In vitro studies on the infection and replication of porcine circovirus type 2 in cells of the porcine immune system.
Porcine circovirus type 2 (PCV2) nucleic acid and/or antigens are consistently observed in cells of monocytic morphology in lesions of pigs affected by post-weaning multisystemic wasting syndrome (PMWS). In this study, PCV2 antigen was detected in the cytoplasm of monocytes, pulmonary macrophages (PMs) and monocyte-derived macrophages exposed to the virus in vitro, by immunofluorescence analysis (IFA) and the phenotype of these cells confirmed by detection of monocytic cell surface markers using flow cytometry. Viral antigen was not observed in lymphocytic cells. Replication of the virus in PMs was investigated further by comparison to that observed in the continuous pig kidney cell line (PK15A) using quantitative virus titration, quantitative PCR and by the detection of double stranded DNA intermediates of viral replication by Southern blotting analyses. Although increases in viral DNA and levels of infectious virus progeny and the presence of replicative intermediates, indicative of viral replication, were observed in PK15A cells, no such changes were observed in PMs in spite of the fact that infectious virus, viral antigen and viral DNA persisted in the cells for at least the duration of the experiment. These results suggest that in vivo, monocytic cells may not represent the primary target for PCV2 replication.
Gilpin
D F
DF
Department of Veterinary Science, Queens University Belfast, Stoney Road, Stormont, Belfast, BT4 3SD, UK.
McCullough
K
K
Meehan
B M
BM
McNeilly
F
F
McNair
I
I
Stevenson
L S
LS
Foster
J C
JC
Ellis
J A
JA
Krakowka
S
S
Adair
B M
BM
Allan
G M
GM
D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D015139:Blotting, Southern / Q000662:veterinary; D002455:Cell Division / Q000276:immunology; D018173:Circoviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D005434:Flow Cytometry / Q000662:veterinary; D019085:Fluorescent Antibody Technique, Direct / Q000662:veterinary; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000821:virology*; D016676:Macrophages, Alveolar / Q000166:cytology / Q000276:immunology / Q000821:virology*; D016133:Polymerase Chain Reaction; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000821:virology*; D014779:Virus Replication; D019282:Wasting Syndrome / Q000276:immunology / Q000662:veterinary* / Q000821:virology
D000956:Antigens, Viral; D004279:DNA, Viral