Eradication of established hepatic human neuroblastoma metastases in mice with severe combined immunodeficiency by antibody-targeted interleukin-2

Pancook, James D.; Becker, Jürgen C.; Gillies, Stephen D.; Reisfeld, Ralph A.

doi:10.1007/s002620050256

Cited by 35 publications

(15 citation statements)

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“…Prior to analysis of tumor lesions, experiments were carried out to demonstrate that each amplified TCRBV product was obtained with the expected size (see Table 1) and no spurious amplification products were observed. To validate the semiquantitative reverse transcriptase-PCR methodology, serially diluted cDNA was PCR amplified for different numbers of cycles (26,28,30,32, and 34 cycles) using the primers BV 1-18 in combination with the constant region primer BC end labeled with [␥- 33 P]. The data from these experiments (data not shown) were used to determine the amount of cDNA and the number of cycles through which the specific PCR products accumulated exponentially, enabling determination of the relative abundance of each TCRBV region.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, we have tried to overcome these problems by developing antibody-IL-2 fusion proteins that combine the unique targeting ability of antibodies with the multifunctional activity of cytokines. The therapeutic effectiveness of such constructs for the treatment of established hepatic, pulmonary, and subcutaneous melanoma metastases has been documented in a number of different murine tumor models (9,10,(26)(27)(28). Detailed characterization in various tumor models revealed the superiority of antibody-IL-2 fusion proteins over comparable amounts of IL-2, the parental antibody, the combination of both or nonspecific antibody-IL-2 fusion proteins (9).…”

Section: Figmentioning

confidence: 99%

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Activation of preexisting T cell clones by targeted interleukin 2 therapy

Straten

Guldberg

Seremet

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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The induction of an immunological antitumor response capable of eradicating metastatic tumors is the ultimate goal of immunotherapy. We have recently shown that this can be achieved by interleukin 2 (IL-2) therapy directed Melanoma is a highly malignant tumor but several lines of evidence suggest that it is capable of eliciting a specific immune response, i.e., a number of melanoma-associated antigens have been identified and the presence of clonotypic T cells has been demonstrated in melanoma lesions (1-4). Therefore, several immunomodulatory therapeutic approaches were initiated to improve the prognosis of melanoma patients. Interleukin 2 (IL-2) is one of the most potent antitumor cytokines known quote (5), and was recently approved for treatment of metastatic melanoma. However, objective responses induced by systemic IL-2 therapy are still insufficient, and the associated side effects are severe (6). These findings are due to the fact that a systemic application of IL-2 disregards the paracrine nature of this cytokine under physiological conditions (7).As a means to target IL-2 directly to the tumor site, we have recently shown that human IL-2 can be genetically engineered as a fusion protein with the chimeric mouse-human mAb 14.18 which recognizes the ganglioside GD 2 , retaining both antigen binding and cytokine activity (8). Furthermore, we have shown that treatment with this antibody-IL-2 fusion protein can eradicate human hepatic and pulmonary melanoma metastases in severe combined immunodeficient mice (9) as well as autologous murine B16 melanomas (10). Although it was shown in these studies that tumor eradication was dependent on CD8 ϩ T cells, it is not known whether tumor clearance is associated with a clonal expansion of T cells. Furthermore, it remains to be established whether such a clonal expansion would be due to a de novo induction or to the activation and expansion of preexisting T cell clones. Here, we demonstrate both the overexpression of certain T cell receptor variable ␤ regions (TCRBV) as well as the clonal expansion of T cells in melanoma lesions subsequent to targeted IL-2 therapy. However, clonally expanded T cells were also detectable prior to therapy, suggesting that antibody-IL-2-targeted therapy acts as an activator rather than an inducer of an antitumor T cell response.

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Section: Resultsmentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

Activation of preexisting T cell clones by targeted interleukin 2 therapy

Straten

Guldberg

Seremet

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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“…Treatment with ICs results in activation of T cell-mediated antitumor cytotoxicity [24] and in NK cell-mediated control of tumor growth [25, 26]. The antibody component of the IC, upon binding to antigens on the tumor cells, activates NK cells via CD16-Fc interaction [27].…”

Section: Introductionmentioning

confidence: 99%

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

Gubbels

Gadbaw

Buhtoiarov

et al. 2011

Cancer Immunol Immunother

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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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“…1 Studies in mice of antidisialoganglioside (anti-GD2)/interleukin-2 and anti-GD2/lymphotoxin-␣ fusion proteins (immunocytokines) demonstrated the eradication of hepatic metastases of neuroblastoma 2,3 and the induction of protective immunity against melanoma. 4 These and other immunocytokine strategies have been based on the stimulation of NK-cell-or T-cell-mediated responses.…”

Section: Introductionmentioning

confidence: 99%

Antidisialoganglioside/granulocyte macrophage–colony-stimulating factor fusion protein facilitates neutrophil antibody-dependent cellular cytotoxicity and depends on FcγRII (CD32) and Mac-1 (CD11b/CD18) for enhanced effector cell adhesion and azurophil granule exocytosis

Metelitsa

Gillies

Super

et al. 2002

Blood

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Polymorphonuclear leukocytes (PMNs) mediate antibody-dependent cellular cytotoxicity (ADCC), which is increased by the addition of granulocyte-macrophage colony-stimulating factor (GM-CSF). We sought to determine whether PMN ADCC also would be increased by the addition of an antibody/GM-CSF fusion protein and whether this would be associated with the up-regulation and activation of Mac-1 (CD11b/CD18) and with azurophil granule exocytosis. ADCC against LA-N-1 human neuroblastoma cells was evaluated with 4-hour calcein acetoxymethyl ester (calcein-AM) microcytotoxicity assay, electron microscopy, and multiparameter flow cytometry. With the calcein-AM assay, LA-N-1 cell survival was 10%, 55%, and 75% when PMN ADCC was mediated by the antidisialoganglioside (anti-GD2) immunocytokine hu14. IntroductionAntibody-cytokine fusion proteins combine the targeting ability of antibodies with the immune stimulation of cytokines for cancer immunotherapy. 1 Studies in mice of antidisialoganglioside (anti-GD2)/interleukin-2 and anti-GD2/lymphotoxin-␣ fusion proteins (immunocytokines) demonstrated the eradication of hepatic metastases of neuroblastoma 2,3 and the induction of protective immunity against melanoma. 4 These and other immunocytokine strategies have been based on the stimulation of NK-cell-or T-cell-mediated responses. Polymorphonuclear leukocytes (PMNs) constitute the largest population of white blood cells, and they can mediate antibody-dependent cellular cytotoxicity (ADCC) against tumor cells. [5][6][7][8][9] In vitro studies demonstrated the strong facilitation of PMN ADCC against neuroblastoma cells by mixing granulocytemacrophage colony-stimulating factor (GM-CSF) with an anti-GD2 antibody and by a human-mouse chimeric anti-GD2/GM-CSF immunocytokine. 5,10 Fc receptors (FcR) involved in PMN ADCC may include Fc␥RII (CD32) and Fc␥RIII (CD16), which are constitutively expressed, and Fc␥RI (CD64), which is induced by interferon-␥ and G-CSF. 11,12 In addition, Fc␣RI of PMNs may be activated by constructing bispecific monoclonal antibodies (mAbs) that recognize a target cell molecule and Fc␣RI. 13 Binding of an antibody-cytokine fusion protein to FcR may be affected by the physical presence of the cytokine or by the cytokinemodulating FcR expression. 14,15 FcRs used by PMNs in ADCC with anti-GD2/GM-CSF immunocytokines have not yet been determined.␤ 2 -Integrin receptors, particularly Mac-1 (␣ m ␤ 2 , CD11b/CD18, and complement receptor type 3), have been reported to have an essential role in PMN ADCC against neuroblastoma, lymphoma, and breast cancer cell lines. 5,13,16 Mac-1 is a major PMN ␤ 2 -integrin receptor that, on activation, acquires the ability to bind multiple ligands and to mediate several adhesion-dependent processes, including phagocytosis, superoxide production, and degranulation. 17 It recently has been demonstrated that Mac-1 function is necessary for PMN spreading on antibody-coated targets and for tumor cytolysis. 13 FcR and GM-CSF receptors activate Mac-1 For personal use only. on May 12, 2018. by ...

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Eradication of established hepatic human neuroblastoma metastases in mice with severe combined immunodeficiency by antibody-targeted interleukin-2

Cited by 35 publications

References 0 publications

Activation of preexisting T cell clones by targeted interleukin 2 therapy

Activation of preexisting T cell clones by targeted interleukin 2 therapy

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

Antidisialoganglioside/granulocyte macrophage–colony-stimulating factor fusion protein facilitates neutrophil antibody-dependent cellular cytotoxicity and depends on FcγRII (CD32) and Mac-1 (CD11b/CD18) for enhanced effector cell adhesion and azurophil granule exocytosis

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