Potent inhibition of OKT3-induced T cell proliferation and suppression of CD147 cell surface expression in HeLa cells by scFv-M6-1B9

Intasai, Nutjeera; Tragoolpua, Khajornsak; Pingmuang, Prakitnavin; Khunkaewla, Panida; Moonsom, Seangdeun; Kasinrerk, Watchara; Lieber, André; Tayapiwatana, Chatchai

doi:10.1016/j.imbio.2008.12.006

Cited by 15 publications

(26 citation statements)

References 67 publications

(81 reference statements)

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“…N18E2 could therefore be used in a general technology for BV-display of bioactive molecules such as scFv. In previous studies, we provided evidence that scFv-M6-1B9 was biologically active as an intrabody, and could be used to diminish the expression of CD147 at the surface of human cells [38,39]. The data presented here showed that scFv-M6-1B9 was also functional when displayed on the BV vector envelope.…”

Section: Resultssupporting

confidence: 54%

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Baculovirus display of single chain antibody (scFv) using a novel signal peptide

et al. 2010

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BackgroundCells permissive to virus can become refractory to viral replication upon intracellular expression of single chain fragment variable (scFv) antibodies directed towards viral structural or regulatory proteins, or virus-coded enzymes. For example, an intrabody derived from MH-SVM33, a monoclonal antibody against a conserved C-terminal epitope of the HIV-1 matrix protein (MAp17), was found to exert an inhibitory effect on HIV-1 replication.ResultsTwo versions of MH-SVM33-derived scFv were constructed in recombinant baculoviruses (BVs) and expressed in BV-infected Sf9 cells, N-myristoylation-competent scFvG2/p17 and N-myristoylation-incompetent scFvE2/p17 protein, both carrying a C-terminal HA tag. ScFvG2/p17 expression resulted in an insoluble, membrane-associated protein, whereas scFvE2/p17 was recovered in both soluble and membrane-incorporated forms. When coexpressed with the HIV-1 Pr55Gag precursor, scFvG2/p17 and scFvE2/p17 did not show any detectable negative effect on virus-like particle (VLP) assembly and egress, and both failed to be encapsidated in VLP. However, soluble scFvE2/p17 isolated from Sf9 cell lysates was capable of binding to its specific antigen, in the form of a synthetic p17 peptide or as Gag polyprotein-embedded epitope. Significant amounts of scFvE2/p17 were released in the extracellular medium of BV-infected cells in high-molecular weight, pelletable form. This particulate form corresponded to BV particles displaying scFvE2/p17 molecules, inserted into the BV envelope via the scFv N-terminal region. The BV-displayed scFvE2/p17 molecules were found to be immunologically functional, as they reacted with the C-terminal epitope of MAp17. Fusion of the N-terminal 18 amino acid residues from the scFvE2/p17 sequence (N18E2) to another scFv recognizing CD147 (scFv-M6-1B9) conferred the property of BV-display to the resulting chimeric scFv-N18E2/M6.ConclusionExpression of scFvE2/p17 in insect cells using a BV vector resulted in baculoviral progeny displaying scFvE2/p17. The function required for BV envelope incorporation was carried by the N-terminal octadecapeptide of scFvE2/p17, which acted as a signal peptide for BV display. Fusion of this peptide to the N-terminus of scFv molecules of interest could be applied as a general method for BV-display of scFv in a GP64- and VSV-G-independent manner.

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Section: Resultssupporting

confidence: 54%

“…The ligand of scFv-M6-1B9 is M6, also called CD147 [38,39], a transmembrane glycoprotein highly expressed in various types of malignant cells [57] and tumors, e.g. nasopharyngeal carcinoma [58].…”

Section: Discussionmentioning

confidence: 99%

Baculovirus display of single chain antibody (scFv) using a novel signal peptide

et al. 2010

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“…This is consistent with our previous study in HeLa cells that showed no difference in MMP-2 and MMP-9 activities. 16 Importantly, the proteolytic activity of MMPs can be also regulated by the endogenous TIMPs. 42,43 To clarify, we further analyzed the expression of TIMP-1 and TIMP-2 and the endogenous tissue inhibitors of MMPs, 44 and the data supported that the functional properties of MMPs and TIMPs are still balanced and scFv-M6-1B9 intrabodies could not influence the enzyme-inhibitor interaction.…”

Section: Discussionmentioning

confidence: 99%

“…Our work demonstrated the usefulness of an adenoviral vector system in transferring intrabodies into HeLa cells to improve our intrabody gene transfer efficiency. 16 Furthermore, switching Ad fibers from serotype 5-35, which uses CD46 as a receptor, increased the transduction efficiency and showed more advantages than Ad5 vector for gene therapy. Previously, modification of Ad5-based vector with Ad35 tropism also improved gene transfer in primary malignant hematopoietic cells, which were refractory to the Ad5 vector.…”

Section: Discussionmentioning

confidence: 99%

“…12 We previously showed that an adenovirus serotype 5 vector encoding scFv-M6-1B9 intrabody gene against EMMPRIN (Ad5-scFv-M6-1B9) efficiently decreased the surface expression of EMMPRIN on 293A cells 15 and HeLa cells. 16 It retained EMMPRIN in the ER with an ER retention signal (KDEL peptide), and intrabody-antigen complexes are considered to be degraded via different mechanisms. 17,18 Serotype 5 Ad vectors infect cells through the coxsackie and adenovirus receptor (CAR).…”

Section: Introductionmentioning

confidence: 99%

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EMMPRIN reduction via scFv-M6-1B9 intrabody affects α3β1-integrin and MCT1 functions and results in suppression of progressive phenotype in the colorectal cancer cell line Caco-2

et al. 2014

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Extracellular matrix metalloproteinase inducer (EMMPRIN) exhibits overexpression in various cancers and promotes cancer progression and metastasis via the interaction with its associated molecules. The scFv-M6-1B9 intrabody has a potential ability to reduce EMMPRIN cell surface expression. However, the subsequent effect of scFv-M6-1B9 intrabody-mediated EMMPRIN abatement on its related molecules, α3β1-integrin, MCT1, MMP-2 and MMP-9, is undefined. Our results demonstrated that the scFv-M6-1B9 intrabody efficiently decreased α3β1-integrin cell surface expression levels. In addition, intracellular accumulation of MCT1 and lactate were increased. These results lead to suppression of features characteristic for tumor progression, including cell migration, proliferation and invasion, in a colorectal cancer cell line (Caco-2) although there was no difference in MMP expression. Thus, EMMPRIN represents an attractive target molecule for the disruption of cancer proliferation and metastasis. An scFv-M6-1B9 intrabody-based approach could be relevant for cancer gene therapy.

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Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

Chupradit,

Muneekaew,

Wattanapanitch

2024

Cancer Immunol Immunother

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Chimeric antigen receptor (CAR) T cell therapy has shown promising results in hematologic malignancies, but its effectiveness in solid cancers remains challenging. Macrophages are immune cells residing within the tumor microenvironment. They can phagocytose tumor cells. Recently, CAR macrophages (CAR-M) have been a promising candidate for treating solid cancers. One of the common cancer antigens overexpressed in various types of cancer is CD147. CAR-T and NK cells targeting CD147 antigen have shown significant efficacy against hepatocellular carcinoma. Nevertheless, CAR-M targeting the CD147 molecule has not been investigated. In this study, we generated CAR targeting the CD147 molecule using the THP-1 monocytic cell line (CD147 CAR-M). The CD147 CAR-M exhibited typical macrophage characteristics, including phagocytosis of zymosan bioparticles and polarization ability toward M1 and M2 phenotypes. Furthermore, the CD147 CAR-M demonstrated enhanced anti-tumor activity against K562 and MDA-MB-231 cells without exhibiting off-target cytotoxicity against normal cells. Our research provides valuable insights into the potential of CD147 CAR-M as a promising platform for cancer immunotherapy, with applications in both hematologic malignancies and solid cancers. Graphical abstract

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Potent inhibition of OKT3-induced T cell proliferation and suppression of CD147 cell surface expression in HeLa cells by scFv-M6-1B9

Cited by 15 publications

References 67 publications

Baculovirus display of single chain antibody (scFv) using a novel signal peptide

Baculovirus display of single chain antibody (scFv) using a novel signal peptide

EMMPRIN reduction via scFv-M6-1B9 intrabody affects α3β1-integrin and MCT1 functions and results in suppression of progressive phenotype in the colorectal cancer cell line Caco-2

Engineered CD147-CAR macrophages for enhanced phagocytosis of cancers

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