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. (October 2020)

• Record Type: Journal Article
• Authors: Ferrari, Luca; Martelli, Paolo; Saleri, Roberta; De Angelis, Elena; Ferrarini, Giulia; Cavalli, Valeria; Passeri, Benedetta; Bazzoli, Gianluca; Ogno, Giulia; Magliani, Walter; Borghetti, Paolo
• Is Part Of: Comparative immunology, microbiology and infectious diseases. Volume 72(2020)
• Publication Date: 2020-10
• Subjects: Ab/s antibody/ies -- ADCC antibody-dependent cell-mediated cytotoxicity -- ADV Aujeszky's disease virus -- A-10-S (AKVTMTCSAS) alanine-lysine-valine-threonine-methionine-threonine-cysteine-serine-alanine-serine (killer peptide, KP) -- APC antigen presenting cells -- ASFV African swine fever virus -- BSA bovine serum albumin -- Cap capsid -- CD cluster of differentiation -- CD172α CD172 alpha -- CD8α CD8 alpha -- CD8β CD8 beta -- CD8αβ CD8 alpha/beta (heterodimer) -- cRPMI complete RPMI -- CSFV classical swine fever virus -- CTL cytotoxic T lymphocytes -- Da Dalton -- DP CTL double positive (CD4+CD8α+) cytotoxic T lymphocytes -- DC dendritic cells -- DMSO dimethylsulfoxide -- ELISA enzyme-linked immunosorbent assay -- ELISPOT enzyme-linked immunospot -- F(ab) fragment antigen-binding -- F(ab')2 divalent fragment antigen-binding -- FBS foetal bovine serum -- Fc fragment crystallizable -- FcRIII Fc receptor 3 -- FITC fluorescein isothiocyanate -- FMO fluorescence-minus-one -- Fmoc 9-fluorenylmethoxycarbonyl -- hi heat-inactivated -- HCV hepatitis C virus -- HIV-1 human immunodeficency virus type 1 -- HSV-1 herpes simplex virus type 1 -- IFN-γ interferon-gamma -- Ig immunoglobulin -- IL interleukin -- κ kappa light chain -- KP killer peptide -- KT killer toxin -- LAK lymphokine-activated killer -- MHC-II major histocompatibility complex type 2 -- MLV modified live virus -- MOI multiplicity of infection -- MSTAVSKCAT methionine-serine-threonine-alanine-valine-serine-lysine-cysteine-alanine-threonine (scramble peptide, SP) -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide -- MW molecular weight -- NK natural killer -- N10K NQVSLTCLVK (asparagine-glutamine-valine-serine-leucine-threonine-cysteine-leucine-valine-lysine) -- NKT natural killer T -- OD optical density -- PBMC peripheral blood mononuclear cells -- PBS phosphate buffer saline -- PE phycoerythrin -- PE/Cy7 phycoerythrin/cyanin 7 -- PCV2b porcine circovirus type 2b -- PHA phytohemagglutinin -- pI isoelectrical point (IEP) -- PRRSV-1 porcine reproductive and respiratory syndrome virus type 1 (European) -- PRRSV-2 porcine reproductive and respiratory syndrome virus type 2 (North American) -- qPCR quantitative polymerase chain reaction -- RNA ribonucleic acid -- RPMI Roswell Park Memorial Institute -- RT reverse transcription -- SD standard deviation -- SDS sodium dodecyl sulfate -- SIGN-R1 specific ICAM‐3 (intercellular adhesion molecule 3) grabbing non‐integrin receptor 1 -- SP scramble peptide (MSTAVSKCAT) -- Th1 T helper 1 -- VHCDR3 variable heavy chain complementary-determining region 3; Killer peptide (KP) -- Pro-inflammatory monocytes -- Natural killer (NK) T cells -- Double positive cytotoxic T lymphocytes (DP CTL) -- Cytokines -- TNF-α -- IFN-γ secreting cells -- Viral cross-reactivity -- Porcine reproductive and respiratory syndrome virus (PRRSV) -- Porcine circovirus type 2 (PCV2); Communicable diseases in animals -- Periodicals; Veterinary immunology -- Periodicals; Veterinary microbiology -- Periodicals; Immunology -- Periodicals; Microbiology -- Periodicals; Communicable diseases -- Periodicals; Communicable Diseases -- immunology -- Periodicals; Communicable Diseases -- veterinary -- Periodicals; Allergy and Immunology -- Periodicals; Microbiology -- Periodicals; Veterinary Medicine -- Periodicals; Immunologie -- Périodiques; Microbiologie -- Périodiques; Maladies infectieuses -- Périodiques; Communicable diseases; Immunology; Microbiology; Electronic journals; Periodicals; 636.08969
• DOI: 10.1016/j.cimid.2020.101523
• Languages: English
• ISSNs: 0147-9571

Abstract

Highlights: KP early induces pro-inflammatory CD172α+CD14 +high monocytes and CD3 + CD16 + NKT cells. KP triggers CD8α and CD8β expression on CTL and CD4 + CD8α + Th memory cells. KP induces two CD4 + CD8α + CTL subsets associated with increased TNF-α and IFN-γ secretion. KP boosts PRRSV-1- and PCV2b-specific IFN-γ SC reactivity and cross-reactivity. KP is promising as a Th1 response immunomodulator or vaccine adjuvant in pigs. 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.