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At least 19 recordsLinked to original sources

Experimental anterior uveitis after subcutaneous injection of feline sarcoma virus.

Feline sarcoma virus (FeSV) is a naturally occurring virus that causes spontaneous tumors in cats. The immunologic and morphologic characteristics of these tumors have been studied extensively. It was recently observed in experiments undertaken to induce systemic malignancy with this virus, that severe uveitis and clinical blindness occurred. An investigation of the ophthalmologic changes was undertaken. A fulminent anterior uveitis was produced in cats by a series of subcutaneous injections of live FeLV-FeSV. This intraocular inflammation occurred in five of six animals using high viral titers, and four of seven with lower titers, resulting from the freeze thaw process. On histopathologic examination, most animals demonstrated dysplastic changes of the ciliary body in addition to the iridocyclitis. The remainder of the eye was unaffected. These animals developed systemic tumors unaccompanied by local inflammation, many of which spontaneously regressed. Notable features of this potential model for uveitis are that (1) direct injection into the eye is unnecessary, and (2) intravenous administration inducing immune tolerance with antigenic overload presented to the spleen is avoided. This inflammatory reaction seems to be specific to the iris and ciliary body. Levels of live virus detected in the aqueous humor exceeded those in the serum. These results suggest that the virus may be actively secreted by the ciliary epithelium, or may preferentially proliferate within the eye.

Animals↗

The use of immunohistochemistry and the polymerase chain reaction for detection of feline leukemia virus and feline sarcoma virus in six cases of feline ocular sarcoma.

Ocular sarcoma was diagnosed by light microscopic examination in enucleated globes (n = 4), orbital tissue biopsy (n = 1) and ocular evisceration contents (n = 1) from six cats. To determine if feline leukemia virus (FeLV) or a replication-defective FeLV, feline sarcoma virus (FeSV), was present in these ocular sarcomas, immunohistochemistry (IHC) and polymerase chain reaction (PCR) for FeLV were utilized. Immunohistochemical staining for FeLV glycoprotein 70 (gp70) was performed on all six formalin-fixed, paraffin-embedded tumors using an avidin-biotin complex technique. DNA was extracted from each specimen and a 166 bp region of the FeLV long-terminal repeat (LTR) was amplified by PCR. All tumors were composed primarily of spindle cells; two neoplasms had PAS-positive basement membrane enveloping areas of spindle cells. All tumors involved the uvea and five of six tumors showed transcleral extension, one of which invaded the optic nerve. Immunohistochemical staining for FeLV gp 70 was negative. PCR to amplify a portion of the FeLV LTR was negative. Based on these findings of these limited number of cases, FeLV/FeSV may not play a role in the tumorigenesis of feline ocular sarcomas. However, additional tumors representing all morphological subtypes should be investigated for the presence of viral antigen and DNA. It is important to determine the etiology and pathogenesis of these malignant ocular sarcomas. If the cell of origin and pathogenesis involve ocular and lenticular injury, and FeLV/FeSV is not present, then the clinical management of cases of feline ocular trauma, uveitis and glaucoma may prevent the development of this tumor.

Journal Article↗

Susceptibility of human cell lines to feline leukemia virus and feline sarcoma virus.

The susceptibility of human fibroblasts and human lymphoid cell lines to feline leukemia virus (FeLV) and feline sarcoma virus (FeSV) was investigated. Human cells were highly sensitive to infection by FeLV subgroups B and C and FeSV but resistant to infection by FeLV subgroup A. The cells became infected, produced infectious virus, and displayed no differences in morphology or viability. T-cell lines appeared to be more sensitive to FeLV and FeSV infection and to produce more virus than did autochthonous B-cells. B-cell lines with membrane IgG appeared more resistant to infection than did those with no membrane IgG.

B-Lymphocytes↗

Lack of detection of feline leukemia and feline sarcoma viruses in diffuse iris melanomas of cats by immunohistochemistry and polymerase chain reaction.

Diffuse iris melanoma was confirmed by light-microscopic examination in 10 formalin-fixed, paraffin-embedded globes from 10 cats. To determine if feline leukemia virus or a replication defective feline leukemia virus, feline sarcoma virus, was present in these anterior uveal melanomas, immunohistochemistry and polymerase chain reaction for feline leukemia virus were utilized. Immunohistochemical staining for feline leukemia virus glycoprotein 70 was performed on all 10 tumors using an avidin-biotin complex technique. The DNA was extracted from each specimen and a 166-base pair region of the feline leukemia virus long terminal repeat was targeted by polymerase chain reaction. Immunohistochemical staining for feline leukemia virus glycoprotein 70 and polymerase chain reaction amplification of a feline leukemia virus long terminal repeat region were negative in all cases. Feline leukemia virus/feline sarcoma virus was not detected in any neoplasms and therefore was unlikely to play a role in the tumorigenesis of these feline diffuse iris melanomas.

Animals↗

In vitro transformation of murine pre-B lymphoid cells by Snyder-Theilen feline sarcoma virus.

Snyder-Theilen feline sarcoma virus (ST-FeSV) codes for a protein kinase with specificity for tyrosine residues (Barbacid et al., Proc. Natl. Acad. Sci. U.S.A. 77:5158-5163, 1980), properties analogous to those of the transforming gene product of Abelson murine leukemia virus (Witte et al., Nature (London) 283:826-831, 1980). In the present report, ST-FeSV was demonstrated to transform murine hematopoietic cells under in vitro assay conditions which detect lymphoid cell transformation by Abelson murine leukemia virus. Bone marrow colony formation was shown to require ST-FeSV, follow single-hit kinetics, and require the presence of mercaptoethanol in the agar medium. ST-FeSV-induced colonies could be established in culture as continuous cell lines that demonstrated unrestricted self-renewal capacity and leukemogenicity in vivo. The hematopoietic blast cells transformed by ST-FeSV in culture appeared to be at an early stage of B cell differentiation. They possessed Lyb 2 surface antigens, were dependent on mercaptoethanol for growth, and contained only low levels of terminal deoxynucleotidyl transferase. Moreover, a large fraction of the lines synthesized immunoglobulin mu chain in the absence of light chains. Thus, the phenotype of ST-FeSV hematopoietic transformants was indistinguishable from that of the pre-B lymphoblast transformants induced by Abelson murine leukemia virus. These findings indicate that the in vitro functional similarities in the onc gene products of ST-FeSV and Abelson murine leukemia virus may reflect a common pathway by which they exert their oncogenic potential.

Animals↗

Structure and origins of the HZ2-feline sarcoma virus.

The HZ2-feline sarcoma virus (HZ2-FeSV) is a replication-defective acute transforming feline retrovirus with oncogene homology to Abelson murine leukemia virus (A-MuLV) (P. Besmer, W.D. Hardy,Jr., E. E. Zuckerman, P. J. Bergold, L. Lederman, and H. W. Snyder, Jr. (1983) Nature (London) 303, 825-828). In contrast to A-MuLV which was isolated from a hematopoietic tumor, the HZ2-FeSV derives from a multicentric fibrosarcoma. We have molecularly cloned the HZ2-FeSV provirus from mink HZ2-FeSV nonproducer cells. The molecularly cloned HZ2-FeSV provirus is biologically active upon transfection of NIH 3T3 indicator cells. The genetic structure of the HZ2-FeSV provirus was determined by EM heteroduplex and Southern blot analysis. The HZ2-FeSV has a 6.8 kb-viral genome with the structure: 5' delta gag-abl-delta pol-delta env 3'. The abl insert, which is 1.4 kb, is located 1.9 kb from the 5' end and 3.5 kb from the 3' end of the viral genome. The 5' 1.9 kb in the HZ2-FeSV are colinear with 5' FeLV sequences, and the 3' 3.5 kb are colinear with 3' FeLV sequences, with the exception of a 0.85-kb deletion in the env gene. HZ2-FeSV v-abl and A-MuLV v-abl share 1.2 kb of abl sequences which are known to specify the protein kinase domain of the abl gene product and are necessary for fibroblast transformation in vitro. The DNA from several tumor tissues of cat 3590 from which the HZ2-FeSV was obtained was found to contain several HZ2-FeSV-related proviruses including the HZ2-FeSV. The variant HZ2-FeSVs have indistinguishable 5' gag-abl sequences; however, they differ in 3' sequences which likely do not include any abl sequences. The DNAs from fibrosarcomas obtained by inoculation of kittens with tumor extract were found to contain variant HZ2-FeSV proviruses as well. Taken together these results indicate a role for the HZ2-FeSVs in sarcomagenesis.

Abelson murine leukemia virus↗

Feline uveal melanoma model induced with feline sarcoma virus.

This paper described the first animal model of a virally induced uveal melanoma. Tumors developed following the injection of an RNA tumor virus, i.e., Gardner strain feline sarcoma virus, into the anterior chamber of newborn kittens. Histologically, the tumors were found to be iris and ciliary body melanomas, many of which showed invasion. The histology and ultrastructure of those tumors are described.

Animals↗

Preparation of rat monoclonal antibodies to epitopes encoded by the viral oncogene (v-fms) of McDonough feline sarcoma virus.

The McDonough strain of feline sarcoma virus (SM-FeSV) contains a viral oncogene, v-fms, transduced from cat cellular genetic sequences designated c-fms. Monoclonal antibodies reactive to antigenic determinants encoded by v-fms were prepared by immunizing rats with live, syngeneic SM-FeSV-transformed cells, and fusing splenic lymphocytes from a tumor-bearing animal with cultured rat myeloma cells. Culture supernatants from hybrids producing antibodies to epitopes encoded by v-fms were identified by immunoprecipitation of radiolabeled polypeptides from SM-FeSV-transformed mink cells. Four positive hybrids were cloned twice in soft agar, established as stable lines, and grown in defined serum-free medium to facilitate purification of homogeneous antibodies. The monoclonal antibodies were used to assay SM-FeSV-specific products by "immunoblotting" of electrophoretically separated proteins, and by fixed-cell immunofluorescence.

Animals↗

Feline leukemia virus-and feline sarcoma virus-related polypeptides released by virus producer and nonproducer cells.

Polypeptides specific for feline leukemia virus (FeLV) have been identified in the media of cells that produce FeLV as well as in nonproducer cells transformed by feline sarcoma viruses (FeSV). Cat fibroblasts that were persistently infected with FELV release the major virus envelope glycoprotein, whereas cultured cat lymphoma cells shed both glycopeptides related to the virus core gene (gag) and glycopeptides related to the virus envelope gene (env). Mink cells and cat cells transformed by FeSV secrete polypeptides of a wide range of sizes that cross-react with the major virus core protein p27. Differences in the classes of p27-related proteins produced may be related to the strain of virus and the cell type. Cat cells transformed by FeSV release a glycopeptide that appears to be processed differently from those identified in the media of FeSV-transformed mink cells. The possibility that such FeLV-related secretory proteins may interfere with the immune response of the host is discussed.

Animals↗

McDonough feline sarcoma virus: characterization of the molecularly cloned provirus and its feline oncogene (v-fms).

The genetic structure of the McDonough strain of feline sarcoma virus (SM-FeSV) was deduced by analysis of molecularly cloned, transforming proviral DNA. The 8.2-kilobase pair SM-FeSV provirus is longer than those of other feline sarcoma viruses and contains a transforming gene (v-fms) flanked by sequences derived from feline leukemia virus. The order of genes with respect to viral RNA is 5'-gag-fms-env-3', in which the entire feline leukemia virus env gene and an almost complete gag sequence are represented. Transfection of NIH/3T3 cells with cloned SM-FeSV proviral DNA induced foci of morphologically transformed cells which expressed SM-FeSV gene products and contained rescuable sarcoma viral genomes. Cells transformed by viral infection or after transfection with cloned proviral DNA expressed the polyprotein (P170gag-fms) characteristic of the SM-FeSV strain. Two proteolytic cleavage products (P120fms and pp55gag) were also found in immunoprecipitates from metabolically labeled, transformed cells. An additional polypeptide, detected at comparatively low levels in SM-FeSV transformants, was indistinguishable in size and antigenicity from the envelope precursor (gPr85env) of feline leukemia virus. The complexity of the v-fms gene (3.1 +/- 0.3 kilobase pairs) is approximately twofold greater than the viral oncogene sequences (v-fes) of Snyder-Theilen and Gardner-Arnstein FeSV. By heteroduplex, restriction enzyme, and nucleic acid hybridization analyses, v-fms and v-fes sequences showed no detectable homology to one another. Radiolabeled DNA fragments representing portions of the two viral oncogenes hybridized to different EcoRI and HindIII fragments of normal cat cellular DNA. Cellular sequences related to v-fms (designated c-fms) were much more complex than c-fes and were distributed segmentally over more than 40 kilobase pairs in cat DNA. Comparative structural studies of the molecularly cloned proviruses of Synder-Theilen, Gardner-Arnstein, and SM-FeSV showed that a region of the feline-leukemia virus genome derived from the pol-env junction is represented adjacent to v-onc sequences in each FeSV strain and may have provided sequences preferred for recombination with cellular genes.

Animals↗

Biochemical characterization of cells transformed via transfection by feline sarcoma virus proviral DNA.

Murine fibroblasts transformed by transfection with DNA from mink cells infected with the Snyder-Theilen strain of feline sarcoma virus and subgroup B feline leukemia virus were analyzed for the presence of integrated proviral DNA and the expression of feline leukemia virus- and feline sarcoma virus-specific proteins. The transformed murine cells harbored at least one intact feline sarcoma virus provirus, but did not contain feline leukemia virus provirus. The transformed murine cells expressed an 85,000-dalton protein that was precipitated by antisera directed against feline leukemia virus p12, p15, and p30 proteins. No feline oncornavirus-associated cell membrane antigen reactivity was detected on the surfaces of the transformed murine cells by indirect membrane immunofluorescence techniques. The 85,000-dalton feline sarcoma virus-specific protein was also found in feline cells transformed by transfection. However, these cells also contained env gene products. The results of this study demonstrate that the feline sarcoma virus genome is sufficient to transform murine cells and that expression of the 85,000-dalton gag-x protein is associated with transformation of both murine and feline cells transformed by transfection.

Animals↗

Cytotoxic immune response of puppies to feline sarcoma virus induced tumors.

The cytotoxic immune response of puppies to feline sarcoma virus induced tumors was studied. Neonatal puppies were compared with adolescent dogs. Three different types of cytotoxicity were investigated: complement dependent cytotoxicity, T-cell-mediated cytotoxicity and antibody dependent cellular cytotoxicity. The relationship between the spontaneous regression of the sarcoma and the development of the immune system of the puppies is discussed.

Animals↗

Monoclonal antibodies to the v-fes product and to feline leukemia: virus P27 interspecies-specific determinants encoded by feline sarcoma viruses.

Monoclonal antibodies to p27 gag and v-fes specific determinants on the gag-onc poly-protein encoded by Snyder-Theilen feline sarcoma virus (ST-FeSV) were prepared. In order to obtain hybridoma clones specific to the antigenic determinants encoded by the FeSV genome, Lou rats were immunized with ST-FeSV-transformed, virus-nonproducing syngeneic cells, and boosted with either the same cells or affinity-purified feline leukemia virus (FeLV) p27. Three distinct clones reactive to both FeLV p27 and p85gag-fes, and one clone specific for a p85fes determinant were established. The anti-p27 monoclonal antibodies also reacted with the polyproteins p95gag-fes and p83gag-fgr, from Gardner-Arnstein (GA) and Theilen-Pedersen (TP1) FeSV, respectively. The anti-p27 monoclonal antibodies reacted with MuLV p30 and RD114 p28 but not with RSV, MMTV, or BLV. These results indicated that the part of the p27 gag gene that is preserved in ST-, GA, and TP1-FeSV encodes interspecies-specific p27 determinants.

Animals↗

Differential requirements of gag and gamma-actin domains for transforming potential of Gardner-Rasheed feline sarcoma virus.

The oncogene of Gardner-Rasheed feline sarcoma virus (GR-FeSV) encodes the 70-kilodalton protein containing gag(p15), gamma-actin, and fgr domains. To determine the role of these domains in the biological activity of P70gag-actin-fgr, we have constructed in-frame deletion and insertion mutants of GR-FeSV. We found, first, that the gamma-actin region could be deleted without affecting the transforming ability of these constructs, although an insertion mutant in the middle of the gamma-actin domain (map position 671) was partially defective in transformation and specifically had a reduced level of in vitro autophosphorylation activity. Second, mutations affecting the C-terminal third of the gag region appeared to abolish the ability to transform NIH 3T3 cells and autophosphorylation activity. These results suggest that the gamma-actin domain is not essentially required for the transforming activity of GR-FeSV but that it may take part in maintaining the conformational integrity of P70gag-actin-fgr and that the gag(p15) domain might have a critical role in modulating the function of P70gag-actin-fgr.

Actins↗

Chromosomal assignment of the human homologues of feline sarcoma virus and avian myeloblastosis virus onc genes.

Retroviral transforming genes, v-onc genes, are derived from normal cellular sequences that are called cellular onc (c-onc) genes. DNA from mouse-human somatic cell hybrids that have selectively lost human chromosomes was used in Southern blots to map the chromosomal location of two human onc genes. Cloned human homologues of retroviral onc genes were used as probes. Because the human c-fes gene, which is homologous to feline sarcoma virus, segregates concordantly with human chromosome 15, and the human c-myb gene, which is homologous to avian myeloblastosis virus onc genes, segregates concordantly with human chromosome 6, we have assigned the c-fes and the c-myb genes to human chromosomes 15 and 6, respectively. Nonrandom chromosomal defects involving these human chromosomes have been observed in neoplasms. These studies should be valuable in determining whether specific rearrangements involving these chromosomes result in the abnormal expression of these onc genes in human malignancies.

Avian Leukosis Virus↗

Structural analysis of a variant clone of Snyder-Theilen feline sarcoma virus.

A variant clone of Snyder-Theilen feline sarcoma virus (ST-FeSV) encoding a polyprotein with a molecular weight of approximately 104 kDa (P104) was compared to the P85 encoding prototype clone of ST-FeSV. Analysis of chimeric genes constructed with the viral oncogenes of the two clones indicated that the variant clone coded for a larger polyprotein than the prototype clone because of genetic differences in its 3' portion. Comparative DNA sequence analysis revealed that one nucleotide just upstream of the termination condon TGA in the prototype proviral DNA was deleted from the variant clone resulting in a 468-bp larger open reading frame. Furthermore, it appeared that the U3 regions of the long terminal repeats (LTRs) of the variant clone contained an insertion of 71 bp as compared to the LTRs of the prototype clone. In addition, both clones differed also from each other with respect to genetic sequences deleted from their env gene regions.

Base Sequence↗