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Induction of tumor-specific T cell immunity by anti-DR5 antibody therapy.

Takeda K, Yamaguchi N, Akiba H, Kojima Y, Hayakawa Y, Tanner JE, Sayers TJ, Seki N, Okumura K, Yagita H, Smyth MJ - J. Exp. Med. (2004)

Bottom Line: Here we report that a monoclonal antibody (mAb) against the mouse TRAIL receptor, DR5, exhibited potent antitumor effects against TRAIL-sensitive tumor cells in vivo by recruiting Fc receptor-expressing innate immune cells, with no apparent systemic toxicity.Administration of the agonistic anti-DR5 mAb also significantly inhibited experimental and spontaneous tumor metastases.These results suggested that the antibody-based therapy targeting DR5 is an efficient strategy not only to eliminate TRAIL-sensitive tumor cells, but also to induce tumor-specific T cell memory that affords a long-term protection from tumor recurrence.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Juntendo University School of Medicine, 2-1-1 Hongo, Bukyou-ku, Tokyo 113-8421, Japan. ktakeda@med.juntendo.ac.jp

ABSTRACT
Because tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) preferentially induces apoptosis in tumor cells and plays a critical role in tumor surveillance, its receptor is an attractive target for antibody-mediated tumor therapy. Here we report that a monoclonal antibody (mAb) against the mouse TRAIL receptor, DR5, exhibited potent antitumor effects against TRAIL-sensitive tumor cells in vivo by recruiting Fc receptor-expressing innate immune cells, with no apparent systemic toxicity. Administration of the agonistic anti-DR5 mAb also significantly inhibited experimental and spontaneous tumor metastases. Notably, the anti-DR5 mAb-mediated tumor rejection by innate immune cells efficiently evoked tumor-specific T cell immunity that could also eradicate TRAIL-resistant variants. These results suggested that the antibody-based therapy targeting DR5 is an efficient strategy not only to eliminate TRAIL-sensitive tumor cells, but also to induce tumor-specific T cell memory that affords a long-term protection from tumor recurrence.

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Cytotoxic activity of MD5-1 in the presence of FcR-bearing cells. (A) FcR-mediated cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of fresh or fixed effector cells. CMA was added to inhibit perforin-dependent cytotoxicity and 2.4G2 was used to block FcγRII and FcγRIII. (B) Cytotoxic activity of MD5-1 against R331-mock and R331-FLIP tumor cells in the presence of various effector cells. (C) Cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of rabbit or mouse complement.
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fig2: Cytotoxic activity of MD5-1 in the presence of FcR-bearing cells. (A) FcR-mediated cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of fresh or fixed effector cells. CMA was added to inhibit perforin-dependent cytotoxicity and 2.4G2 was used to block FcγRII and FcγRIII. (B) Cytotoxic activity of MD5-1 against R331-mock and R331-FLIP tumor cells in the presence of various effector cells. (C) Cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of rabbit or mouse complement.

Mentions: Next, we examined the ability of FcR-expressing effector cells including fresh macrophages and NK cells to mediate MD5-1 cytotoxicity in vitro because these innate immune cells are well-known mediators of antibody-dependent cellular cytotoxicity (ADCC) in vivo (31, 32). Macrophages induced MD5-1 cytotoxicity against 4T1 cells more effectively than NK cells, and both were inhibited by anti-FcR mAb (Fig. 2 A). MD5-1 cytotoxicity was also induced by NK cells in the presence or absence of a perforin inhibitor, CMA, suggesting that MD5-1 did not trigger the perforin-dependent ADCC by NK cells in vitro (Fig. 2 A). Consistent with this notion, macrophages and NK cells from perforin−/− mice or FasL mutant gld mice induced MD5-1 cytotoxicity against 4T1 cells at a comparable level to that induced by wild-type cells (unpublished data). Importantly, A20 cells that only express FcγRII, macrophages isolated from FcRγ−/− mice, and paraformaldehyde-fixed FcR+ cells, all triggered MD5-1–mediated cytotoxicity, confirming that cross-linking was sufficient to induce DR5- and caspase-mediated cell death (Fig. 2, A and B). However, live (fresh) effector cells always induced higher levels of MD5-1 cytotoxicity than those observed with fixed effector cells, suggesting some contribution of ADCC, especially by macrophages (Fig. 2 A). All effector cells also triggered cytotoxicity against R331-mock cells in the presence of MD5-1, however, R331-FLIP cells were rather refractory (Fig. 2 B). Some residual MD5-1 cytotoxicity against R331-FLIP cells in the presence of macrophages or NK cells suggested that these effector cells were capable of mediating a small part of MD5-1–dependent cell death independently of caspase activation. Collectively, these results indicated that MD5-1 was capable of triggering FcR-dependent cytolysis of tumor cells in vitro by cross-linking DR5 and FcR (even FcγRII), inducing both DR5-mediated caspase-dependent cell death and caspase-independent ADCC. Notably, MD5-1 did not kill tumor cells in the presence of rabbit or mouse complement (Fig. 2 C). The fact that macrophages and NK cells effectively mediated MD5-1 cytotoxicity via their FcR implied a possible cytotoxic effect of MD5-1 in vivo.


Induction of tumor-specific T cell immunity by anti-DR5 antibody therapy.

Takeda K, Yamaguchi N, Akiba H, Kojima Y, Hayakawa Y, Tanner JE, Sayers TJ, Seki N, Okumura K, Yagita H, Smyth MJ - J. Exp. Med. (2004)

Cytotoxic activity of MD5-1 in the presence of FcR-bearing cells. (A) FcR-mediated cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of fresh or fixed effector cells. CMA was added to inhibit perforin-dependent cytotoxicity and 2.4G2 was used to block FcγRII and FcγRIII. (B) Cytotoxic activity of MD5-1 against R331-mock and R331-FLIP tumor cells in the presence of various effector cells. (C) Cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of rabbit or mouse complement.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2211825&req=5

fig2: Cytotoxic activity of MD5-1 in the presence of FcR-bearing cells. (A) FcR-mediated cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of fresh or fixed effector cells. CMA was added to inhibit perforin-dependent cytotoxicity and 2.4G2 was used to block FcγRII and FcγRIII. (B) Cytotoxic activity of MD5-1 against R331-mock and R331-FLIP tumor cells in the presence of various effector cells. (C) Cytotoxic activity of MD5-1 against 4T1 tumor cells in the presence of rabbit or mouse complement.
Mentions: Next, we examined the ability of FcR-expressing effector cells including fresh macrophages and NK cells to mediate MD5-1 cytotoxicity in vitro because these innate immune cells are well-known mediators of antibody-dependent cellular cytotoxicity (ADCC) in vivo (31, 32). Macrophages induced MD5-1 cytotoxicity against 4T1 cells more effectively than NK cells, and both were inhibited by anti-FcR mAb (Fig. 2 A). MD5-1 cytotoxicity was also induced by NK cells in the presence or absence of a perforin inhibitor, CMA, suggesting that MD5-1 did not trigger the perforin-dependent ADCC by NK cells in vitro (Fig. 2 A). Consistent with this notion, macrophages and NK cells from perforin−/− mice or FasL mutant gld mice induced MD5-1 cytotoxicity against 4T1 cells at a comparable level to that induced by wild-type cells (unpublished data). Importantly, A20 cells that only express FcγRII, macrophages isolated from FcRγ−/− mice, and paraformaldehyde-fixed FcR+ cells, all triggered MD5-1–mediated cytotoxicity, confirming that cross-linking was sufficient to induce DR5- and caspase-mediated cell death (Fig. 2, A and B). However, live (fresh) effector cells always induced higher levels of MD5-1 cytotoxicity than those observed with fixed effector cells, suggesting some contribution of ADCC, especially by macrophages (Fig. 2 A). All effector cells also triggered cytotoxicity against R331-mock cells in the presence of MD5-1, however, R331-FLIP cells were rather refractory (Fig. 2 B). Some residual MD5-1 cytotoxicity against R331-FLIP cells in the presence of macrophages or NK cells suggested that these effector cells were capable of mediating a small part of MD5-1–dependent cell death independently of caspase activation. Collectively, these results indicated that MD5-1 was capable of triggering FcR-dependent cytolysis of tumor cells in vitro by cross-linking DR5 and FcR (even FcγRII), inducing both DR5-mediated caspase-dependent cell death and caspase-independent ADCC. Notably, MD5-1 did not kill tumor cells in the presence of rabbit or mouse complement (Fig. 2 C). The fact that macrophages and NK cells effectively mediated MD5-1 cytotoxicity via their FcR implied a possible cytotoxic effect of MD5-1 in vivo.

Bottom Line: Here we report that a monoclonal antibody (mAb) against the mouse TRAIL receptor, DR5, exhibited potent antitumor effects against TRAIL-sensitive tumor cells in vivo by recruiting Fc receptor-expressing innate immune cells, with no apparent systemic toxicity.Administration of the agonistic anti-DR5 mAb also significantly inhibited experimental and spontaneous tumor metastases.These results suggested that the antibody-based therapy targeting DR5 is an efficient strategy not only to eliminate TRAIL-sensitive tumor cells, but also to induce tumor-specific T cell memory that affords a long-term protection from tumor recurrence.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Juntendo University School of Medicine, 2-1-1 Hongo, Bukyou-ku, Tokyo 113-8421, Japan. ktakeda@med.juntendo.ac.jp

ABSTRACT
Because tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) preferentially induces apoptosis in tumor cells and plays a critical role in tumor surveillance, its receptor is an attractive target for antibody-mediated tumor therapy. Here we report that a monoclonal antibody (mAb) against the mouse TRAIL receptor, DR5, exhibited potent antitumor effects against TRAIL-sensitive tumor cells in vivo by recruiting Fc receptor-expressing innate immune cells, with no apparent systemic toxicity. Administration of the agonistic anti-DR5 mAb also significantly inhibited experimental and spontaneous tumor metastases. Notably, the anti-DR5 mAb-mediated tumor rejection by innate immune cells efficiently evoked tumor-specific T cell immunity that could also eradicate TRAIL-resistant variants. These results suggested that the antibody-based therapy targeting DR5 is an efficient strategy not only to eliminate TRAIL-sensitive tumor cells, but also to induce tumor-specific T cell memory that affords a long-term protection from tumor recurrence.

Show MeSH
Related in: MedlinePlus