CD30‐specific AB1‐AB2‐AB3 internal image antibody network: Potential use as anti‐idiotype vaccine against Hodgkin's lymphoma

Abstract

The tumor-associated CD30 antigen is presently under study as a target for active specific immunotherapy of Hodgkin's lymphoma with anti-idiotypic antibodies. Internal image antibodies (Ab2 beta) 9G10 and 14G9 against the CD30-specific antibody HRS-4 (Ab1) have been described, which induce a CD30-specific T- and B-cell response in BALB/c mice and New Zealand white rabbits. In extension of this work, murine monoclonal anti-idiotypic Ab2 beta 9G10, mimicking structures of the nominal CD30 antigen, was used to generate monoclonal Ab3 in mice and polyclonal Ab3 in rabbits with specificity for CD30. The Ab2 beta 9G10-specific murine monoclonal Ab3 4A4 bound specifically to the 120-kDa band of CD30 present on Hodgkin cell lines and Hodgkin tumor tissue, and effectively inhibited binding of Ab1 HRS-4 to Ab2 9G10 as well as to CD30+ cells. Monoclonal Ab3 4A4 was cytotoxic for CD30+ cell lines in vitro and effectively prevented the s.c. growth of L540 cell tumors after passive i.v. administration in a SCID mouse tumor model. While this cytotoxic effect of the IgM subclass monoclonal Ab3 4A4 was due to complement activation, the murine monoclonal Ab1 HRS-4 and a polyclonal Ab3 preparation of IgG-subclass from New Zealand white rabbits were cytotoxic by an antibody-dependent cell-mediated mechanism in vitro. In conclusion, Ab2 beta 9G10 is able to induce a CD30-specific cytotoxic IgG and IgM response. Cytotoxicity was shown to be mediated by complement activation and antibody-dependent cell-mediated cytotoxicity in vitro and in vivo and across species barriers. Thus, the CD30-like Ab2 beta 9G10 may hold promise for effective active specific immunotherapy of human Hodgkin's lymphoma.

T cell activation by recombinant FcεRI γ-chain immune receptors: an extracellular spacer domain impairs antigen-dependent T cell activation but not antigen recognition

“…To test whether the receptor configuration alters the idiotope of the antigen binding domain, we analyzed the idiotypic profile of the HRS3-scFv-␥ and HRS3-scFv-Fc-␥ receptor, respectively, utilizing a panel of four antiidiotypic mAbs (9G10, 12D3, 14B12, 14G9) that define at least three different idiotopes of the HRS3 mAb binding domain. [10][11][12] Figure 2b summarizes binding of the four anti-idiotypic mAbs to HRS3-scFv-␥ and HRS3-scFv-Fv-␥ receptor grafted MD45 cells as monitored by FACS analysis utilizing saturating antibody concentrations (6 g/ml). The HRS3-scFv-␥ and HRS3-scFv-Fc-␥ receptor reacted in a similiar pattern to the tested antiidiotypic idiotypic antibodies indicating that the idiotypic profile is unlikely to be influenced by the receptor configurations.…”

“…The anti-CD30 mAb HRS3, the anti-HRS3 idiotypic mAbs 9G10, 12D3, 14G9, 14B12 and the antiidiotypic mAb BW2064/399 with specificity for an anti-CEA mAb have been described elsewhere. 11,12,16,17 Chimeric receptors The generation of the CD30-specific HRS3-scFv-␥ receptor has been recently described, 7,18 the HRS3-scFv-Fc-␥ receptor was obtained by replacing the scFv domain within the BW431/26-scFv-Fc-␥ receptor 9 by the HRS3-scFv domain. Briefly, the HRS3-scFv DNA was flanked by XbaI (5Ј) and BamHI (3Ј) restriction sites, respectively, by PCR techniques utilizing the following primer oligonucleotides: 5Ј-GCG GCC CAG TCT AGA ATG GCC CAG-3Ј (sense); 5Ј-ACC TGG ATC CGC CCG TTT GAT TTC-3Ј (antisense) (restriction sites underlined).…”

A chimeric receptor that selectively targets membrane-bound carcinoembryonic antigen (mCEA) in the presence of soluble CEA

“…MD45 T cell clones transfected with the humBW431/26-CH2/CH3-␥ receptor and untransfected MD45 cells, respectively, were incubated in microtiter plates coated with the anti-BW431/26 idiotypic mAb BW2064/36 13 or, for control reasons, with the anti-idiotypic mAb 9G10 with specificity for the anti-CD30 mAb HRS3. 15 As shown in Figure 2a, incubation of receptor grafted cells with immobilized anti-idiotypic BW2064/36 antibody resulted in increased IL-2 secretion whereas incubation with an immobilized, isotype matched anti-idiotypic control antibody did not. Incubation of the anti-CEA receptor- bation with CEA + or with CEA − tumor cells.…”

Adoptive immunotherapy with genetically engineered T cells: modification of the IgG1 Fc ‘spacer’ domain in the extracellular moiety of chimeric antigen receptors avoids ‘off-target’ activation and unintended initiation of an innate immune response

“…The anti-HRS3 idiotypic antibodies 9G10, 12D3, 14B9 and 14G9 carrying internal and non-internal image epitopes were described elsewhere. 18 The phycoerythrin (PE)-conjugated F(ab 0 )2 goat antihuman IgG antibody was purchased from Southern Biotechnology (Birmingham, Alabama, USA), the fluorescein isothiocyanate-conjugated anti-CD3 mAb UCHT-1, the fluorescein isothiocyanate-conjugated anti-CD32 mAb KB61 and the PE-conjugated anti-CD64 mAb 10.1 from Dako, Glostrup, Denmark. The allophycocyanin (APC)-conjugated anti-CD3 mAb UCHT-1, the PE-conjugated anti-CD16 mAb3G8, the fluorescein isothiocyanate-conjugated anti-CD107a mAb H4A3 and the fluorochromeconjugated isotype controls were purchased from BD Bioscience, San Diego, CA, USA; the PE-labelled anti-CD30 mAb (Ki-2) and Fc-blocking reagent from Miltenyi Biotech (Bergisch Gladbach, Germany).…”