Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells

The cytotoxic T lymphocyte-antigen 4 (CTLA-4) has been considered an IC exclusively expressed on T cells, where it counteracts the co-stimulatory CD28 receptor, by competing for its binding to CD-80 and CD-86. We recently found that it is expressed also on tumor and NK cells, suggesting other possib...

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Main Authors: Margherita Passariello, Cinzia Vetrei, Emanuele Sasso, Guendalina Froechlich, Chiara Gentile, Anna Morena D’Alise, Nicola Zambrano, Elisa Scarselli, Alfredo Nicosia, Claudia De Lorenzo
Format: Article
Language:English
Published: MDPI AG 2020-08-01
Series:Cancers
Subjects:
Online Access:https://www.mdpi.com/2072-6694/12/8/2204
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spelling doaj-0cdd6be6eb2a488b9448e2597c892e642020-11-25T03:18:25ZengMDPI AGCancers2072-66942020-08-01122204220410.3390/cancers12082204Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK CellsMargherita Passariello0Cinzia Vetrei1Emanuele Sasso2Guendalina Froechlich3Chiara Gentile4Anna Morena D’Alise5Nicola Zambrano6Elisa Scarselli7Alfredo Nicosia8Claudia De Lorenzo9Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Via Pansini 5, 80131 Napoli, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Via Pansini 5, 80131 Napoli, ItalyNouscom srl, Via di Castel Romano 100, 00128 Rome, ItalyCeinge—Biotecnologie Avanzate S.C. A.R.L., via Gaetano Salvatore 486, 80145 Naples, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Via Pansini 5, 80131 Napoli, ItalyNouscom srl, Via di Castel Romano 100, 00128 Rome, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Via Pansini 5, 80131 Napoli, ItalyNouscom srl, Via di Castel Romano 100, 00128 Rome, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Via Pansini 5, 80131 Napoli, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Via Pansini 5, 80131 Napoli, ItalyThe cytotoxic T lymphocyte-antigen 4 (CTLA-4) has been considered an IC exclusively expressed on T cells, where it counteracts the co-stimulatory CD28 receptor, by competing for its binding to CD-80 and CD-86. We recently found that it is expressed also on tumor and NK cells, suggesting other possible unknown roles of CTLA-4. To shed light on these novel aspects of CTLA-4, we used Ipilimumab, the first FDA approved human antibody targeting CTLA-4, in parallel studies with two novel human mAbs we isolated by using an efficient phage display selection strategy on live activated lymphocytes and purified mouse and human CTLA-4. The selection for cross-reactive mAbs was guaranteed by a high throughput sequencing to identify the sequences commonly enriched by two parallel pannings on human and mouse CTLA-4. Two isolated antibodies were found to bind with high affinity to both human and mouse CTLA-4 and lymphocytes, showing nanomolar or sub-nanomolar Kd values. They were able to kill Treg cells by ADCC, and to activate both human and mouse PBMCs, by strongly increasing cytokines secretion. Interestingly, they activated NK cells, exhibited cytotoxicity against cancer cells by inducing ADCC and inhibited tumor cell growth by affecting CTLA-4 downstream pathways in a similar fashion to CD-80 and CD-86 ligands and differently from Ipilimumab. Moreover, the novel mAbs showed a reduced ability to interfere in the binding of CD-80 ligands to CTLA-4 on T cells with respect to Ipilimumab, suggesting that they could allow for anti-tumor effects without the irAEs associated with the potent antagonistic activity of Ipilimumab.https://www.mdpi.com/2072-6694/12/8/2204CTLA-4immune checkpointimmunomodulatory mAbscancer immunotherapynatural killer cells
collection DOAJ
language English
format Article
sources DOAJ
author Margherita Passariello
Cinzia Vetrei
Emanuele Sasso
Guendalina Froechlich
Chiara Gentile
Anna Morena D’Alise
Nicola Zambrano
Elisa Scarselli
Alfredo Nicosia
Claudia De Lorenzo
spellingShingle Margherita Passariello
Cinzia Vetrei
Emanuele Sasso
Guendalina Froechlich
Chiara Gentile
Anna Morena D’Alise
Nicola Zambrano
Elisa Scarselli
Alfredo Nicosia
Claudia De Lorenzo
Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells
Cancers
CTLA-4
immune checkpoint
immunomodulatory mAbs
cancer immunotherapy
natural killer cells
author_facet Margherita Passariello
Cinzia Vetrei
Emanuele Sasso
Guendalina Froechlich
Chiara Gentile
Anna Morena D’Alise
Nicola Zambrano
Elisa Scarselli
Alfredo Nicosia
Claudia De Lorenzo
author_sort Margherita Passariello
title Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells
title_short Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells
title_full Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells
title_fullStr Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells
title_full_unstemmed Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells
title_sort isolation of two novel human anti-ctla-4 mabs with intriguing biological properties on tumor and nk cells
publisher MDPI AG
series Cancers
issn 2072-6694
publishDate 2020-08-01
description The cytotoxic T lymphocyte-antigen 4 (CTLA-4) has been considered an IC exclusively expressed on T cells, where it counteracts the co-stimulatory CD28 receptor, by competing for its binding to CD-80 and CD-86. We recently found that it is expressed also on tumor and NK cells, suggesting other possible unknown roles of CTLA-4. To shed light on these novel aspects of CTLA-4, we used Ipilimumab, the first FDA approved human antibody targeting CTLA-4, in parallel studies with two novel human mAbs we isolated by using an efficient phage display selection strategy on live activated lymphocytes and purified mouse and human CTLA-4. The selection for cross-reactive mAbs was guaranteed by a high throughput sequencing to identify the sequences commonly enriched by two parallel pannings on human and mouse CTLA-4. Two isolated antibodies were found to bind with high affinity to both human and mouse CTLA-4 and lymphocytes, showing nanomolar or sub-nanomolar Kd values. They were able to kill Treg cells by ADCC, and to activate both human and mouse PBMCs, by strongly increasing cytokines secretion. Interestingly, they activated NK cells, exhibited cytotoxicity against cancer cells by inducing ADCC and inhibited tumor cell growth by affecting CTLA-4 downstream pathways in a similar fashion to CD-80 and CD-86 ligands and differently from Ipilimumab. Moreover, the novel mAbs showed a reduced ability to interfere in the binding of CD-80 ligands to CTLA-4 on T cells with respect to Ipilimumab, suggesting that they could allow for anti-tumor effects without the irAEs associated with the potent antagonistic activity of Ipilimumab.
topic CTLA-4
immune checkpoint
immunomodulatory mAbs
cancer immunotherapy
natural killer cells
url https://www.mdpi.com/2072-6694/12/8/2204
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