Interactions of poult enteritis and mortality syndrome-associated reovirus with various cell types in vitro

Abstract

An avian reovirus, ARV-CU98, has recently been isolated from poults experiencing poult enteritis and mortality syndrome (PEMS). To further understand ARV-CU98 and its role in PEMS, the current study investigates interactions of ARV-CU98 with various cell types in vitro. When macrophages, B cells, T cells, and liver cells of chicken or turkey origin were co-incubated with ARV-CU98, only cells of liver origin demonstrated cytopathic effects, the presence of viral antigen, and reduced metabolic activity over time. Furthermore, distinctive pockets of viral particles were evident in electron microscopic examination of a chicken hepatocellular carcinoma (LMH) cell line, but not in a chicken macrophage cell line (MQ-NCSU) co-incubated with virus. Additional evidence of viral replication in LMH, cells but not MQ-NCSU cells was demonstrated by the presence of two viral bands (43 and 145 kD size) in cell lysates from LMH cells exposed to ARV-CU98. Although not capable of being infected by ARV-CU98, MQ-NCSU cells do appear to be activated by the virus since IL-1 mRNA expression is increased in MQ-NCSU cells 2 h after addition of the virus. LMH cells exposed to the virus demonstrate a decrease in IL-1 mRNA expression by 8 to 10 h after addition of the virus, perhaps corresponding to the initiation of infection by the virus. In conclusion, this study demonstrates that ARV-CU98 actively infects and replicates in LMH cells, but not in lymphocytes or macrophages, suggesting that the liver may be a target and site of replication of ARV-CU98 in poults experiencing PEMS.

The pathogenesis of turkey origin reoviruses in turkeys and chickens

“…Reovirus infections in turkeys, often in flocks experiencing enteritis, have been reported previously (Simmons et al ., 1972;Reynolds et al ., 1987;Lozano et al ., 1989;Heggen-Peay et al ., 2002). However, the role of reovirus in disease induction has not been clearly established as the virus can be found in specimens from healthy flocks and it is not uncommon to detect other enteric viruses in the same specimens (Dees et al ., 1972).…”

“…While most reports focus on disease in chickens, clinical disease in turkeys has been reported to be similar to what is observed in chickens, and includes viral arthritis (Page et al ., 1982) and poult enteritis (Dees et al ., 1972;Simmons et al ., 1972;Gershowitz & Wooley, 1973;Reynolds et al ., 1987;Lozano et al ., 1989;Heggen-Peay et al ., 2002). Historically, it has been assumed that all avian reoviruses from chickens and turkeys are closely related.…”

Characterization of interleukin-1β mRNA expression in chicken macrophages in response to avian reovirus

“…Moreover, enhanced IL-1b activity was detected in culture supernatant of macrophages from poults suffering from poult enteritis and mortality syndrome (PEMS) caused by a reovirus infection (Heggen et al, 2000). Increases of IL1b mRNA expression level have also been detected in a chicken macrophage cell line treated with the PEMSassociated reovirus (Heggen-Peay et al, 2002). These results suggest that IL-1b is an important factor in controlling the pathogenesis of many diseases.…”

Genomic characterization of a turkey reovirus field strain by Next-Generation Sequencing

“…Virus isolation and propagation were conducted in LMH (Leghorn Male-chicken Hepatocellular-carcinoma, ATCC CRL-2113) cell cultures as routine cell culture procedures (Heggen-Peay et al, 2002). Viral RNA extraction from the TARV infected cell culture fluid was carried out with a total RNA extraction Kit (QIAGEN, Valencia, CA, USA) following manufacturer's instructions.…”