Breaking Tolerance in a Mouse Model of Multiple Myeloma by Chemoimmunotherapy

Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel.
Advances in Cancer Research (Impact Factor: 5.32). 12/2010; 107:1-37. DOI: 10.1016/S0065-230X(10)07001-6
Source: PubMed


A unique mouse model of multiple myeloma (MM), namely 5T2MM-bearing mouse, was useful for elucidating the pathophysiological mechanisms underlying the disease. Increased accumulation of suppressive CD4(+)CD25(High)Foxp3(+) regulatory T cells (Tregs) was observed in the thymus and lymphoid peripheral organs during disease progression. Adoptive transfer of Tregs, but not other thymocytes, from 5T2MM-bearing mice led to increased progression of disease manifestations in young syngeneic mice. Depletion of Tregs, a proposed strategy in cancer immunotherapy, was tested using cyclophosphamide (CYC), an alkylating agent with selective cytotoxicity. Both low- and high-dose CYC, administered to sick mice with hind limb paralysis, caused the paralysis to disappear, the plasma tumor cells in the bone marrow (BM) cavity to be replaced by normal cell populations, and the survival of the mice to be significantly prolonged. Low-dose CYC, which selectively depletes Tregs, decreased MM incidence, in contrast to high-dose CYC, which was generally cytotoxic, and did not reduce MM incidence. In contrast, low-dose CYC induced Tregs to become susceptible to apoptosis by downregulating Bcl-xL and CTLA-4 in these cells, and by decreasing the production of IL-2 by effector CD4 cells. This treatment consequently triggered the recovery of IFN-gamma-producing natural killer T cells and the maturation of dendritic cells. Transient gradual depletion of Tregs in low-dose CYC-treated 5T2MM mice was maintained beyond 45 days. Thus, less frequent injections of low-dose CYC enabled us to recruit compatible immune-derived cells that would reduce tumor load and delay or prevent tumor recurrence, hence breaking immune tolerance toward MM tumor cells.

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    • "A promising alternative strategy is the development of specific immunotherapies selectively targeting myeloma cells10,11,12,13. However, a major problem in this area is the immune tolerance to tumor cells and tumor-associated antigens14,15. To overcome this problem, this study examined the potential of improving the antigenicity of myeloma through metabolic engineering of its cell surface carbohydrate antigens. "
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    ABSTRACT: Aim: To study the antitumor effect of anti-NprPSA monoclonal antibody (mAb) in combination with ManNPr, a precursor of N-propionyl PSA, in multiple myeloma (MM), and to explore the mechanisms of the action. Methods: Human multiple myeloma cell line RPMI-8226 was tested. The cells were pre-treated with ManNPr (1, 2, and 4 mg/mL), and then incubated with anti-NprPSA mAb (1 mg/mL). Cell apoptosis in vitro was detected using MTT assay and flow cytometry. BALB/c nude mice were inoculated sc with RPC5.4 cells. On 5 d after the injection, the mice were administered sc with anti-NprPSA mAb (200 μg/d) and ManNPr (5 mg/d) for 8 d. The tumor size and body weight were monitored twice per week. TUNEL assay was used for detecting apoptosis in vivo. The apoptotic pathway involved was examined using Western blot analysis and caspase inhibitor. Results: Treatment of RPMI-8226 cells with anti-NprPSA mAb alone failed to inhibit cell growth in vitro. In RPMI-8226 cells pretreated with ManNPr, however, the mAb significantly inhibited the cell proliferation, decreased the viability, and induced apoptosis, which was associated with cleavage of caspase-3, caspase-8, caspase-9, and poly(ADP-ribose) polymerase. In the mouse xenograft model, treatment with the mAb in combination with ManNPr significantly inhibited the tumor growth, and induced significant apoptosis as compared to treatment with the mAb alone. Moreover, apoptosis induced by the mAb in vivo resulted from the activation of the caspases and poly(ADP-ribose) polymerase. Conclusion: The anti-NprPSA mAb in combination with ManNPr is an effective treatment for in vitro and in vivo induction of apoptosis in multiple myeloma.
    Full-text · Article · Nov 2012 · Acta Pharmacologica Sinica
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    • "Modeling of naïve T cells as a separate entity will also allow us to study the effects of adoptive transfers, which are currently under clinical investigation for several cancer types. One could also incorporate immune cells such as T regulatory cells which are implicated in inducing tolerance and model the recovery of responsive TCD8 cells (Sharabi and Ghera, 2010). Moreover, the case of uncontrolled T cell and Tag-expressing cell growth is similar to autoimmune response and though it is not a focus of the current study, the model can be used to study the relationship between tumor and autoimmunity since tolerance (Schuetz et al., 2010) and dysregulation of immune responses (Reeves et al., 2009) are implicated in both diseases. "
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    ABSTRACT: CD8+ T cells have the potential to influence the outcome of cancer pathogenesis, including complete tumor eradication or selection of malignant tumor escape variants. The Simian virus 40 large T-antigen (Tag) oncoprotein promotes tumor formation in Tag-transgenic mice and also provides multiple target determinants (sites) for responding CD8+ T cells in C57BL/6 (H-2(b)) mice. To understand the in vivo quantitative dynamics of CD8+ T cells after encountering Tag, we constructed a dynamic model from in vivo-generated data to simulate the interactions between Tag-expressing cells and CD8+ T cells in distinct scenarios including immunization of wild-type C57BL/6 mice and of Tag-transgenic mice that develop various tumors. In these scenarios the model successfully reproduces the dynamics of both the Tag-expressing cells and antigen-specific CD8+ T cell responses. The model predicts that the tolerance of the site-specific T cells is dependent on their apoptosis rates and that the net growth of CD8+ T cells is altered in transgenic mice. We experimentally validate both predictions. Our results indicate that site-specific CD8+ T cells have tissue-specific apoptosis rates affecting their tolerance to the tumor antigen. Moreover, the model highlights differences in apoptosis rates that contribute to compromised CD8+ T cell responses and tumor progression, knowledge of which is essential for development of cancer immunotherapy.
    Full-text · Article · Jul 2011 · Frontiers in Physiology
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    • "Continuous low-dose CYC treatments at 45-day intervals were shown to be most effective in reducing and/or preventing MM development [4] [59]. Moreover, even when the initial injection of a high-dose CYC was chosen, further repeated injections of low-dose CYC at 45-day intervals prevented the disease [59]. Less frequent administrations of low-dose CYC enabled recruitment of compatible immune-derived cells that reduced tumor load and prolonged survival with minimal residual disease. "
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    ABSTRACT: Multiple myeloma (MM) is a progressive B-lineage neoplasia characterized by clonal proliferation of malignant plasma cells. Increased numbers of regulatory T cells (Tregs) were determined in mouse models and in patients with MM, which correlated with disease burden. Thus, it became rational to target Tregs for treating MM. The effects of common chemotherapeutic drugs on Tregs are reviewed with a focus on cyclophosphamide (CYC). Studies indicated that selective depletion of Tregs may be accomplished following the administration of a low-dose CYC. We report that continuous nonfrequent administrations of CYC at low doses block the renewal of Tregs in MM-affected mice and enable the restoration of an efficient immune response against the tumor cells, thereby leading to prolonged survival and prevention of disease recurrence. Hence, distinctive time-schedule injections of low-dose CYC are beneficial for breaking immune tolerance against MM tumor cells.
    Full-text · Article · Apr 2011
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