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Quantification of camelid cytokine mRNA expression in PBMCs by microfluidic qPCR technology Rodon, Jordi Te, Nigger Ballester, Maria Segalés, Joaquim Serra Gironella, Joan Bensaid, Albert Producció Animal Genètica i Millora Animal Sanitat Animal 619 Camelids are economically and socially important in several parts of the world and might carry pathogens with epizootic or zoonotic potential. However, biological research in these species is limited due to lack of reagents. Here, we developed RT-qPCR assays to quantify a panel of camelid innate and adaptive immune response genes, which can be monitored in a single run. The assays were validated with PHA, PMA-ionomycin, and Poly I:Cstimulated PBMCs from alpaca, dromedary camel and llama, including normalization by multiple reference genes. Further, comparative gene expression analyses for the different camelid species were performed by a unique microfluidic qPCR assay. Compared to unstimulated controls, PHA and PMA-ionomycin stimulation elicited robust Th1 and Th2 responses in PBMCs from camelid species. Additional activation of type I and type III IFN signalling pathways was described exclusively in PHA-stimulated dromedary lymphocytes, in contrast to those from alpaca and llama. We also found that PolyI:C stimulation induced robust antiviral response genes in alpaca PBMCs. The proposed methodology should be useful for the measurement of immune responses to infection or vaccination in camelid species. This study was performed as part of the Zoonotic Anticipation and Preparedness Initiative (ZAPI project) [Innovative Medicines initiative (IMI) grant 115760] and the Veterinary Biocontained research facility Network (VetBioNet) project (EU Grant Agreement INFRA-2016-1 Nº731014), with assistance and financial support from IMI and the European Commission and contributions from EFPIA partners. J.R. was partially supported by the VetBioNet project. IRTA is supported by CERCA Programme/Generalitat de Catalunya. info:eu-repo/semantics/acceptedVersion 2023-09-17 info:eu-repo/semantics/article Rodón, Jordi, Nigeer Te, María Ballester, Joaquím Segalés, Júlia Vergara‐Alert, and Albert Bensaïd. 2023. “Quantification of Camelid Cytokine mRNA Expression in PBMCs by Microfluidic qPCR Technology.” Developmental and Comparative Immunology 149: 105061. doi:10.1016/j.dci.2023.105061. 0145-305X http://hdl.handle.net/20.500.12327/2761 https://doi.org/10.1016/j.dci.2023.105061 eng Developmental and Comparative Immunology EC/PF7/115760/EU/Zoonotic Anticipation and Preparedness Initiative/ZAPI EC/H2020/731014/EU/Veterinary Biocontained facility Network for excellence in animal infectiology research and experimentation/VetBioNet Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess 44 Elsevier