PMID- 29364214
OWN - NLM
STAT- MEDLINE
DCOM- 20180619
LR  - 20191226
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 130
DP  - 2017 Dec 26
TI  - Assessment of Antibody-based Drugs Effects on Murine Bone Marrow and Peritoneal 
      Macrophage Activation.
LID - 10.3791/56689 [doi]
LID - 56689
AB  - Macrophages are phagocytic innate immune cells, which initiate immune responses 
      to pathogens and contribute to healing and tissue restitution. Macrophages are 
      equally important in turning off inflammatory responses. We have shown that 
      macrophages stimulated with intravenous immunoglobulin (IVIg) can produce high 
      amounts of the anti-inflammatory cytokine, interleukin 10 (IL-10), and low levels 
      of pro-inflammatory cytokines in response to bacterial lipopolysaccharides (LPS). 
      IVIg is a polyvalent antibody, primarily immunoglobulin Gs (IgGs), pooled from 
      the plasma of more than 1,000 blood donors. It is used to supplement antibodies 
      in patients with immune deficiencies or to suppress immune responses in patients 
      with autoimmune or inflammatory conditions. Infliximab, a therapeutic anti-tumor 
      necrosis factor alpha (TNFalpha) antibody, has also been shown to activate 
      macrophages to produce IL-10 in response to inflammatory stimuli. IVIg and other 
      antibody-based biologics can be tested to determine their effects on macrophage 
      activation. This paper describes methods for derivation, stimulation, and 
      assessment of murine bone marrow macrophages activated by antibodies in vitro and 
      murine peritoneal macrophages activated with antibodies in vivo. Finally, we 
      demonstrate the use of western blotting to determine the contribution of specific 
      cell signaling pathways to anti-inflammatory macrophage activity. These protocols 
      can be used with genetically modified mice, to determine the effect of a specific 
      protein(s) on anti-inflammatory macrophage activation. These techniques can also 
      be used to assess whether specific biologics may act by changing macrophages to 
      an IL-10-producing anti-inflammatory activation state that reduces inflammatory 
      responses in vivo. This can provide information on the role of macrophage 
      activation in the efficacy of biologics during disease models in mice, and 
      provide insight into a potential new mechanism of action in people. Conversely, 
      this may caution against the use of specific antibody-based biologics to treat 
      infectious disease, particularly if macrophages play an important role in host 
      defense against that infection.
FAU - Kozicky, Lisa
AU  - Kozicky L
AD  - British Columbia Children's Hospital Research Institute, University of British 
      Columbia; lkozicky@bcchr.ca.
FAU - Sly, Laura M
AU  - Sly LM
AD  - British Columbia Children's Hospital Research Institute, University of British 
      Columbia.
LA  - eng
GR  - CIHR2016-LS/CIHR/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20171226
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Immunoglobulins, Intravenous)
RN  - 0 (Lipopolysaccharides)
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/*drug effects/immunology
MH  - Immunoglobulins, Intravenous/*pharmacology
MH  - Lipopolysaccharides/*pharmacology
MH  - Macrophage Activation/*drug effects/immunology
MH  - Macrophages, Peritoneal/*drug effects/immunology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Models, Animal
PMC - PMC5908398
EDAT- 2018/01/25 06:00
MHDA- 2018/06/21 06:00
PMCR- 2019/12/26
CRDT- 2018/01/25 06:00
PHST- 2018/01/25 06:00 [entrez]
PHST- 2018/01/25 06:00 [pubmed]
PHST- 2018/06/21 06:00 [medline]
PHST- 2019/12/26 00:00 [pmc-release]
AID - 56689 [pii]
AID - 10.3791/56689 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Dec 26;(130):56689. doi: 10.3791/56689.