1][2][3]. This bacterium is also known to be part of the skin microbiota [4,5].…”

Diagnosis of Staphylococcus aureus Infection in Bovine Mastitis using its Affinity Purified Fraction

“…Based on proteomic analysis, the current study introduces two affinity purified protein bands isolated from S. aureus extract by affinity column chromatography (CNBr-Sepharose 4B) to be used in the bovine mastitis diagnosis via indirect ELISA. Adoption of proteomics in identification and characterization of S.aureus proteins was previously proved (Santos et al, 2021). Use of CNBr-Sepharose 4B for protein purification of S. aureus is unique and, in most cases, resulted in purified immunogenic components (Forman et al, 1987).…”

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

“…This observation encouraged us to study their effect on T-cell-mediated immunity which is considered more and more crucial for solid protection against S. aureus infection [ 21 ]. Furthermore, some studies have encouraged the use of a granulocyte-macrophage colony-stimulating factor (GM-CSF) DNA vaccine as a powerful immunoadjuvant [ 22 , 23 ] that could be related to T-cell-derived IL-17 production [ 24 , 25 ].…”

“…It should be highlighted that there is growing evidence that T cell immunity, especially T-cell-derived interleukin (IL)-17, plays a pivotal role in protection against S. aureus infections [ 24 , 26 , 27 ]; therefore, a fine-tuned characterization of T cell immunity elicited by the S. aureus proteins above is proposed here. Thus, in the present study, we investigate the immune response induced by three recombinant proteins of S. aureus , EF-G, ENO and PGK in combination with GM-CSF DNA vaccine in mice for future development of use in the cow.…”