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Biomedical subjects

J Zeuthen

Publications and source records attributed to J Zeuthen.

At least 19 recordsLinked to original sources
PubMed1979

Characterization of human chromosomal unit fibers.

A structural component of mitotic chromosomes that partially explains the compaction of DNA within mitotic chromosomes is suggested on the basis of the occurrence of long, regular cylindrical structures in preparations of isolated human chromosomes. These structures, "unit fibers", of a rather constant diameter of about 4,000 A have been postulated to be formed by coiling of the 250-300 A solenoid chromatin fiber that itself is formed by coiling of the 100 A string of nucleosome fiber. The human chromatid would thus be composed by a hierarchy of helices with contraction ratios for DNA at each level of coiling of 7 (string of nucleosomes), 5 (solenoid) and 40 (4,000 A "unit fiber" or "super-solenoid") which results in an overall contraction ratio for DNA in the "unit fiber" structures of about 1,400, which is approximately 5-fold less than the final contraction of DNA in intact chromatids of condensed metaphase chromosomes. The present report concerns more detailed studies with respect to the dimensions and cytochemical properties of the "unit fiber" structures observed in preparations of isolated human mitotic chromosomes that provide direct and indirect evidence in support of their "super-solenoid" structure and relate to known properties of human mitotic chromosomes.

Cells, Cultured↗
PubMed1979

Chromosomal unit fibers in Drosophila.

Chromosomal unit fibers consisting of long, regular fibers of about 0.40 micron diameter were obtained from disintegrated, isolated chromosomes of two Drosophila melanogaster cell lines. In one cell line with an essentially normal karyotype, three clearly defined size classes of 15,13, and 11 micron length were observed corresponding to the three larger chromosomes of Drosophila. In a cell line carrying an additional translocation between the two largest chromosomes a 19 micron fiber derived from the translocation chromosome was observed. Direct determinations of the DNA content per micron length of Drosophila unit fibers show that DNA is contracted by a factor of about 1400x in agreement with calculations based on the length of the unit fibers and the known DNA content of the individual Drosophila chromosomes. These findings support our previously proposed model for the unit fiber sub-structure of chromosomes as being derived by a hierarchy of coiling with the corresponding contraction ratios being 7 (100 A string of nucleosomes), 5 to 6 (250-300 A thick nucleohistone fiber), and about 40 (unit fiber), resulting in a total contraction of DNA in unit fibers in the order of 1400x.

Animals↗
PubMed1977

Somatic cell hybrids between human lymphoma lines. III. Surface markers.

Hybrids between two human lymphoma lines, Raji and Daudi (8A) and Raji BJAB (83) were examined for genetically determined and/or differentiation-related surface markers. HL-A B ce-l alloantigens, Fc and complement receptors, EBV receptors and beta2 microglobulin showed an autonomous ("co-dominant") expression in the hybrid. This is in contrast to most previous studies on other differentiation markers, involving as a rule crosses between cells of different lineages, where the differentiated pattern usually became "eclipsed" in the hybrid. Staining of activated complement and complement consumption tests showed intermediate or partially suppressed expression in the hybrids. This may be viewed in relation to the fact that these reactions do not merely depend on complement binding to the receptor, but also on subsequent activation and binding of the activated complement. A more complex interaction is also suggested for immunoglobulin production. Surface immunoglobulin showed a suppressive or intermediate pattern in both hybrids, whereas intracellular kappa chain production showed an amplification in the 83 hybrid. The beta2 microglobulin deficiency of the Daudi parent was corrected in the Raji/Daudi hybrid. Two new HL-A specificities,A10 and BW17, appeared on this hybrid which were not present on the parental lines. This suggests that the HL-A deficiency of the Daudi cell is due to its lack of beta2 microglobulin.

B-Lymphocytes↗
PubMed1977

Double immunoglobulin production in cloned somatic cell hybrids between two human lymphoid cell lines.

Several clones of independently established somatic cell hybrids between two human lymphoid cell lines, Raji and Namalwa, were examined for surface immunoglobulin expression. Double-antibody radioimmunoassays were established for kappa and lambda light chains. Immunoglobulins were detergent-extrated by Triton X-100 and quantified by radioimmunoassay. The Raji parent expressed small amounts of kappa chains on its surface, and the Namalwa parent a 10 fold greater amount of lambda chains. We show that the majority of the hybrid clones co-express both parental phenotypes.

Burkitt Lymphoma↗
PubMed1977

Spontaneous and induced patterns of the Epstein-Barr virus (EBV) cycle in a new set of somatic cell hybrids.

A somatic cell hybrid line and its subclones obtained by fusing two Burkitt lymphoma lines (Raji and Namalwa) were examined for the expression of EBV-specific antigens both spontaneously and after induction. The hybrids retained spontaneous early antigen (EA) production at the level characteristic of Raji. Similarly the more permissive Raji pattern dominated the induction of EA by IUDR treatment or P3HR-1 virus superinfection. These findings accord with our previous results on independently derived Raji/Namalwa hybrids. Virus capsid antigen was induced in the hybrids by P3HR-1 virus superinfection though at a lower level than in the Raji parental cell.

Antigens, Viral↗
PubMed1977

Higher-order structure of human mitotic chromosomes.

From observations on the partial disintegration of isolated human metaphase chromosomes we propose that human metaphase chromatids have a rather simple organization based on the folding and coiling of a long, regular, hollow cylindrical structure with a diameter of about 4000 A. This cylindrical structure, the unit fiber, is postulated to be a super-solenoid formed by the coiling of a 300 A solenoid, itself composed by coiling the basic string of nucleosomes. The structure of a human chromatid would thus be a hierarchy of helices, the contraction ratio of each coil, in ascending order of size, being approximately 7, 6, 40, and 5. This model appears to explain the estimated mass/unit length and accounts for many of the known features of human mitotic chromatids.

Chromosomes↗
PubMed1976

Inducibility of the Epstein-Barr virus (EBV) cycle and surface marker properties of EBV-negative lymphoma lines and their in vitro EBV-converted sublines.

Two EBV-negative lymphoma lines of human B-cell origin, BJAB and Ramos, were compared with altogether six of their in vitro EBV-converted, EBNA- and EBV-DNA-carrying sublines (four of Ramos and two of BJAB derivation). All converted lines closely resembled the parental line with regard to karyotype and HL-A and B antigen typing. Induction of EBV antigens (EA and VCA) by P3HR-1 virus superinfection was either similar in the converted and the negative lines, or somewhat increased in certain converted lines. These findings argue against a simple, virally determined repressor model and emphasize the role or cellular controls in restricting the EBV cycle in virus-carrying B-lymphocyte lines of human origin. IUdR inducibility varied in the different converted lines. There was a possible relationship between average number of EBV-genome equivalents per cell and inducibility. Converted sublines did not differ from the original negative lines with regard to surface immunoglobulin and Fc receptors. There was a dramatic increase in complement-consuming ability, however, following EBV conversion. Among the EBV-positive lines, there was a linear relationship between complement-consuming and EBV-receptor activity, the latter measured by a quantitative absorption test.

Antigens, Viral↗
PubMed1976

Properties of the K562 cell line, derived from a patient with chronic myeloid leukemia.

The K562 cell line derived from a CML patient in blast crisis was examined for properties of B and T lymphocytes and cell lines. K562 lacks the B markers of immunoglobulins, Epstein-Barr virus (EBV) genome and associated nuclear antigen, and receptors for EBV. A low proportion of cells from rosettes with sheep erythrocytes, the frequency of which is considerably increased after neuraminidase treatment. Unlike B lines but like T lines, K562 cells are lysed rapidly by C'/Fc receptor-positive human blood leukocytes and do not stimulate MLC reactions. On the other hand, K562 lacks T antigen, high radiosensitivity and sensitivity to growth inhibition by thymidine. The cells do not contain N-APase, an enzyme found in all lines derived from lymphoid cells and in lymphoproliferative diseases. By scanning electron microscopy, K562 cells were seen to be rounded and relatively smooth, with small numbers of short microvilli resembling undifferentiated leukemic cells. A few cells had narrow ridge-like profiles and small ruffles similar to granulocytic leukemic cells. K562 is strongly positive for immunoglobuln Fc receptors and pinocytosis, but does not phagocytose or mediate antibody-dependent phagocytosis or cytolysis. Among histochemical stains, K562 is positive for esterase, lipid, and acid phosphatase. There seems to be no doubt that K562 is not a B cell line. While it has some T cell properties, these are not exclusive. Some of its characteristics indicate that it is probably not lymphoid. Due to its low level of differentiation, its nature cannot be stated with certainty. On the basis of the possible presence of the cellular marker of chronic myeloid leukemia, the Ph chromosome, it may be regarded as belonging to the granulocytic series of cells.

Alkaline Phosphatase↗
PubMed1976

Somatic cell hybrids between human lymphoma lines. II. Spontaneous and induced patterns of the Epstein-Barr virus (EBV) cycle.

The regulation of spontaneous, IUDR-induced and P3HR-1 virus-induced EA and VCA production patterns was studied in two new somatic hybrids between human lymphoma lines. The hybrid 8A was derived from the crossing of the non-producer Raji with the spontaneous producer Daudi line. The second hybrid, 83, was produced by the fusion of Raji with the EBV-genome-negative B-lymphoma line, BJAB. The studies suggest the following EBV regulation patterns: (1) the spontaneous production of EA and VCA appears to be regulated by controls that differ from the regulators of P3HR-1 virus-induced or IUDR-induced EA synthesis. While spontaneous producer status was dominant over non-producer status, the level of EA inducibility was set by one of the parental cells, Raji ATG, and could either raise (in the previously studied Raji/Namalwa hybrid, cf Nyormoi et al. 1973) or depress (in Raji/Daudi) the level of relative EA inducibility found in the partner cell. (2) Although EA production is a prerequisite for VCA synthesis, the latter is under its own restriction mechanisms, quite independent of those that regulate the level of EA synthesis. (3) Inducibility of EA synthesis by P3HR-1 virus and by IUDR appear to be under the influence of at least partially identical controls. (4) EBV-negative lymphoma cells, exemplified by BJAB, may exert a "complementation" effect on the EA inducibility of their EBV-positive fusion partner, in spite of their own restrictivity against virus-induced EA synthesis. In more general terms, it is obvious that the EBV cycle is under the influence of multiple regulatory mechanisms in the human lymphoid cell. Depending on the parental cell and viral genomes that are allowed to interact, somatic cell hybrids may display a variety of patterns. At this time, cell hybridization is one of the few pathways that permit an approach to this complex and completely unknown world.

Antigens, Viral↗
PubMed1976

Hybridization of a human myeloma permanent cell line with mouse cells.

A population of hybrid cells derived from the fusion of a permanent human myeloma cell line, which secretes complete IgE, and a subline of mouse L cells, did not secrete IgE as evidenced by sensitive immunosorbent tests. Also, the hybrid cells were observed not to contain intracellular IgE (epsilon or lambda chains) in amounts to be detectable by fluorescent antibody techniques. The doubling times and cell cycle parameters of the hybrid cells were found to be similar to those of the slow-growing parental human myeloma cells, in addition, the growth of the hybrid cells was characterized by a higher degree of contact inhibition than the parent mouse cells.

Animals↗
PubMed1976

Expression of immunoglobulin synthesis in human myeloma x non-lymphoid cell heterokaryons: evidence for negative control.

Heterokaryons formed between human myeloma cells and various types of mouse and human non-lymphoid cells loose their cytoplasmic content of lambda light chains, a component of IgE produced by the myeloma parent. This loss of immunoglobulin content was observed regardless of the species origin (mouse or human) of the non-lymphoid partner cell, suggesting that the factors responsible for extinction of this differentiated function are not specific for a species. The kinetics of the loss of immunoglobulin content was essentially identical in the different experiments, since all myeloma X non-lymphoid cell heterokaryons were scored as negative after immunofluorescence staining for lambda chains 4-6 hr after infusion. Myeloma cells treated with inhibitors of protein synthesis (puromycin and cycloheximide) also lost their cytoplasmic content of immunoglobulin after 4 hr. These results indicate that the fusion of myeloma cells with non-lymphoid cells results in an immediate inhibition of immunoglobulin synthesis.

Animals↗