Growth Of L929 Cells Research Articles

Growth and morphology of anchorage-dependent animal cells in a liquid/liquid interface system.

In general, anchorage-dependent animal cells cultivated on a solid culture substrate, such as polystyrene, are collected by trypsin treatment. This treatment may have detrimental effects such as the proteolysis of the cell membrane proteins. To avoid these effects, cell cultivation using a liquid/liquid interface system has been investigated. In this cultivation method, the cells grow at the interface between a culture medium and a hydrophobic liquid. In this study, various fluorocarbons (FC-40, FC-70, KPF-91, KPF-102, and KPF-142) were used as substrates for the interface, and the cultivation of fibroblast cells (L-929; the mouse-derived cell line) at the interfaces was investigated. Early in the cultivation period, the growth of L-929 cells depended on the substrate type. Although cell cultivation at the interfaces was possible, it was slower than that at the polystyrene surface. Cell spreading at the interfaces was relatively small, which indicates that cell adhesion at the interfaces may be weak. In particular, the cells at the MEM/FC-70 interface anchored with one another and formed multicellular hemispherical aggregations shaped like spheroids. The difference in the adhesions to the interfaces appears to be dependent on the contaminants contained in the fluorocarbons because the physical properties of the fluorocarbon did not affect the cell growth and adhesion. Moreover, subcultivation from the interfaces to the same interface was possible without trypsin treatment. In this case, the delay of the growth at the interfaces did not occur because the cells were not affected by trypsin treatment.

The effect of oligosaccharides on the production of tumor necrosis factor-alpha by macrophage-like cell line J774/JA-4.

Stimulation and modulation of tumor necrosis factor-alpha (TNF-alpha) production during treatment of the murine macrophage-like cell line J774/JA-4 with 25 oligosaccharides were studied. Direct stimulation of TNF-alpha production by oligosaccharides was measured with a cytotoxic assay using the L929 cell line. Twelve samples showed a significantly higher production (P < or = 0.01) of TNF-alpha than the controls. Modulation of TNF-alpha production by treatment with oligosaccharides, followed by stimulation with lipopolysaccharide (LPS) from E. coli, was examined using the L929 bioassay system. In three samples TNF-alpha production increased significantly, but in four samples, production was reduced significantly (P < or = 0.01). No samples showed modulation of growth or viability of L929 cells within the first 26 hr. The present results are useful in the application of these oligosaccharides which is potentially applicable in medical and food technology.

Induction of neutralizing antibody in mice by immunization with recombinant 56 kDa protein of Orientia tsutsugamushi

Anti-oriential antibody inhibits Orientia tsutsugamushi attachment to, and penetration of, host cells. However, O. tsutsugamushi antigens that induce the production of a neutralizing antibody have not been identified. The authors immunized mice and rabbits with the recombinant 56 kDa protein of O. tsutsugamushi fused to the maltose binding protein of Escherichia coli (MBP-Bor56) and analysed their effect on O. tsutsugamushi attachment to or penetration of L929 cells. O. tsutsugamushi attachment and penetration were measured by using an indirect immunofluorescent antibody assay (IFA). O. tsutsugamushi growth in L929 cells was determined by [ 3H]thymidine uptake assay. By IFA, we observed a 96% reduction of attachment or penetration of O. tsutsugamushi treated with rabbit anti-MBP-Bor56 sera. [ 3H]thymidine uptake showed that mouse anti-MBP-Bor56 sera caused a 91% reduction in O. tsutsugamushi growth, when compared to mouse anti-MBP sera. These results suggest that the 56 kDa protein of O. tsutsugamushi plays an important role in O. tsutsugamushi attachment to or penetration of cells.

Characteristics of silica-chitosan complex membrane and their relationships to the characteristics of growth and adhesiveness of L-929 cells cultured on the biomembrane

The relationships between the chemical and physical characteristics of a silica-chitosan complex membrane (SiCM) and the characteristics of growth and adhesion of fibroblast-like L-929 cells cultured on the surface of SiCM were investigated for development of a novel implant biomaterial for use as artificial skin. Silicon ethoxide was used to cross-link the chitosan complex membrane to enhance its oxygen permeability. The composition of the SiCM was stepwise controlled by adjustment of the mixing ratio between silica and chitosan, for silica contents of 20, 33, 43, and 50 wt%. The zeta potential of the SiCM varied negatively according to the silica content, from +20 mV for chitosan membrane (0%-SiCM) to −0.6 mV for 43%-SiCM. Similarly, the wet contact angle decreased from 68° to 41° according to the increase in the silica content up to 43%. However, significant increases in the wet contact angle and the zeta potential occurred upon further increase of the silica content to 50%. The adhesion and growth characteristics of L-929 cells on SiCM were found to be related to the surface properties of the SiCMs. The cell proliferation rate was higher on the 0, 20 and 50%-SiCMs than on a typical polystyrene culture dish. The strongest cell adhesion was obtained on 50%-SiCM among the complex membranes. Thus, the surface wet contact angle and the zeta potential of 50%-SiCM, which is highly permeable to oxygen, were found to be suitable for L-929 cell adhesion and growth, and therefore to promote tissue culture on the SiCM in vitro.

Prostaglandin regulation of gene expression and growth in normal and malignant tissues.

Prostaglandins bind with cell surface receptors and cause an activation of adenylate cyclase and generation of adenosine 3′5′ cyclic monophosphate (cAMP) (1,2). C-AMP is thought to play a central role in the regulation of cell growth since the growth of many types of cultured cells is arrested by adding exogenous dibutyryl adenosine 3′5′ cyclic monophosphate (dbcAMP) (3,4). In 1971, a report by Pastan and Johnson demonstrated that growth of L-929 cells was inhibited by prostaglandin E1 (PGE1). Like epinephrine and glucagon, many prostaglandins are able to trigger the adenylate cyclase enzyme in the cell membrane and stimulate a rise in cAMP; it was therefore postulated that the block in cell growth caused by PGE1 was solely due to the increased intracellular concentration of cAMP (5). Since that time many investigators have shown that prostaglandins with an α,β unsaturated ketone ring structure inhibit cell growth (1,6–10) in normal and malignant cells. In this paper, we demonstrate using cell mutants that the growth inhibitory prostaglandins do not require cAMP or protein kinase mediated mechanism. In addition, we discuss the finding that inhibition of endogenous prostaglandin synthesis causes inhibition of osteoblast cell growth.KeywordsAdenylate CyclaseProstaglandin SynthesisCyclic MonophosphateEarly Gene ExpressionOsteoblast GrowthThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Fretting corrosion resistance and fretting corrosion product cytocompatibility of ferritic stainless steel.

To avoid nickel ion release from SUS317L as an implant material, a new type of nickel, commercially free, of high purity, and high chromium ferritic stainless steel, was developed. The new stainless steel (FJ) was studied for aspects of fretting corrosion and cytocompatibility compared with SUS317L. A pin-on-plate fretting corrosion test in an artificial physiologic solution, and cell culture in media with the addition of the artificial physiologic solution used for fretting was conducted. Resistance to the fretting induced crevice corrosion of FJ was higher than that of SUS317L because of the favorable electrochemical stability of the FJ alloy. The amount of iron ion or colloidal fine particles released from FJ was about a quarter of that from SUS317L, although the weight loss of a pin of FJ was almost 5/3 that of SUS317L. The artificial physiologic solution used for SUS317L fretting was more harmful to the growth of L929 and MC3T3-E1 cells than that used for FJ fretting. FJ was therefore superior to SUS317L as a biomaterial, judging from the resistance to fretting-induced crevice corrosion, electrochemical stability, and the cytocompatibility of fretting corrosion products.

Attachment and growth of cultured fibroblast cells on silk protein matrices

The attachment and growth of L-929 cells on films made of Bombyx mori silk proteins--fibroin and sericin and their mixtures--was studied by a cell culture method. Both cell attachment and growth were dependent on a minimum of around 90% sericin in the mixture. The results from electron micrography as well as from the DSC measurements supported the notion that the mixture of the two proteins fibroin and sericin has a phase-separated structure in the solid state. The observed minimum of sericin in the cell attachment and growth is thought to be a result of this phase-separated structure. Films of pure component proteins (i.e., 100% fibroin or sericin) exhibited as high a cell attachment and growth as collagen, a widely used mammalian cell culture substrate. However, a morphological study of the attached cells revealed that the cells attached to silk fibroin were extended and had a spindle shape, just like the cells attached to collagen, while the cells attached to the silk sericin had a different shape. It is concluded, therefore, that the attachment condition on silk fibroin is ideal for the viability, growth and function of the cells.

Cytokine sensitivity and methylation of lysine in Rickettsia prowazekii EVir and interferon-resistant R. prowazekii strains.

Modified Rickettsia prowazekii strains have been derived from the avirulent Madrid E strain by passage in the lungs of white mice (strain EVir) or by selection for resistance to gamma interferon (IFN-gamma) (strains 427-19 and 87-17) or alpha/beta interferon (IFN-alpha/beta) (strains 83-2P, 60P, 103-2P, and 110-1P). Compared with the Madrid E strain, strain EVir has increased virulence (N. M. Balayeva and V. N. Nikolskaya, J. Hyg. Epidemiol. Microbiol. Immunol. 17:11-20, 1973) and a different lysine methylation profile in its surface protein antigen (A. V. Rodionov, M. E. Eremeeva, and N. M. Balayeva, Acta Virol. 35:557-565, 1991). The other six strains differ from the Madrid E strain in their resistance to IFN and their ability to grow well in untreated macrophagelike RAW264.7 cells. In the present study, to determine which properties are shared by these strains, we examined R. prowazekii EVir for the following: (i) the sensitivity of its growth in L929 cells to the cytokines IFN-alpha/beta, IFN-gamma, tumor necrosis factor alpha (TNF-alpha), and IFN-gamma plus TNF-alpha; (ii) the ability to grow in untreated RAW264.7 cells; and (iii) the ability to induce interferon in L929 cell cultures; we also evaluated strains 83-2P and 87-17 for lysine methylation. Multiplication of strain EVir in growing L929 cells was not markedly inhibited by either IFN-alpha/beta or IFN-gamma. In X-irradiated L929 cells, growth of strain EVir was slightly inhibited (11%) by TNF-alpha alone, somewhat inhibited (38%) by IFN-gamma alone, and markedly inhibited (87%) by IFN-gamma plus TNF-alpha. Nitrite production was induced in X-irradiated, strain EVir-infected L929 cell cultures treated with TNF-alpha alone or IFN-gamma alone; however, more nitrite was produced in infected cultures treated with IFN-gamma plus TNF-alpha. Nitrite production, the dramatic inhibitory effect of IFN-gamma plus TNF-alpha, and the modest inhibitory effect of IFN-gamma on the growth of strain EVir in X-irradiated L929 cells were all alleviated by the addition of the nitric oxide synthase inhibitor NG-methyl-L-arginine. Strain EVir grew very well in untreated macrophagelike RAW264.7 cells and appeared defective in the ability to induce IFN in L929 cell cultures. All strains grown in L929 cells in the presence of radiolabeled lysine had similar percentages of their radioactivity as methylated lysines.(ABSTRACT TRUNCATED AT 400 WORDS)

Analysis of the peptidoglycan of Rickettsia prowazekii.

In the present study, peptidoglycan from Rickettsia prowazekii, an obligate intracellular bacterium, was purified. The rickettsial peptidoglycan is like that of gram-negative bacteria; that is, it is sodium dodecyl sulfate insoluble, lysozyme sensitive, and composed of glutamic acid, alanine, and diaminopimelic acid in a molar ratio of 1.0:2.3:1.0. The small amount of lysine found in the peptidoglycan preparation suggests that a peptidoglycan-linked lipoprotein(s) may be present in the rickettsiae. D-Cycloserine, a D-alanine analog which inhibits the biosynthesis of bacterial cell walls, prevented rickettsial growth in mouse L929 cells at a high concentration and altered the morphology of the rickettsiae at a low concentration. These effects were prevented by the addition of D-alanine. This suggests that R. prowazekii contains D-alanine in the peptidoglycan and has D-Ala-D-Ala ligase and alanine racemase activities.

Interferon‐α/β and Rickettsia prowazekii: Induction and Sensitivitya

Rickettsia prowazekii Madrid E established persistent infections in cultures of growing L-929 cells. Although some L-929 cells died, the cultures survived, remained infected with rickettsiae, and continued to grow. R. prowazekii Madrid E also induced interferon in L-929 cell cultures, and this interferon modulated rickettsial growth. Production of interferon (anti-viral activity) by cultures of R. prowazekii-infected L-929 cells was directly related to the initial rickettsial infection and was blocked by erythromycin. The media collected from R. prowazekii-infected L-929 cells suppressed not only the replication of vesicular stomatitis virus but also the growth of R. prowazekii in fresh L-929 cells. Both anti-viral and anti-rickettsial activities in the media were neutralized by antibodies against murine interferons-alpha and -beta, but not by antibodies against murine interferon-gamma. In addition, a commercial preparation of virus-induced interferons-alpha and -beta also suppressed rickettsial growth in L-929 cells. The combination of treating L-929 cells with this virus-induced interferon and infecting them with R. prowazekii killed some of the L-929 cells.

Biocompatibility of wound management products: the effect of various monosaccharides on L929 and 2002 fibroblast cells in culture.

The effects of various monosaccharides on the growth of human 2002 and mouse L929 fibroblast cultures have been investigated. Eleven monosaccharides having acidic, neutral, and basic characteristics were evaluated in a bioassay procedure developed for the investigation of biocompatibility of wound management materials. Rate of growth in both cell lines was inhibited by D-galacturonic acid and by D-glucuronic acid. Although most neutral sugars produced no significant change in the growth rate or in the morphology of the cells, galactose produced a significant increase in the growth rate of both cell lines whilst L-fucose caused a significant decrease in growth of the L929 cells but did not significantly affect the growth of 2002 cells; xylose increased the growth rate of L929 but not 2002 cells. D-Glucosamine, a basic sugar, produced inhibition of growth which followed a different pattern from that produced by the acidic sugars; N-acetylglucosamine produced a species specific increase in cell growth of L929 cells. The results show that the effects produced by the monosaccharides on the cultured fibroblasts are related to their chemical structure and to cell line, and suggest that the use of galactose as a possible aid to wound healing should be investigated.

The intracellular release of methotrexate from a synthetic drug carrier system targeted to fc receptor-bearing cells

Methotrexate was conjugated to trinitrophenyl (TNP)-labeled poly ( d -lysine) either directly, or through a disulfide or a triglycine spacer. Conjugates were complexed with heparin and anti-TNP antiserum and tested for their growth inhibitory effects in cultured cells. When tested in Fc receptor-positive WEHI-3 cells, the anti-TNP immune complex of the conjugate with the disulfide spacer was more effective than that with the triglycine spacer. It had no effect on the growth of Fc receptor-negative L-929 cells. Without spacer, the direct conjugate was ineffective in both cell lines. The growth inhibitory effect of the drug-containing immune complexes was partially abolished in presence of an irrelevant, drug-free immune complex. NH 4Cl at 3 m M did not increase the cytotoxicity of the disulfide conjugate complexes, and decreased the effect of the triglycine conjugate complexes. These findings suggest that: (1) hapten-polymer conjugates can be used as drug carriers for targeting Fc receptor-bearing cells when given with an anti-hapten antibody, (2) disulfide spacers are effectively cleaved following Fc receptor-mediated endocytosis of a drug-carrying immune complex, (3) disulfide spacers are most likely to be cleaved in compartments other than endosomes or lysosomes.

Relationship between intracellular concentration of S-adenosylhomocysteine and inhibition of vaccinia virus replication and inhibition of murine L-929 cell growth.

9-(trans-2',trans-3'-Dihydroxycyclopent-4'-enyl)-adenine (compound 1) and -3-deazaadenine (compound 2), which are specific inhibitors of S-adenosylhomocysteine (AdoHcy) hydrolase, were reported earlier by our laboratory (M. Hasobe, J. G. McKee, D. R. Borcherding, and R. T. Borchardt, Antimicrob. Agents Chemother. 31:1849-1851, 1987) to have anti-vaccinia virus activity with reduced murine L-929 cell toxicity compared with the prototype compound neplanocin A. In this study, we showed that the antiviral and cytotoxic effects of compounds 1 and 2 can be related to intracellular concentrations of AdoHey, which are elevated in cells treated with these inhibitors of AdoHcy hydrolase. For example, concentrations of analogs 1 and 2 that produce 50% inhibition of vaccinia virus replication caused only slight elevations in intracellular levels of AdoHcy (from 50 [controls] to 100 to 125 [drug-treated cells] pmol/mg of protein) and elevations in the ratios of AdoHcy/S-adenosylmethionine (from 0.05 to 0.1 [controls] to 0.15 to 0.19 [drug-treated cells]). In contrast to the extreme susceptibility of virus replication to slight elevations in intracellular AdoHcy, cell viability was quite tolerant to higher levels of this metabolite. For example, concentrations of analogs 1 and 2 that produced 50% inhibition of L-929 cell replication caused significant increases in intracellular levels of AdoHcy (to 825 to 950 pmol/mg of protein) and elevations in AdoHcy/S-adenosylmethionine ratios (approximately 1.3). These data make it possible to assign a therapeutic index of 7 to 8 to these compounds on the basis of the comparison of intracellular levels of AdoHcy that caused 50% inhibition of vaccinia virus replication with those that caused 50% inhibition of L-929 cell replication.

Effects of injection routes of growing tumors and PSK or OK-432 on antiproliferative activity of mouse serum.

The present study examined the effects of various treatments on the antiproliferative activity of mouse serum. Its activity was estimated against the growth of EL4 tumor cells and L929 cells and against splenic blastogenesis in culture. The activity varied among mouse strains tested and among individuals in any strain. However, normal outbred NIH Swiss mice showed the highest activity among the strains and the least variation among individuals. The activity of serum from NIH Swiss mice constantly decreased 7 or 14 days after an injection of 10(6) Ehrlich or sarcoma 180 tumor cells subcutaneously in the right-hind footpad, intradermally in the right side of the chest or into the palm. Other routes, such as intraperitoneal, intravenous in the tail vein, subcutaneous in the right side of the chest and intramuscular in the left thigh, however, hardly affected the activity. The activity also decreased 7 days after an injection into the footpad of a biological response modifier such as PSK or OK-432. The antiproliferative activity of mouse serum seem to depend on macrophages but not natural killer-cell activity, because treatment with silica but not anti-(asialo-GM1) antibody totally reduced the activity. The active fraction was heat-stable (100 degrees C, 30 min) and its molecular mass was 127-140 kDa.

Gamma-interferon-induced inhibition of the growth of Rickettsia prowazekii in fibroblasts cannot be explained by the degradation of tryptophan or other amino acids.

We examined the role of amino acid deprivation in gamma-interferon-induced (IFN-gamma) suppression of the growth of Rickettsia prowazekii in mouse L929 cells and human fibroblasts by measuring the amino acid pools in untreated and IFN-gamma-treated cells. In recombinant IFN-gamma-treated cultures of human fibroblasts, tryptophan was undetectable in both the intracellular pool and the extracellular medium. In contrast, tryptophan was not depleted from the intracellular pool or the extracellular medium of L929 cells treated with recombinant IFN-gamma or crude mouse lymphokines. None of the other amino acids measured was severely depleted in IFN-gamma-treated L929 cells and human fibroblasts. Extracts prepared from IFN-gamma-treated human fibroblasts exhibited indoleamine 2,3-dioxygenase activity, converting tryptophan into products that cochromatographed with N-formylkynurenine and kynurenine; however, extracts prepared from untreated human fibroblasts, untreated L929 cells, recombinant IFN-gamma-treated L929 cells, and mouse lymphokine-treated L929 cells did not degrade tryptophan. Human HeLa cells resembled the human fibroblasts in that they degraded tryptophan after IFN-gamma treatment. Similarly, mouse 3T3-A31 cells and mouse embryo fibroblasts resembled mouse L929 cells in that they did not degrade tryptophan. Supplementation of the extracellular medium with additional tryptophan reconstituted the tryptophan pool in mock-infected and R. prowazekii-infected, X-irradiated, IFN-gamma-treated human fibroblasts to values greater than those observed in untreated control cultures. However, reconstitution of the tryptophan pool did not relieve IFN-gamma-induced inhibition of rickettsial growth. Addition of kynurenine or N-formylkynurenine to rickettsia-infected human fibroblasts at concentrations four times the usual tryptophan concentration did not inhibit growth of R. prowazekii. We conclude that neither tryptophan depletion nor depletion of the other amino acids studied explains the inhibitory effect of IFN-gamma on rickettsial growth in mouse L929 cells. In IFN-gamma-treated human fibroblasts, either tryptophan depletion is not involved in the inhibition of rickettsial growth or tryptophan depletion and some other mechanism(s) together contribute to the inhibition of rickettsial growth.

5-Fluoroorotate: a new liposome-dependent cytotoxic agent

The potency of 5-fluoroorotate for inhibition of L929 or CV1-P cell growth is increased by encapsulation in negatively charged liposomes. The optimal liposome composition is dipalmitoylphosphatidylglycerol: cholesterol, 67:33. Unextruded large unilamellar liposomes are the optimal size for delivery. This compound is the second transport-negative drug which we have found to exhibit liposome-dependent delivery.

Regulation of Prolyl Hydroxylase Activity in L-929 Cells by Mechanisms Unrelated to Glycolytic Metabolism

Previous studies showing that prolyl hydroxylase is activated by addition of lactate to logarithmic cultures of L-929 and 3T6 mouse cell lines and that the enzyme normally increases in dense cultures which accumulate lactic acid, have led to the hypothesis that regulation is mediated through glycolytic metabolism. We present evidence that the extent of lactate production plays a minor role in controlling the density-dependent activation of prolyl hydroxylase in L-929 cells. The initial rate of glycolysis in stationary phase cells was lower than that in the logarithmic phase so that intracellular levels of glycolytic intermediates would not be related to activation of the enzyme. Nevertheless, because of increased numbers of cells in stationary phase, higher concentrations of lactate occur in the culture medium. Eliminating accumulation of lactate during the growth of L-929 cells by reducing the normal glucose concentration (5.5 mM) to lowglucose (0.5 mM) did not significantly affect the growth rate and still allowed a 10-fold activation of prolyl hydroxylase when cells reached stationary phase. Growth in normal glucose resulted in only an additional 2-fold increase in specific enzyme activity. These results imply that density-dependent activation of prolyl hydroxylase occurs mainly through a mechanism independent of lactate production. The elevated enzyme activity in dense cells was not decreased when lactate was removed and its production prevented by replacement of high-glucose medium with low-glucose medium for 24 hours. This result differed markedly from the dramatic decrease in enzyme activity observed within several hours after replating stationary phase cells in fresh medium, but at a lower cell density. The conclusion that there are at least two mechanisms regulating prolyl hydroxylase activity in L-929 cells was further supported by the observation that the maximal extent of enzyme activation by ascorbate or lactate in logarithmic phase L-929 cells was much less than the extent of density-dependent activation.

Anti-steroid action in cultured L-929 mouse fibroblasts

L-929 mouse fibroblast growth is arrested by the glucocorticosteroid dexamethasone (dex), which also decreases adhesiveness to culture plates. Both dex effects were abolished when RU 486, a new synthetic anti-hormonal steroid, was added to the culture medium. Using [ 3H]RU 486, binding studies revealed that RU 486 bound ~ 25,000 sites/cell of the glucocorticosteroid receptor (GR) with affinity higher than that of dex and translocated GR to the nucleus. However, the distribution of steroid-receptor complexes between cytosol and nuclei was different for the agonist and the antagonist, with more nuclear accumulation in the case of dex. Estradiol increases L-929 cell growth and adhesiveness to culture plates, but not if the anti-estrogen tamoxifen (tam) was added. These observations initially made in serum containing medium, were confirmed in serum-free, chemically defined culture medium (SF). In SF medium, tam (1 μM) provoked death of most L-929 cells after 10 days of culture, leading to the selection of a variant clone LB of tam-resistant cells. Tam binds to the estrogen receptor (ER), but with less affinity than estradiol. ER concentration, estimated by the binding of 4-hydroxytamoxifen (OH-tam) and of estradiol was lower in LB cells than in original tam-sensitive L-929 cells. The concentration of specific anti-estrogen binding sites in the particulate fraction of the cells, measured by OH-tam binding, non competed by estradiol, was also significantly diminished in tam-resistant LB cells.

In vivo and in vitro effect of progesterone on the growth of some mouse and human tumours.

The effect of progesterone on the growth of tumours of different morphological and cytokinetic characteristics and origin has been investigated, such as mammary aplastic carcinoma, Ehrlich ascitic and solid tumour, fibrosarcoma, melanoma B-16, myeloid leukaemia and three cell lines, HeLa, HEp-2 and L929. The administration of progesterone has been proved to stimulate the growth of aplastic carcinoma of the breast in vivo. Directly implemented into cell culture of the same tumour it increased the incorporation of 3H-thymidine into DNA. In HeLa and HEp-2 cell cultures progesterone stimulated the population growth of these cells. However, progesterone has shown no effect at all on the growth of fibrosarcoma, melanoma, Ehrlich tumour, myeloid leukaemia and L929 cells, fibroblasts of C3H mice.

Inhibition of the growth of Rickettsia prowazekii in cultured fibroblasts by lymphokines.

The effect of lymphokine treatment of mouse and human fibroblast cell lines on the growth of Rickettsia prowazekii within the fibroblasts was studied. Treatment of mouse L929 cells with concanavalin A- or antigen-induced mouse lymphokines both before and after infection with R. prowazekii led to clearance of the rickettsiae from a substantial proportion of the cells and suppression of rickettsial growth in those cells which remained infected. Similar but less dramatic anti-rickettsial effects were observed in L929 cells treated with mouse lymphokines either only before or after infection with rickettsiae. Mouse lymphokine treatment of L929 cells had similar anti-rickettsial effects on the avirulent E strain and the virulent Breinl strain of R. prowazekii. Addition of cycloheximide or emetine to L929 cells at the same time as the lymphokines markedly suppressed the inhibition of rickettsial growth by the lymphokines. Mouse lymphokine treatment inhibited rickettsial survival and growth in mouse 3T3-A31 cells as well as in mouse L929 cells, but had no effect on rickettsial survival and growth in human foreskin fibroblasts. Conversely, concanavalin A-induced human lymphokines inhibited rickettsial survival and growth in human foreskin fibroblasts but had no effect on rickettsial survival and growth in mouse L929 cells. The rickettsia inhibitory activity in concanavalin A-induced mouse lymphokines was destroyed by heating the lymphokines at 80 degrees C for 10 min or by holding the lymphokines at pH 2 for 24 h but was retained after heating at 56 degrees C for 30 min.