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Additional file 3: Figure S2. a Sequence alignment among the deduced SBP-4 amino acid sequences from Egypt and USA B. caballi isolates (1, 2) and SBP4 from Babesia bovis (3). Consensus sequences are shown in the bottom row. bIn silico prediction of trans-membrane domains and signal peptides in Egyptian B. caballi SBP-4 using TMHMM2.

Additional file 5: Figure S4. a Anti-histidin antibodies recognize a 37 kDa antigen in a cell lysate of recombinant E. coli transformed with TOPO® TA plasmid containing the full-length Bcsbp4 gene. Lane 1: (+ ve) cultures induced with L-Arabinose; Lane 2: (- ve) non-Arabinose induced cultures. b Specific recognition of rBcSBP4 protein by B. caballi positive serum in ELISA (a 1:100 dilution of serum was serially diluted by two-fold) using: serum from an equid derived from a horse experimentally infected with B. caballi (blue columns); field horse positive for B. caballi infection (orange columns); and horse that tested negative to infection with B. caballi (grey columns).

Combined_supplemental_materials-Delaney_et_al for Avoidance of the NLRP3 Inflammasome by the Stealth Pathogen, Coxiella burnetii by Martha A. Delaney, Andreas den Hartigh, Samuel J. Carpentier, Timothy P. Birkland, Donald P. Knowles, Brad T. Cookson and Charles W. Frevert in Veterinary Pathology

Additional file 2: Figure S1. Features of the BcSBP4 protein: the BcSBP4 was predicted to be secreted. The location of the predicted signal peptide and the regions containing the two peptides selected for antibody production are indicated by a green rectangle. The predictions were performed using online software at http://wlab.ethz.ch/protter .

Additional file 6: Figure S5. Specificity of the anti BcSBP4 peptide rabbit antibodies: western blot analysis against (i) normal horse RBC lysate, (ii) B. caballi-infected RBC lysate, (iii) recombinant B. caballi Spb4 and (iv) recombinant B. bovis Hap2 using serum indicated above. Abbreviation: M, size markers.

Additional file 1: Dataset S1. Amino acid sequence of BcSPB4.

Additional file 4: Figure S3. Secondary structure prediction of B. caballi SBP4 ( http://minnou.cchmc.org/ ).

Background Transmission of arthropod-borne apicomplexan parasites that cause disease and result in death or persistent infection represents a major challenge to global human and animal health. First described in 1901 as Piroplasma equi, this re-emergent apicomplexan parasite was renamed Babesia equi and subsequently Theileria equi, reflecting an uncertain taxonomy. Understanding mechanisms by which apicomplexan parasites evade immune or chemotherapeutic elimination is required for development of effective vaccines or chemotherapeutics. The continued risk of transmission of T. equi from clinically silent, persistently infected equids impedes the goal of returning the U. S. to non-endemic status. Therefore comparative genomic analysis of T. equi was undertaken to: 1) identify genes contributing to immune evasion and persistence in equid hosts, 2) identify genes involved in PBMC infection biology and 3) define the phylogenetic position of T. equi relative to sequenced apicomplexan parasites. Results The known immunodominant proteins, EMA1, 2 and 3 were discovered to belong to a ten member gene family with a mean amino acid identity, in pairwise comparisons, of 39%. Importantly, the amino acid diversity of EMAs is distributed throughout the length of the proteins. Eight of the EMA genes were simultaneously transcribed. As the agents that cause bovine theileriosis infect and transform host cell PBMCs, we confirmed that T. equi infects equine PBMCs, however, there is no evidence of host cell transformation. Indeed, a number of genes identified as potential manipulators of the host cell phenotype are absent from the T. equi genome. Comparative genomic analysis of T. equi revealed the phylogenetic positioning relative to seven apicomplexan parasites using deduced amino acid sequences from 150 genes placed it as a sister taxon to Theileria spp. Conclusions The EMA family does not fit the paradigm for classical antigenic variation, and we propose a novel model describing the role of the EMA family in persistence. T. equi has lost the putative genes for host cell transformation, or the genes were acquired by T. parva and T. annulata after divergence from T. equi. Our analysis identified 50 genes that will be useful for definitive phylogenetic classification of T. equi and closely related organisms. Published version

Natural sheep scrapie is a prion disease characterized by the accumulation of PrP(Sc) in brain and lymphoid tissues. Previous studies suggested that lymph node macrophages and follicular dendritic cells (FDC) accumulate PrP(Sc). In this study, lymph nodes were analyzed for the presence of PrP(Sc) and macrophage or FDC markers using dual immunohistochemistry. A monoclonal antibody (mAb) to the C-terminus of PrP reacted with CD172a+ macrophages and CD21+ FDC processes in secondary follicles. However, a PrP N-terminus-specific mAb reacted with CD21+ FDC processes but not CD172a+ macrophages in secondary follicles. Neither the PrP N-terminus nor C-terminus-specific mAb reacted with CD172a+ macrophages in the medulla. These results indicate that lymph node follicular macrophages acquire PrP(Sc) by phagocytosis of CD21+ FDC processes. The results also suggest that follicular macrophages have proteases that process full-length PrP(Sc) to N-terminally truncated PrP(Sc).