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Biomedical subjects

D H Moore

Publications and source records attributed to D H Moore.

At least 181 records · Page 10Linked to original sources

Biologic characteristics of some mouse mammary tumor viruses.

The mammary tumor virus (MuMTV) in the milks of 7 mouse strains and substrains was titrated for infectivity in 4 strains. The data indicated that: 1) Each strain shed a different MuMTV and some genetic strains carried two MuMTV's, each discernible by its mouse strain preference in infectivity tests. 2) Less than 5% of RIIIf and about 10% of Af mice shed detectable MuMTV antigen in their milks after the third parturition. After the sixth parturition, 33% of RIIIf and 50% of Af, and after the ninth parturition, 60% of RIIIf and 90% of Af mice shed viral antigen in their milks. The MuMTV's in milks of high-parity mothers were most infectious in mouse strains different from those most susceptible to MuMTV in RIII and A milks of low-parity mothers. Therefore, RIII and A mice each harbored two viruses, one that was removed by foster-nursing and the other that was not. 3) The susceptibility incidence of RIIIfC57BL mice to RIII virus changed gradually from about 10% in 1970 to about 70% in 1975. Susceptibility of C57BL mice to RIII virus did not change appreciably over this period, and the natural tumor incidence in RIIIfC57BL remained unchanged (about 10%). In addition to their susceptibility to RIII virus, C57BL mice were also susceptible to GR virus; they were relatively resistant to other strains tested. They were especially resistant to RIIIf virus, to which Af and BALB/c mice were very susceptible. 4) Approximately 90% of C3HfC57BL and C3HfBALB/c mice shed antigen in their milks after the third parturition, although the tumor incidence was less and occurred later than in C3H mice. No clear-cut differences could be detected in infectivities between low-parity C3H milk and high-parity C3Hf milks tested in several assay strains.

Adenocarcinoma↗

Murine mammary tumor virus: characterization of infection of nonmurine cells.

Murine mammary tumor virus (MuMTV) was used to productively infect feline and mink cells. MuMTV "proviral" DNA could be detected in the infected cells by molecular hybridization using radioactive MuMTV complementary DNA as a probe. Kinetic analysis of MuMTV proviral DNA synthesis after infection showed that maximum MuMTV DNA synthesis was achieved by 8 h; however, this was followed by a decline in detectable proviral DNA and eventual stabilization at a lower level. MuMTV synthesis in feline cells was greatly stimulated by the synthetic glucocorticoid, dexamehtasone. On the other hand, MuMTV synthesis in mink cells was relatively at a much higher level in absence of dexamethasone and the stimulation with dexamethasone was not as marked as in the case with infected feline cells. Thermal denaturation of hybrids between MuMTV complementary DNA and infected mink cell RNA revealed no difference from homologous hybrids.

Animals↗

Purification of the chromosomes of the Indian muntjac by flow sorting.

Metaphase chromosomes were isolated from a male Indian muntjac cell line, were stained with ethidium bromide and were analyzed by flow microfluorometry to establish a deoxyribonucleic acid (DNA)-based karyotype. Five major peaks were evident on the chromosomal DNA distribution corresponding to the five chromosome types in this species. The amount of DNA in each chromosome was confirmed by cytophotometric measurements of intact metaphase spreads. The five chromosome types were separated by flow sorting at rates up to several hundred chromosomes per second. The sorted chromosomes were identified by morphology and by Giemsa banding patterns. The automsomes, Numbers 1, 2 and 3, and the X + 3 composite chromosome were separated with a high degree of purity (90%). The centromere region of the X + 3 chromosome was fragile to mechanical shearing, and during isolation a small proportion of these chromosomes broke into four segiments: the long arm, the short arm, the short arm plus centromere and the centromere region. A large fraction of the constitutive heterochromatin of this species is present in the centromere region of the X + 3 chromosome and in the Y chromosome; these two regions possess similar amounts of DNA and therefore sort together. Chromosome flow sorting is rapid, reproducible and precise; it allows the collection of microgram quantities of purified chromosomes.

Animals↗

Further immunization studies with mammary tumor virus.

A single i.m. dose of formalin-inactivated murine mammary tumor virus greatly reduces viral expression and mammary tumorigenesis in Af (tumor incidence, 39%) and RIIIf (tumor incidence, 11%) mice, which carry only endogenous, gamete-transmitted virus. In C57BL mice, 1 mug of vaccine in Freund's complete adjuvant protects against later challenge with RIII virus.

Animals↗

Experimental infection of a cat kidney cell line with the mouse mammary tumor virus.

A cat kidney cell line, CRFK-F2, was successfully inoculated in suspension and in monolayer culture with a purified mouse mammary tumor virus derived from RIII milk. The virus produced by the infected cells was identified by immunogluorescence, electron microscopy, and RNA-directed DNA polymerase assays; it was a B-type virion that did not cross-react with mouse or feline leukemia-sarcoma viruses, had spikes on its envelope, and had a RNA-directed DNA polymerase reaction that was typical of mouse mammary tumor virus. The producing cells were identified as cat cells by chromosome number, cytotoxic assays, and isoenzyme migratory patterns. A standardized method for the in vitro inoculation of cat cells is described that presently permits highly reproducible results. For the first time, the mouse mammary tumor virus is seen replicating in cells from another species, thus offering an opportunity to study the kinetics of infection of that virus.

Animals↗

Record-keeping for a colony of 15,000 mice without use of a computer.

An economical and easy to learn method of record-keeping was developed which utilizes standard office supply 3 5 cards for cage cards and central office records. The method developed has been used to maintain all records of a mouse colony of 18 inbred strains and substrains for 9 yr. With this system, detailed pedigrees, postmortem data, tumor incidence, and average age are available in minutes.

Animals↗

Prognostically favorable immunogens of human breast cancer tissue: antigenic similarity to murine mammary tumor virus.

A leukocyte migration procedure was utilized to test cellular hypersensitivity of breast cancer patients' leukocytes to autologous and homologous breast cancer tissues and to murine milk containing murine mammary tumor virus (MuMTV). The in vitro responsiveness of the leukocytes and the antigenicity of breast cancer tissues were compared with in vivo prognostically favorable lymphoreticuloendothelial (L-RE) responses seen microscopically at the time of mastectomy and with the results of skin window tests of cellular hypersensitivity. The data suggest that immunogens appear in the in situ phase of the disease and provoke prognostically favorable L-RE responses. These immunogens possess antigenic similarity to some component(s) of MuMTV. Progression of the disease is associated with or preceded by a loss of tissue immunogenicity and/or diminished specific cellular hypersensitivity. The findings are pertinent to investigations of human mammary carcinogenesis and immunoprophylaxis.

Animals↗

Chromosome measurement and sorting by flow systems.

A flow microfluorometer was used to measure metaphase chromosomes in suspension at rates up to 100,000 per min. Chromosomes from cells of the Chinese hamster M3-1 cell line were isolated, stained for DNA with the fluorescent dye ethidium bromide, and analyzed for DNA content. Nine distinct peaks were resolved that correspond well with independent chromosomal DNA measurements made with a high-resolution scanning cytophotometer. Chromosomes were sorted from each peak by an electronic cell sorter. Visual examination of each fraction indicated the purity of the sorted chromosomes. This novel technology allows separation of purified populations of individual chromosomes suitable for biochemical and biological characterizations.

Animals↗

High-speed quantitative karyotyping by flow microfluorometry.

We describe a new flow-system technique for high-speed quantitative karyotyping. Metaphase chromosomes are isolated from cells, stained with a DNA-specific fluorochrome, and individually measured for stain content at rates up to 10(5)/min in a flow microfluorometer. The resulting distribution contains quantitative information about the relative frequency of occurrence and the chromosomal DNA content of the measured chromosomes. Such analyses of Chinese hamster chromosomes are in good agreement with independent results from scanning cytophotometry and with cytogenetic observations of the banded chromosomes. Analyses of two karyotypically different clones of Chinese hamseter M3-1 lines provide the quantitative characterization of two stable translocations. Initial data on human chromosomes are presented and agree well with results of scanning cytophotometry and suggest potential clinical applications as preparative and instrumental techniques improve.

Animals↗

Identification of the virions in the in vitro L1210(V) leukemia cell lines by morphological, virological, and immunological techniques.

In vitro L1210 (V) cell lines contained abundant intracytoplasmic A-particles, numerous C-type particles, a small number of B-type particles, and occasional intracisternal A-particles. The intracytoplasmic A-particles were incorporated into both spiked (B-type) and smooth-surfaced (C-type) particles formed at the budding site. Both B-and C-type particles also developed by gradual accululation of neucleooid material. The particles, particularly the C-type, exhibited a wide range of densities. The cells showed strong surface immunofluoresence for both murine mammary tumor virus and Gross murine leukemia virus antigens and variable degrees of cytoplasmic immunoflurescence for the protein antigens (p1 to p6) of Rauscher leukemia virus. The cells, the culture supernatant, and the purified virus each gave positive reactions with murine mammary tumor virus and murine leukemia virus antisera by immunodiffusion. The viral particles failed to infect C57BL, C57BL/6 X DBA/2F1 (hereafter called BD2F1), BALB/c, Af,and RIIIf mice. Howver, the cells were highly tumorigenic in BD1F-1 mice, moderately tumorigenic in BALB/c mice, but not tumorigenic in C57BL, Af, and RIIIf mice.

AKR murine leukemia virus↗

Separation of B and C type virions by centrifugation in gentle density gradients.

Murine type B particles were separated from type C (Rauscher leukemia virus) by means of gentle (low-increment rate) density gradients. The best separation was obtained when the density ranged from 1.13 to 1.20 g/cm(3) when sucrose was used and from 1.12 to 1.28 g/cm(3) with CsCl. The buoyant densities of the B and C particle bands in sucrose were 1.18 and 1.16 g/cm(3), respectively. The CsCl gradient gave a better separation with the B particles banding at a density of 1.20 g/cm(3) and with the C particle density little different from its value in sucrose.

Animals↗