Protease Inhibition-An Established Strategy to Combat Infectious Diseases.

Therapeutic agents with novel mechanisms of action are urgently needed to counter the emergence of drug-resistant infections. Several decades of research into proteases of disease agents have revealed enzymes well suited for target-based drug development. Among them are the three recently validated proteolytic targets: proteasomes of the malarial parasite Plasmodium falciparum, aspartyl proteases of P. falciparum (plasmepsins) and the Sars-CoV-2 viral proteases. Despite some unfulfilled expectations over previous decades, the three reviewed targets clearly demonstrate that selective protease inhibitors provide effective therapeutic solutions for the two most impacting infectious diseases nowadays-malaria and COVID-19.

Temporal analysis of the bovine lymph node transcriptome during cattle tick (Rhipicephalus microplus) infestation.

Livestock production is a fundamental source of revenue and nutrition, wherein cattle-farming constitutes one of the major agricultural industries. Vectors and vector-borne diseases constitute one of the major factors that decrease the livelihood of all farming communities, more so in resource-poor communities and developing countries. Understanding the immunological responses during tick infestation in cattle is instrumental in the development of novel and improved tick control strategies, such as vaccines. In this study, gene expression patterns were compared within the lymph nodes of three cattle breeds at different life stages of the cattle tick, Rhipicephalus microplus. For Bonsmara (5/8Bos taurus indicus × 3/8B. t. taurus) cattle specifically, some 183 genes were found to be differentially expressed within the lymph nodes during larval and adult tick feeding, relative to uninfested cattle. Overall, the data provides evidence for a transcriptional regulatory network that is activated during immature tick infestation, but is down-regulated towards basal transcriptional levels when adult ticks are feeding. Specific processes in the lymph nodes of Bonsmara cattle were found to be differentially regulated on a transcriptional level. These include: (1) Leukocyte recruitment to the lymph node via chemokines and chemotaxis, (2) Trans-endothelial and intranodal movement on the reticular network, (3) Active regulation of cellular transcription and translation in the lymph node (including leukocyte associated cellular regulatory networks) and (4) Chemokine receptors regulating the movement of cells out of the lymph node. This work provides a first transcriptome analysis of bovine lymph node responses in tick-infested cattle. Findings show a dynamic immune response to tick infestation for the Bonsmara cattle breed, and that suppression of the maturation of the cattle hosts' immunity is especially evident during the larval feeding stages.

Comparison of the differential regulation of T and B-lymphocyte subsets in the skin and lymph nodes amongst three cattle breeds as potential mediators of immune-resistance to Rhipicephalus microplus.

Although varying natural resistance to ticks between highly resistant Brahman (Bos taurus indicus), resistant Bonsmara (5/8 B. t. indicus x 3/8 B. t. taurus) and susceptible Holstein-Friesian (B. t. taurus) breeds is documented in skin and blood, little information is available describing draining lymph nodes. To elucidate the cellular dynamics during Rhipicephalus microplus induced immune responses, this study analysed immune factors from these cattle breeds using histology, immunohistochemistry and flow cytometry. Following the collection of skin and lymph node samples before artificial tick infestation, cattle were infested with R. microplus larvae. Subsequent sampling coincided with the tick larvae and adult developmental stages. A significant influx of CD20+ B-lymphocytes in the dermis all cattle breeds was observed while CD3+ T-lymphocytes were significantly increased for more tick resistant breeds. Eosinophil infiltration in germinal centres of lymph nodes was significant for all cattle breeds while tingible body macrophages were significantly increased for adult infested Brahman animals. A negligible fluctuation in CD20+ and CD79α+ B-lymphocyte numbers was present in the lymph node of more resistant cattle breeds, while susceptible animals showed a decrease in B-lymphocytes after infestation, followed by an increase between larvae to adult infested time points. Increased variability of γd T-lymphocyte populations in lymph nodes was correlated with tick susceptibility. In addition, a more stable T helper lymphocyte population was identified in the lymph nodes for the Brahman cattle breed. Results suggest the association of tick susceptibility with differential B-lymphocyte regulation in lymph node tissues, increased variability of WC1+ γδ T-lymphocyte populations in the lymph node as well as a decrease in T helper lymphocytes in the lymph node.

Bovine Immune Factors Underlying Tick Resistance: Integration and Future Directions.

The mechanisms underlying tick resistance within and between cattle breeds have been studied for decades. Several previous papers on bovine immune parameters contributing to tick resistance discussed findings across DNA, RNA, protein, cellular, and tissue levels. However, the differences between bovine host species, tick species and the experimental layouts were not always taken into account. This review aims to (a) give a comprehensive summary of studies investigating immune marker differences between cattle breeds with varying degrees of tick resistance, and (b) to integrate key findings and suggest hypotheses on likely immune-regulated pathways driving resistance. Experimental issues, which may have skewed conclusions, are highlighted. In future, improved experimental strategies will enable more focused studies to identify and integrate immune markers and/or pathways. Most conclusive thus far is the involvement of histamine, granulocytes and their associated pathways in the tick-resistance mechanism. Interestingly, different immune markers might be involved in the mechanisms within a single host breed in contrast to between breeds. Also, differences are evident at each tick life stage, limiting the level to which datasets can be compared. Future studies to further elucidate immune molecule dynamics across the entire tick life cycle and in-depth investigation of promising markers and pathways on both molecular and cellular level are in dire need to obtain a scientifically sound hypothesis on the drivers of tick resistance.

Genetic diversity, acaricide resistance status and evolutionary potential of a Rhipicephalus microplus population from a disease-controlled cattle farming area in South Africa.

The Southern cattle tick, Rhipicephalus microplus is a hematophagous ectoparasite of great veterinary and economic importance. Along with its adaptability, reproductive success and vectoring capacity, R. microplus has been reported to develop resistance to the major chemical classes of acaricides currently in use. In South Africa, the Mnisi community in the Mpumalanga region offers a unique opportunity to study the adaptive potential of R. microplus. The aims of this study therefore included characterising acaricide resistance and determining the level and pattern of genetic diversity for R. microplus in this region from one primary population consisting of 12 communal dip-stations. The level of acaricide resistance was evaluated using single nucleotide polymorphisms (SNPs) in genes that contribute to acaricide insensitivity. Additionally, the ribosomal internal transcribed spacer 2 (ITS2) gene fragments of collected individuals were sequenced and a haplotype network was constructed. A high prevalence of alleles attributed to resistance against formamidines (amitraz) in the octopamine/tyramine (OCT/Tyr) receptor (frequency of 0.55) and pyrethroids in the carboxylesterase (frequency of 0.81) genes were observed. Overall, the sampled tick population was homozygous resistant to pyrethroid-based acaricides in the voltage-gated sodium channel (VGS) gene. A total of 11 haplotypes were identified in the Mnisi R. microplus population from ITS2 analysis with no clear population structure. From these allele frequencies it appears that formamidine resistance in the Mnisi community is on the rise, as the R. microplus populations is acquiring or generating these resistance alleles. Apart from rearing multi-resistant ticks to commonly used acaricides in this community these ticks may pose future problems to its surrounding areas.