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Cancer Immunology, Immunotherapy
Published in: Cancer Immunology, Immunotherapy 12/2011

01-12-2011 | Original article

Ab-IL2 fusion proteins mediate NK cell immune synapse formation by polarizing CD25 to the target cell-effector cell interface

Authors: Jennifer A. A. Gubbels, Brian Gadbaw, Ilia N. Buhtoiarov, Sachi Horibata, Arvinder K. Kapur, Dhara Patel, Jacquelyn A. Hank, Stephen D. Gillies, Paul M. Sondel, Manish S. Patankar, Joseph Connor

Published in: Cancer Immunology, Immunotherapy | Issue 12/2011

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Abstract

The huKS-IL2 immunocytokine (IC) consists of IL2 fused to a mAb against EpCAM, while the hu14.18-IL2 IC recognizes the GD2 disialoganglioside. They are under evaluation for treatment of EpCAM+ (ovarian) and GD2+ (neuroblastoma and melanoma) malignancies because of their proven ability to enhance tumor cell killing by antibody-dependent cell-mediated cytotoxicity (ADCC) and by antitumor cytotoxic T cells. Here, we demonstrate that huKS-IL2 and hu14.18-IL2 bind to tumor cells via their antibody components and increase adhesion and activating immune synapse (AIS) formation with NK cells by engaging the immune cells’ IL-2 receptors (IL2R). The NK leukemia cell line, NKL (which expresses high affinity IL2Rs), shows fivefold increase in binding to tumor targets when treated with IC compared to matching controls. This increase in binding is effectively inhibited by blocking antibodies against CD25, the α-chain of the IL2R. NK cells isolated from the peritoneal environment of ovarian cancer patients, known to be impaired in mediating ADCC, bind to huKS-IL2 via CD25. The increased binding between tumor and effector cells via ICs is due to the formation of AIS that are characterized by the simultaneous polarization of LFA-1, CD2 and F-actin at the cellular interface. AIS formation of peritoneal NK and NKL cells is inhibited by anti-CD25 blocking antibody and is 50–200% higher with IC versus the parent antibody. These findings demonstrate that the IL-2 component of the IC allows IL2Rs to function not only as receptors for this cytokine but also as facilitators of peritoneal NK cell binding to IC-coated tumor cells.
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Literature
1.
Richards JM (1989) Therapeutic uses of interleukin-2 and lymphokine-activated killer (LAK) cells. Blood Rev 3:110–119PubMedCrossRef
2.
Curiel TJ, Coukos G, Zou L, Alvarez X, Cheng P, Mottram P, Evdemon-Hogan M, Conejo-Garcia JR, Zhang L, Burow M, Zhu Y, Wei S, Kryczek I, Daniel B, Gordon A, Myers L, Lackner A, Disis ML, Knutson KL, Chen L, Zou W (2004) Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med 10:942–949PubMedCrossRef
3.
Nelson BH (2008) The impact of T-cell immunity on ovarian cancer outcomes. Immunol Rev 222:101–116PubMedCrossRef
4.
Bubenik J, Indrova M, Perlmann P, Berzins K, Mach O, Kraml J, Toulcova A (1985) Tumour inhibitory effects of TCGF/IL-2/-containing preparations. Cancer Immunol Immunother 19:57–61PubMedCrossRef
5.
Bubenik J, Perlmann P, Indrova M, Simova J, Jandlova T, Neuwirt J (1983) Growth inhibition of an MC-induced mouse sarcoma by TCGF (IL 2)-containing preparations. Preliminary report. Cancer Immunol Immunother 14:205–206PubMedCrossRef
6.
Rosenberg SA, Lotze MT, Muul LM, Chang AE, Avis FP, Leitman S, Linehan WM, Robertson CN, Lee RE, Rubin JT et al (1987) A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high-dose interleukin-2 alone. N Engl J Med 316:889–897PubMedCrossRef
7.
Lotze MT, Matory YL, Rayner AA, Ettinghausen SE, Vetto JT, Seipp CA, Rosenberg SA (1986) Clinical effects and toxicity of interleukin-2 in patients with cancer. Cancer 58:2764–2772PubMedCrossRef
8.
Lode HN, Xiang R, Becker JC, Gillies SD, Reisfeld RA (1998) Immunocytokines: a promising approach to cancer immunotherapy. Pharmacol Ther 80:277–292PubMedCrossRef
9.
Reisfeld RA, Becker JC, Gillies SD (1997) Immunocytokines: a new approach to immunotherapy of melanoma. Melanoma Res 7(Suppl 2):S99–S106
10.
Connor JP, Felder M, Hank J, Harter J, Gan J, Gillies SD, Sondel P (2004) Ex vivo evaluation of anti-EpCAM immunocytokine huKS-IL2 in ovarian cancer. J Immunother 27:211–219PubMedCrossRef
11.
Neal ZC, Yang JC, Rakhmilevich AL, Buhtoiarov IN, Lum HE, Imboden M, Hank JA, Lode HN, Reisfeld RA, Gillies SD, Sondel PM (2004) Enhanced activity of hu14.18-IL2 immunocytokine against murine NXS2 neuroblastoma when combined with interleukin 2 therapy. Clin Cancer Res 10:4839–4847PubMedCrossRef
12.
Osenga KL, Hank JA, Albertini MR, Gan J, Sternberg AG, Eickhoff J, Seeger RC, Matthay KK, Reynolds CP, Twist C, Krailo M, Adamson PC, Reisfeld RA, Gillies SD, Sondel PM (2006) A phase I clinical trial of the hu14.18-IL2 (EMD 273063) as a treatment for children with refractory or recurrent neuroblastoma and melanoma: a study of the Children’s Oncology Group. Clin Cancer Res 12:1750–1759PubMedCrossRef
13.
Johnson EE, Lum HD, Rakhmilevich AL, Schmidt BE, Furlong M, Buhtoiarov IN, Hank JA, Raubitschek A, Colcher D, Reisfeld RA, Gillies SD, Sondel PM (2008) Intratumoral immunocytokine treatment results in enhanced antitumor effects. Cancer Immunol Immunother 57:1891–1902PubMedCrossRef
14.
Wagner K, Schulz P, Scholz A, Wiedenmann B, Menrad A (2008) The targeted immunocytokine L19-IL2 efficiently inhibits the growth of orthotopic pancreatic cancer. Clin Cancer Res 14:4951–4960PubMedCrossRef
15.
Schliemann C, Palumbo A, Zuberbuhler K, Villa A, Kaspar M, Trachsel E, Klapper W, Menssen HD, Neri D (2009) Complete eradication of human B-cell lymphoma xenografts using rituximab in combination with the immunocytokine L19-IL2. Blood 113:2275–2283PubMedCrossRef
16.
Wong JY, Chu DZ, Williams LE, Liu A, Zhan J, Yamauchi DM, Wilczynski S, Wu AM, Yazaki PJ, Shively JE, Leong L, Raubitschek AA (2006) A phase I trial of (90)Y-DOTA-anti-CEA chimeric T84.66 (cT84.66) radioimmunotherapy in patients with metastatic CEA-producing malignancies. Cancer Biother Radiopharm 21:88–100PubMedCrossRef
17.
Wong JYC, Chu DZ, Yamauchi DM, Williams LE, Liu A, Wilczynski S, Wu AM, Shively JE, Doroshow JH, Raubitschek AA (2000) A phase I radioimmunotherapy trial evaluating 90yttrium-labeled anti-carcinoembryonic antigen (CEA) chimeric T84.66 in patients with metastatic CEA-producing malignancies. Clin Cancer Res 6:3855–3863PubMed
18.
Lyu MA, Kurzrock R, Rosenblum MG (2008) The immunocytokine scFv23/TNF targeting HER-2/neu induces synergistic cytotoxic effects with 5-fluorouracil in TNF-resistant pancreatic cancer cell lines. Biochem Pharmacol 75:836–846PubMedCrossRef
19.
Gillies SD, Reilly EB, Lo KM, Reisfeld RA (1992) Antibody-targeted interleukin 2 stimulates T-cell killing of autologous tumor cells. Proc Natl Acad Sci USA 89:1428–1432PubMedCrossRef
20.
Xiang R, Lode HN, Dolman CS, Dreier T, Varki NM, Qian X, Lo KM, Lan Y, Super M, Gillies SD, Reisfeld RA (1997) Elimination of established murine colon carcinoma metastases by antibody-interleukin 2 fusion protein therapy. Cancer Res 57:4948–4955PubMed
21.
Ko YJ, Bubley GJ, Weber R, Redfern C, Gold DP, Finke L, Kovar A, Dahl T, Gillies SD (2004) Safety, pharmacokinetics, and biological pharmacodynamics of the immunocytokine EMD 273066 (huKS-IL2): results of a phase I trial in patients with prostate cancer. J Immunother 27:232–239PubMedCrossRef
22.
Shusterman S, London W, Gillies SD, Hank JA, Voss S, Seeger RC, Reynolds CP, Kimball J, Albertini MA, Wagner B, Gan J, Eickhoff J, DeSantes KD, Cohn SL, Hecht T, Gadbaw B, Reisfeld RA, Maris JM, Sondel PM (2010) Anti-tumor activity of hu14.18-IL2 in relapsed/refractory neuroblastoma patients: a Children’s Oncology Group (COG) phase II study. J Clin Oncol 28:4969–4975PubMedCrossRef
23.
Yu AL, Gilman A, Ozkaynak MF, London WB, Kreissman S, Chen H, Smith M, Anderson B, Villablanca J, Matthay KK, Shimada H, Grupp SA, Seeger R, Reynolds CP, Buxton A, Reisfeld RA, Gillies SD, Cohn SL, Maris JM, Sondel PM (2010) Chimeric Anti-GD2 antibody with GM-CSF, IL2 and 13-cis retinoic acid for high-risk neuroblastoma: a children’s oncology group (COG) phase 3 study. N Engl J Med 363:1324–1334
24.
Becker JC, Pancook JD, Gillies SD, Furukawa K, Reisfeld RA (1996) T cell-mediated eradication of murine metastatic melanoma induced by targeted interleukin 2 therapy. J Exp Med 183:2361–2366PubMedCrossRef
25.
Pancook JD, Becker JC, Gillies SD, Reisfeld RA (1996) Eradication of established hepatic human neuroblastoma metastases in mice with severe combined immunodeficiency by antibody-targeted interleukin-2. Cancer Immunol Immunother 42:88–92PubMedCrossRef
26.
Becker JC, Pancook JD, Gillies SD, Mendelsohn J, Reisfeld RA (1996) Eradication of human hepatic and pulmonary melanoma metastases in SCID mice by antibody-interleukin 2 fusion proteins. Proc Natl Acad Sci USA 93:2702–2707PubMedCrossRef
27.
Gillies SD, Lan Y, Lo KM, Super M, Wesolowski J (1999) Improving the efficacy of antibody-interleukin 2 fusion proteins by reducing their interaction with Fc receptors. Cancer Res 59:2159–2166PubMed
28.
Neal ZC, Imboden M, Rakhmilevich AL, Kim KM, Hank JA, Surfus J, Dixon JR, Lode HN, Reisfeld RA, Gillies SD, Sondel PM (2004) NXS2 murine neuroblastomas express increased levels of MHC class I antigens upon recurrence following NK-dependent immunotherapy. Cancer Immunol Immunother 53:41–52PubMedCrossRef
29.
Gillies SD, Lan Y, Williams S, Carr F, Forman S, Raubitschek A, Lo KM (2005) An anti-CD20-IL-2 immunocytokine is highly efficacious in a SCID mouse model of established human B lymphoma. Blood 105:3972–3978PubMedCrossRef
30.
Lustgarten J, Marks J, Sherman LA (1999) Redirecting effector T cells through their IL-2 receptors. J Immunol 162:359–365PubMed
31.
Buhtoiarov IN, Neal Z, Jan J, Buhtoiarova TN, Hank JA, Yamane B, Rakhmilevich AL, Patankar MS, Gubbels JAA, Reisfeld RA, Gillies SD, Sondel PM (2010) Differential internalization of hu14.18-IL2 by human NK cells and human tumor cells: influence on conjugate formation, cytotoxicity and infiltration into tumors. J Leukoc Biol 89:625–638
32.
Belisle JA, Gubbels JA, Raphael CA, Migneault M, Rancourt C, Connor JP, Patankar MS (2007) Peritoneal natural killer cells from epithelial ovarian cancer patients show an altered phenotype and bind to the tumour marker MUC16 (CA125). Immunology 122:418–429PubMedCrossRef
33.
Davis DM, Chiu I, Fassett M, Cohen GB, Mandelboim O, Strominger JL (1999) The human natural killer cell immune synapse. Proc Natl Acad Sci USA 96:15062–15067PubMedCrossRef
34.
Joshi T, Ganesan LP, Cheney C, Ostrowski MC, Muthusamy N, Byrd JC, Tridandapani S (2009) The PtdIns 3-kinase/Akt pathway regulates macrophage-mediated ADCC against B cell lymphoma. PLoS ONE 4:e4208PubMedCrossRef
35.
Gubbels JA, Belisle J, Onda M, Rancourt C, Migneault M, Ho M, Bera TK, Connor J, Sathyanarayana BK, Lee B, Pastan I, Patankar MS (2006) Mesothelin-MUC16 binding is a high affinity, N-glycan dependent interaction that facilitates peritoneal metastasis of ovarian tumors. Mol Cancer 5:50PubMedCrossRef
36.
Gubbels JA, Felder M, Horibata S, Belisle JA, Kapur A, Holden H, Petrie S, Migneault M, Rancourt C, Connor JP, Patankar MS (2010) MUC16 provides immune protection by inhibiting synapse formation between NK and ovarian tumor cells. Mol Cancer 9:11PubMedCrossRef
37.
Zheng X, Wang Y, Wei H, Sun R, Tian Z (2009) LFA-1 and CD2 synergize for the ERK1/2 activation in the NK cell immunological synapse. J Biol Chem 284:21280–21287
38.
Robertson MJ, Cochran KJ, Cameron C, Le JM, Tantravahi R, Ritz J (1996) Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. Exp Hematol 24:406–415PubMed
39.
Lanier LL, Ruitenberg JJ, Phillips JH (1988) Functional and biochemical analysis of CD16 antigen on natural killer cells and granulocytes. J Immunol 141:3478–3485PubMed
40.
Lanier LL (2008) Up on the tightrope: natural killer cell activation and inhibition. Nat Immunol 9:495–502PubMedCrossRef
41.
42.
Rosenstein M, Yron I, Kaufmann Y, Rosenberg SA (1984) Lymphokine-activated killer cells: lysis of fresh syngeneic natural killer-resistant murine tumor cells by lymphocytes cultured in interleukin 2. Cancer Res 44:1946–1953PubMed
43.
Hatakeyama M, Minamoto S, Taniguchi T (1986) Intracytoplasmic phosphorylation sites of Tac antigen (p55) are not essential for the conformation, function, and regulation of the human interleukin 2 receptor. Proc Natl Acad Sci USA 83:9650–9654PubMedCrossRef
44.
Rickert M, Wang X, Boulanger MJ, Goriatcheva N, Garcia KC (2005) The structure of interleukin-2 complexed with its alpha receptor. Science 308:1477–1480PubMedCrossRef
45.
Wang X, Rickert M, Garcia KC (2005) Structure of the quaternary complex of interleukin-2 with its alpha, beta, and gammac receptors. Science 310:1159–1163PubMedCrossRef
46.
47.
Greene WC, Leonard WJ (1987) Human interleukin 2 receptor (Tac antigen). Methods Enzymol 150:682–700PubMedCrossRef
48.
49.
Lode HN, Xiang R, Dreier T, Varki NM, Gillies SD, Reisfeld RA (1998) Natural killer cell-mediated eradication of neuroblastoma metastases to bone marrow by targeted interleukin-2 therapy. Blood 91:1706–1715PubMed
50.
Gestwicki JE, Cairo CW, Strong LE, Oetjen KA, Kiessling LL (2002) Influencing receptor-ligand binding mechanisms with multivalent ligand architecture. J Am Chem Soc 124:14922–14933PubMedCrossRef
51.
Orange JS, Harris KE, Andzelm MM, Valter MM, Geha RS, Strominger JL (2003) The mature activating natural killer cell immunologic synapse is formed in distinct stages. Proc Natl Acad Sci USA 100:14151–14156PubMedCrossRef
52.
van Spriel AB, Leusen JH, van Egmond M, Dijkman HB, Assmann KJ, Mayadas TN, van de Winkel JG (2001) Mac-1 (CD11b/CD18) is essential for Fc receptor-mediated neutrophil cytotoxicity and immunologic synapse formation. Blood 97:2478–2486PubMedCrossRef
53.
Krzewski K, Chen X, Strominger JL (2008) WIP is essential for lytic granule polarization and NK cell cytotoxicity. Proc Natl Acad Sci USA 105:2568–2573PubMedCrossRef
54.
Krzewski K, Chen X, Orange JS, Strominger JL (2006) Formation of a WIP-, WASp-, actin-, and myosin IIA-containing multiprotein complex in activated NK cells and its alteration by KIR inhibitory signaling. J Cell Biol 173:121–132PubMedCrossRef
55.
Lai P, Rabinowich H, Crowley-Nowick PA, Bell MC, Mantovani G, Whiteside TL (1996) Alterations in expression and function of signal-transducing proteins in tumor-associated T and natural killer cells in patients with ovarian carcinoma. Clin Cancer Res 2:161–173PubMed
56.
Patankar MS, Yu J, Morrison JC, Belisle JA, Lattanzio FA, Deng Y, Wong NK, Morris HR, Dell A, Clark GF (2005) Potent suppression of natural killer cell response mediated by the ovarian tumor marker CA125. Gynecol Oncol 99:704–713PubMedCrossRef
57.
Delgado DC, Hank J, Kolesar J, Lorentzen D, Gan J, Seo S, Kim KM, Shusterman S, Gillies SD, Reisfeld RA, Yang R, Gadbaw B, DeSantes KD, London WB, Seeger RC, Maris J, Sondel PM (2010) The role for genotypes of killer Ig-like receptors (KIRs), their ligands, and Fcγ receptors on responses of neuroblastoma patients to Hu14.18-IL2: a children’s oncology group report. Cancer Res 70:9554–9561PubMedCrossRef
58.
Cartron G, Dacheux L, Salles G, Solal-Celigny P, Bardos P, Colombat P, Watier H (2002) Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor FcgammaRIIIa gene. Blood 99:754–758PubMedCrossRef
59.
Weng WK, Levy R (2003) Two immunoglobulin G fragment C receptor polymorphisms independently predict response to rituximab in patients with follicular lymphoma. J Clin Oncol 21:3940–3947PubMedCrossRef
Metadata
Title
Ab-IL2 fusion proteins mediate NK cell immune synapse formation by polarizing CD25 to the target cell-effector cell interface
Authors
Jennifer A. A. Gubbels
Brian Gadbaw
Ilia N. Buhtoiarov
Sachi Horibata
Arvinder K. Kapur
Dhara Patel
Jacquelyn A. Hank
Stephen D. Gillies
Paul M. Sondel
Manish S. Patankar
Joseph Connor
Publication date
01-12-2011
Publisher
Springer-Verlag
Published in
Cancer Immunology, Immunotherapy / Issue 12/2011
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI
https://doi.org/10.1007/s00262-011-1072-9

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