Establishment of a Cell Line from the Hematophagous Bat Desmodus Rotundus Susceptible to Lyssavirus Rabies

Desmodus rotundus bats are the main reservoir for Lyssavirus rabies (RABV) in the Americas, however, knowledge of virus-host interactions in this species is very limited due to challenges associated with establishing in vivo experimental infections. In this context, cell culture becomes a valuable tool for expanding knowledge on the dynamics of RABV infection in its natural host in the Americas. This study aimed to develop and characterize a cell line from D. rotundus bat and to evaluate its susceptibility to RABV. Primary cultures of D. rotundus fetal kidney cells were immortalized using the plasmid pSV1 containing the Large T and Small T antigen gene sequences of Simian Virus 40 (SV40). The obtained clones were selected by limiting dilution and transfection was confirmed by PCR for Large T SV40 gene. The resulting cell line was named FKDR (Fetal Kidney Desmodus Rotundus). The growth curve, senescence assay, and karyotype analysis of the primary and FKDR cell cultures were performed. The susceptibility of FKDR cells to RABV was determined through direct fluorescent antibody test (dFAT), and compared with that of BHK-21, Tb1Lu, and CaroPe cells. Once immortalized, the cell adopted a fusiform appearance and showed the absence of senescence markers, chromosomal anomalies, and accelerated cell growth, compared to the primary fetal cell. The FKDR and the other cell lines used for RABV infections exhibited positive staining by immunofluorescence but no cytopathic effect. The cell line described here holds potential for studies of RABV infections in D. rotundus bats.