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1.
Radioimmunoassay for Avian C-Type Virus Group-Specific Antigen: Detection in Normal and Virus-Transformed Cells 总被引:14,自引:10,他引:4
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A radioimmunoassay has been developed for detection of avian C-type virus (30,000 mol wt) group-specific (gs) antigen. The method is 10- to 1,000-fold more sensitive than immunological methods previously available. By the radioimmunoassay technique, normal chicken embryo cells, which have previously been classified as gs negative or weakly gs positive, contain clearly detectable amounts of gs antigen. The assay has been used to study the effect of chemical induction and superinfection by mammalian C-type viruses on the expression of avian gs antigen in mammalian cells nonproductively transformed by avian sarcoma viruses. 相似文献
2.
Avian Oncornavirus Group-Specific Antigen: Detection and Quantification by Radioimmunoassay 总被引:5,自引:2,他引:3
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A double-antibody competitive-inhibition radioimmunoassay for avian group-specific antigen is described. A viral protein preparation, consisting primarily of gs-1, was labeled with radioiodine. Hamster and rabbit antisera were reacted with the labeled antigen, and the resultant antibody-antigen complexes were precipitated with the appropriate antiglobulin. Standard curves based on the inhibition of binding of labeled antigen to antibody after preincubation with unlabeled antigen were prepared. These showed the lower limit of sensitivity to be 2 to 10 ng of unlabeled protein when labeled antigen of 2,000 to 5,000 counts per min per ng of specific activity was used in the assay. Extracts of avian oncornavirus-infected cells, likewise, were able to inhibit binding of labeled antigen. This technique will be very useful for the quantification of small amounts of oncornavirus group-specific protein. 相似文献
3.
The XC rat cell line was found to support the replication of a strain of the Moloney murine sarcoma-leukemia virus. In growth curve experiments cytopathology was paralleled by the production of murine sarcoma virus and leukemia virus progeny having the biologic, antigenic, and biophysical properties of the infecting virus. 相似文献
4.
Characterization of Intracellular Ribonucleic Acid Specific for the Murine Sarcoma-Leukemia Virus Complex 总被引:9,自引:5,他引:4
Nilambar Biswal Michael B. Grizzard Robert M. McCombs Matilda Benyesh-Melnick 《Journal of virology》1968,2(11):1346-1352
Virus particles were continuously produced by a cell line (78A1) of rat embryo fibroblasts that had been transformed by the murine sarcoma-leukemia virus complex. Since most of the mature virions were found in the extracellular fluid and were not cell-associated, a measurable quantity of viral ribonucleic acid (RNA) could not be extracted from these cells. Cycloheximide, a protein inhibitor, was successfully used to accumulate viral RNA within the cells. This ribonuclease-sensitive RNA, with a sedimentation coefficient of 71S, had the same base composition as the high molecular weight RNA (S(20,w) = 71) isolated from purified virions released by the transformed cells. 相似文献
5.
Immunological Studies on Viral Polypeptide Synthesis in Cells Replicating Murine Sarcoma-Leukemia Virus 总被引:13,自引:8,他引:5
G. Shanmugam Giancarlo Vecchio Domenica Attardi Maurice Green 《Journal of virology》1972,10(3):447-455
Antibodies to disrupted murine sarcoma-leukemia virus (MSV[MLV]) were used to study the synthesis of viral polypeptides in the transformed, virus-producing rat cell line 78A1. When cultures were labeled for 10 min with radioactive amino acids, about 9% of the total labeled proteins were precipitated with antiserum against purified MSV(MLV), and 3 to 4% were precipitated with the same antiserum after it had been absorbed with an extract from uninfected rat cells. The difference is due to the presence in the unabsorbed antiserum of antibodies to cellular proteins that are present in purified virus preparations. Intracellular viral proteins labeled with radioactive amino acids were isolated by immunoprecipitation and analyzed by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. The mobilities of intracellular viral polypeptides were identical to those of the purified virion. However, labeled polypeptides having electrophoretic mobilities lower than that of the major virion polypeptide, the group-specific antigen of molecular weight 31,000, were present in higher proportion in the total cell extract and in the membrane fraction than in the virion. These polypeptides appear to be of cellular origin for they were present only in minute amounts in the immunoprecipitates obtained with the absorbed serum. After a 10-min labeling period, radioactive proteins were assembled into extracellular virions rapidly for the first 4 hr followed by a slower rate. More than 2% of the total proteins of the cell labeled in a 10-min pulse were assembled into virions at the completion of a 24-hr chase. The high-molecular-weight polypeptides with the same mobilities as those detected in the immunoprecipitate of intracellular proteins were found in virions released from cells after a 10-min pulse. A larger proportion of these high-molecular-weight proteins was detected in virions released after short chase periods (30-120 min) than after longer chase periods (6-24 hr). Two possible interpretations of these data are that the high-molecular-weight cell-derived polypeptides (i) have a turnover rate higher than that of the major virion polypeptides or (ii) are cleaved proteolytically from the virions during long incubation in the culture media. 相似文献
6.
Focus Formation by a Murine Sarcoma-Leukemia Virus Complex: I. Theoretical Analysis 总被引:1,自引:1,他引:0
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The transformation of cells in culture by a murine sarcoma-leukemia virus complex leads to the formation of identifiable foci. Titration patterns of focus counts vs. virus dilutions have been explained as being due to the interactions of three virions: leukemia, defective sarcoma, and competent hybrid. This interaction is analyzed quantitatively and equations are derived which permit determination of the composition of a sarcoma virus pseudotype as well as use of the technique for the assay of exogeneous leukemia virus. The analysis is based on the statistical distribution of virions in the culture cells and assumes that all virions have equal probability of infecting a cell and that transformation of any one cell will lead to focus formation. Limitations of the assumptions and expected measurement accuracy are discussed. 相似文献
7.
8.
Robert H. Purcell Doris C. Wong Harvey J. Alter Paul V. Holland 《Applied microbiology》1973,26(4):478-484
A micro-solid-phase radioimmunoassay (micro-SPRIA) for hepatitis B antigen (HB Ag) was developed for use with microtiter serological equipment. Radiolabeled immunoglobulin G was prepared from human and animal sera containing hepatitis B antibody (HB Ab); it was not necessary to isolate specific HB Ab by immunochemical means. A micro-SPRIA prepared with guinea pig reagents was approximately as sensitive as the AusRIA radioimmunoassay, but, like the AusRIA test, yielded false positive results. A micro-SPRIA prepared with human reagents was slightly less sensitive but did not yield false positive results. These micro-SPRIA tests offer several advantages, including conservation of reagents, adaptability to other antigen-antibody systems, ease of performance (especially when testing large numbers of specimens), and economy. 相似文献
9.
Detection and Assay of Avian Tumor Virus Group-Specific Antigen and Antibody by the Paired Radioiodine-Labeled Antibody Technique 总被引:2,自引:1,他引:1
The paired radioiodine-labeled antibody technique (PRILAT) was applied to the detection and quantitation of avian tumor virus group-specific (gs) antigens and antibody. The technique proved to be specific, repeatable, and appreciably more sensitive than the microcomplement-fixation test for avian leukosis (COFAL). The PRILAT facilitated direct measurement of comparative antigen content of several types of transformed, neoplastic, or virus-infected cells and the magnitude of nonspecific antibody binding by appropriate control cells. The versatility of the technique was illustrated by application to the detection and quantitation of gs antibody content of chicken, turkey, pigeon, and hamster sera. Antibodies were detected in COFAL-negative sera from hamsters bearing tumors induced by the Schmidt-Ruppin strain of Rous sarcoma virus. Sera from chickens bearing similar tumors were not positive for gs antibodies, although sera from turkeys and chickens immunized with avian leukosis virus did contain gs antibodies. 相似文献
10.
Focus Formation by a Murine Sarcoma-Leukemia Virus Complex: II. Quantitative Aspects of the Interaction Between Radiation Leukemia Virus and Its Murine Sarcoma Virus Pseudotype in Strain C57BL Mouse Embryo Cells 总被引:6,自引:3,他引:3
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A quantitative study has been made of the interactions between radiation leukemia virus (RadLV), its murine sarcoma virus pseudotype, and their C57BL host cells. The elimination of interference phenomena by delayed infection of cells with RadLV made possible the quantitative determination of the pseudotype in terms of defective sarcoma and endogenous RadLV particles. This in turn permitted the quantitative assessment of RadLV helper activity and of the various factors which influence the accuracy and sensitivity of the helper assay. 相似文献
11.
Infectivity and RNA Patterns as Functions of High- and Low-Dilution Passage of Murine Sarcoma-Leukemia Virus: Evidence for Autointerference Within an Oncornavirus Population 总被引:6,自引:6,他引:0
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Heterogeneity of buoyant density and RNA content of virions of Moloney murine leukemia-sarcoma complex [MSV (MLV)] was the result of passage at low dilution. Heterogeneous stocks revealed two major RNA components in the population, with the smaller component, apparent mol wt 4 x 10(6) to 5 x 10(6), becoming predominant upon serial passage at low dilution. Concomitantly, infectivity titers of both MLV and MSV decreased upon serial passage at low dilution. MSV (MLV) passaged at high dilution retained high titers and a rather homogeneous high-molecular-weight RNA population characteristic of high-buoyant-density virions. Interference of both MLV and MSV replication was demonstrated by employing mixed inocula containing both low- and high-dilution passage stocks of MSV (MLV). In contrast to results with MSV (MLV), MLV freed of MSV by limit dilution did not show heterogeneity of buoyant density or of RNA when propagated at low dilution. 相似文献
12.
DNA Polymerase of Murine Sarcoma-Leukemia Virus: Lack of Detectable RNase H and Low Activity With Viral RNA and Natural DNA Templates 总被引:14,自引:12,他引:2
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Kirsten murine sarcoma-leukemia virus (Ki-MSV[MLV]) was found to contain less RNase H per unit of viral DNA polymerase than avian Rous sarcoma virus (RSV). Upon purification by chromatography on Sephadex G-200 and subsequent glycerol gradient sedimentation the avian DNA polymerase was obtained in association with a constant amount of RNase H. By contrast, equally purified DNA polymerase of Ki-MSV(MLV) and Moloney [Mo-MSV(MLV)] lacked detectable RNase H if assayed with two homopolymer and phage fd DNA-RNA hybrids as substrates. On the basis of picomoles of nucleotides turned over, the ratio of RNase H to purified avian DNA polymerase was 1:20 and that of RNase H to purified murine DNA polymerase ranged between <1:2,800 and 5,000. Based on the same activity with poly (A).oligo(dT) the activity of the murine DNA polymerase was 6 to 60 times lower than that of the avian enzyme with denatured salmon DNA template or with avian or murine viral RNA templates assayed under various conditions (native, heat-dissociated, with or without oligo(dT) and oligo(dC) and at different template enzyme ratios). The template activities of Ki-MSV(MLV) RNA and RSV RNA were enhanced uniformly by oligo(dT) but oligo(dC) was much less efficient in enhancing the activity of MSV(MLV) RNA than that of RSV RNA. It was concluded that the purified DNA polymerase of Ki-MSV(MLV) differs from that of Rous sarcoma virus in its lack of detectable RNase H and in its low capacity to transcribe viral RNA and denatured salmon DNA. Some aspects of these results are discussed. 相似文献
13.
Gilbert R. Irwin Alfred M. Allen William H. Bancroft Milton Willhight Philip K. Russell 《Applied microbiology》1974,28(4):600-604
Sera from a survey of 6,026 people were tested for hepatitis B surface antigen by using radioimmunoassay and counterelectrophoresis. Forty-eight sera (0.79%) were positive by counterelectrophoresis and 152 sera (2.52%) were positive by radioimmunoassay, using the most liberal of the recommended criteria for positivity (i.e., counts 3 standard deviations above the mean). Absorption tests performed on the 152 radioimmunoassay-positive sera showed that 10 (6.6%) were false-positive reactions to guinea pig protein, 74 (48.6%) were due to false-positive reaction(s) with other protein(s) in the test system, and 68 (44.8%) were true positives. There was a strong correlation between the degree of elevation of radioactive counts and the proportions of sera that were true positives; all 49 sera with counts >50 standard deviation units above the mean were true positives, but only 19 (18.4%) of the 103 sera with counts <50 standard deviation units were true positives. A few sera with high counts required absorption with type-specific (type D) antisera. The following conclusions were reached from this study: (i) absorption tests should be run on all radioimmunoassay-positive, counterelectrophoresis-negative sera; (ii) most (about 90%) false positives are not due to anti-guinea-pig protein reactions; and (iii) radioimmunoassay, in combination with absorption tests, yields a modest increase (about 35%) in detection of true positives over use of counterelectrophoresis alone. 相似文献
14.
THE possibility of an oncogenic virus in human breast cancer has been increased by recent findings. Virus-like particles resembling the mammary tumour virus (MTV) were found in electron micrographs of human breast cancer tissue1 and particles physically and morphologically similar to MTV particles have been seen in human milk2. These particles were found more frequently in the milk of American women with a history of breast cancer in their immediate families and in the milk of Parsi women in Bombay than in the milk of nonselected American women3. Parsi women are three times more likely to have breast cancer than other women in Bombay3. The detection of RNA-dependent DNA-polymer-ase activity in such particles isolated from human milk emphasized the possibility that these particles represent an oncogenic RNA virus4. In addition to the physical and morphological resemblance of human milk particles and MTV an immunological relationship between these two kinds of particles seems probable; sera from breast cancer patients neutralize the biological activity of MTV whereas normal human sera did not do so5. We report data supporting the hypothesis of an immunological cross relationship between MTV and human breast cancer. 相似文献
15.
Rabies Group-Specific Ribonucleoprotein Antigen and a Test System for Grouping and Typing of Rhabdoviruses 总被引:13,自引:9,他引:13
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L. G. Schneider B. Dietzschold R. E. Dierks W. Matthaeus P.-J. Enzmann K. Strohmaier 《Journal of virology》1973,11(5):748-755
Cell-associated ribonucleoprotein (RNP) was isolated from BHK-21 cells infected with several strains of rabies and rabies-related viruses. The RNP-antigen from rabies and related viruses induced the formation of complement-fixing, precipitating, and immunofluorescent antibodies, and proved to be the group-specific antigen common to all rabies viruses. Antigens of the envelope which induce virus-neutralizing antibodies are apparently determinative for the serotype of a virus as evidenced by two-way neutralization tests. A combination of these methods seems to be a useful approach to the serological grouping and typing of rhabdoviruses. 相似文献
16.
Cell Surface Antigen Induced by Friend Murine Leukemia Virus Is also in the Virion 总被引:8,自引:3,他引:5
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Intact particles of Friend leukemia virus derived from infectious mouse serum absorb only trace amounts of cytotoxic anti-FMR antibodies, but physical disruption of the virions by freezing and thawing, by ether extraction or by detergent treatment releases large amounts of FMR antigenic activity. Thus this antigen, previously considered to occur mainly as a neo-antigen on the surfaces of virus-infected cells and as a soluble substance in the serum of infected mice, may be primarily a virion component. 相似文献
17.
Immunoelectrophoretic Analysis of Avian Ribonucleic Acid Tumor Virus Group-Specific Antigens 总被引:4,自引:4,他引:0
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Tumors induced in pigeons by inoculation with the Schmidt-Ruppin strain of Rous sarcoma virus regressed after about 6 weeks. Sera from these pigeons, taken 8 weeks after inoculation, had complement-fixing group-specific antibody titers of 1:2 to 1:256. In immunoelectrophoresis with the pigeon serum, disrupted BAI strain A (myeloblastosis) avian tumor virus showed at least five precipitin arcs. The pattern of precipitin lines was dependent in part on the means used for virus disruption, and ethyl ether and nonionic detergents appeared to be both effective and relatively mild reagents. Immunoelectrophoretic comparison of pigeon serum with serum from a tumor-bearing hamster and that from virus-inoculated rabbits yielded similar, though not identical, results. 相似文献
18.
Origin and Structure of the Group-Specific, Complement-Fixing Antigen of Rickettsia rickettsii 总被引:9,自引:0,他引:9
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Rickettsia rickettsii was treated with ether and examined by negative-contrast electron microscopy. Group-specific complement-fixing antigen was seen to be originating from the cell wall. The antigen was composed predominately of round particles 10 to 60 nm in diameter. Intact R. rickettsii and antigen from ether-treated organisms were purified by density gradient centrifugation and analyzed by polyacrylamide gel electrophoresis. The whole rickettsial cell was composed of a minimum of 30 proteins which ranged in molecular weight from about 23,000 to 155,000. The "soluble" antigen contained nine proteins ranging in molecular weight from about 28,000 to 150,000. 相似文献
19.
Miriam N?rder Pablo D. Becker Ingo Drexler Claudia Link Volker Erfle Carlos A. Guzmán 《PloS one》2010,5(6)
Background
Modified vaccinia virus Ankara (MVA), a highly attenuated strain of vaccinia virus, has been used as vaccine delivery vector in preclinical and clinical studies against infectious diseases and malignancies. Here, we investigated whether an MVA which does not encode any antigen (Ag) could be exploited as adjuvant per se.Methodology/Principal Findings
We showed that dendritic cells infected in vitro with non-recombinant (nr) MVA expressed maturation and activation markers and were able to efficiently present exogenously pulsed Ag to T cells. In contrast to the dominant T helper (Th) 1 biased responses elicited against Ags produced by recombinant MVA vectors, the use of nrMVA as adjuvant for the co-administered soluble Ags resulted in a long lasting mixed Th1/Th2 responses.Conclusions/Significance
These findings open new ways to potentiate and modulate the immune responses to vaccine Ags depending on whether they are co-administered with MVA or encoded by recombinant viruses. 相似文献20.
JOHNG S. RHIM C. R. LENGEL H. Y. CHO K. K. TAKEMOTO H. C. TURNER R. J. HUEBNER R. V. GILDEN 《Nature: New biology》1972,235(58):188-190
WE reported accelerated transformation by DNA viruses (SV40 and polyoma) in rat embryo (RE) cells chronically infected with a C-type RNA virus1,2. Recently we found in RE cells transformed by polyoma virus a new complement-fixing (CF) antigen detectable by rat antisera having broad reactivity with the various intraspecies and interspecies antigens of the RNA tumour viruses3–8; this antigen, however, was distinct from the murine intraspecies and interspecies group-specific (gs) antigens both immunologically and by virtue of other properties. It is also distinct from the polyoma virion (capsid) and tumour (“T”) antigens. 相似文献