Research
Dominance of highly divergent feline leukemia virus A progeny variants in a cat with recurrent viremia and fatal lymphoma
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* Corresponding author: A Katrin Helfer-Hungerbuehler khungerbuehler@vetclinics.uzh.ch
1 Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland
2 Clinic for Small Animal Internal Medicine, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland
3 Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland
4 Institute of Veterinary Pathology, University of Giessen, Giessen, Germany
5 Institute of Biomedical Engineering, University of Zurich and ETH, Zurich, Switzerland
Retrovirology 2010, 7:14 doi:10.1186/1742-4690-7-14
Published: 19 February 2010Additional files
Additional file 1:
Multicentric lymphoma. Lymphoma detected upon necropsy. A) Kidney. B) Spleen. C) Histology of sternal lymph node: diffuse proliferation of mainly medium sized lymphatic cells with round nuclei, coarsely stippled chromatin and one to multiple medium sized nucleoli. Hematoxylin and Eosin, bar = 10 μm. D) Liver: positive immunohistochemical labeling of periportal infiltrating neoplastic cells for CD45R. Avidin-biotin complex method; Papanicolaou's hematoxylin counterstain, bar = 50 μm.
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Additional file 2:
Amino acid alignment of FeLV env sequences. Amino acid alignment of the env coding region from three FeLV subtypes (FeLV-A/Glasgow-I [GenBank: M12500], FeLV-B/Gardner-Arnstein [GenBank: K01209] and FeLV-C/Sarma [GenBank: M14331]) and the three env variants (KI261-I, KI261-II and SP261-III). The start of the SU region, the transmembrane domain (TM), the variable regions VRA, VRB and VRC, the PRR and the C2 disulfide-bonded loop (S-S) are labeled (according to [52]). Circles and stars represent amino acid sequences containing Asn-X-Ser/Thr, which indicate possible sites of N-glycosylation, as previously described [53,83]. Potential N-linked glycosylation sites that are conserved in FeLV-A/Glasgow-1 and all env variants are represented by filled circles. New potential N-linked glycosylation sites in the env variants that were not present in the challenge strain FeLV-A/Glasgow-1 are labeled with filled stars; those that were present in FeLV-A/Glasgow-1 but lost in the env variants are marked with empty stars. Dots represent identical residues, and dashes represent spaces, which were introduced for proper alignment.
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Additional file 3:
Comparison of nucleotide sequences of the FeLV U3 region. Nucleotide sequence comparison of the U3 region from FeLV-A/Glasgow-1 [GenBank: M12500] and the progeny virus variants retrieved from cat #261. Sequences found in multiple clones were depicted once. KI261-I was found in clones from the kidney. SP261-III, pKH11.8 and pKH11.9 were found in clones from the spleen. All other listed sequences were found in at least two different tissues, including the kidney, liver, spleen, and bone marrow. Enhancer elements and their corresponding nucleotide sequences are marked in the reference strain FeLV-A/Glasgow-1 [GenBank: M12500]. Nucleotides differing from the originally inoculated strain are indicated. Primer sequences for PCR [41] were located at positions -415 to -392 and 9 to 32 (not included in the figure). Previously described mutations are indicated by numbers in brackets: (1) Jackson et al., 1996; (2) Nishigaki et al., 1997; (3) Fulton et al., 1990 and (4) Matsumoto et al., 1992. Numbers at the top of the sequence indicate nucleotide positions relative to the presumptive RNA cap site in the FeLV/Glasgow-1 LTR [53]. Grey dashed lines indicate unknown sequences.
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Additional file 4:
FeLV-A/Glasgow-1 and env variant provirus and viral loads in the tissues from cat #261. A) Provirus loads of FeLV-A/Glasgow-1 and the env variants. B) Viral (cDNA) loads of FeLV-A/Glasgow-1 and env variants. Viral tissue loads were normalized to GAPDH (top) and to RPS7 cDNA copy numbers (bottom). Tissues with apparent lymphoma are indicated by shaded areas.
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Additional file 5:
FeLV viral loads in tissues from cat #261 normalized to RPS7. A) Total FeLV viral (cDNA) loads (U3 region PCR) in tissues with and without apparent lymphoma (analogous to Fig. 3C). B) Viral (cDNA) loads of FeLV-A/Glasgow-1 and env variants (analogous to Fig. 5C). C) Viral (cDNA) loads of env variants in tissues with and without apparent lymphoma (analogous to Fig. 5D). D) Viral (cDNA) loads of FeLV-A/Glasgow-1 in tissues with and without apparent lymphoma. Viral loads were normalized to RPS7 cDNA copy numbers determined by TaqMan real-time PCR, as described [27]. Viral loads were tested for statistically significant differences using the Mann-Whitney U-test (pMWU as indicated).
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