PMID- 37318241
OWN - NLM
STAT- MEDLINE
DCOM- 20230619
LR  - 20231116
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 195
DP  - 2023 May 19
TI  - Determination of Vaccine Immunogenicity Using Bovine Monocyte-Derived Dendritic 
      Cells.
LID - 10.3791/64874 [doi]
AB  - Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs) within 
      the immune system. They patrol the organism looking for pathogens and play a 
      unique role within the immune system by linking the innate and adaptive immune 
      responses. These cells can phagocytize and then present captured antigens to 
      effector immune cells, triggering a diverse range of immune responses. This paper 
      demonstrates a standardized method for the in vitro generation of bovine 
      monocyte-derived dendritic cells (MoDCs) isolated from cattle peripheral blood 
      mononuclear cells (PBMCs) and their application in evaluating vaccine 
      immunogenicity. Magnetic-based cell sorting was used to isolate CD14(+) monocytes 
      from PBMCs, and the supplementation of complete culture medium with interleukin 
      (IL)-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) was used to 
      induce the differentiation of CD14(+) monocytes into naive MoDCs. The generation 
      of immature MoDCs was confirmed by detecting the expression of major 
      histocompatibility complex II (MHC II), CD86, and CD40 cell surface markers. A 
      commercially available rabies vaccine was used to pulse the immature MoDCs, which 
      were subsequently co-cultured with naive lymphocytes. The flow cytometry analysis 
      of the antigen-pulsed MoDCs and lymphocyte co-culture revealed the stimulation of 
      T lymphocyte proliferation through the expression of Ki-67, CD25, CD4, and CD8 
      markers. The analysis of the mRNA expression of IFN-gamma and Ki-67, using 
      quantitative PCR, showed that the MoDCs could induce the antigen-specific priming 
      of lymphocytes in this in vitro co-culture system. Furthermore, IFN-gamma secretion 
      assessed using ELISA showed a significantly higher titer (**p < 0.01) in the 
      rabies vaccine-pulsed MoDC-lymphocyte co-culture than in the non-antigen-pulsed 
      MoDC-lymphocyte co-culture. These results show the validity of this in vitro MoDC 
      assay to measure vaccine immunogenicity, meaning this assay can be used to 
      identify potential vaccine candidates for cattle before proceeding with in vivo 
      trials, as well as in vaccine immunogenicity assessments of commercial vaccines.
FAU - Liaqat, Fatima
AU  - Liaqat F
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency.
FAU - Kangethe, Richard Thiga
AU  - Kangethe RT
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency.
FAU - Pichler, Rudolf
AU  - Pichler R
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency.
FAU - Liu, Bo
AU  - Liu B
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency.
FAU - Huber, Johann
AU  - Huber J
AD  - Teaching and Research Farm Kremesberg, Clinical Unit for Herd Health Management 
      in Ruminants, Department for Farm Animals and Veterinary Public Health, 
      University of Veterinary Medicine, Vienna.
FAU - Wijewardana, Viskam
AU  - Wijewardana V
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency.
FAU - Cattoli, Giovanni
AU  - Cattoli G
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency.
FAU - Porfiri, Luca
AU  - Porfiri L
AD  - Animal Production and Health Section, Joint Food and Agriculture Organization 
      (FAO)/International Atomic Energy Agency (IAEA) Centre of Nuclear Techniques in 
      Food and Agriculture, International Atomic Energy Agency; l.porfiri@iaea.org.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20230519
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Ki-67 Antigen)
RN  - 0 (Rabies Vaccines)
RN  - 0 (Antigens)
SB  - IM
MH  - Cattle
MH  - Animals
MH  - *Monocytes
MH  - Leukocytes, Mononuclear
MH  - Dendritic Cells
MH  - Ki-67 Antigen/metabolism
MH  - *Rabies Vaccines
MH  - Immunogenicity, Vaccine
MH  - Antigens/metabolism
MH  - Cell Differentiation
MH  - Cells, Cultured
EDAT- 2023/06/15 13:07
MHDA- 2023/06/19 13:08
CRDT- 2023/06/15 09:23
PHST- 2023/06/19 13:08 [medline]
PHST- 2023/06/15 13:07 [pubmed]
PHST- 2023/06/15 09:23 [entrez]
AID - 10.3791/64874 [doi]
PST - epublish
SO  - J Vis Exp. 2023 May 19;(195). doi: 10.3791/64874.