Comparative analysis of innate immune responses to viral infection among animals

Bats harbour various viruses without severe symptoms and act as natural reservoirs. This tolerance of bats toward viral infections is assumed to be originated from the uniqueness of their immune system. However, how the innate immune response varies between primates and bats remains unclear. To illuminate differences in innate immune responses among animal species, we performed a comparative single-cell RNA-sequencing analysis on peripheral blood mononuclear cells (PBMCs) from four species including Egyptian fruit bats inoculated with various infectious stimuli. Overall design: PBMCs were obtained from human (Homo sapiens), chimpanzee (Pan troglodytes), rhesus macaque (Macaca mulatta), and Egyptian fruit bat (Rousettus aegyptiacus). PBMCs were treated with herpes simplex virus type 1 (HSV-1) (strain F; GenBank accession number: GU734771), Sendai virus (SeV) (strain Cantrell, clone cCdi; GenBank accession number: AB855654) or lipopolysaccharide (LPS). On one day post infection, these infected/stimulated PBMCs (HSV-1, SeV, and LPS) were collected. Unstimulated PBMCs (Mock) were also prepared. A total of 16 samples (4 species × 4 stimuli) were loaded into the Next GEM chip G using a Chromium controller. Reverse transcription were performed in each GEM and barcoded cDNA were produced. After cDNA amplification, enzymatic fragmentation end repair, A-tailing, adaptor ligation and index PCR were processed.