Possible two-stage mechanism in experimental leukemogenesis

The augmenting influence of urethan on leukemogenesis by x-radiation in mice has been found to operate when the urethan treatment follows the radiation, but not when the sequence is reversed. The result is in keeping with the idea that urethan acts as a promoting factor in leukemogenesis, as defined by the twostage mechanism hypothesis of carcinogenesis. It may also have a practical bearing on leukemia development in man.     

Leukemogenesis by urethan in C57B1 mice bearing isologous tissues from x-irradiated mice

A single exposure of mice of a low-leukemic strain (C57B1/6) to 400 r of total-body x-irradiation caused the early appearance of a factor which, though not itself capable of inducing leukemia in adult C57B1 recipients, did cause its appearance when the recipients also received urethan by injection. Positive results were obtained with seven different tissues from irradiated animals, whether urethan was injected 1 day or 30 days after the irradiation. No such effect was obtained in either series in the absence of urethan treatment. When urethan was injected into normal C57B1 mice, or into mice previously implanted with tissues from nonirradiated mice, only marginal leukemogenic activity was evinced. The possible involvement of a precursor virus is discussed.     

Diethyl pyrocarbonate: formation of urethan in treated beverages

Isotope dilution analyses with tritium-labeled diethyl pyrocarbonate show that the carcinogen urethan is formed in orange juice, white wine, and beer. Commercial use of the antimicrobial food additive diethyl pyrocarbonate can result in urethan concentrations of 0.1 to 0.2 milligram per liter in orange juice and of the order of 1 milligram per liter in white wine and beer.     

Latent meiotic anomalies related to an ancestral exposure to a mutagenic agent

When urethan is administered to cytologically normal F(2) progeny descended from grandsires exposed to ethyl methanesulfonate, meiotic anomalies in the forms of site-specific X-chromosome deletions and bivalent associations are noted in spermatocytes of male Armenian hamsters examined 6 and 8 days after treatment, respectively. These latent anomalies are initiated and retained in a premutated state for two generations after the ancestral exposure to ethyl methanesulfonate, the additional impetus required to complete the mutational processes being supplied by urethan.     

DISSOCIATION OF HOMOGRAFT RESPONSE TO ALLOGENEIC VERSUS XENOGENEIC SKIN GRAFTS IN IRRADIATED MICE

Lethally x-irradiated (C3H x DBA/2)F(1) mice were protected with syngeneic (isogenic) bone marrow and/ or prior treatment with urethan and skin grafted at various times after irradiation. The ability to reject a first-set xenogeneic (heterologous) skin graft with normal vigor returned within 92 days; the first-set response to allogeneic (homologous) grafts was still impaired 350 days after irradiation.     

Mouse leukemia virus activation by chemical carcinogens

The induction of lymphomas in C57BL mice by methylcholanthrene, urethan, or diethylnitrosamine was accompanied by the development of murine leukemia viral antigen in most of the lymphoid tumors. The cell-free transmission of lymphomas induced by methylcholanthrene and the development of antibody to murine leukemia virus prior to the detection of overt lymphoma in these mice suggest that unmasking of a latent leukemia virus is an indigenous actuating cause of the lymphomas.     

An early event in the interferon-induced transmembrane signaling process

Human interferon stimulates a transient two- to threefold increase in the concentration of diacylglycerol and inositol tris-phosphate within 15 to 30 seconds of cell exposure to interferon. Antibodies to interferon inhibit this effect. The stimulation was measurable in isolated cell membranes exposed to interferon. Human alpha and beta, but not gamma, interferon stimulate this increase in cells containing the appropriate interferon receptor. The effect was proportional to the number of interferon receptors. Both the diacylglycerol increase and antiviral effects induced by interferon could be correlated in terms of dose dependence. Thus, a transient diacylglycerol increase is an early event in the interferon-induced transmembrane signaling process.     

Homograft tolerance in mice: use of urethan and sublethal irradiation

Adult mice [CBA/J (H-2(k))], which received either a single sublethal dose of x-radiation (500 rad) or urethan plus 500 rad, were given intravenous injections of C3H/HeJ (H-2(k)) spleen or bone marrow cells (18 to 42 x 10(6) cells per mouse) or both, for 3 days. C3H/HeJ tail-skin homografts were retained (over 130 days) by these mice, whereas BALB/cJ (H-2(d)) homografts all were rejected within 33 days. Similarly irradiated or urethan-treated controls (or controls treated with a combination of both), which did not receive C3H cells, rejected both homografts. Specific homograft tolerance is induced in adult mice by this procedure.     

Interferon production and action in mouse, hamster and somatic hybrid mouse-hamster cells

A hybrid mouse-hamster cell line was developed from a mouse cell line which produces a high titer of interferon and is sensitive to its action, and a hamster cell line which produces little interferon and is relatively insensitive to its action. Parental cell lines demonstrated complete species specificity with respect to interferon production and action. The hybrid cells produced interferon (or interferons) effective when tested on the mouse cell line and primary hamster cells; the hybrids were sensitive to the action of both mouse and hamster interferons. Hybrid cells produced ten times more hamster interferon than the parent hamster cell line and were eight times more sensitive to hamster interferon than the parent hamster cell line.     

Cell-to-cell transfer of interferon-induced antiproliferative activity

Interferon-treated cells rapidly and efficiently transferred the antiproliferative activity of interferon to untreated cells. This phenomenon was not due to the carry-over of interferon by the interferon-treated cells. Thus, to evoke an antiproliferative state, interferon did not directly contact each cell in a population. The results suggest a novel mechanism by which interferon may indirectly regulate cell growth, and suggests that cells other than those of the immune system may play a role in controlling tumor growth in tissue where cell-to-cell contact occurs.     

Interferon inducers in vitro: difference in sensitivity to inhbitiros of RNA and protein synthesis

Interferon can be induced by diverse agents in a variety of mammalian cell cultures through apparently two mechanisms. One results in an early (2 to 10 hours) appearance of interferon and is relatively resistant to inhibition by actinomycin, puromycin, or fluorophenylalanine. A second mechanism results in a late (18 to 24 hours) appearance of interferon and is more sensitive to inhibition by these inhibitors. The molecular basis for each mechanism is unclear. Since each interferon inducer may have multiple effects on the cell, the differences observed may not necessarily reflect a fundamental difference in the mechanism of interferon stimulation.     

Immune specific induction of interferon production in cultures of human blood lymphocytes

Human blood lymphocytes stimulated with nonviral antigens in vitro produce an antiviral substance with the biological and biochemical characteristics of interferon. The induced response was specific for cells obtained from immune donors. Cells from nonimmune donors did not produce interferon on exposure to these substances. The quantity of interferon produced by antigen stimulation was related to concentration of antigen over a relatively narrow range; with higher concentrations induction was decreased. Interferon production was maximum during days 4 to 7 in culture. In contrast, phytohemagglutinin-induced interferon was primarily produced during the first 4 days in culture.     

鸡γ-干扰素基因在大肠杆菌中的表达

构建鸡γ-干扰素基因(lFN-γ)的融合表达质粒,并在原核系统表达。根据GenBank发表的鸡γ-干扰素核苷酸序列,使用prim er5设计一对特异性引物,通过RT-PCR技术从ConA诱导培养的鸡脾脏淋巴细胞中克隆出鸡γ-干扰素基因并对其进行测序。测序结果表明,鸡γ-干扰素基因全长495bp,具有一个完整的开放阅读框,编码164个氨基酸,与国外发表的序列同源性为100%。将序列连接到原核表达载体pET28 a(+)上,转化大肠杆菌BL21(DE3),经IPTG诱导后进行SDS-PAGE电泳分析。结果表明鸡γ-干扰素表达蛋白大小为20.8KD,并以包涵体形式表达。为鸡γ-干扰素生物制剂或疫苗佐剂的开发奠定基础。     

Molecular weight of human gamma interferon is similar to that of other human interferons

The molecular weight (as determined by molecular sieve chromatography) of human gamma interferon, formerly referred to as immune or type II interferon, is between 40,000 and 70,000. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, gamma interferon activity was recovered mainly from two regions of the gels corresponding to molecular weights of 20,000 and 25,000. The results suggest that in native form human gamma interferon may be aggregated.     

Interferon: production by chick erythrocytes activated by cell fusion

In the presence of Sendai virus inactivated with ultraviolet light, nucleated chick erythrocytes can be fused with several types of human cells to form heterokaryons. Although chick erythrocytes alone cannot be stimulated by Sendai virus to produce interferon, fusion with a human cell (AH-1) which itself may produce human interferon results in heterokaryons in which the erythrocyte genome is activated and chick interferon is produced. When nucleated chick erythrocytes are fused with another type of human cell (HeLa clone S-3) which does not produce human interferon when stimulated, no chick interferon is detectable, despite morphologic changes suggestive of activation of the erythrocyte nuclei.     

Suppression of gamma interferon production by inactivated feline leukemia virus

Supernatants from cultures of normal feline lymphocytes stimulated with Staphylococcus enterotoxin A showed antiviral activity, characterized as a gamma-like interferon. With the addition of inactivated feline leukemia virus, markedly less interferon was produced. The reduction in interferon production was not attributable to lowered lymphocyte viability or reduced mitogenic properties of Staphylococcus enterotoxin A and appears to be a direct retroviral effect. This finding may reflect clinically relevant events that may contribute to the development of the feline or human states of acquired immunodeficiency.     

Interferon binding: the first step in establishment of antiviral activity

Chick cells incubated at 1 degrees C with interferon fail to develop antiviral activity, but this activity appears subsequent to a 7-hour incubation at 37 degrees C after removal of interferon by repeated washings. Treatment with actinomycin D blocks the development of the latter activity. Cells incubated with interferon at 1 degrees C for up to 1 hour and then washed and incubated for 2 hours at 37 degrees C develop a degree of antiviral activity proportional to the concentration of interferon at initial incubation; at any concentration, the antiviral activity increased with the duration of initial incubation at 1 degrees C, but a maximal response was reached at 10 or 20 minutes. Treatment with trypsin after incubation with interferon at 1 degrees C inhibited development of antiviral activity. Interferon is rapidly bound to a superficial cell site, and this binding is necessary for development of antiviral activity in chick cells.     

应用干扰素治疗肝炎研究进展

干扰素联合用药是治疗慢性乙型肝炎和丙型肝炎中最常用的治疗方案,联合用药不但可以提高干扰素的治疗效果,还可减轻干扰素的不良反应.但是,干扰素联合用药也存在着一定问题,如何改善现有的治疗方案和利用基因工程手段寻找新的治疗肝炎药物是此后一段时期该研究领域学者将要面临的新的挑战.