1 Department of Medical Oncology, Disease Center for Hematologic Neoplasia, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA, USA; 2 Molecular Tumorbiology and Tumorimmunology, Department for Hematology/Oncology, University of Cologne, Germany
Received 15 August 2003; revised 23 November 2003; accepted 23 December 2003
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ABSTRACT |
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Key words: antigen presentation, CD40, CD154, CLL, immunotherapy
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Introduction |
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Molecular and immunological characterization of B-CLL |
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CLL cells not only lack costimulatory molecules, they also actively secrete immune inhibitory cytokines including IL-10 and TGF-ß, which suppress antigen-specific responses in advanced cancer and granulomatous diseases, such as sarcoidos [9, 10]. Furthermore, T-cells in CLL patients are often dysfunctional as demonstrated by poor IL-2 secretion and proliferative responses to mitogens or alloantigens [11].
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CD40 in normal B-cells and B-CLL |
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Anti-apoptotic effects of CD40 signaling |
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CD40 activation induces APC function in B-CLL cells |
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Another approach to upregulate accessory and costimulatory molecules on B-CLL cells is adenoviral transfection of CLL cells [31]. Initial studies showed technical difficulties, suggesting that transfection of the human molecule might be more difficult than the murine homolog and that activation might be a prerequisite of successful transfection with CD40L (CD154) [32, 33]. Using an optimized adenovirus-free packaging system Wendtner et al. first demonstrated efficient infection of resting CLL cells using recombinant adeno-associated virus (rAAV) vectors encoding CD40 ligand (MOI 100; efficiency 97%). This resulted in upregulation of CD80 in infected and noninfected bystander cells, inducing proliferation of allogeneic T-cells [34]. This effect can be amplified by transduction of leukemic cells with the interleukin-2 gene [35]. Briones et al. demonstrated the potential of this approach in a murine lymphoma model: vaccination using A20 lymphoma cells adenovirally transduced with CD40-L successfully induced rejection of a lethal dose of parental cells [36]. Furthermore, recent observations suggest that difficulty in CD40L transfection can be over-come by simple coculture of CLL cells with CD40L-transfected bystander cells [37].
To better quantify the effect of CD40 ligation on B-CLL cells and to optimize the approach for vaccination strategies we dissected the effects of CD40 activation on CLL cells by quantifying the density of adhesion, costimulatory and MHC molecules. B-CLL cells activated via CD40 in the presence of IL-4 and INF- upregulate the adhesion molecule CD58, CD80 and MHC-II molecules to levels comparable with immature dendritic cells (iDC), while MHC class-I molecules were induced to levels higher than iDC. Correspondingly, presentation of alloantigen by CD40-CLL cells was comparable to normal CD40-activated B-cells and strongest when activated in the presence of IL-4 and INF-
(Figure 1). Non-activated B-CLL cells secreted little IL-6 or IL-12. Similar to normal B-cells, IL-12 but not IL-6 secretion could be upregulated when CD40-CLL cells were cultured in the presence of INF-
, suggesting that this feedback mechanism is still intact in B-CLL cells [38]. While CD40 activation and INF-
also induced IL-10 and TGF-ß secretion by B-CLL cells, the addition of IL-4 antagonized this effect. Furthermore, CD40-activated B-CLL cells induced T-cells to secrete GM-CSF and INF-
. This secretion is reduced when B-CLL cells were cocultured with IL-10 and partially restored when INF-
was present in the B-CLL culture. Thus generation of CD40-CLL cells for vaccination purposes might be optimized by co-activation with IL-4 and/or INF-
and simultaneous blockade of IL-10 (Schultze et al., unpublished results).
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Use of normal CD40-activated B-cells for the presentation of tumor-derived antigens |
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Vaccination using B-CLL cells transfected with CD40L |
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Future directions |
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Treatment at early stages or post-transplantation will more likely be settings in which induction of anti-tumor immunity could prove beneficial. It remains to be determined, if cellular adjuvants can be substituted by FDA-approved reagents that can be produced on a large scale.
Taken together, there is a clear incentive to further develop CD40-based strategies into novel therapies for CLL. However, several biological questions need to be addressed and in light of increasing regulatory difficulties for cellular therapies and the fast development of alternative approaches, it needs to be carefully evaluated whether these exciting but complex vaccines have the chance to become standard care for CLL.
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FOOTNOTES |
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