Abstract
Feline coronaviruses (FCoVs) infect both wild and domestic cat populations world-wide. FCoVs present as two main biotypes: the mild feline enteric coronavirus (FECV) and the fatal feline infectious peritonitis virus (FIPV). FIPV develops through mutations from FECV during a persistence infection. So far, the molecular mechanism of FECV-persistence and contributing factors for FIPV development may not be studied, since field FECV isolates do not grow in available cell culture models. In this work, we aimed at establishing feline ileum and colon organoids that allow the propagation of field FECVs. We have determined the best methods to isolate, culture and passage feline ileum and colon organoids. Importantly, we have demonstrated using GFP-expressing recombinant field FECV that colon organoids are able to support infection of FECV, which were unable to infect traditional feline cell culture models. These organoids in combination with recombinant FECVs can now open the door to unravel the molecular mechanisms by which FECV can persist in the gut for a longer period of time and how transition to FIPV is achieved.
Highlights
The diverse family of the Coronaviridae causes infections in a wide range of mammals, birds and humans
Feline coronaviruses (FCoVs) occur in two different biotypes: feline enteric coronavirus (FECV) and feline infectious peritonitis virus (FIPV)
Tissue-derived organoids rely on the ability to isolate stem cell containing crypts. These crypts are grown in the presence of differentiation factors (Wnt3a, R-Spondin, Noggin and EGF), allowing them to grow into three-dimensional mini-gut organoids [33]. As these complex cultures more closely resemble the multi-cell types found in their natural tissue counterparts, they often contain factors, which are required for the replication and propagation of viruses that are missing in standard cell cultures
Summary
The diverse family of the Coronaviridae causes infections in a wide range of mammals, birds and humans. These crypts are grown in the presence of differentiation factors (Wnt3a, R-Spondin, Noggin and EGF), allowing them to grow into three-dimensional mini-gut organoids [33] As these complex cultures more closely resemble the multi-cell types found in their natural tissue counterparts, they often contain factors, which are required for the replication and propagation of viruses that are missing in standard cell cultures. To determine if these model systems could be used to support the relevant serotype I FECV growth, we established a cat intestinal organoid culture system and show that it is capable of supporting infection with GFP-expressing recombinant serotype I FECV generated by reverse genetics. This model will open the doors to study the molecular mechanism of serotype I FECV-persistence in its natural enteric environment
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