Peptide vaccines designed with the aid of immunoinformatic against Caseous Lymphadenitis promotes humoral and cellular response induction in mice

Abstract

Caseous Lymphadenitis (CLA) is a chronic disease that affects also small ruminants. CLA is caused by Corynebacterium pseudotuberculosis and is responsible for high economic losses due to the formation of superficial and visceral granulomas, the latter is considered as asymptomatic CLA causing high levels of dissemination. Several vaccination strategies, in which the use of synthetic peptides stands out. Thus, this work aimed to evaluate the protective potential of peptide vaccines designed to determine the immunodominant epitopes of CP40 against CLA in mice. The animals were divided into eight groups separated in controls (G1—PBS, G2—Saponin and G9—rCP40) and experimental (G3—pep1, G4- pep2, G5-pep3, G6-pep4, G7-pep5 and G8-pep6), these were vaccinated on days 0 and 15 by a subcutaneous route. 60 days after the first immunization, all animals were challenged with C. pseudotuberculosis. On days 0, 15, 60, and 120 after the first immunization, blood samples were taken to measure immunoglobulins. On the same day of the challenge, the splenocytes were isolated and assayed for the production of IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17, and IL-10. After vaccinations, the animals were challenged and all of them were affected by the disease which led to their death. The G6 and G8 groups provided 10% protection and the G7 provided 20%. The G3 and G4 groups provided 30% and 40% protection respectively. The peptides showed the production of Total IgG antibodies and cytokines (IL-2, IL-4, IL-6, IFN-γ, and TNF-α), indicating a possible activation of the Th1 type response. However, groups G3, G5, G6, and G8 showed production of IL-17. None of the study groups showed IL-10 production. The immunogenicity of the peptides was not enough to protect these animals and it is believed that the use of adjuvants based on PAMPs may improve the immune response offered by these peptides.

Evaluation of the Association of Recombinant Proteins NanH and PknG from Corynebacterium pseudotuberculosis Using Different Adjuvants as a Recombinant Vaccine in Mice

“…Previous studies have already shown that IgG2a levels associated with the Th1 response highly correlate with phagocytic capacity [ 26 ]. Several studies using recombinant C. pseudotuberculosis proteins to compose vaccine formulations also reported significant levels of IgG2a production leading to the Th1-type immune response and generating protection in challenged animals [ 9 , 24 , 29 ], corroborating the results found here. Finally, even though the G2 and G3 groups had the same number of animals protected against the challenge when comparing IgG levels, the G2 group elicited the highest levels of antibodies, and that can be attributed to the use of the saponin adjuvant.…”

“…Furthermore, several studies have already used the saponin adjuvant associated with C. pseudotuberculosis antigens. The study by Droppa-Almeida et al (2021) showed that the saponin adjuvant was associated with peptides from the CP40 protein of C. pseudotuberculosis [ 24 ]. Moreover, in the studies by Droppa-Almeida et al (2016) and Silva et al (2014), vaccines for CLA containing recombinant antigens associated with saponin protected 90% and 100% of immunized animals, respectively [ 19 , 21 ].…”

Bioinformatic Approach of B and T Cell Epitopes of PLD and CP40 Proteins of Corynebacterium pseudotuberculosis ovis Mexican Isolate 2J-L towards a Peptide-Based Vaccine

“…Although all groups were affected during the challenge with a virulence strain, they also showed the production of total IgG antibodies and cytokines (IL-2, IL-4, IL-6, IFN-γ, and TNF-α), indicating a possible activation of the Th1 type response. The authors suggest the use of adjuvants based on PAMPs to improve the immune response offered by these peptides [ 32 ]. Since peptide sequences are also included on some predicted epitopes in our work, we consider it interesting to evaluate a multi-epitope vaccine with the combination of all peptides in the same formulation to enhance levels of protection.…”

Fasudil and SR1001 synergistically protect against sepsis-associated pancreatic injury by inhibiting RhoA/ROCK pathway and Th17/IL-17 response

“…Splenocytes were harvested and distributed into 96-well plates as described previously [ 24 ]. CD4+T cells were isolated using immunomagnetic beads (MACS beads, USA) and examined by the flow cytometry for purity.…”