In situ molecular hybridization for detection of Aleutian mink disease parvovirus DNA by using strand-specific probes: identification of target cells for viral replication in cell cultures and in mink kits with virus-induced interstitial pneumonia

Abstract

Strand-specific hybridization probes were utilized in in situ molecular hybridization specifically to localize replicative form DNA of Aleutian mink disease parvovirus (ADV). Throughout in vitro infection, duplex replicative form DNA of ADV was located in the cell nuclei. Single-stranded virion DNA and capsid proteins were present in the nuclei early in infection, but were later translocated to the cytoplasm. In neonatal mink, ADV causes acute interstitial pneumonia, and replicative forms of viral DNA were found predominantly in alveolar type II cells of the lung. Viral DNA was also found in other organs, but strand-specific probes made it possible to show that most of this DNA represented virus sequestration. In addition, glomerular immune complexes containing intact virions were detected, suggesting that ADV virions may have a role in the genesis of ADV-induced glomerulonephritis.

Violet Mink Develop an Acute Disease after Experimental Infection with Aleutian Disease Virus (ADV) Isolate ADV SL3

“…The nucleic acid hybridization data are interesting if compared to recent findings after natural (HAAS et al, 1988) and experimental infection (ALEXANDERSEN et al, 1987BLOOM et al, 1985), respectively. In both systems ADV displays a lymphotropism, with mesenteric lymph node and spleen as the major replication sites.…”

“…In situ hybridization was done with deep-frozen and paraffin-embedded tissue sections, respectively, as previously described (ALEXANDERSEN et al, 1987). Specificity control was assured by including ADV-infected as well as non-infected CCC clone 81 cells.…”

Tissue distribution, intracellular localization and proteolytic processing of rat 4‐hydroxyphenylpyruvate dioxygenase

“…The thorax cavity was opened and the animals were perfused through the left ventricle with 300 ml physiological saline containing 0.0005% (w/v) heparin followed by 450 ml 0.1% (v/v) glutaraldehyde or 4% (w/v) paraformaldehyde in phosphate buffered saline (PBS); 137 mM NaCl, 2.7 mM KCl, 4.3 mM Na 2 HPO 4 ·7H 2 O, KH 2 PO 4, pH 7.3. For in situ hybridization organs were immersion-fixed in a freshly prepared fixative containing periodate-lysine-paraformaldehydeglutaraldehyde (PLPG) for 4 h at 4 (C as described previously (Alexandersen et al, 1987). For immunohistochemical examinations fixation was carried out in 4% (w/v) paraformaldehyde for 4 h at 4 (C, and for immunoelectron microscopy fixation was performed in 0.1% (v/v) glutaraldehyde for 1 h at 4 (C.…”

Immune mechanisms in the pathogenesis of viral diseases: a review

“…Mink homozygous for the recessive Aleutian coat color gene (a) are particularly susceptible, but severity and outcome of the disease are also influenced by the virus strain. ADV replicates predominantly in B or pre-B lymphocytes of lymphatic tissues and in bone marrow cells (Roth et al, 1984;Kaaden et al, 1986Kaaden et al, ,1990Alexandersen et al, 1987;Haas et al, 1990;Wohlsein et al, 1990). Except in newborn mink, virus replication does not directly damage host cells.…”

“…Tissue injury and death of the infected animal are caused by a marked immune response to viral antigens with subsequent deposition of pathogenic IC in certain target tissues. In contrast, ADVinfected newborn mink from antibody-free mothers develop an acute and fatal interstitial pneumonia, and the type II pneumocyte in the lung is the major target cell of virus replication (Alexandersen, 1986;Alexandersen et al, 1987 ).…”