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M P Colombo

Publications and source records attributed to M P Colombo.

At least 145 records · Page 8Linked to original sources
PubMed1985

Adoptive immunotherapy of a BALB/c lymphoma by syngeneic anti-DBA/2 immune lymphoid cells: characterization of the effector population and evidence for the role of the host's non-T cells.

It has been previously shown that the BALB/c lymphoma YC8 is susceptible to lysis by syngeneic anti-DBA/2 lymphocytes and that YC8-bearing BALB/c mice can be cured by adoptive transfer of such immune effectors. In this study in vivo and in vitro functions of the curative immune lymphocytes have been evaluated together with the role of the host immune system in the mechanism of tumor eradication. It was found that the curative anti-DBA/2 lymphocytes were not directly cytotoxic to YC8 cells although they developed into YC8-lytic cells after in vitro restimulation with YC8. In vivo, the immune lymphocytes were able to mediate a tumor-specific delayed type hypersensitivity reaction against YC8 but had a low tumor-neutralizing activity in the Winn assay. Proliferation of infused BALB/c anti-DBA/2 lymphocytes was necessary for the in vivo therapeutic effect, since irradiation of effector cells or treatment of the donor immune lymphocytes with vinblastine abolished their curative capacity. Immunodepression of the T cell compartment of the prospective tumor-bearing animals by thymectomy plus irradiation or its abrogation in B mice (thymectomized, lethally irradiated, and reconstituted with fetal liver cells) did not interfere with the therapeutic effect of the transferred anti-DBA/2 lymphocytes. Blocking the macrophage functions of the host by carrageenan, however, abolished the therapeutic effect of immune lymphocytes. These data indicate that a radiation-resistant, non-T cell is involved in the tumor eradication induced by anti-DBA/2 lymphocytes. It was also shown that cured mice, tested 90 days after therapy, become resistant to 5 X 10(3) LD80 YC8 cells and that this resistance was due to the presence of memory cells derived from the transferred and not from the host lymphocyte population.

Animals↗
PubMed1985

In vivo reexpression of H-2 antigens in B16 melanoma cells.

We have previously shown that B16-A (H-2b) murine melanoma cells, when cultured in vitro for more than ten passages, have an undetectable or reduced expression of H-2Kb and Db antigens, respectively. We have now studied the possibility to restore H-2 expression (measured by quantitative antisera absorption) in B16-A cells either by a limited (30 days) period of in vivo growth or by treatment with immune interferon. In vivo transplants in allogeneic H-2k or H-2d mice and in H-2-compatible but Mls and multiple non-H-2 loci incompatible mice restored the normal expression of Kb and Db antigens. Cells obtained from tumors grown in syngeneic or in minor histocompatibility antigens-allogeneic mice showed only a weak increase in Db antigens. Such induction of H-2 expression appears to be mediated by the host's immune system, since (1) cells obtained from tumors grown in allogeneic BALB/c nude mice expressed much lower levels of H-2 antigens than those from tumors grown in normal BALB/c mice, and (2) it was possible to induce H-2 expression by growing B16 cells in syngeneic C57Bl/6 mice previously allostimulated with unrelated BALB/c tumor. In vitro treatment with immune interferon restored the expression of both Kb and Db antigens. We hypothesize that H-2 reexpression on B16 cells grown in allogeneic hosts could take place via the nonspecific components of the immune response, such as immune interferon.

Animals↗
PubMed1984

Passive adoptive immunotherapy of low-immunogenic BALB/c lymphoma by syngeneic alloimmune T lymphocytes.

The virus-induced BALB/c lymphoma YC8 is known to be lysed in vitro by syngeneic lymphoid cells immune to non-H-2 antigens of B10.D2 and DBA/2 backgrounds. This tumor is weakly immunogenic in vivo and kills 100% of syngeneic mice with 1 X 10(3) cells given either intravenously (i.v.) or intraperitoneally (i.p.). We show here that i.v.-injected YC8 cells grow preferentially in the liver, where colonies become microscopically visible after 7-10 days, and, less frequently, in the kidneys and spleen but not in the lung. Passive adoptive immunotherapy of this tumor was carried out with alloimmune BALB/c anti-B10.A, anti-pool (donors were immunized with lymphocytes from 5 different strains), anti-A and anti-DBA/2 splenic and lymph node cells. When administered i.p. 1, 3 or 5 days after tumor cells had been given i.p. and with a schedule of 5 subsequent daily inocula, anti-DBA/2 lymphocytes cured 100%, 80% and 60% of animals respectively. A weaker effect was obtained with anti-pool immune cells whereas anti-B10.A and anti-A lymphoid cells had not therapeutic effects. When given i.v., the anti-DBA/2 immune lymphocytes were able to cure both i.v. and i.p. tumor-injected mice. A significant effect was observed also when the onset of immunotherapy was delayed until 7 or 10 days after tumor injection. By depleting the BALB/c anti-DBA/2 immune cells with appropriate monoclonal antibodies and complement, it was found that Lyt 1+ 2-cells played the major role in eradicating the neoplasm. in vitro phenotypic and functional analysis showed that the immune cell population included 70% of Thy 1+, 38% of Lyt 1+ and 18-20% of Lyt 2+ cells. Immune lymphocytes were not cytotoxic in vitro to YC8 or DBA/2 targets whereas they proliferated after restimulation with DBA/2 but only weakly with YC8 cells. This shows that it is possible to cure mice bearing a disseminated lymphoma which expresses non-immunogenic antigens recognized by BALB/c anti-DBA/2 immune T lymphocytes. These immune lymphocytes had no cytotoxic activity in vitro and their major effector cell subpopulation displayed the Thy 1+, Lyt 1+ phenotype.

Animals↗
PubMed1983

In vitro detection of cell-mediated immunity to individual tumor-specific antigens of chemically induced BALB/c fibrosarcomas.

In this study we have analyzed the in vitro cell-mediated cytotoxicity of immune peritoneal exudate cells (PEC), elicited in syngeneic mice against the MCA-induced, TSTA-bearing BALB/c fibrosarcoma CA-2, GI-17 and C-3. The 4 h 51Cr-release assay showed the immune PEC effectors to be specifically cytotoxic to fibrosarcoma used for the immunization, but not to other syngeneic MCA-induced tumors or normal fibroblasts. Cold target inhibition experiments on CA-2 cells confirmed the specificity of the reaction. When PEC, lymph-node and spleen cells from BALB/c anti-CA-2 mice were compared for anti-tumor activity, only PEC were found to kill tumor cells significantly. PEC effectors did not have a significant level of NK or NC activities since they were unable to destroy YAC-1 or WEHI-164 tumor cells. PEC anti-CA-2 were analyzed for the expression of T-cell markers by anti-Thy 1.2, anti-Ly 1.2 and Ly 2.2 monoclonal antibodies. Anti-tumor specific effector cells were identified as mature T cells since they were not adherent to plastic and showed Thy 1.2+, Ly 1.2- and Ly 2.2+ phenotypes. In addition, anti-H-2Kd but not anti-H-2Dd alloantiserum added to target cells, blocked CA-2 tumor lysis, thus supporting the conclusion that the T-cell response against TSTA is H-2 restricted.

Animals↗
PubMed1983

Laser oncology in Italy: experimental research and clinical applications at the National Cancer Institute, Milan.

CO2, Nd:YAG, and argon lasers have been used for 5 years for experimental and clinical procedures at the National Cancer Institute of Milan under the auspices of the National Research Council. Experimental data regarding tumor cell spread through lymphatic and hematic channels and cell viability in the fumes after CO2 laser irradiation are reported. New advantages in the combined treatment modalities have been demonstrated, since the synergism between the low power argon laser beam and chemotherapeutic agents (anthracyclines) was assessed on cell cultures and the least toxic and the most effective doses of the drugs were identified. Due to the optical properties of the laser beam, physical measurements of the delivered energy and mathematical calculation of the energy per tissue volume unit to be ablated increase the precision in microsurgery especially when conservative managements are indicated. Main applications in surgical oncology, and comparison versus conventional methods, both with vaporization and excisional techniques, according to specific therapeutic protocols, in general surgery, otolaryngology, gynecology, and endoscopy, even on an outpatient basis, are finally illustrated in a series of 600 cases.

Animals↗
PubMed1983

Expression of H-2 and viral antigens and resistance to the antitumor lysis of tunicamycin-treated MBL-2 lymphoma cells.

The role of protein glycosylation in the tumor lysis mediated by effector cells derived from Moloney-sarcoma-virus(MSV)-immune mice was studied. Treatment of the Moloney-virus-induced H-2b lymphoma target cells, MBL-2, with tunicamycin (TM), an inhibitor of the protein-N-linked glycosilation, was found to cause a loss of susceptibility to lysis by MSV-immune syngeneic effectors cells, while the same target cells remained fully sensitive to the lytic action of anti-H-2b-immune lymphocytes. Examination of MBL-2 cell surface by lactoperoxidase, 125I iodination, and immunoprecipitation by antiviral protein sera revealed that env but not gag viral gene-encoded products were expressed on the surface of this lymphoma. The TM-induced alteration of cell surface expression of H-2Db, H-2Kb, and gp70 antigens was examined by a combined approach of serological and biochemical techniques. The results were concordant in indicating that (1) after 16 h of TM treatment the cells showed a decreased expression of the three glycoproteins, (2) H-2Db (the restriction element in this system) resulted more affected by the treatment than its counterpart H-2Kb (75% vs 50% reduction as compared to untreated cells), (3) an additional lighter form of H-2Kb was found on the surface of TM-treated cells. In the context of an "associative recognition' of Db and gp70 by MSV-immune effector cells, our results may explain the loss of susceptibility to antitumor effectors of TM-treated MBL-2 cells by a quantitative reduction in the expression of both molecules which interact to create the target structure of syngeneic effectors.

Animals↗
PubMed1983

Impaired H-2 expression in B16 melanoma variants.

We studied the expression of H-2b alloantigens in three different B16 melanoma lines cultures in vitro. Cell lines were B16-F1 and two cell cultures (named B16-A and B16-B) newly derived from two different in vivo sublines of B16 melanoma. The assays used were in vivo tumour growth in allogeneic (BALB/c and B10.BR) as compared to syngeneic mice, in vitro cell-mediated cytotoxicity by anti-H-2b immune lymphocytes and absorption of anti-H-2b antisera activity. The B16-F1 line was able to efficiently kill allogeneic hosts, could not be lysed by anti-H-2b cytotoxic effectors and did not express any serologically detectable amount of H-2b alloantigens. The B16-A line was H-2 positive during the early in vitro passages, then, at the 8th-10th passages, it acquired the capacity to kill allogeneic hosts, lost the sensitivity to anti-H-2b cytotoxic effectors and the H-2Kb antigens became undetectable The expression of H-2Db was reduced, although at a lower degree. Similar data were obtained with B16-B cells, which after 10 in vitro passages grew and killed allogeneic hosts, showed a decreased sensitivity to cytotoxic anti-H-2b effectors and a very low expression of the K region antigens. The results indicate that H-2 expression is altered in B16 melanoma lines and this may influence the different metastatic capacity of such cells.

Animals↗
PubMed1983

H-2K restriction of the T cell-mediated lysis of a chemically-induced BALB/c fibrosarcoma.

Previous work has shown that a cytotoxic T lymphocyte (CTL) immune response of syngeneic mice immunized with a chemically-induced BALB/c (H-2d) fibrosarcoma was directed against an individual tumour-associated antigen. To see whether this reaction was restricted by products of the major histocompatibility complex (MHC), anti-H-2 alloantisera to K or D antigens were used to interfere with the CTL-mediated immune response. Antisera to Kd but not to Dd antigens inhibited the lytic activity of CTL against fibrosarcoma cells. In addition, the study of the CTL response in F1 leads to P antitumour immunized chimeric mice showed that antitumour cytotoxicity developed only when F1 and parental host shared the Kd region. Both experiments strongly indicate that recognition of the individual tumour-associated antigen of the BALB/c fibrosarcoma is restricted by the products of H-2Kd genes.

Animals↗
PubMed1981

Importance of non-H-2 genes in the control of the in vivo growth of the virus induced lymphoma YC8.

The genetic control of the in vivo growth of the Moloney virus-induced BALB/c lymphoma YC8 was studied in F1 hybrids between BALB/c and several strains differing at the MHC and/or at the level of non-H-2 genes. Parental strains of the B10 and C3Hf but not of A, BALB/c or DBA/2 backgrounds introduced a significant resistance to the growth of 10(2) YC8 cells (a dose able to kill 100% of BALB/c mice) in semisyngeneic hosts. This resistance appeared to be due to non-H-2 genes although a modulation of the tumour growth by genes encoded by the MHC was also evident. The study of backcrosses between susceptible BALB/c and resistant (BALB/C X B10.BR)F1 crosses revealed that 83% of animals developed lethal tumours after injection of 10(2) YC8 cells. This high frequency of tumour takes was not linked to genes of the MHC. Adult thymectomy plus sublethal irradiation was able to abrogate the resistance of (BALB/c x B10.BR) or (BALB/c x B10.RIII)F1 mice to YC8 growth. Since the injection of silica also impaired the resistance to YC8, we tentatively concluded that the genetic control of resistance to YC8 is mediated both by T cells and macrophage-like cells.

Animals↗
PubMedDate not supplied

Adoptive immunotherapy of cancer with immune and activated lymphocytes: experimental and clinical studies.

Recent studies of passive adoptive immunotherapy of experimental tumors indicate that histologically different neoplasms can be cured by this procedure in mice, rats and guinea pigs. In this paper two main approaches of adoptive immunotherapy with lymphocytes are considered. One which makes use of specific tumor-immune cells and is applicable to immunogenic tumors, and the other which uses activated (allostimulated and/or IL-2-activated) lymphocytes and is applicable to immunogenic and non-immunogenic neoplasms. Experimental models of both approaches and results provided by them are reviewed. These studies indicate that transfer of tumor-reactive lymphocytes with or without the combined administration of IL-2 into syngeneic tumor-bearing animals can lead to the eradication of a disseminated neoplasia when certain conditions are met. In particular, it was found that high tumor burdens, delay of treatment and low number of transferred lymphocytes can adversely affect the results. It has also been shown that the therapeutic effect of treatment with anti-cancer drugs or irradiation may be significantly improved by the addition of adoptive immunotherapy. The successful treatment of immunogenic tumors often requires the inhibition of suppressor lymphocytes by Cy or irradiation. Non-immunogenic tumors can be successfully treated only by providing activated lymphocytes and high doses of IL-2. Recent findings of few available human studies of adoptive immunotherapy are also reviewed, and the problems of toxicity and possible therapeutic effects of infusion of autologous, activated lymphocytes and IL-2 are discussed.

Animals↗
PubMedDate not supplied

In vitro analysis of the melanoma/endothelium interaction increasing the release of soluble intercellular adhesion molecule 1 by endothelial cells.

Melanoma cells constitutively release intercellular adhesion molecule 1 (ICAM-1) as soluble ICAM-1 (sICAM-1), and its levels are elevated in melanoma patients and correlate with disease progression. However, this correlation is not absolute, suggesting that specific characteristics of neoplastic cells and/or ICAM-1-positive non-neoplastic cells may influence the amounts of circulating sICAM-1. In this study, we found a weak correlation (r = 0.55; r2 = 0.3) between sICAM-1 release by 40 metastatic melanomas (36 primary cultures and 4 cell lines), and ICAM-1 expression on neoplastic cells. In addition, melanoma-secreted interleukin-1alpha (IL-1alpha) (1/40) but not vascular endothelial growth factor (VEGF) (29/40), significantly (P < 0.05) up-regulated the shedding of sICAM-1 by human umbilical vein endothelial cells (HUVEC). This was completely abolished by IL-1alpha/beta neutralizing antibodies both at the protein and mRNA level. Altogether, our results suggest that (i) the extent of sICAM-1 release is distinctive for individual melanomas and can be independent of ICAM-1 expression; (ii) tumor endothelia may sustain levels of sICAM-1 in selected melanomas; (iii) melanoma-released VEGF does not affect ICAM-1 expression and sICAM-1 release by HUVEC. Melanoma-derived sICAM-1 inhibits cell-mediated cytotoxicity of melanoma cells; therefore, constitutive levels of sICAM-1 release and IL-1alpha secretion by individual melanomas can differentially influence tumor progression and the clinical effectiveness of cytotoxic-cell-based vaccines.

Cells, Cultured↗
PubMedDate not supplied

Immunotherapy. I: Cytokine gene transfer strategies.

The cytokine approach to gene therapy of cancer stems from early studies of direct, repeated injection of recombinant cytokines at the tumor site, and extension of the bystander effect that enables a few cytokine gene transduced cells in a tumor to bring about its total destruction. This effect can be extended through the immune system, since cytokine-activated regression of a small mass of tumor cells can afford systemic protection. Transduced cells used as a vaccine provide a local concentration of both cytokine and tumor antigens. Cytokines sustain antigen uptake and presentation by increasing the immunogenic potential of the environment through the recruitment of antigen presenting cells and leukocytes, and activation of a cascade of events which amplify and tone up the efficacy of a vaccine. The promises and difficulties of this approach are discussed by considering what is still missing from experimental studies and what can best be done as soon as possible in animals and humans to reach compelling conclusions.

Animals↗
PubMedDate not supplied

DNA vaccination against the ovarian carcinoma-associated antigen folate receptor alpha (FRalpha) induces cytotoxic T lymphocyte and antibody responses in mice.

Human folate receptor alpha (FRalpha) is a folate-binding protein that is selectively overexpressed in ovarian carcinoma and has been regarded as a suitable target antigen for immunotherapy purposes. To study the possible use of this antigen in DNA vaccination, FRalpha cDNA was ligated into the VR1012 (Vical) expression vector under the transcriptional control of the cytomegalovirus promoter. A total of 100 microg of purified plasmid DNA was injected intramuscularly in BALB/c mice three times at 14-day intervals. At 10 days after the second injection, the sera of the animals (100%) displayed significant antibody titers (by indirect immunofluorescence and fluorescence-activated cell sorter analysis) against syngeneic C26 cells transduced with FRalpha, but not against unmodified C26 cells. Immunoglobulin G2a was the predominant isotype. In addition, specific cytotoxic T lymphocyte activity against FRalpha-transduced C26 cells could be detected in splenocytes from all immunized animals. Coinjection of a plasmid containing interleukin-2 cDNA increased both antibody titers and cytotoxic T lymphocyte activity. Challenge by subcutaneous injection with FRalpha-transduced C26 cells (performed 10 days after the third injection) showed a statistically significant delay in tumor growth. Vaccination with the FRalpha and interleukin-2 cDNA mixture, which was performed after an intravenous injection of FRalpha-transduced cells, enhanced the mean survival time and reduced the number of lung metastases, thus suggesting that such vaccination is effective even against preexisting tumor cells.

Animals↗
PubMedDate not supplied

Prevention by delay: nonspecific immunity elicited by IL-12 hinders Her-2/neu mammary carcinogenesis in transgenic mice.

As a natural consequence of the expression of the activated transforming rat Her-2/neu oncogene all mammary glands of female transgenic BALB/c (BALB-neuT) mice develop atypical epithelial hyperplasia which progresses to invasive carcinoma. A lobular carcinoma is palpable in all mammary glands of 33-week-old BALB-neuT mice. This progression is markedly delayed by systemic administration of IL-12. In a series of studies the best administration schedule, the lowest dose and the most effective administration time have been defined. The cellular and molecular mechanisms resulting in the delay of carcinogenesis have been established. By means of a series of downstream mediators IL-12 inhibits the angiogenic burst that goes along with the passage from preneoplastic to neoplastic and invasive lesions; it also recruits lymphoid cells in the mammary pad and activates their cytotoxicity towards neoplastic cells and newly formed vessels; and furthermore, it induces lymphoid cells to trigger antiangiogenic activities in neoplastic epithelial cells. Effective, low-dose and non-toxic IL-12 treatments may thus be envisaged as a possible option in the management of preneoplastic mammary lesions and in mammary cancer prevention.

Animals↗
PubMedDate not supplied

Cytokine gene transduction in tumor cells: interleukin (IL)-2 or IL-4 gene transfer in human melanoma cells.

Cytokine gene transfer into mouse tumor cells has been shown to stimulate a strong immune response resulting in the rejection of the transduced tumor when injected in vivo. Therefore, retroviral vectors containing the human interleukin (IL)-2 or IL-4 gene have been constructed to transduce human melanoma cells to explore whether their immunogenicity can be increased both in vitro and in vivo. Our preliminary results indicate that retroviral vectors can efficiently transduce the IL-2 and or IL-4 gene into melanoma clones, inducing production of either cytokine in the range of 0.5-2 ng/ml/10(5) cells in 48-72 h. No modifications of the growth rate, morphology and antigenicity of the transduced tumor cells were found.

Cytotoxicity Tests, Immunologic↗