@article{amaral_mcqueen_bellingham-johnstun_poston_darville_nagarajan_laplante_kaser_2021, title={Host-Pathogen Interactions of Chlamydia trachomatis in Porcine Oviduct Epithelial Cells}, volume={10}, ISSN={["2076-0817"]}, url={https://www.mdpi.com/2076-0817/10/10/1270}, DOI={10.3390/pathogens10101270}, abstractNote={Chlamydia trachomatis (Ct) causes the most prevalent bacterial sexually transmitted disease leading to ectopic pregnancy and infertility. Swine not only have many similarities to humans, but they are also susceptible to Ct. Despite these benefits and the ease of access to primary tissue from this food animal, in vitro research in swine has been underutilized. This study will provide basic understanding of the Ct host–pathogen interactions in porcine oviduct epithelial cells (pOECs)—the counterparts of human Fallopian tube epithelial cells. Using NanoString technology, flow cytometry, and confocal and transmission-electron microscopy, we studied the Ct developmental cycle in pOECs, the cellular immune response, and the expression and location of the tight junction protein claudin-4. We show that Ct productively completes its developmental cycle in pOECs and induces an immune response to Ct similar to human cells: Ct mainly induced the upregulation of interferon regulated genes and T-cell attracting chemokines. Furthermore, Ct infection induced an accumulation of claudin-4 in the Ct inclusion with a coinciding reduction of membrane-bound claudin-4. Downstream effects of the reduced membrane-bound claudin-4 expression could potentially include a reduction in tight-junction expression, impaired epithelial barrier function as well as increased susceptibility to co-infections. Thereby, this study justifies the investigation of the effect of Ct on tight junctions and the mucosal epithelial barrier function. Taken together, this study demonstrates that primary pOECs represent an excellent in vitro model for research into Ct pathogenesis, cell biology and immunity.}, number={10}, journal={PATHOGENS}, publisher={MDPI AG}, author={Amaral, Amanda F. and McQueen, Bryan E. and Bellingham-Johnstun, Kimberly and Poston, Taylor B. and Darville, Toni and Nagarajan, Uma M. and Laplante, Caroline and Kaser, Tobias}, year={2021}, month={Oct} }
 @article{kick_wolfe_amaral_cortes_almond_crisci_gauger_pittman_kaeser_2021, title={Maternal Autogenous Inactivated Virus Vaccination Boosts Immunity to PRRSV in Piglets}, volume={9}, ISSN={["2076-393X"]}, url={https://doi.org/10.3390/vaccines9020106}, DOI={10.3390/vaccines9020106}, abstractNote={Maternal-derived immunity is a critical component for the survival and success of offspring in pigs to protect from circulating pathogens such as Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV-2). The purpose of this study is to investigate the transfer of anti-PRRSV immunity to piglets from gilts that received modified-live virus (MLV) alone (treatment (TRT) 0), or in combination with one of two autogenous inactivated vaccines (AIVs, TRT 1+2). Piglets from these gilts were challenged with the autogenous PRRSV-2 strain at two weeks of age and their adaptive immune response (IR) was evaluated until 4 weeks post inoculation (wpi). The systemic humoral and cellular IR was analyzed in the pre-farrow gilts, and in piglets, pre-inoculation, and at 2 and 4 wpi. Both AIVs partially protected the piglets with reduced lung pathology and increased weight gain; TRT 1 also lowered piglet viremia, best explained by the AIV-induced production of neutralizing antibodies in gilts and their transfer to the piglets. In piglets, pre-inoculation, the main systemic IFN-γ producers were CD21α+ B cells. From 0 to 4 wpi, the role of these B cells declined and CD4 T cells became the primary systemic IFN-γ producers. In the lungs, CD8 T cells were the primary and CD4 T cells were the secondary IFN-γ producers, including a novel subset of porcine CD8α−CCR7− CD4 T cells, potentially terminally differentiated CD4 TEMRA cells. In summary, this study demonstrates that maternal AIV vaccination can improve protection of pre-weaning piglets against PRRSV-2; it shows the importance of transferring neutralizing antibodies to piglets, and it introduces two novel immune cell subsets in pigs—IFN-γ producing CD21α+ B cells and CD8α−CCR7− CD4 T cells.}, number={2}, journal={VACCINES}, publisher={MDPI AG}, author={Kick, Andrew R. and Wolfe, Zoe C. and Amaral, Amanda E. and Cortes, Lizette M. and Almond, Glen W. and Crisci, Elisa and Gauger, Phillip C. and Pittman, Jeremy and Kaeser, Tobias}, year={2021}, month={Feb} }
 @article{kick_amaral_frias-de-diego_cortes_fogle_crisci_almond_käser_2021, title={The Local and Systemic Humoral Immune Response Against Homologous and Heterologous Strains of the Type 2 Porcine Reproductive and Respiratory Syndrome Virus}, volume={12}, ISSN={1664-3224}, url={http://dx.doi.org/10.3389/fimmu.2021.637613}, DOI={10.3389/fimmu.2021.637613}, abstractNote={The humoral immune response plays a crucial role in the combat and protection against many pathogens including the economically most important, highly prevalent, and diverse pig pathogen PRRSV – the Porcine Reproductive and Respiratory Syndrome Virus. In addition to viremia and viral shedding analyses, this study followed the local and systemic humoral immune response of pigs for 63 days upon inoculation with one of three types of Type-2 PRRSV (PRRSV-2) strains – one modified live virus (MLV) vaccine strain, and two lineage 1 PRRSV-2 strains, NC134 and NC174. The local response was analyzed by quantifying immunoglobulin (Ig)A in nasal swabs. The systemic response was studied by the quantification of IgG with ELISA and homo- and heterologous neutralizing antibodies (NAs) utilizing a novel method of flow cytometry. In all PRRSV-2 inoculated groups, viral nasal shedding started at 3 dpi, peaked between 3 and 7 days post inoculation, and was cleared at 28–35 dpi with sporadic rebounds thereafter. The local IgA response started 4–7 days after viral shedding occurred and showed a bi-phasic course with peaks at 14 dpi and at 28–35 dpi. Of note, the NC134 and NC174 strains induced a much stronger local IgA response. As reported earlier, main viremia lasted from 7 dpi to 28 dpi (NC174), 42 dpi (NC134) or until the end of the study (MLV). Similar to the local IgA response, the systemic IgG response started 4–7 days after viremia; but in contrast to viremia, serum IgG levels stayed high for all PRRSV-2 inoculated groups until the end of the study. A significant finding was that while the serum NA response in the MLV group was delayed by 28 days, serum NAs in pigs infected with our two NC134 and NC174 strains could be detected as early as 7 dpi (NC134) and 14 dpi (NC174). Compared to homologous NA responses, the NA responses against heterologous strains was strong but slightly delayed between our lineage 1 one strains or non-existent between the MLV and lineage 1 strains. This study improves our understanding of the relationship between local and systemic infections and the humoral immune response induced by PRRSV-2 infection or MLV vaccination. Our data also provide novel insights into the timeline of the development of homologous and heterologous NA levels – by both MLV vaccination or infection with two strains from the currently prevalent PRRSV-2 lineage 1.}, journal={Frontiers in Immunology}, publisher={Frontiers Media SA}, author={Kick, Andrew R. and Amaral, Amanda F. and Frias-De-Diego, Alba and Cortes, Lizette M. and Fogle, Jonathan E. and Crisci, Elisa and Almond, Glen W. and Käser, Tobias}, year={2021}, month={Mar} }
 @article{amaral_rebelatto_klein_takeuti_filho_mores_cardoso_barcellos_2019, title={Antimicrobial susceptibility profile of historical and recent Brazilian pig isolates of Pasteurella multocida}, volume={39}, ISSN={["1678-5150"]}, DOI={10.1590/1678-5150-PVB-5810}, abstractNote={ABSTRACT: Pasteurella (P.) multocida is the causative agent of pneumonic pasteurellosis in swine, which is commonly associated with the final stages of enzootic pneumonia or porcine respiratory disease complex. Although this syndrome is one of the most common and important diseases of pigs, data on antimicrobial susceptibility of P. multocida isolates are uncommon in Brazil. Therefore, the present study was carried out to determine and to compare antimicrobial susceptibility profile of Brazilian P. multocida isolated from pigs with lesions of pneumonia or pleuritis during two-time periods. Historical isolates (period of 1981 to 1997; n=44) and recent isolates (period of 2011 to 2012; n=50) were used to determine the MIC of amoxicillin, enrofloxacin, florfenicol and tetracycline by microbroth dilution. Florfenicol had the lowest level of resistance for both historical and recent isolates (0% and 6%, respectively), while tetracycline had the highest (20.5% and 34%, respectively). Multi-drug resistance (MDR) to amoxicillin/florfenicol/tetracycline was observed in 6% of recent isolates. There was a significant increase (p˂0.05) in resistance for amoxicillin and enrofloxacin in recent isolates compared with historic isolates (3.8% and 18%, respectively), most likely due to the selective pressure of antimicrobial usage to treat and prevent P. multocida infections. The results of this study showed an increase of isolates resistant to important drugs used in treatment of P. multocida infections in pigs, demonstrating the need for the implementation of rational use of antimicrobials in Brazilian swine industry.}, number={2}, journal={PESQUISA VETERINARIA BRASILEIRA}, author={Amaral, Amanda F. and Rebelatto, Raquel and Klein, Cada S. and Takeuti, Karine Ludwig and Filho, Joao X. O. and Mores, Nelson and Cardoso, Marisa R. and Barcellos, David E. S. N.}, year={2019}, month={Feb}, pages={107–111} }