PMID- 29863666
OWN - NLM
STAT- MEDLINE
DCOM- 20180730
LR  - 20200518
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 135
DP  - 2018 May 18
TI  - Development and Functional Characterization of Murine Tolerogenic Dendritic 
      Cells.
LID - 10.3791/57637 [doi]
LID - 57637
AB  - The immune system operates by maintaining a tight balance between coordinating 
      responses against foreign antigens and maintaining an unresponsive state against 
      self-antigens as well as antigens derived from commensal organisms. The 
      disruption of this immune homeostasis can lead to chronic inflammation and to the 
      development of autoimmunity. Dendritic cells (DCs) are the professional 
      antigen-presenting cells of the innate immune system involved in activating naive 
      T cells to initiate immune responses against foreign antigens. However, DCs can 
      also be differentiated into TolDCs that act to maintain and promote T cell 
      tolerance and to suppress effector cells contributing to the development of 
      either autoimmune or chronic inflammation conditions. The recent advancement in 
      our understanding of TolDCs suggests that DC tolerance can be achieved by 
      modulating their differentiation conditions. This phenomenon has led to 
      tremendous growth in developing TolDC therapies for numerous immune disorders 
      caused due to break in immune tolerance. Successful studies in preclinical 
      autoimmunity murine models have further validated the immunotherapeutic utility 
      of TolDCs in the treatment of autoimmune disorders. Today, TolDCs have become a 
      promising immunotherapeutic tool in the clinic for reinstating immune tolerance 
      in various immune disorders by targeting pathogenic autoimmune responses while 
      leaving protective immunity intact. Although an array of strategies has been 
      proposed by multiple labs to induce TolDCs, there is no consistency in 
      characterizing the cellular and functional phenotype of these cells. This 
      protocol provides a step-by-step guide for the development of bone marrow-derived 
      DCs in large numbers, a unique method used to differentiate them into TolDCs with 
      a synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic 
      acid-difluoro-propyl-amide (CDDO-DFPA), and the techniques used to confirm their 
      phenotype, including analyses of essential molecular signatures of TolDCs. 
      Finally, we show a method to assess TolDC function by testing their 
      immunosuppressive response in vitro and in vivo in a preclinical model of 
      multiple sclerosis.
FAU - Wei, Hsi-Ju
AU  - Wei HJ
AD  - Department of Biochemistry, School of Medicine, Case Western Reserve University.
FAU - Letterio, John J
AU  - Letterio JJ
AD  - Department of Pediatrics, Division of Pediatric Hematology/Oncology, Case Western 
      Reserve University; Angie Fowler Cancer Institute, Rainbow Babies & Children's 
      Hospital, University Hospitals, Cleveland.
FAU - Pareek, Tej K
AU  - Pareek TK
AD  - Department of Pediatrics, Division of Pediatric Hematology/Oncology, Case Western 
      Reserve University; Angie Fowler Cancer Institute, Rainbow Babies & Children's 
      Hospital, University Hospitals, Cleveland; tkp5@case.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Video-Audio Media
DEP - 20180518
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Dendritic Cells/*immunology
MH  - Immune Tolerance/*genetics
MH  - Mice
PMC - PMC6101271
EDAT- 2018/06/05 06:00
MHDA- 2018/07/31 06:00
PMCR- 2020/05/18
CRDT- 2018/06/05 06:00
PHST- 2018/06/05 06:00 [entrez]
PHST- 2018/06/05 06:00 [pubmed]
PHST- 2018/07/31 06:00 [medline]
PHST- 2020/05/18 00:00 [pmc-release]
AID - 57637 [pii]
AID - 10.3791/57637 [doi]
PST - epublish
SO  - J Vis Exp. 2018 May 18;(135):57637. doi: 10.3791/57637.