In situ PCR-associated immunohistochemistry identifies cell types harbouring the Maedi-Visna virus genome in tissue sections of sheep infected naturally.
Maedi-Visna virus (MVV) is a non-oncogenic ovine lentivirus whose main targets are the lung, mammary gland, central nervous system and joints. Cells of the monocyte-macrophage lineage are the major viral target in vivo; other cell types are infected as well, as indicated by several studies, largely based on the examination of animals infected experimentally or on the in vitro infection of cultured cells. Aim of this study was to investigate the cell types harbouring the viral genome in lungs and mammary glands of animals infected naturally by using in situ PCR-associated immunohistochemistry. Several types of cells were infected: in the lung type I and II pneumocytes, interstitial and alveolar macrophages, endothelial cells and fibroblast-like cells. Epithelial cells, macrophages, endothelial cells and fibroblast-like cells were infected also in the mammary gland. These results indicate that the in situ PCR, a powerful technique which combines the high sensitivity of the conventional PCR with the ability to localise the cellular targets within a tissue, can be improved further by its association with the immunohistochemistry. This can be especially advantageous when the presence and localisation of the target sequence are investigated in the context of a tissue with its complex cellular organisation.
Carrozza
Maria Luisa
ML
Scuola Normale Superiore, Pisa, Italy.
Mazzei
Maurizio
M
Bandecchi
Patrizia
P
Arispici
Mario
M
Tolari
Francesco
F
D000818:Animals; D007150:Immunohistochemistry; D008168:Lung / Q000166:cytology* / Q000821:virology*; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000821:virology*; D011021:Pneumonia, Progressive Interstitial, of Sheep / Q000821:virology*; D016133:Polymerase Chain Reaction; D012756:Sheep; D012757:Sheep Diseases / Q000821:virology; D014790:Visna-maedi virus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism