An engineered anti-idiotypic antibody-derived killer peptide (KP) early activates swine inflammatory monocytes, CD3+CD16+ natural killer T cells and CD4+CD8α+ double positive CD8β+ cytotoxic T lymphocytes associated with TNF-α and IFN-γ secretion

Luca Ferrari; Paolo Martelli; Roberta Saleri; Elena De Angelis; Giulia Ferrarini; Valeria Cavalli; Benedetta Passeri; Gianluca Bazzoli; Giulia Ogno; Walter Magliani; Paolo Borghetti

doi:10.1016/j.cimid.2020.101523

An engineered anti-idiotypic antibody-derived killer peptide (KP) early activates swine inflammatory monocytes, CD3⁺CD16⁺ natural killer T cells and CD4⁺CD8α⁺ double positive CD8β⁺ cytotoxic T lymphocytes associated with TNF-α and IFN-γ secretion

Comp Immunol Microbiol Infect Dis. 2020 Oct:72:101523. doi: 10.1016/j.cimid.2020.101523. Epub 2020 Jul 23.

Authors

Affiliations

¹ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: luca.ferrari@unipr.it.
² Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: paolo.martelli@unipr.it.
³ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: roberta.saleri@unipr.it.
⁴ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: elena.deangelis@unipr.it.
⁵ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: giulia.ferrarini@studenti.unipr.it.
⁶ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: valeria.cavalli@unipr.it.
⁷ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: benedetta.passeri@unipr.it.
⁸ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: gianluca.bazzoli@studenti.unipr.it.
⁹ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: giulia.ogno@studenti.unipr.it.
¹⁰ Department of Medicine and Surgery, University of Parma, Via Gramsci, 14 - 43126, Parma, Italy. Electronic address: walter.magliani@unipr.it.
¹¹ Department of Veterinary Science, University of Parma, Strada del Taglio, 10 - 43126, Parma, Italy. Electronic address: paolo.borghetti@unipr.it.

PMID: 32758800
DOI: 10.1016/j.cimid.2020.101523

Abstract

This study evaluated the early modulation of the phenotype and cytokine secretion in swine immune cells treated with an engineered killer peptide (KP) based on an anti-idiotypic antibody functionally mimicking a yeast killer toxin. The influence of KP on specific immunity was investigated using porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) as ex vivo antigens. Peripheral blood mononuclear cells (PBMC) from healthy pigs were stimulated with KP and with a scramble peptide for 20 min, 1, 4 and 20 h or kept unstimulated. The cells were analyzed using flow cytometry and ELISA. The same time-periods were used for KP pre-incubation/co-incubation to determine the effect on virus-recalled interferon-gamma (IFN-γ) secreting cell (SC) frequencies and single cell IFN-γ productivity using ELISPOT. KP induced an early dose-dependent shift to pro-inflammatory CD172α⁺CD14^+high monocytes and an increase of CD3⁺CD16⁺ natural killer (NK) T cells. KP triggered CD8α and CD8β expression on classical CD4^-CD8αβ⁺ cytotoxic T lymphocytes (CTL) and double positive (DP) CD4⁺CD8α⁺ Th memory cells (CD4⁺CD8α^+low CD8β^+low). A fraction of DP cells also expressed high levels of CD8α. The two identified DP CD4⁺CD8α^+high CD8β^+low/+high CTL subsets were associated with tumor necrosis factor alpha (TNF-α) and IFN-γ secretion. KP markedly boosted the reactivity and cross-reactivity of PRRSV type-1- and PCV2b-specific IFN-γ SC. The results indicate the efficacy of KP in stimulating Th1-biased immunomodulation and support studies of KP as an immunomodulator or vaccine adjuvant.

Keywords: Cytokines; Double positive cytotoxic T lymphocytes (DP CTL); IFN-γ secreting cells; Killer peptide (KP); Natural killer (NK) T cells; Porcine circovirus type 2 (PCV2); Porcine reproductive and respiratory syndrome virus (PRRSV); Pro-inflammatory monocytes; TNF-α; Viral cross-reactivity.

MeSH terms

Animals
Antibodies, Anti-Idiotypic / immunology*
Interferon-gamma / immunology*
Monocytes / immunology*
Natural Killer T-Cells / immunology*
Peptides / immunology
Swine
T-Lymphocytes, Cytotoxic / immunology*
Tumor Necrosis Factor-alpha / metabolism*

Substances

Antibodies, Anti-Idiotypic
Peptides
Tumor Necrosis Factor-alpha
Interferon-gamma