PMID- 34126553
OWN - NLM
STAT- MEDLINE
DCOM- 20211115
LR  - 20211115
IS  - 1873-2534 (Electronic)
IS  - 0165-2427 (Linking)
VI  - 238
DP  - 2021 Aug
TI  - New mAbs facilitate quantification of secreted equine TNF-alpha and flow cytometric 
      analysis in monocytes and T cells.
PG  - 110284
LID - S0165-2427(21)00102-1 [pii]
LID - 10.1016/j.vetimm.2021.110284 [doi]
AB  - Tumor necrosis factor-alpha (TNF-alpha) is a pleiotropic cytokine, that is involved in 
      acute inflammation and is employed as a biomarker of inflammatory diseases in 
      several species for which reliable quantification is available. We aimed to 
      develop suitable tools to quantify TNF-alpha in equine samples. We generated two new 
      mAbs against equine TNF-alpha (clones 48 and 292), evaluated their specificity for 
      this cytokine, and confirmed detection of native TNF-alpha in stimulated equine PBMC. 
      The TNF-alpha mAbs were paired in a fluorescent bead-based assay for quantification 
      of equine TNF-alpha. The TNF-alpha assay had a wide quantification range of 12 pg/mL - 
      38.4 ng/mL. In addition, TNF-alpha mAb 48 was used for a detailed analysis of TNF-alpha 
      production in PBMC by intracellular staining and flow cytometry. TNF-alpha was 
      expressed by CD14(+) monocytes after LPS stimulation and by monocytes and 
      lymphocytes after polyclonal stimulation with PMA and ionomycin in vitro. TNF-alpha 
      expressing lymphocytes consisted mainly of CD4(+) T cells. CD8(+) T cells and 
      other lymphocytes also expressed TNF-alpha. The new mAbs evaluated here for soluble 
      and intracellular TNF-alpha will enable the detailed analysis of this important 
      pro-inflammatory cytokine during equine immune responses and inflammatory 
      diseases of the horse.
CI  - Copyright (c) 2021 The Authors. Published by Elsevier B.V. All rights reserved.
FAU - Schnabel, Christiane L
AU  - Schnabel CL
AD  - Department of Population Medicine and Diagnostic Sciences, College of Veterinary 
      Medicine, Cornell University, Ithaca, NY, USA; Institute of Immunology, College 
      of Veterinary Medicine, Leipzig University, Leipzig, Germany(1).
FAU - Babasyan, Susanna
AU  - Babasyan S
AD  - Department of Population Medicine and Diagnostic Sciences, College of Veterinary 
      Medicine, Cornell University, Ithaca, NY, USA.
FAU - Freer, Heather
AU  - Freer H
AD  - Department of Population Medicine and Diagnostic Sciences, College of Veterinary 
      Medicine, Cornell University, Ithaca, NY, USA.
FAU - Larson, Elisabeth M
AU  - Larson EM
AD  - Department of Population Medicine and Diagnostic Sciences, College of Veterinary 
      Medicine, Cornell University, Ithaca, NY, USA.
FAU - Wagner, Bettina
AU  - Wagner B
AD  - Department of Population Medicine and Diagnostic Sciences, College of Veterinary 
      Medicine, Cornell University, Ithaca, NY, USA. Electronic address: 
      bw73@cornell.edu.
LA  - eng
PT  - Journal Article
DEP - 20210610
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Tumor Necrosis Factor-alpha)
SB  - IM
MH  - Animals
MH  - *Antibodies, Monoclonal
MH  - CHO Cells
MH  - Cloning, Molecular
MH  - Cricetinae
MH  - Cricetulus
MH  - Female
MH  - Flow Cytometry
MH  - Gene Expression Regulation
MH  - *Horses
MH  - Immunomagnetic Separation
MH  - Lymphocyte Activation/immunology
MH  - Male
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Monocytes/*physiology
MH  - T-Lymphocytes/*physiology
MH  - Tumor Necrosis Factor-alpha/*immunology/*metabolism
OTO - NOTNLM
OT  - Bead-based assay
OT  - Cytokine
OT  - Flow cytometry
OT  - Horse
OT  - Inflammation
EDAT- 2021/06/15 06:00
MHDA- 2021/11/16 06:00
CRDT- 2021/06/14 20:28
PHST- 2021/03/09 00:00 [received]
PHST- 2021/06/07 00:00 [revised]
PHST- 2021/06/08 00:00 [accepted]
PHST- 2021/06/15 06:00 [pubmed]
PHST- 2021/11/16 06:00 [medline]
PHST- 2021/06/14 20:28 [entrez]
AID - S0165-2427(21)00102-1 [pii]
AID - 10.1016/j.vetimm.2021.110284 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2021 Aug;238:110284. doi: 10.1016/j.vetimm.2021.110284. 
      Epub 2021 Jun 10.