Breeding Saguinus and Callithrix species of marmosets under laboratory conditions.
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Biomedical subjects
Publications and source records attributed to L G Wolfe.
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Simian sarcoma virus, type 1 (SSV-1)-transformed non-producer cell lines were established by infection of normal marmoset fibroblast cells (HF) with limiting dilutions of SSV-1. Four focus-derived cell lines were identified as non-producers by assay of culture fluids for focus-forming activity and by the mixed culture cytopathogenicity test with XC cells. Further studies failed to detect production of type-C virus by 3H-uridine labelling, reverse transcriptase assay or electron microscopy. The non-producer cell lines, designated HF/SSV-NPI, IV, V, and VI, appeared morphologically transformed and cloned in soft agar with the same efficiency as HF/SSV-1 virus-producing transformed cells. Expression of either cytoplasmic or cell surface virus-related antigens was not detected by immunofluorescence or serum cytotoxicity tests. The presence of sarcoma genome in the transformed non-producer cell lines was demonstrated by rescue of focus-forming activity following superinfection with non-transforming helper virus or by cocultivation with helper-virus-producing cell lines. The SSV-1-transformed non-producer primate cells provide a useful tool for future studies.
Type C virus produced by dog thymus cells (A7573) that were infected with virus (HL-23V), isolated from cultured leukocytes of an acute myelogenous leukemia patient, transformed marmoset and horse cells in vitro and induced virus-producing fibromas in marmosets. The tumors and transformed foci were indistinguishable morphologically from those induced by simian sarcoma virus, type 1 (SSV-1/SSAV-1). HL-23V was indistinguishable from SSV-1/SSAV-1 by immunofluorescence and neutralization tests, and the nontransforming virus associated with HL-23V completely inhibited SSV-1 focus induction in interference tests. Cell cultures established from a marmoset fibroma produced transforming and nontransforming virus biologically and antigenically indistinguishable from HL-23V and SSV-1/SSAV-1.
Adult common marmosets, inoculated with either of 2 oncogenic Herpesvirus saimiri (HVS) strains, developed fatal lymphoproliferative disease within 23-25 days post inoculation (PI). The disease was identical to HVS-induced lymphoma in cotton-topped and white-lipped marmosets. Common marmosets inoculated with an attenuated HVS strain developed persistent infection; virus has been recovered from cocultivated lymphocytes of these animals for more than 384 days PI.
Three lymphoblastoid cell lines were established from splenic lymphocytes of a lymphomatous baboon (Papio hamadryas) by co-cultivation of the lymphocytes with X-irradiated cells of marmoset or baboon lymphoblastoid cell cultures; the baboon splenic lymphocytes failed to grow when cultured alone. A herpesvirus, associated with each cell line, was identified by immunofluorescence, molecular hybridization and electron microscopy. Antigenic comparison with Epstein-Barr virus (EBV) showed that the baboon herpesvirus and EBV shared cross-reacting viral capsid antigens (VCA): 20 of 20 (100%) anti-VCA (EBV)-positive human sera and 55 of 62 (89%) baboon sera reacted with the baboon lymphoblastoid cells and baboon sera stained EBV VCA in P3HR-1 and EB-3 cells. No nuclear antigen, as assayed by anti-complement immunofluorescence tests, was detected in baboon lymphoblastoid cells when human or baboon anti-VCA positive sera were used. Baboon anti-VCA-positive sera also failed to stain EBV nuclear antigens (EBNA) in Raji or P3HR-1 cells. Preliminary molecular hybridization studies showed only approximately 40% homology between viral DNA of baboon cell lines and DNA of EBV derived from P3HR-1 cells.
Two strains of feline fibrosarcoma virus (ST-FeSV and GA-FeSV) were found to induce tumors in cats and marmosets, and to transform feline and marmoset cells in vitro after primary inoculation. A third strain (SM-FeSV) failed to induce tumors or transform marmoset cells after primary inoculation; however, when SM-FeSV-injected marmoset cultures were passed 26 times in vitro, the cell cultures released infectious virus which transformed marmoset fibroblasts but still failed to induce tumors in marmosets. ST-FeSV induced mainly round-cell type transformation (r foci), GA-FeSV induced predominantly mixed round-fusiform cell type transformation (fr foci), and SM-FeSV induced r and fr type foci with a higher proportion of fusiform cells in the fr foci than seen with GA-FeSV. Transforming virus was obtained from r or mixed r/fr foci of ST-FeSV but not from fr foci; heat treatment changed the virus from producing almost exclusively r type foci to inducing an increased number of fr foci. Passage of FeSV in cat cells yielded viruses with a higher ratio of infectivity for feline vs marmoset cells, while passage of FeSV in marmoset cells yielded virus with a relatively higher infectivity ratio for marmoset cells; the three strains differed in the degree of change in the infectivity ratio. Despite the alteration of host range of SM-FeSV propagated in marmoset fibroblasts, the virus retained feline P-30 antigen by CF and FA assays. Neutralization tests did not indicate but also did not exclude an alteration of the surface antigens of ST-FeSV or SM-FeSV propagated in marmoset fibroblasts. The alteration of the relative infectivity of FeSV during passage in marmoset cells may be due to: (1) the selection of a variant present in the original heterogenous uncloned population; (2) mutation; or (3) recombination with some marmoset genetic material, possibly an as yet unidentified endogenous marmoset virus.
The Subhuman Primate Pregnancy Test kit was used for diagnosis of pregnancy in 17 marmosets. This hemagglutination inhibition test for urinary chorionic gonadotropin accurately indicated conception by positive responses in 72 and 76 specimens collected between the 4th and 10th wk following the estimated time of fertilization. The rate of false positive results did not exceed 1%. A radioimmunoassay system was used to quantify chorionic gonadotropin concentrations in urine. Detectable levels were present by the 2nd week of pregnancy. Peak chorionic gonadotropin levels were found between the 7th to 9th wk after conception. Chorionic gonadotropin excretion rates declined to undetectable levels during the final month of gestation.
Two adult and two neonatal cotton-topped marmosets and two neonatal white-lipped marmosets (Saguinus species) were inoculated with 10(7) plaque-forming units of cytomegalovirus (Colburn strain). No overt clinical disease developed in four marmosets during observation for eight months; one adult and one neonatal cotton-topped marmoset died from nonspecific causes 63 and 259 days after inoculation, respectively. By days 7-16 after inoculation, all marmosets developed plasma antibodies, which were detectable by neutralization and immunofluorescence assays (peak titers, 1:128-1:256 and 1:64-1:256, respectively). Attempts to isolate virus from whole blood, peripheral lymphocytes, oropharyngeal swabs, or vaginal swabs by cocultivation with permissive cell cultures were unsuccessful. Virus was recovered, however, by cocultivation from the kidney tissues of the adult marmoset that died. Immunosuppressive treatment with azathioprine resulted in a fourfold increase in antibody levels in plasma of two of three marmosets.
The response of cotton-topped (CT) or white-lipped (WL) marmosets, inoculated with material containing Epstein-Barr virus (EBV), was studied. Twenty-five marmosets inoculated with 10(6)-10(8) cells from human or simian EBV-carrying lymphoblastoid cell cultures developed no detectable clinical disease. Transitory antibody titres to VCA were detected in sera from several of these marmosets. Thirty-eight marmosets inoculated with P3 virus (10(9) particles) and seven marmosets inoculated with 10(5) infectious units of HR-1 virus likewise developed no recognizable clinical or haematological disease over several years of observation. Fourteen marmosets were inoculated with 10(2)-10(4) transforming units (TFU) of EBV strain B95-8; five of 11 animals inoculated with 10(3)-10(4) TFU of virus developed malignant lymphoma and died 31-110 days PI. Antibodies to one or more EBV-specified antigens were detected in sera from nine of the inoculated marmosets. EBV-carrying lymphoblastoid cell cultures were established from the spleen and lymph-nodes of one animal that died with malignant lymphoma and from another animal that died 19 days PI from undetermined causes.
Reproductive performances of wild-caught and laboratory-born marmosets maintained in a paired-cage system were summarized. Wild-caught, white-lipped (WL) marmosets (Saguinus fuscicollis, S nigricollis) averaged 2.6 live births per pair per yr compared with 1.6 for cotton-topped (CT) marmosets (S oedipus); S fuscicollis illigeri pairs were the best producers. Of first and second generation laboratory-born animals, crossbred or hybrid WL pairs were the best producers; first generation pairs averaged 2.7 live births per pair per yr, comparable to the production of wild-caught WL pairs. In contrast, laboratory-born Sf illigeri pairs produced only 0.8 live births per pair per yr. No significant differences in reproductive performance were observed between first generation outbred vs inbred and between parent-reared vs hand-reared pairs. Based on limited data, the reproductive capabilities of wild-caught and laboratory-born Callithrix jacchus appeared superior to both WL and CT marmosets. Ten examples of marmoset models in biomedical research were discussed briefly.
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Explore the source record for details and available documents.
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Explore the source record for details and available documents.
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