Quantification of camelid cytokine mRNA expression in PBMCs by microfluidic qPCR technology

• Published in: Developmental and comparative immunology Vol. 149; p. 105061
• Main Authors: Rodon, Jordi, Te, Nigeer, Ballester, Maria, Segalés, Joaquim, Vergara-Alert, Júlia, Bensaid, Albert
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01.12.2023
• Subjects: Camelid species; Cytokines; Fluidigm biomark; Immune responses; PBMCs; RT-qPCR; Immune responses; Cytokines; PBMCs; RT-qPCR; Camelid species; Fluidigm biomark
• ISSN: 0145-305X; 1879-0089
• DOI: 10.1016/j.dci.2023.105061

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:C-stimulated 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. •Development of reagents and assays to monitor immune responses in camelid species.•Assays were validated using stimulated cells from alpaca, dromedary camel and llama.•A unique microfluidic qPCR allows interspecies comparative gene expression analyses.•PHA and PMA-ionomycin provoke robust Th1 and Th2 responses in camelid cells.•PolyI:C enhances the transcription of antiviral response genes in alpaca cells.