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At least 73 records · Page 4Linked to original sources
PubMed1989

Modification of foot-and-mouth disease virus after serial passages in the presence of antiviral polyclonal sera.

Foot-and-mouth disease virus (FMDV) shows a remarkable antigenic variability. Like other RNA viruses, this virus has a high rate of mutation. It has been proposed that selection exerted by the host's antibodies could play a major role in the rapid evolution of FMDV. The present work reports the selection of FMDV antibody-resistant populations (Nr), after serial passages of cloned FMDV A24 Cruzeiro strain on secondary monolayers of bovine fetal kidney cells in the presence of subneutralizing antiviral polyclonal sera (APS). After a limited number of passages under selective pressure, the virus population showed the following characteristics: (1) increased resistance to neutralization by APS; (2) altered electrophoretic mobility of structural viral proteins (VP1); (3) remarkable plaque size reduction, (4) a pronounced thermosensitivity (ts); and (5) decreased pathogenicity for mice, in both uncloned and cloned small plaque size populations. This indicates that FMDV populations under antibody pressure in vitro, have acquired, in addition to expected characteristics of natural FMDV variants (resistance to neutralization and altered viral structural proteins), phenotypic markers which correspond to attenuated, less virulent variants.

Antibodies, Viral↗
PubMed1987

Podospora anserina does not senesce when serially passaged in liquid culture.

A procedure was developed for the prolonged growth of the ascomycete fungus Podospora anserina in liquid culture to determine the effects of such growth on the senescence phenotype. Senescence in P. anserina, which is maternally inherited and associated with the excision and amplification of specific mitochondrial plasmids, occurs when this species is grown on solid medium. In two independent experiments no evidence of senescence was observed as mycelia were serially passaged in liquid culture. Further, when separable mycelial masses, termed puff balls, from the liquid cultures were plated on solid medium, a significant increase in their average longevity was observed. The apparent immortality of P. anserina in liquid culture was not dependent upon mitochondrial DNA rearrangements, nor was it affected by the presence of a previously described senescence plasmid, alpha senDNA. Evidence was obtained indicating that growth in liquid culture exerts selective pressure to maintain the wild-type mitochondrial genome.

Ascomycota↗
PubMed1990

Serial passage of infectious transcripts of a cucumber mosaic virus satellite RNA clone results in sequence heterogeneity.

Populations of the D-satellite RNA (D-sat) of cucumber mosaic virus (CMV) have a major site of heterogeneity at which a significant proportion of the molecules vary from each other. The generation of this heterogeneity was investigated by using infectious transcript RNA of a prototype D-sat clone as an inoculum of uniform sequence to initiate serial passages in five different host plant species. RNase protection assays indicated that heterogeneity at the same site seen in natural D-sat RNA populations developed within two to four passages in most of the hosts tested, and the proportion of the population exhibiting this heterogeneity accumulated to a much higher level in tobacco than in the other hosts. Characterization of the heterogeneity which developed in tobacco showed it to be qualitatively different from that in the original D-sat population. This analysis of a CMV satellite RNA is the first example in which infectious transcript RNA from a clone has been used to investigate the rapid generation and selection of heterogeneity in a genomic RNA population replicating in plants.

Base Sequence↗
PubMed1979

Tumor-associated antigens of chemically-induced murine tumors; the emergence of MuLV and fetal antigens after serial passage in culture.

Using radioiodinated Staphylococcus aureus protein A [125I]SPA to measure syngeneic, allogeneic and heterogeneic IgG bound to murine tumor cells, we performed a serological analysis of surface antigens of 8 solid tumors and 2 leukemias of BALB/c mice (3 chemically-induced colon carcinomas, 3 chemically-induced sarcomas, 1 murine leukemia virus (MuLV) induced leukemia, 1 irradiation induced leukemia, 1 spontaneous melanoma and 1 spontaneous sarcoma). We were able to detect and distinguish between at least five separate antigenic specificities on these tumors. Unique tumor-associated antigens were found on 3 of the tumors, MuLV related antigens on 8 tumors, fetal antigens on 7 tumors and two distinct common antigens on 7 tumors (common antigen 1 (CA-1) on 5 tumors and common antigen 2 (CA-2) on 2 tumors). Neither of the common antigens was found to be sarcoma, carcinoma or tissue-tupe specific. A number of tumors which did not originally express either MuLV or fetal antigens in primary cultures expressed these antigens after several serial passages in vitro.

Animals↗
PubMed1995

Encapsidation and serial passage of a poliovirus replicon which expresses an inactive 2A proteinase.

The multiple roles of the viral proteinase 2A in poliovirus replication have been difficult to assess because, to date, it has not been possible to isolate and characterize a viral genome with an inactive 2Apro. We have previously reported that a poliovirus replicon containing an inactive 2Apro by virtue of a change at amino acid 109 from a cysteine to a serine (C109S) was replication competent when transfected into cells previously infected with vaccinia virus (R. Pal-Ghosh and C. D. Morrow, J. Virol. 67:4621-4629, 1993). To further develop this system, we have used a poliovirus replicon which contains the human immunodeficiency virus type 1 (HIV-1) gag gene positioned between nucleotides 1174 and 2470 of the poliovirus genome and have engineered a second mutation within this replicon to change the codon for amino acid 109 of the 2Apro from cysteine to serine (2AC109S). Transfection of this replicon into cells previously infected with vaccinia virus results in the replication and expression of a protein with a molecular mass consistent with that of a P1-HIV-1 Gag-2A fusion protein. Using a recently described complementation system which relies on the capacity of a recombinant vaccinia virus (VV-P1) to provide the capsid precursor (P1) in trans (D. C. Ansardi, D. C. Porter, and C. D. Morrow, J. Virol. 67:3684-3690, 1993; and D. C. Porter, D. C. Ansardi, W. S. Choi, and C. D. Morrow, J. Virol. 67:3712-3719, 1993), we have encapsidated this replicon containing the 2AC109S mutation. By using reverse transcription PCR, we demonstrated that after 15 serial passages the encapsidated replicon still contained the 2AC109S mutation. Infection of cells with a stock of encapsidated replicon, either in the presence or in the absence of vaccinia virus, resulted in the expression of the P1-HIV-1 Gag-2A fusion protein. Expression of the P1-HIV-1 Gag fusion protein in cells infected with the encapsidated replicon containing the 2AC109S mutation was reduced compared with the expression of P1-HIV-1 Gag in those cells infected with a replicon containing a wild type 2A gene. The protein expression and replication of the replicon RNA in cells containing the 2AC109S mutation was maintained for a longer period of time than for the replicons containing the wild-type 2A gene, possibly because of a reduced cytopathic effect.(ABSTRACT TRUNCATED AT 400 WORDS)

Base Sequence↗
PubMed1984

Selection of attenuated dengue 4 viruses by serial passage in primary kidney cells. I. Attributes of uncloned virus at different passage levels.

Attempts were made to attenuate prototype dengue (DEN) 4 (H-241) virus. The original viremic human serum was passed once in a susceptible monkey and twice in Aedes albopictus mosquitoes and then serially passed in primary dog kidney (PDK) and African green monkey kidney (GMK) cells. Weekly transfers of undiluted virus were carried to the 50th passage in both primary cell cultures. Biological markers were studied at passages 7, 15, 30 and 50. Parental DEN-4 phenotype characteristics included large plaque formation in LLC-MK2 cells, plaque formation in GMK cells, cytopathic effect in LLC-MK2 cells, growth in human monocyte cultures, growth at 39 degrees C, consistent production of viremia in monkeys and short-incubation neurovirulence in mice. At the seventh passage in both PDK and GMK cell cultures, DEN-4 viruses exhibited reduced plaque-size in LLC-MK2, and failed to plaque in GMK, to produce cytopathic effect in LLC-MK2, or to grow in human monocytes. Serial passage in PDK, as opposed to GMK, resulted in a graduated loss of monkey virulence. Rhesus monkeys inoculated with the PDK 50 strain failed to develop detectable viremia and only 1 of 4 developed an antibody response. Also, replication of PDK 50 was completely shut-off at 39 degrees C. The graduated change in biological properties noted, particularly those in PDK cells, provide a range of potential vaccine candidates for evaluation in human beings.

Aedes↗
PubMed1989

[The effect of serial passage of Coxiella burnetii in chick embryos on pathogen phase variability].

Coxiella burnetii has been investigated in different egg passages (EP-17, 163). Coxiella in EP 163 was completely eliminated from the suspension during the purification but not that in EP 17. It means that the coxiella of the two EP with a seroreaction of phase II are different, and phase II positivity of EP 17 can't be the result of presence of two populations in the suspension or of one mutation during the multiplication in yolk sacs but of a change on the surface of C. burnetii.

Animals↗
PubMed1984

Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract.

A serum-free medium with bovine pituitary extract as the only undefined supplement has been developed for long-term culture of human mammary epithelial cells. This medium supports serial subculture of normal cells for 10-20 passages (1:10 splits) without conditioning or special substrates, and it supports rapid clonal growth with plating efficiencies up to 35%. It consists of an optimized basal nutrient medium, MCDB 170, supplemented with insulin, hydrocortisone, epidermal growth factor, ethanolamine, phosphoethanolamine, and bovine pituitary extract. Replacement of pituitary extract with prostaglandin E1 and ovine prolactin yields a defined medium that supports rapid clonal growth and serial subculture for three or four passages. Cultures initiated in these media from normal reduction mammoplasty tissue remain diploid and maintain normal epithelial morphology, distribution of cell-associated fibronectin, expression of keratin fibrils, and a low level of expression of milk fat globule antigen. Large cell populations can now be generated and stored frozen, permitting multiple experiments over a period of time with cells from a single donor. These media greatly extend the range of experiments that can be performed both conveniently and reproducibly with cultured normal and tumor-derived human mammary epithelial cells.

Animals↗
PubMed1977

Establishment of an Epstein-Barr virus-negative B-cell lymphoma line from a Japanese Burkitt's lymphoma and its serial passage in hamsters.

An Epstein-Barr virus (EBV)-negative lymphoma line (JBL) was established in vitro from pleural effusion of an EBV-seropositive 29-year-old Japanese female with Burkitt's lymphoma. JBL cells as well as her original lymphoma cells bore monoclonal surface IgM with lambda light chains. The JBL line grew in single cell suspension with a doubling time of 30 hours. Attempts were made to serially transplant JBL cells in antilymphocyte serum-treated newborn hamsters; intraperitoneal implantation of 1-3 X 10(7) cells gave rise to invasive tumors in all recipients with death after 10 to 14 days. The hamster-passage line, now in the 9th passage, has been converted to an ascitic form with progression to leukemia in some animals. A "starry sky" pattern closely resembling the human tumor material was preserved in every tumor through serial animal passage.

Animals↗
PubMed1980

Effects of serial passage on the endocrine response and steroid metabolism of a rat mammary carcinoma.

A rat mammary carcinoma induced by 7-12 dimethylbenzanthracene was serially transplanted into successive generations of thymectomized host animals. After its 2nd passage, the growth of the tumour appeared hormone-dependent, regressing after oophorectomy and regrowing with administration of oestradiol 17 beta to the host. Third-generation transplanted tumours, however, showed only a transient regression after oophorectomy, and the growth of tumours after further passages appeared ovary-independent. Loss in hormone dependency was not accompanied by histological changes. There was however a progressive increase with successive transplantation in the ability of tumours to metabolize 7 alpha [3H] testosterone in vitro. This was accounted for by raised conversion to 5 alpha androstanediol.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1988

Chemically induced murine T lymphomas: continued rearrangement within the T-cell receptor beta-chain gene during serial passage.

The first constant region of the Tcrb gene was completely deleted from the DNA of 8/10 mouse cell lines established from 3-methylcholanthrene-induced RF/J thymic lymphomas, but 6/7 primary lymphomas contained the first constant region sequences. DNA from RF/J thymic lymphomas induced by N-methyl-N-nitrosourea was then examined serially as the tumors were passaged in vivo and adapted to growth in culture as uncloned and, in some cases, cloned lines. Patterns of Tcrb-specific restriction fragments from most tumors changed extensively during continued propagation. Analysis of the patterns often suggested that initial DNA rearrangements within the Tcrb complexes of monoclonal tumors had been followed by further rearrangements within the same genes. However, these different patterns may alternatively have represented successive outgrowth of separate lineages from lymphomas that were polyclonal in origin.

Animals↗
PubMed1978

Serial passages of larval Echinococcus granulosus from equine origin in mice. I. Infection with protoscolices.

Starting with protoscolices from Echinococcus granulosus cysts of equine origin it was possible to isolate and maintain several "strains" of the parasite in mice through successive transfers of protoscolices. The most advanced strain is now in its 7th generation. Several criteria to evaluate the results are presented and discussed. It is concluded that serial echinococcosis of E. granulosus through passages with protoscolices is a practical method to maintain different strains for fundamental and applied comparative studies.

Animals↗
PubMedDate not supplied

Increased response to cisplatin after long-term serial passage of a squamous cell carcinoma xenograft.

We have retrospectively investigated the response to cisplatin of a squamous cell carcinoma of the head and neck xenografted to nude mice during nine years of serial transplantation. Tumour growth rate decreased gradually. After nine years and over 100 passages, there was a sudden increase in cisplatin sensitivity. Histopathological examination showed that, of two histopathologically different subpopulations present in earlier passages, the predominant one was no longer detectable. The DNA-index did not change.

Animals↗
PubMed1981

Temperature-sensitive alteration in fusion activity of subacute sclerosing panencephalitis virus during serial passages in vitro and expression of hemagglutinin on the infected cells.

The Biken strain of subacute sclerosing panencephalitis (SSPE) virus, a maturation-defective variant of measles virus, was serially passed in human embryonic lung (HEL) cells at 37 C. The strain formed syncytial giant cells (GC) at both 37 C and 39 C, but the surface of infected cells did not show hemadsorption at early passages of the strain. However, GC-forming activity of the strain diminished at 39 C after 25 passages or more and hemadsorption on the infected cells became positive at both 37 C and 39 C after 40 passages or more of the strain. Hardly any infectious cell-free virus was detected in the culture fluid even after hemadsorption became positive. Possible mechanisms for the defect of SSPE virus were discussed.

Cell Fusion↗