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Combined allogeneic tumor cell vaccination and systemic interleukin 12 prevents mammary carcinogenesis in HER-2/neu transgenic mice.

Nanni P, Nicoletti G, De Giovanni C, Landuzzi L, Di Carlo E, Cavallo F, Pupa SM, Rossi I, Colombo MP, Ricci C, Astolfi A, Musiani P, Forni G, Lollini PL - J. Exp. Med. (2001)

Bottom Line: IL-12 treatment strongly increased the cell vaccine efficacy.Specific anti-HER-2/neu antibodies were produced and a nonpolarized activation of CD4(+) and CD8(+) cells secreting IL-4 and interferon (IFN)-gamma were evident.In conclusion, our data show that an allogeneic HER-2/neu-expressing cell vaccine combined with IL-12 systemic treatment can prevent the onset of genetically determined tumors.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Section, Department of Experimental Pathology, University of Bologna, Italy.

ABSTRACT
Transgenic Balb/c mice expressing the transforming rat HER-2/neu oncogene develop early and multifocal mammary carcinomas. Within the first 5 months of life the tissue-specific expression of HER-2/neu causes a progression in all their 10 mammary glands from atypical hyperplasia to invasive carcinoma. It was previously observed that chronic administration of interleukin (IL)-12 increased tumor latency, but every mouse eventually succumbed to multiple carcinomas. A significant improvement in tumor prevention was sought by administering allogeneic mammary carcinoma cells expressing HER-2/neu combined with systemic IL-12. This treatment reduced tumor incidence by 90% and more than doubled mouse lifetime. For the maximum prevention p185(neu) antigen must be expressed by allogeneic cells. IL-12 treatment strongly increased the cell vaccine efficacy. The mammary glands of mice receiving the combined treatment displayed a markedly reduced epithelial cell proliferation, angiogenesis, and HER-2/neu expression, while the few hyperplastic foci were heavily infiltrated by granulocytes, macrophages, and CD8(+) lymphocytes. Specific anti-HER-2/neu antibodies were produced and a nonpolarized activation of CD4(+) and CD8(+) cells secreting IL-4 and interferon (IFN)-gamma were evident. A central role for IFN-gamma in the preventive effect was proven by the lack of efficacy of vaccination in IFN-gamma gene knockout HER-2/neu transgenic Balb/c mice. A possible requirement for IFN-gamma is related to its effect on antibody production, in particular on IgG2a and IgG2b subclasses, that were not induced in IFN-gamma knockout HER-2/neu mice. In conclusion, our data show that an allogeneic HER-2/neu-expressing cell vaccine combined with IL-12 systemic treatment can prevent the onset of genetically determined tumors.

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Release of cytokines by spleen cells restimulated or not in vitro with Neu/H-2q cells. Spleen cells from untreated control mice (white bar); from mice treated with IL-12 (gray bar); from mice treated with Neu/H-2q cells (cross-hatched bar); and from mice treated with Neu/H-2q cells plus IL-12 (black bar). Asterisks denote a significant difference (P < 0.05 at least by Student's t test) with untreated controls. Mean ± SEM of groups of 3–5 Balb-neuT mice.
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fig6: Release of cytokines by spleen cells restimulated or not in vitro with Neu/H-2q cells. Spleen cells from untreated control mice (white bar); from mice treated with IL-12 (gray bar); from mice treated with Neu/H-2q cells (cross-hatched bar); and from mice treated with Neu/H-2q cells plus IL-12 (black bar). Asterisks denote a significant difference (P < 0.05 at least by Student's t test) with untreated controls. Mean ± SEM of groups of 3–5 Balb-neuT mice.

Mentions: At the fifteenth week of age, after the second course, groups of 3–5 control and treated mice were killed to evaluate spleen cell reactivity. Both total spleen cell yield and the absolute number of CD4+ cells were significantly higher in mice treated with Neu/H-2q cells plus IL-12. However, CTL assays against HER-2/neu-positive or -negative targets showed only ∼10% of specific lysis of HER-2/neu-positive targets by spleen cells from Balb-neuT mice vaccinated with Neu/H-2q cells plus IL-12. These spleen cells did not display any protective activity when they were admixed with a tumorigenic dose of Neu/H-2d syngeneic mammary tumor cells in a Winn-type neutralization assay in Balb-neuT mice (data not shown). On the other hand, an increased spontaneous proliferation was displayed in vitro by the CD4+ lymphocyte subpopulation. It was further increased in an antigen-specific manner by the addition of mitomycin-blocked syngeneic Neu/H-2d cells. Moreover, total spleen cells and CD8+ lymphocytes of mice repeatedly treated with Neu/H-2q cells plus IL-12 spontaneously released more IFN-γ than leukocytes from control or IL-12–treated mice (Fig. 6) . Large amounts of IFN-γ were released by total and CD8+ spleen cells after restimulation by Neu/H-2q cells. IL-4 was secreted spontaneously or after restimulation by both total spleen cells and CD4+ lymphocytes (but not by CD8+ lymphocytes) from mice treated with Neu/H-2q cells plus IL-12. This cytokine release pattern was triggered by the recognition of both H-2q allogeneic MHC glycoproteins and p185neu since IFN-γ and IL-4 were produced not only in response to p185neu plus alloantigens (Neu/H-2q) but also by the presentation of p185neu on syngeneic cells (Neu/H-2d) as well as by alloantigens alone (Neuneg/H-2q) (Table II).


Combined allogeneic tumor cell vaccination and systemic interleukin 12 prevents mammary carcinogenesis in HER-2/neu transgenic mice.

Nanni P, Nicoletti G, De Giovanni C, Landuzzi L, Di Carlo E, Cavallo F, Pupa SM, Rossi I, Colombo MP, Ricci C, Astolfi A, Musiani P, Forni G, Lollini PL - J. Exp. Med. (2001)

Release of cytokines by spleen cells restimulated or not in vitro with Neu/H-2q cells. Spleen cells from untreated control mice (white bar); from mice treated with IL-12 (gray bar); from mice treated with Neu/H-2q cells (cross-hatched bar); and from mice treated with Neu/H-2q cells plus IL-12 (black bar). Asterisks denote a significant difference (P < 0.05 at least by Student's t test) with untreated controls. Mean ± SEM of groups of 3–5 Balb-neuT mice.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195980&req=5

fig6: Release of cytokines by spleen cells restimulated or not in vitro with Neu/H-2q cells. Spleen cells from untreated control mice (white bar); from mice treated with IL-12 (gray bar); from mice treated with Neu/H-2q cells (cross-hatched bar); and from mice treated with Neu/H-2q cells plus IL-12 (black bar). Asterisks denote a significant difference (P < 0.05 at least by Student's t test) with untreated controls. Mean ± SEM of groups of 3–5 Balb-neuT mice.
Mentions: At the fifteenth week of age, after the second course, groups of 3–5 control and treated mice were killed to evaluate spleen cell reactivity. Both total spleen cell yield and the absolute number of CD4+ cells were significantly higher in mice treated with Neu/H-2q cells plus IL-12. However, CTL assays against HER-2/neu-positive or -negative targets showed only ∼10% of specific lysis of HER-2/neu-positive targets by spleen cells from Balb-neuT mice vaccinated with Neu/H-2q cells plus IL-12. These spleen cells did not display any protective activity when they were admixed with a tumorigenic dose of Neu/H-2d syngeneic mammary tumor cells in a Winn-type neutralization assay in Balb-neuT mice (data not shown). On the other hand, an increased spontaneous proliferation was displayed in vitro by the CD4+ lymphocyte subpopulation. It was further increased in an antigen-specific manner by the addition of mitomycin-blocked syngeneic Neu/H-2d cells. Moreover, total spleen cells and CD8+ lymphocytes of mice repeatedly treated with Neu/H-2q cells plus IL-12 spontaneously released more IFN-γ than leukocytes from control or IL-12–treated mice (Fig. 6) . Large amounts of IFN-γ were released by total and CD8+ spleen cells after restimulation by Neu/H-2q cells. IL-4 was secreted spontaneously or after restimulation by both total spleen cells and CD4+ lymphocytes (but not by CD8+ lymphocytes) from mice treated with Neu/H-2q cells plus IL-12. This cytokine release pattern was triggered by the recognition of both H-2q allogeneic MHC glycoproteins and p185neu since IFN-γ and IL-4 were produced not only in response to p185neu plus alloantigens (Neu/H-2q) but also by the presentation of p185neu on syngeneic cells (Neu/H-2d) as well as by alloantigens alone (Neuneg/H-2q) (Table II).

Bottom Line: IL-12 treatment strongly increased the cell vaccine efficacy.Specific anti-HER-2/neu antibodies were produced and a nonpolarized activation of CD4(+) and CD8(+) cells secreting IL-4 and interferon (IFN)-gamma were evident.In conclusion, our data show that an allogeneic HER-2/neu-expressing cell vaccine combined with IL-12 systemic treatment can prevent the onset of genetically determined tumors.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Section, Department of Experimental Pathology, University of Bologna, Italy.

ABSTRACT
Transgenic Balb/c mice expressing the transforming rat HER-2/neu oncogene develop early and multifocal mammary carcinomas. Within the first 5 months of life the tissue-specific expression of HER-2/neu causes a progression in all their 10 mammary glands from atypical hyperplasia to invasive carcinoma. It was previously observed that chronic administration of interleukin (IL)-12 increased tumor latency, but every mouse eventually succumbed to multiple carcinomas. A significant improvement in tumor prevention was sought by administering allogeneic mammary carcinoma cells expressing HER-2/neu combined with systemic IL-12. This treatment reduced tumor incidence by 90% and more than doubled mouse lifetime. For the maximum prevention p185(neu) antigen must be expressed by allogeneic cells. IL-12 treatment strongly increased the cell vaccine efficacy. The mammary glands of mice receiving the combined treatment displayed a markedly reduced epithelial cell proliferation, angiogenesis, and HER-2/neu expression, while the few hyperplastic foci were heavily infiltrated by granulocytes, macrophages, and CD8(+) lymphocytes. Specific anti-HER-2/neu antibodies were produced and a nonpolarized activation of CD4(+) and CD8(+) cells secreting IL-4 and interferon (IFN)-gamma were evident. A central role for IFN-gamma in the preventive effect was proven by the lack of efficacy of vaccination in IFN-gamma gene knockout HER-2/neu transgenic Balb/c mice. A possible requirement for IFN-gamma is related to its effect on antibody production, in particular on IgG2a and IgG2b subclasses, that were not induced in IFN-gamma knockout HER-2/neu mice. In conclusion, our data show that an allogeneic HER-2/neu-expressing cell vaccine combined with IL-12 systemic treatment can prevent the onset of genetically determined tumors.

Show MeSH
Related in: MedlinePlus