Enhanced Antitumor Immunity Elicited by Dendritic Cell Vaccines Is a Result of Their Ability to Engage Both CTL and IFNγ-producing NK Cells

Activation of cytotoxic T lymphocytes (CTLs) is a primary goal of many cancer vaccination therapies. We have evaluated two vaccination platforms, dendritic cells (DCs) and recombinant adenoviruses (rAds), for their ability to elicit CTL response and antitumor protection. Although rAd was more potent in CTL priming, DC vaccination provided greater protective and therapeutic antitumor activity. Subsequent analyses ruled out the possibility that the two vaccines elicit qualitatively distinct CTL, and demonstrated instead that DCs could better engage natural killer (NK) cells as an additional effector mechanism. We demonstrated that, although both DCs and rAd can stimulate rapid NK expansion, only DC-activated NK cells are able to produce interferon-γ (IFNγ) and mediate antitumor protection. Moreover, our data showed that exogenously delivered DCs preferentially engaged the Mac-1highCD27high NK subset, thereby suggesting that this NK population plays a predominant role in NK:DC interaction. Interestingly, at least 3 days were required for DC-triggered NK cells to acquire effector functions, indicating that a similar priming process operates between T cells and NK cells. Our results suggest that the nature of the vaccine platform can determine the relative involvement of NK and T cells in antitumor immunity, and that methods to augment NK function should be included in vaccination strategies in order to complement CTL-mediated control of tumor growth. Activation of cytotoxic T lymphocytes (CTLs) is a primary goal of many cancer vaccination therapies. We have evaluated two vaccination platforms, dendritic cells (DCs) and recombinant adenoviruses (rAds), for their ability to elicit CTL response and antitumor protection. Although rAd was more potent in CTL priming, DC vaccination provided greater protective and therapeutic antitumor activity. Subsequent analyses ruled out the possibility that the two vaccines elicit qualitatively distinct CTL, and demonstrated instead that DCs could better engage natural killer (NK) cells as an additional effector mechanism. We demonstrated that, although both DCs and rAd can stimulate rapid NK expansion, only DC-activated NK cells are able to produce interferon-γ (IFNγ) and mediate antitumor protection. Moreover, our data showed that exogenously delivered DCs preferentially engaged the Mac-1highCD27high NK subset, thereby suggesting that this NK population plays a predominant role in NK:DC interaction. Interestingly, at least 3 days were required for DC-triggered NK cells to acquire effector functions, indicating that a similar priming process operates between T cells and NK cells. Our results suggest that the nature of the vaccine platform can determine the relative involvement of NK and T cells in antitumor immunity, and that methods to augment NK function should be included in vaccination strategies in order to complement CTL-mediated control of tumor growth. IntroductionMost cancer vaccines have aimed at eliciting CD8+ cytotoxic T lymphocyte (CTL) activity because CTLs have the ability to directly eliminate tumor cells that express specific antigens. In this regard, both the quantity and quality (i.e., functional avidity) of antigen-specific CTLs are critical to the efficacy of antitumor responses.1Yang S Tsang KY Schlom J Induction of higher-avidity human CTLs by vector-mediated enhanced costimulation of antigen-presenting cells.Clin Cancer Res. 2005; 11: 5603-5615Crossref PubMed Scopus (33) Google Scholar,2Zeh HJ 3rd Perry-Lalley D Dudley ME Rosenberg SA Yang JC High avidity CTLs for two self-antigens demonstrate superior in vitro and in vivo antitumor efficacy.J Immunol. 1999; 162: 989-994PubMed Google Scholar However, tumors often express low levels of major histocompatibility complex class I molecules, thereby enabling their escape from CTL-mediated elimination. Ultimate control of tumor growth will therefore require additional effector mechanisms.3Gilboa E How tumors escape immune destruction and what we can do about it.Cancer Immunol Immunother. 1999; 48: 382-385Crossref PubMed Scopus (123) Google Scholar Studies have shown that natural killer (NK) cells can spontaneously detect and lyse transformed cells, possibly as a result of low major histocompatibility complex class I expression, thereby controlling tumor initiation and spread.4Mocikat R Braumuller J Gumy A Egeter O Ziegler H Reusch U et al.Natural killer cells activated by MHC class I(low) targets prime dendritic cells to induce protective CD8 T cell responses.Immunity. 2003; 19: 561-569Abstract Full Text Full Text PDF PubMed Scopus (342) Google Scholar,5Glas R Franksson L Une C Eloranta M Ohlen C Orn A et al.Recruitment and activation of natural killer (NK) cells in vivo determined by the target cell phenotype. An adaptive component of NK cell-mediated responses.J Exp Med. 2000; 191: 129-138Crossref PubMed Scopus (132) Google Scholar Additionally, NK cells are able to kill tumors that express normal levels of major histocompatibility complex class I, by activating the receptors that recognize specific ligands expressed by many tumor cells.6Cerwenka A Baron JL Lanier LL Ectopic expression of retinoic acid early inducible-1 gene (RAE-1) permits natural killer cell-mediated rejection of a MHC class I-bearing tumor in vivo.Proc Natl Acad Sci USA. 2001; 98: 11521-11526Crossref PubMed Scopus (493) Google Scholar Furthermore, upon activation, NK cells secrete cytokines such as interferon-γ (IFNγ) that can modulate both innate and adaptive antitumor activities.7Mailliard RB Son YI Redlinger R Coates PT Giermasz A Morel PA et al.Dendritic cells mediate NK cell help for Th1 and CTL responses: two-signal requirement for the induction of NK cell helper function.J Immunol. 2003; 171: 2366-2373Crossref PubMed Scopus (309) Google Scholar,8Martin-Fontecha A Thomsen LL Brett S Gerard C Lipp M Lanzavecchia A et al.Induced recruitment of NK cells to lymph nodes provides IFN-gamma for T(H)1 priming.Nat Immunol. 2004; 5: 1260-1265Crossref PubMed Scopus (1100) Google Scholar Taken together, these data suggest important roles for both CTL and NK cells in antitumor immunity.Two vaccination platforms, dendritic cells (DCs) and recombinant adenoviruses (rAds), have been intensively evaluated in animal models as as in A C therapeutic vaccination Immunol. 2005; PubMed Scopus Google S of dendritic cells as a to induce Immunol. Google Scholar DCs are important for the induction of a immune because have a to cell and CD8+ T cells. can mediate expression in a of cell in thereby to antigen-specific CD8+ S of dendritic cells as a to induce Immunol. Google J of the CD8+ response 2003; PubMed Scopus Google Scholar Moreover, have shown that both DCs and can engage NK cells of the tumor the for cancer vaccines to target effector H Yang H et and cells to tumor in the Immunol. PubMed Scopus Google A stimulate natural killer cell and demonstrate antitumor in the of 5: Full Text Full Text PDF PubMed Scopus Google E activation of adaptive and innate antitumor Immunol. 2003; 171: PubMed Scopus Google Scholar We have in tumor that vaccines are more potent in eliciting CTL vaccines DC vaccination provides better antitumor J R J cells an a for tumor PubMed Scopus Google J A J dendritic cells prime T cells a of and for cancer vaccines Cancer Res. 2000; Google J C et and antitumor is determined by the tumor the Immunol. 2004; PubMed Scopus Google Scholar was the is by the quality of the CTLs by the two vaccination platforms, as by et E functional and of CTL recombinant Immunol. 2005; PubMed Scopus Google Scholar by the involvement of NK cells as an additional effector a of primary CTL by DC and rAd and a similar quality of CTL However, the two in their to stimulate NK functional DCs and could induce rapid and activation of NK cells in the lymph the of and activated NK cells was and more in as a result of In the of specific tumor antitumor was only in in an analyses that NK cells more and the NK cells thereby suggesting that NK cells by DC have an additional process of functional We that the superior antitumor by DC vaccination was of the of antigen-specific CTL and activated NK and vaccines elicit qualitatively CD8+ the of and in a in were of cells days vaccines provided a of the that were a control DCs tumors in thereby that antitumor was specific However, were of tumor cells by was the of rAd to protective efficacy and data In of the cells and more the superior efficacy of DC vaccination was in a therapeutic the rAd the DC vaccine was days of cells that the levels of antitumor by and DCs be by in the levels of CTL both and that the of the antigen-specific CD8+ response by was greater that by In vivo CTL demonstrated that of target cells was greater in the was to to the quantity of CTL However, that the CD8+ T cells by both vaccines to the of in of thereby suggesting functional Moreover, the of CTL response was in and this ruled out the possibility that CD8+ T cell activated by DCs more of of antigen-specific CD8+ T cells from provided control of tumor growth Taken together, these data suggest that, in to CTL, to the protective response by DC T lymphocyte (CTL) by dendritic cell and rAd vaccines do in of CD8+ T cells was determined by and interferon-γ (IFNγ) data from are from target cells were for the in vivo CTL were target cell and the of antigen-specific was determined by from at this from were for of and for CD8 and to determine functional avidity of antigen-specific CD8+ T cells. data from days were the of CD8+ T cells in the was determined by of in of CD8+ T cells from the and lymph nodes of were that been cells were tumor and tumor the of were greater and activation of NK cells in the lymph rAd and DC vaccines are of activating NK cells, could the of antitumor H Yang H et and cells to tumor in the Immunol. 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NK cells were by and cells at a of for tumor cells were to and were and of tumor cells acid in was at were and of and was for indicating analyses were out IntroductionMost cancer vaccines have aimed at eliciting CD8+ cytotoxic T lymphocyte (CTL) activity because CTLs have the ability to directly eliminate tumor cells that express specific antigens. In this regard, both the quantity and quality (i.e., functional avidity) of antigen-specific CTLs are critical to the efficacy of antitumor responses.1Yang S Tsang KY Schlom J Induction of higher-avidity human CTLs by vector-mediated enhanced costimulation of antigen-presenting cells.Clin Cancer Res. 2005; 11: 5603-5615Crossref PubMed Scopus (33) Google Scholar,2Zeh HJ 3rd Perry-Lalley D Dudley ME Rosenberg SA Yang JC High avidity CTLs for two self-antigens demonstrate superior in vitro and in vivo antitumor efficacy.J Immunol. 1999; 162: 989-994PubMed Google Scholar However, tumors often express low levels of major histocompatibility complex class I molecules, thereby enabling their escape from CTL-mediated elimination. Ultimate control of tumor growth will therefore require additional effector mechanisms.3Gilboa E How tumors escape immune destruction and what we can do about it.Cancer Immunol Immunother. 1999; 48: 382-385Crossref PubMed Scopus (123) Google Scholar Studies have shown that natural killer (NK) cells can spontaneously detect and lyse transformed cells, possibly as a result of low major histocompatibility complex class I expression, thereby controlling tumor initiation and spread.4Mocikat R Braumuller J Gumy A Egeter O Ziegler H Reusch U et al.Natural killer cells activated by MHC class I(low) targets prime dendritic cells to induce protective CD8 T cell responses.Immunity. 2003; 19: 561-569Abstract Full Text Full Text PDF PubMed Scopus (342) Google Scholar,5Glas R Franksson L Une C Eloranta M Ohlen C Orn A et al.Recruitment and activation of natural killer (NK) cells in vivo determined by the target cell phenotype. An adaptive component of NK cell-mediated responses.J Exp Med. 2000; 191: 129-138Crossref PubMed Scopus (132) Google Scholar Additionally, NK cells are able to kill tumors that express normal levels of major histocompatibility complex class I, by activating the receptors that recognize specific ligands expressed by many tumor cells.6Cerwenka A Baron JL Lanier LL Ectopic expression of retinoic acid early inducible-1 gene (RAE-1) permits natural killer cell-mediated rejection of a MHC class I-bearing tumor in vivo.Proc Natl Acad Sci USA. 2001; 98: 11521-11526Crossref PubMed Scopus (493) Google Scholar Furthermore, upon activation, NK cells secrete cytokines such as interferon-γ (IFNγ) that can modulate both innate and adaptive antitumor activities.7Mailliard RB Son YI Redlinger R Coates PT Giermasz A Morel PA et al.Dendritic cells mediate NK cell help for Th1 and CTL responses: two-signal requirement for the induction of NK cell helper function.J Immunol. 2003; 171: 2366-2373Crossref PubMed Scopus (309) Google Scholar,8Martin-Fontecha A Thomsen LL Brett S Gerard C Lipp M Lanzavecchia A et al.Induced recruitment of NK cells to lymph nodes provides IFN-gamma for T(H)1 priming.Nat Immunol. 2004; 5: 1260-1265Crossref PubMed Scopus (1100) Google Scholar Taken together, these data suggest important roles for both CTL and NK cells in antitumor immunity.Two vaccination platforms, dendritic cells (DCs) and recombinant adenoviruses (rAds), have been intensively evaluated in animal models as as in A C therapeutic vaccination Immunol. 2005; PubMed Scopus Google S of dendritic cells as a to induce Immunol. Google Scholar DCs are important for the induction of a immune because have a to cell and CD8+ T cells. can mediate expression in a of cell in thereby to antigen-specific CD8+ S of dendritic cells as a to induce Immunol. Google J of the CD8+ response 2003; PubMed Scopus Google Scholar Moreover, have shown that both DCs and can engage NK cells of the tumor the for cancer vaccines to target effector H Yang H et and cells to tumor in the Immunol. PubMed Scopus Google A stimulate natural killer cell and demonstrate antitumor in the of 5: Full Text Full Text PDF PubMed Scopus Google E activation of adaptive and innate antitumor Immunol. 2003; 171: PubMed Scopus Google Scholar We have in tumor that vaccines are more potent in eliciting CTL vaccines DC vaccination provides better antitumor J R J cells an a for tumor PubMed Scopus Google J A J dendritic cells prime T cells a of and for cancer vaccines Cancer Res. 2000; Google J C et and antitumor is determined by the tumor the Immunol. 2004; PubMed Scopus Google Scholar was the is by the quality of the CTLs by the two vaccination platforms, as by et E functional and of CTL recombinant Immunol. 2005; PubMed Scopus Google Scholar by the involvement of NK cells as an additional effector a of primary CTL by DC and rAd and a similar quality of CTL However, the two in their to stimulate NK functional DCs and could induce rapid and activation of NK cells in the lymph the of and activated NK cells was and more in as a result of In the of specific tumor antitumor was only in in an analyses that NK cells more and the NK cells thereby suggesting that NK cells by DC have an additional process of functional We that the superior antitumor by DC vaccination was of the of antigen-specific CTL and activated NK cells.

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