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F Colotta

Publications and source records attributed to F Colotta.

At least 91 records · Page 5Linked to original sources
PubMed1987

Expression of c-fos protooncogene in normal human peripheral blood granulocytes.

We have investigated by Northern blot analysis the expression of c-fos protooncogene in human peripheral blood polymorphonuclear leukocytes (PMN). Freshly isolated PMN, unlike highly purified circulating lymphoid cells, showed high levels of c-fos transcripts. Appreciable c-fos mRNA was detected in monocytes, but in lesser amounts compared with PMN. Upon exposure to a series of agents that functionally stimulate granulocytes (PMA, inactivated streptococci, TNF, granulocyte and granulocyte/macrophage colony-stimulating factor), a considerable increase in c-fos transcripts was detected. Expression of c-fos in PMN was superinduced by exposure to cycloheximide. These data indicate that the myelomonocytic differentiation pathway c-fos expression is not peculiar to monocytes/macrophages and that PMN may represent a suitable system with which to investigate the link between protooncogene expression and functional activation.

Cell Differentiation↗
PubMed1987

Protooncogene expression in normal and tumor-infiltrating phagocytes.

Expression of cellular protooncogene was investigated in freshly isolated human leukocytes and in tumor-associated macrophages isolated from murine fibrosarcomas. Normal leukocytes express high levels of c-fos and c-raf-1 transcripts. Expression of c-fos and c-raf-1 in leukocytes differ in many respects; in particular, agents that stimulate leukocyte functions augment c-fos expression but have no effects on c-raf-1 transcription. Tumor-associated macrophages constitutively express c-fos protooncogene at levels higher than peritoneal macrophages. Moreover, in contrast to what happens in peritoneal macrophages, tumor-associated macrophages do not respond to endotoxin with an increase in c-fos expression.

Animals↗
PubMed1986

Metastatic growth of a murine tumor: evidence of dissemination to the lungs in the absence of subcutaneous growth.

Growth of MCA-38/B colon adenocarcinoma was detectable 30-33 days after subcutaneous (s.c.) tumor cell inoculation in mice. Seventy percent of the mice receiving 10(7) tumor cells, 50% of those receiving 10(6), and 15% of the mice given 10(5) cells developed s.c. tumors (mean of 4 experiments, total of 80 mice per group). Metastases in the presence of a primary tumor were observed in 11% of 10(7) and in 10% of 10(6) tumor-cell injected animals. Lung metastases were detected in the absence of tumor growth at the site of s.c. cell injection in 19% of 10(7), in 8% of 10(6) and in 5% of 10(5) and 10(4) tumor-cell inoculated mice. In parallel experiments an intravenous (i.v.) inoculum of tumor cells produced lung colonies in 40% of 10(6) and in 14% of 10(5) tumor-cell injected animals. Smaller inocula did not give rise to lung colonies, thus making it unlikely that accidental i.v. inoculations of tumor cells during the s.c. injections caused the observed metastatic dissemination to the lungs.

Adenocarcinoma↗
PubMed1986

Rapid killing of actinomycin D-treated tumor cells by mononuclear phagocytes: characterization of effector cells in mice.

Pretreatment with Actinomycin D (Act D, 1 microgram/ml for 3 hr) rendered WEHI 164 tumor cells susceptible to killing by mouse resident or peptone-induced peritoneal exudate cells (PEC) in a 6-hr 51Cr release assay. Cytotoxicity was attributed to cells of the monocyte macrophage lineage on the basis of tissue distribution, separation by adherence on plastic and carbonyl iron, membrane antigens, and expression in mice with defective T cell- or NK cell-mediated immunity. Macrophages from four strains of mice (C3H/HeJ, A/J, P/J, C57B1/10 ScCR) previously shown to have defective "classical" nonspecific tumoricidal activity were examined for killing of Act-D-treated WEHI 164 cells. C3H/HeJ peritoneal macrophages had little or no DDCC, whereas cells from A/J, P/J, and C57B1/10 ScCR mice had normal levels of this reactivity. Tumor cells exposed to ActD were heterogenous in their susceptibility to killing by PEC, with five lines showing significant, though variable, lysis, whereas 12 tumor lines, normal fibroblasts, and lymphoblasts were not appreciably killed under the same conditions. Macrophage-mediated DDCC was also detectable in a colony assay. DDCC could explain how macrophages contribute to the antitumor activity of selected chemotherapeutic agents in murine tumor models.

Animals↗
PubMed1986

Dissociation between induction of ornithine decarboxylase and oxidative burst by phorbol esters in a macrophage cell line.

In order to investigate the correlation between stimulation of superoxide generation and induction of ornithine decarboxylase (ODC) by 12-O-tetradecanoylphorbol-13-acetate (TPA) we have used the macrophage cell line J774.16 and a clone derived from this line that, by contrast with the parental line, is unable to generate superoxides in response to TPA. No difference was observed between the normal and the defective cells, with respect to ODC induction by TPA over a wide range of TPA concentrations (0.2-5.0 micrograms/ml). Similar results were obtained comparing resident and caseinate-elicited mouse peritoneal macrophages. Although resident macrophages did not generate superoxides in response to TPA, they did not differ from superoxide-generating, caseinate-elicited macrophages with respect to ODC induction. These data suggest a dissociation between the stimulation of the oxidative burst by TPA and a growth factor-like effect such as ODC induction.

Animals↗
PubMed1986

Involvement of tumour necrosis factor in monocyte-mediated rapid killing of actinomycin D-pretreated WEHI 164 sarcoma cells.

Human and murine monocyte-macrophages kill actinomycin D (ActD)-treated WEHI 164 sarcoma cells in a 6-hr 51Cr-release assay (drug-dependent cellular cytotoxicity, DDCC). In this study, we have investigated the cytotoxic activity of human recombinant tumour necrosis factor (hrTNF) against untreated and ActD-treated WEHI 164 sarcoma cells. Human recombinant TNF when added to the 6-hr 51Cr-release assay killed ActD-treated targets at doses ranging from 33 to 0.33 ng/ml, whereas untreated targets were resistant to lysis. The kinetics of lysis of ActD-treated targets was similar for hrTNF and blood monocytes. The protease inhibitors phenyl-methyl-sulphonyl-fluoride (PMSF) and N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) reduced the DDCC activity of monocytes, monocyte supernatants and hrTNF. Killing of drug-sensitized target cells by monocyte supernatants was totally inhibited by a rabbit anti-TNF serum. These, as well as previous data on the physicochemical properties of the soluble cytotoxic factor released by monocytes, suggest that rapid monocyte-mediated killing of ActD-pretreated WEHI 164 sarcoma cells involves TNF or TNF-like molecules.

Animals↗
PubMed1985

Intraperitoneal administration of interferon beta in ovarian cancer patients.

Eight patients with advanced ovarian carcinomas resistant to conventional chemotherapy were injected with interferon (IFN) beta (3 X 10(6) U) intraperitoneally twice a week. Seven subjects had ascites. Side effects included abdominal pain, fever, and constipation, but no hematologic toxicity was observed. Growth of solid tumor lesions was unaffected by IFN beta, with the possible exception of one patient who had stable disease. IFN beta intraperitoneally inhibited completely the formation of ascites in four of seven patients with effusions. Natural killer (NK) cell activity was measured in peripheral blood and tumor-associated lymphocytes (PBL and TAL). Using stringent criteria that included repeated assessment of baseline activity, a clear cut increase in NK cytotoxicity of TAL was detected in two of six subjects from whom TAL could be purified. Augmentation of NK activity was restricted to the peritoneal compartment with no effect on PBL. Studies on biologic response modifiers encompassing an analysis of events taking place at sites directly involved by neoplasia may provide an opportunity for generating information on the in situ regulation of tumor-associated host defense mechanisms in humans.

Adult↗
PubMed1985

Relationship between large granular lymphocytes and NK-1.2+ cells from normal and poly(inosinic:cytidylic acid) (poly(I:C]-treated mice.

The present work analyzes the relationship between large granular lymphocytes (LGL), NK-1.2+ cells, and natural killer (NK) activity of C3H/HeN mice. Different hematic cell fractions were obtained according to their nylon-wool adherence and density on Percoll gradients. NK-1.2+ cells (8% of nucleated cells) were more numerous than LGL (3% of nucleated cells) in the input blood population. Eighty-five percent of LGL were recovered from the sorted NK-1.2+ cell fraction. After incubation on nylon-wool column, 63% of LGL and 36% of NK-1.2+ were eluted in the nonadherent fraction. Eighteen percent of NK-1.2+ cells were recovered from the most adherent elutable cell fraction. After the discontinuous Percoll gradient most LGL were present in the low-density fractions while 20% of NK-1.2+ cells were recovered from the highest-density fraction. NK activity was significant both in the nylon-wool-nonadherent and -adherent fractions. After the Percoll gradient most NK activity was present in the low-density fractions. In the present experimental conditions treatment poly(inosinic:cytidylic acid) (poly(I:C] did not increase the numbers of LGL and NK-1.2+ cells either in the blood or in the spleen. However it increased significantly the NK activity of the input cell populations and of the nonadherent and low-density fractions. Similarly, exposure of specific pathogen-free (SPF) mice to non-SPF conditions stimulated NK cytotoxicity but did not alter the percentage of LGL in the blood or in the spleen. Poly(I:C) treatment induced a shift of LGL and NK-1.2+ cells toward the low-density fractions. In poly(I:C)-treated mice images of granule secretion from LGL were detected. Taken together, the present results indicate that LGL and NK-1.2+ cell populations do not totally overlap. Moreover subpopulations of LGL and NK-1.2+ cells can differ in NK activity, morphology, density, adherence to nylon wool, and response to poly(I:C).

Animals↗
PubMed1985

Large granular lymphocytes from murine blood and intestinal epithelium: comparison of surface antigens, natural killer activity, and morphology.

Large granular lymphocytes obtained from murine blood (B-LGL) and intestinal epithelium (IE-LGL) are cells associated with natural killer (NK) activity and thought to be a first line of defense against tumors and/or infectious organisms. Since B-LGL and IE-LGL represent circulating and mucosal NK effectors, respectively, we compared their surface markers, NK activity and morphology to define possible differences between NK cells in different anatomical compartments. B-LGL and IE-LGL were purified by Percoll gradient centrifugation from nude, normal, and beige C57BL/6 mice. We have defined the following surface phenotypes. B-LGL: In nude mice most of them expressed T-200 (89%), asialo-GM1 (71%), and NK-1.1 (72%); 15% possessed the Thy-1.2 antigen, few cells expressed Ly-2, and none showed Ly-1 positivity. Beige mouse B-LGL were positive for T-200 and NK-1.1. IE-LGL; Nude IE-LGL compared to nude B-LGL showed a similar expression of T-200 and Thy-1.2. Ly-1+ and Ly-2+ cells were more numerous than in B-LGL, whereas NK-1.1+ and asialo-GM1+ cells were less numerous. Interestingly, Ly-2+ IE-LGL were at least partially Thy-1.2-. In euthymic mice IE-LGL had a phenotype comparable to that of nude IE-LGL. The NK activity of B-LGL from nude and normal mice was considerably higher than that of IE-LGL from the corresponding mice. IE-LGL from nude mice possessed larger cytoplasms, and more numerous and bigger azurophilic granules than B-LGL. Similar findings were obtained in normal mice. In beige mice 95% of B-LGL showed a single granule whereas 80% of IE-LGL contained multiple granules (mean 3/cell). Giant granules were frequently found in beige IE-LGL while they were rare in beige B-LGL. Thus, clear differences exist between B-LGL and IE-LGL and they may reflect either different homing patterns of subpopulations of LGL or different stages of maturation of the same lineage of cells.

Animals↗
PubMed1985

Rapid killing of actinomycin D-treated tumor cells--cytotoxicity of cell-free monocyte supernatants.

Human monocytes kill Actinomycin D-treated WEHI 164 sarcoma cells in a 6 h 51Cr release assay (drug dependent cellular cytotoxicity, DDCC). In the present study we have investigated and characterized the human monocyte production of a cytotoxic factor which mediates DDCC. Cell-free supernatants obtained culturing monocytes for 4-5 h kill Actinomycin D-treated WEHI 164 cells but not untreated tumor cells. A series of antiproteases inhibits the cytotoxic activity of cell-free monocyte supernatants, whereas scavengers of reactive oxygen intermediates were ineffective. The lytic activity was destroyed treating supernatants at 100 degrees C for 5 min or by exposure to acid pH or to proteinase K, whereas it was unaffected by heating at 56 degrees C for 30 min. Upon gel filtration on Sephacryl S200, cytolytic activity eluted in the 33,000 molecular weight range.

Catalase↗
PubMed1985

Effects of inactivated streptococci (OK-432) on macrophage functions in mice.

The effects of inactivated streptococci (OK-432) on murine macrophage functions were investigated. In vivo treatment of peritoneal macrophages with OK-432 augmented the direct cytotoxic activity against TU5 tumor cells in a 48 h tritiated thymidine release assay. OK-432 also stimulated the rapid (6 h, 51Cr release) macrophage-mediated killing of Actinomycin D-sensitized WEHI 164 sarcoma cells. Moreover, the expression of la antigens on peritoneal macrophages was found to be greatly enhanced after in vivo treatment with OK-432. The immunomodulatory effects of OK-432 on macrophages functions may contribute to the antitumor activity of inactivated streptococci.

Animals↗
PubMed1985

Rapid killing of actinomycin D-treated tumor cells by human monocytes. II. Cytotoxicity is independent of secretion of reactive oxygen intermediates and is suppressed by protease inhibitors.

Pretreatment with Actinomycin D (ActD, 1 microgram/ml for 3 hr) rendered WEHI 164 tumor cells susceptible to killing by human monocytes in a 6-hr 51Cr release assay. The present study was designed to elucidate the role of reactive oxygen intermediates (ROI) and of proteolytic enzymes in this reactivity. ActD-treated WEHI 164 cells did not trigger any measurable release of O-2 or H2O2 from monocytes. Monocytes exposed to phorbol-12-myristate-13-acetate, which enhanced release of ROI, did not show augmented killing of ActD-treated tumor cells. Scavengers of oxygen metabolites (catalase, superoxide dismutase, gluthatione, and mannitol), which inhibited ROI-mediated PMA-induced monocyte cytotoxicity against erythrocytes, did not affect monocyte killing of ActD-treated WEHI 164 cells. Enzymatically generated ROI with xanthine/xanthine-oxidase glucose/glucose-oxidase did not show preferential killing of ActD-treated WEHI 164 cells. Two patients with chronic granulomatous disease had normal levels of monocyte cytotoxicity against ActD-treated tumor cells. To determine the possible role of proteolytic enzymes in mediating this reactivity, we studied various antiproteases. Organophosphorous agents (DFP and PMSF), chloromethyl-ketone derivatives of tosylamino acids (TLCK and TPCK), Actinomyces products (pepstatin and chymostatin), and the synthetic protease substrate TAME inhibited monocyte-mediated cytotoxicity against ActD-treated WEHI 164 cells. The macromolecular protease inhibitors alpha-1 antitrypsin, bovine pancreatic trypsin inhibitor (BPTI), soybean trypsin inhibitor, and the synthetic protease substrate ATEE had little effect on monocyte cytotoxicity. When monocytes were preincubated with drugs for 1 hr and washed, TLCK, TPCK, and PMSF inhibited cytolysis, whereas the less effective chymostatin and TAME and the inactive BPTI had no effect under these conditions. Inhibition by preincubation with TLCK, PMSF, and TPCK was completely reversed after 6 hr of culture. Supernatants of monocyte cultures had lytic activity against ActD-treated WEHI 164 but not against untreated cells. Antiproteases inhibited the lytic activity of monocyte supernatants. These results strongly suggest that ROI do not play a critical role in monocyte-mediated rapid killing of drug-treated tumor cells, and that proteolytic enzymes are involved in this reactivity.

Adolescent↗
PubMed1984

Arachidonic acid and leukotriene B4 induce aggregation of human peripheral blood mononuclear leucocytes in vitro.

Peripheral human blood mononuclear leucocytes (MNL) aggregated in response to arachidonic acid (AA) in vitro. This phenomenon was similar to that already described for polymorphonuclear cells (PMN). The effect of AA was concentration-dependent and was shared only by the structurally related di-homo-gamma-linolenic acid among the other fatty acids tested. A number of agents able to induce platelet aggregation such as ADP, collagen, serotonin and a stable prostaglandin analogue all failed to stimulate MNL or PMN aggregation. Inhibitors of cyclo-oxygenase activity such as acetylsalicylic acid and indomethacin not only did not prevent AA-induced aggregation, but even potentiated it. In contrast, both nordihydroguaiaretic acid and BW 755C, two inhibitors of cyclo-oxygenase and lipoxygenase, strongly prevented MNL aggregation. Thus AA seems to aggregate MNL through the mediation of lipoxygenase products. This is supported by the observation that leukotriene B4 (LTB4) also induced MNL aggregation. When highly purified lymphocyte and monocyte preparations were assessed separately, the latter responded to AA similarly to mixed MNL whereas lymphocyte aggregation was inconsistent, small and reversible even at high concentrations of AA. Although the pathophysiological significance of the MNL aggregation described here is still obscure, assembly of these cells--particularly monocytes--at the site of injury might be a crucial event.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗
PubMed1984

Rapid killing of actinomycin D-treated tumor cells by human mononuclear cells. I. Effectors belong to the monocyte-macrophage lineage.

Preincubation of tumor cells with actinomycin D (Act D) rendered various murine and human lines susceptible to killing by unstimulated human peripheral blood mononuclear cells (PBM) in a 6-hr 51Cr-release assay. The murine WEHI 164 sarcoma was selected for analysis of drug-dependent cellular cytotoxicity (DDCC) because high levels of killing were detected with this tumor, and it was considerably resistant to natural killer (NK) cell activity. Optimal conditions for induction of susceptibility to lysis included a 3-hr preincubation with 1 microgram/ml Act D. Effector cells of cytotoxicity against Act D-treated WEHI 164 cells were plastic adherent (greater than 85% monocytes). Cells nonadherent to plastic and nylon wool (less than or equal to 1% monocytes) had no appreciable DDCC activity. In contrast NK activity against K562 cells was mediated by nonadherent cells. When PBM were fractionated on a one step discontinuous gradient of Percoll designed to enrich for monocytes (greater than 90% pure), DDCC activity was found in the monocyte fraction, and the lymphoid cell-enriched fraction had no cytotoxicity against Act D-treated WEHI 164 cells. In contrast, NK activity against K562 was recovered with lymphoid cells, and monocytes had no NK cytotoxicity. Upon fractionation on a six step Percoll gradient designed to enrich for large granular lymphocytes (LGL), the denser lymphocytes (fraction 4-6) and the less dense LGL with NK activity (fraction 2-3) had no cytotoxicity against Act D-treated WEHI 164 sarcoma cells. DDCC activity sedimented in fraction 1 in association with monocytes. PBM were fractionated according to monoclonal antibody-defined surface markers by using a fluorescence-activated cell sorter. Effector cells of DDCC were positive for monocyte markers (Mo2, UCHM1) and were negative for NK cell (B73.1, HNK1), T cell (T11), and B cell (Leu-10) markers. Macrophages obtained by culturing blood monocytes in vitro for 5 to 10 days had DDCC activity. Similarly, peritoneal and bronchoalveolar macrophages had considerable cytotoxicity against Act D-treated target cells, whereas minimal or no NK activity was found at these anatomic sites. Cells of human or murine origin, preincubated with Act D for 3 hr, were heterogeneous in their susceptibility to monocyte killing in a 6-hr 51Cr-release assay. High levels of cytotoxicity were observed with the murine WEHI 164 sarcoma and 3T3 "fibroblast" line and with the human CEM leukemia. Monocytes were weakly (but significantly) cytotoxic against the ALAB breast carcinoma (human) and the 8387 sarcoma (human).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1983

Acute lymphoblastic leukemia hand-mirror cells. Study of nine cases.

Acute lymphoblastic leukemia with hand-mirror cells (HMC) was diagnosed in nine adult patients. Blast HMC were seen only in the bone marrow (12-57% range). Cytochemical studies revealed a positive reaction to tartrate-sensitive acid phosphatase in the tail portion of the cells in seven cases, with a strong, localized cytoplasmic reaction in four. Leukemic cells lacked surface immunoglobulins and were E rosette negative in all cases. Normal levels of adenosine deaminase activity (ADA) were found in five of the seven patients. Electron microscope studies confirmed the hand-mirror shape of the cell. These HMC contained large numbers of mitochondria and microspikes in the handle portion of the cell. The patients failed to respond to initial conventional ALL chemotherapy, but the prolonged survival with passable health of the majority of these, despite their lack of complete remission, is emphasized.

Acid Phosphatase↗
PubMed1983

Effect of a streptococcal preparation (OK432) on natural killer activity of tumour-associated lymphoid cells in human ovarian carcinoma and on lysis of fresh ovarian tumour cells.

The streptococcal preparation OK432 was studied for its effects on natural killer (NK) activity of peripheral blood lymphocytes (PBL) from normal donors and from ovarian cancer patients, and of tumour-associated lymphocytes (TAL) from peritoneal effusions. OK432 augmented NK activity against the susceptible K562 line and induced killing of the relatively resistant Raji line. Freshly isolated ovarian carcinoma cells were relatively resistant to killing by unstimulated PBL and TAL. OK432 induced significant, though low, levels of cytotoxicity against 51Cr-labelled ovarian carcinoma cells. Augmentation of killing of fresh tumour cells by OK432 was best observed in a 20 h assay and both autologous and allogeneic targets were lysed. PBL were separated on discontinuous Percoll gradients. Unstimulated and OK432-boosted activity were enriched in the lower density fractions where large granular lymphocytes (LGL) and activity against K562 were found. Thus, OK432 augments NK activity of PBL and TAL in human ovarian carcinomas and induces low, but significant, levels of killing of fresh tumour cells. Effector cells involved in killing of fresh ovarian tumours copurify with LGL on discontinuous gradients of Percoll.

Adult↗