Dataset Information

Interleukin-15:Interleukin-15 receptor ? scaffold for creation of multivalent targeted immune molecules.

ABSTRACT: Human interleukin-15 (hIL-15) and its receptor ? (hIL-15R?) are co-expressed in antigen presenting cells allowing trans-presentation of the cytokine to immune effector cells. We exploited the high-affinity interactions between hIL-15 and the extracellular hIL-15R? sushi domain (hIL-15R?Su) to create a functional scaffold for the design of multispecific fusion protein complexes. Using single-chain T cell receptors (scTCRs) as recognition domains linked to the IL-15:IL-15R? scaffold, we generated both bivalent and bispecific complexes. In these fusions, the scTCR domains retain the antigen-binding activity and the hIL-15 domain exhibits receptor binding and biological activity. As expected, bivalent scTCR fusions exhibited improved antigen binding due to increased avidity, whereas fusions comprising two different scTCR domains were capable of binding two cognate peptide/MHC complexes. Bispecific molecules containing scTCR and scCD8?? domains also exhibit enhanced binding to peptide/MHC complexes, demonstrating that the IL-15:IL-15R? scaffold displays flexibility necessary to support multi-domain interactions with a given target. Surprisingly, functional heterodimeric molecules could be formed by co-expressing the TCR ? and ? chains separately as fusions to the hIL-15 and hIL-15R?Su domains. Together, these properties indicate that the hIL-15 and hIL-15R?Su domains can be used as versatile, functional scaffold for generating novel targeted immune molecules.

SUBMITTER: Wong RL

PROVIDER: S-EPMC3049345 | biostudies-literature | 2011 Apr

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Interleukin-15:Interleukin-15 receptor α scaffold for creation of multivalent targeted immune molecules.

Wong Richard L RL Liu Bai B Zhu Xiaoyun X You Lijing L Kong Lin L Han Kai-Ping KP Lee Hyung-Il HI Chavaillaz Pierre-Andre PA Jin Moonsoo M Wang Yi Y Rhode Peter R PR Wong Hing C HC

Protein engineering, design & selection : PEDS 20101221 4

Human interleukin-15 (hIL-15) and its receptor α (hIL-15Rα) are co-expressed in antigen presenting cells allowing trans-presentation of the cytokine to immune effector cells. We exploited the high-affinity interactions between hIL-15 and the extracellular hIL-15Rα sushi domain (hIL-15RαSu) to create a functional scaffold for the design of multispecific fusion protein complexes. Using single-chain T cell receptors (scTCRs) as recognition domains linked to the IL-15:IL-15Rα scaffold, we generated ...[more]

PMID: 21177283

Similar Datasets

Project description:Oncolytic type-1 herpes simplex viruses (oHSVs) lacking the ?134.5 neurovirulence gene are being evaluated for treatment of a variety of malignancies. oHSVs replicate within and directly kill permissive cancer cells. To augment their anti-tumor activity, oHSVs have been engineered to express immunostimulatory molecules, including cytokines, to elicit tumor-specific immune responses. Interleukin-15 (IL-15) holds potential as an immunotherapeutic cytokine because it has been demonstrated to promote both natural killer (NK) cell-mediated and CD8(+) T cell-mediated cytotoxicity against cancer cells. The purpose of these studies was to engineer an oHSV producing bioactive IL-15. Two oHSVs were constructed encoding murine (m)IL-15 alone (J100) or with the mIL-15 receptor ? (mIL-15R?, J100D) to determine whether co-expression of these proteins is required for production of bioactive mIL-15 from oHSV. The following were demonstrated: i) both oHSVs retain replication competence and cytotoxicity in permissive tumor cell lines. ii) Enhanced production of mIL-15 was detected in cell lysates of neuro-2a cells following J100D infection as compared to J100 infection, suggesting that mIL-15R? improved mIL-15 production. iii) Soluble mIL-15 in complex with mIL-15R? was detected in supernates from J100D-infected, but not J100-infected, neuro-2a, GL261, and CT-2A cells. These cell lines vary in permissiveness to oHSV replication and cytotoxicity, demonstrating soluble mIL-15/IL-15R? complex production from J100D was independent of direct oHSV effects. iv) The soluble mIL-15/IL-15R? complex produced by J100D was bioactive, stimulating NK cells to proliferate and reduce the viability of syngeneic GL261 and CT-2A cells. v) J100 and J100D were aneurovirulent inasmuch as no neuropathologic effects were documented following direct inoculation into brains of CBA/J mice at up to 1x10(7) plaque forming units. The production of mIL-15/mIL-15R? from multiple tumor lines, as well as the lack of neurovirulence, renders J100D suitable for investigating the combined effects of oHSV and mIL-15/IL-15R? in various cancer models.

Dataset Information

Interleukin-15:Interleukin-15 receptor ? scaffold for creation of multivalent targeted immune molecules.

Publications

Interleukin-15:Interleukin-15 receptor α scaffold for creation of multivalent targeted immune molecules.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets