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Preventive DNA vaccination against CEA-expressing tumors with anti-idiotypic scFv6.C4 DNA in CEA-expressing transgenic mice

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Abstract

Carcinoembryonic antigen (CEA) is expressed during embryonic life and in low level during adult life. Consequently, the CEA is recognized by the immune system as a self-antigen and thus CEA-expressing tumors are tolerated. Previously, we constructed a single chain variable fragment using the 6.C4 (scFv6.C4) hybridoma cell line, which gave rise to antibodies able to recognize CEA when C57/Bl6 mice were immunized. Here, the scFv6.C4 ability to prevent the CEA-expressing tumor growth was assessed in CEA-expressing transgenic mice CEA2682. CEA2682 mice immunized with the scFv6.C4 expressing plasmid vector (uP/PS-scFv6.C4) by electroporation gave rise to the CEA-specific AB3 antibody after the third immunization. Sera from immunized mice reacted with CEA-expressing human colorectal cell lines CO112, HCT-8, and LISP-1, as well as with murine melanoma B16F10 cells expressing CEA (B16F10-CEA). Cytotoxic T lymphocytes (CTL) from uP/PS-scFv6.C4 immunized mice lysed B16F10-CEA (56.7%) and B16F10 expressing scFv6.C4 (B16F10-scFv6.C4) (46.7%) cells, against CTL from uP-immunized mice (10%). After the last immunization, 5 × 105 B16F10-CEA cells were injected into the left flank. All mice immunized with the uP empty vector died within 40 days, but uP/PS-scFv6.C4 vaccinated mice (40%) remained free of tumor for more than 100 days. Splenocytes obtained from uP/PS-scFv6.C4 vaccinated mice showed higher T-cell proliferative activity than those from uP vaccinated mice. Collectively, DNA vaccination with the uP-PS/scFv6.C4 plasmid vector was able to give rise to specific humoral and cellular responses, which were sufficient to retard growth and/or eliminate the injected B16F10-CEA cells.

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Abbreviations

B16F10-CEA:

Murine melanoma cell line B16F10 expressing CEA

B16F10-scFv6.C4:

Murine melanoma cell line B16F10 expressing scFv6.C4

CEA:

Carcinoembryonic antigen

CEA2682:

CEA-expressing transgenic mice

CFSE:

Carboxyfluorescein diacetate succinimidyl ester

FAPESP:

Sao Paulo research foundation

RPMIc:

RPMI 1640 medium with supplements

SB:

Sleeping beauty

scFv:

Single-chain variable fragment

scFv6.C4:

Single-chain variable fragment 6.C4

SD:

Standard deviation

uP/PS-scFv6.C4:

scFv6.C4 expressing plasmid vector

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Acknowledgements

Priscila M. A. Denapoli and Bianca F. Zanetti were recipients of the Brazilian National Council for Scientific and Technological Development (CNPq) and São Paulo Research Foundation (FAPESP) scholarships, respectively.

Funding

This study was funded by FAPESP (Grant Number: 2012/21861-1).

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Authors and Affiliations

  1. Research Center for Gene Therapy, Federal University of Sao Paulo, São Paulo, SP, Brazil

    Priscila M. A. Denapoli, Bianca F. Zanetti, Adara A. dos Santos & Sang W. Han

  2. Department of Biophysics, Federal University of Sao Paulo, Rua Mirassol, 207, São Paulo, SP, 04044-010, Brazil

    Jane Z. de Moraes & Sang W. Han

Authors
  1. Priscila M. A. Denapoli
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  2. Bianca F. Zanetti
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Correspondence to Sang W. Han.

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Denapoli, P.M.A., Zanetti, B.F., dos Santos, A.A. et al. Preventive DNA vaccination against CEA-expressing tumors with anti-idiotypic scFv6.C4 DNA in CEA-expressing transgenic mice. Cancer Immunol Immunother 66, 333–342 (2017). https://doi.org/10.1007/s00262-016-1940-4

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