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Vaccines against bovine babesiosis must, ideally, induce a humoral immune response characterized by neutralizing antibodies against conserved epitopes and a cellular Th1 immune response. In Babesia bovis , proteins such as AMA-1, MSA-2c, and RAP-1 have been characterized and antibodies against these proteins have shown a neutralizing effect, demonstrating the implication of B and T-cell epitopes in the immune response. There is evidence of the existence of B and T-cell epitopes in these proteins, however, it remains to be defined, the presence of conserved peptides in strains from around the world containing B and T-cell epitopes, and their role in the generation of a long-lasting immunity. The aim in this paper was to identify peptides of Babesia bovis AMA-1, MSA-2c, and RAP-1 that elicit a neutralizing and long-lasting Th1 immune response. Peptides containing B-cell epitopes of AMA-1, MSA-2c and RAP-1, were identified. The immune response generated by each peptide was characterized in cattle. All peptides tested induced antibodies that recognized intraerythrocytic parasites, however, only 5 peptides generated neutralizing antibodies in vitro : P2AMA-1 (6.28%), P3MSA-2c (10.27%), P4MSA-2c (10.42%), P1RAP-1 (32.45%), and P4RAP-1 (36.98%). When these neutralizing antibodies were evaluated as a pool, the inhibition percentage of invasion increased to 52.37%. When the T cellular response was evaluated, two peptides: P3MSA2c and P2AMA1 induced a higher percentage (>70%) of activated CD4 +/CD45RO+ T cells than unstimulated cells. Additionally, both peptides induced the production of gamma interferon (IFN−) in PBMCs from vaccinated cattle after one year proving the implication of a long-lasting Th1 immune response. In conclusion, we identified conserved peptides containing B and T-cell epitopes in antigens of B. bovis that elicit a Th1 immune response and showed evidence that peptides from the same protein elicit different immune responses, which has implication for vaccine development in bovine babesiosis. [ABSTRACT FROM AUTHOR]

Anaplasma Marginale, a rickettsial gram-negative bacterium, is responsible for causing bovine anaplasmosis which has widely spread among the different parts of the world causing death among cattle. It is mainly spread through the transmission of infected ticks. The manifestation of infected ticks could be controlled to a limited extent. However, subsequently, it is very much necessary for the cattle to develop strong immunity against A. marginale. It could be possible if a subunit vaccine is created against the rickettsial bacteria and introduced into the blood of the cattle for trials. In this paper, we have conducted a study to develop a multi-epitope subunit vaccine against A.marginale by using reverse vaccinology process. Five highly immunogenic outer membrane proteins of A. marginale were selected such as MSP1a, MSP2, Vir B10, OMP1 and OMP11. These proteins bind to the MHC class I molecules of the epithelial cells of mammals and MHC class II molecules of the antigen-presenting cells such as B-cells. The MHC class I molecules present antigens to the cytotoxic t-lymphocytes whereas the MHC class II molecules present antigens to the helper t-lymphocytes thus, initiating a strong immune response among the bovine species. We constructed three vaccine sequences from which two vaccine constructs V2 and V3, were found to be stable with high solubility and negative allergenicity. We also performed molecular docking of the two refined vaccine constructs with Rp-105 (30% resemblance in sequence to TLR4) and TLR-9. The vaccines interacted with the receptors with high binding energy. The best-designed vaccine construct was then reverse transcribed and adapted for the host E.coli K12 strain. The reverse transcribed sequence was later inserted into the pET28a (+) vector for the cloning and expression of the vaccine. The study could be a better initiative for in vitro and in vivo practice in developing an effective vaccine against A. marginale. • The five Outer surface proteins- MSP1a, MSP2, Vir B10, OMP1, and OMP11 were the most antigenic. • The proposed epitopes displayed high antigenic scores, high conservancy and non allergenicity. • Both the modeled vaccine V2 and V3 interacted with the bovine receptors rp-105 (TLR4) and TLR9 with high binding energy. • The designed construct, V3 was adapted for E. coli (K12) for cloning and expression. • Sequence of vaccine constructs showed no similarity with human proteins. [ABSTRACT FROM AUTHOR]

Zukamu granules (ZKMG), as the preferred drug for the treatment of colds in Uygur medical theory, has been used for 1500 years. It is also widely used in China and included in the National Essential Drugs List (2018 edition). It has unique anti-inflammatory, antitussive and analgesic effects. Aiming at the research of traditional Chinese medicine (TCM) with the characteristics of overall regulation of body diseases and the immune regulation mechanism with the concept of integrity, this paper put forward the integrated application of network composite module analysis and animal experiment verification to study the immune regulation mechanism of TCM. The active components and targets of ZKMG were predicted, and network module analysis was performed to explore their potential immunomodulatory mechanisms. Then acute lung injury (ALI) mice and idiopathic pulmonary fibrosis (IPF) rats were used as pathological models to observe the effects of ZKMG on the pathological conditions of infected ALI and IPF rats, determine the contents of Th1, Th2 characteristic cytokines and immunoglobulins, and study the intervention of GATA3/STAT6 signal pathway. The results of network composite module analysis showed that ZKMG contained 173 pharmacodynamic components and 249 potential targets, and four key modules were obtained. The immunomodulatory effects of ZKMG were related to T cell receptor signaling pathway. The validation results of bioeffects that ZKMG could carry out bidirectional immune regulation on Th1/Th2 cytokines in the stage of ALI and IPF, so as to play the role of regulating immune homeostasis and organ protection. The network composite module analysis and verification method is an exploration to study the immune regulation mechanism of TCM by combining the network module prediction analysis with animal experiments, which provides a reference for subsequent research. [Display omitted] [ABSTRACT FROM AUTHOR]