PMID- 8551044
OWN - NLM
STAT- MEDLINE
DCOM- 19960220
LR  - 20190723
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 188
IP  - 1
DP  - 1995 Dec 15
TI  - Characterization of transendothelial chemotaxis of T lymphocytes.
PG  - 97-116
AB  - We have adapted a chemotaxis assay using human umbilical vein endothelial cell 
      (HUVEC) monolayers on microporous membranes for studying lymphocyte 
      transendothelial chemotaxis in vitro. Supernatants of peripheral blood 
      mononuclear cells stimulated with phytohemagglutinin (PHA) were identified as an 
      excellent source of lymphocyte chemoattractant activity. The activity in PHA 
      supernatant typically caused 2-6% of peripheral blood lymphocytes (PBL) to 
      transmigrate compared to 0.1-0.3% to media control. Checkerboard analysis 
      demonstrated that transmigration was directional and not attributable to random 
      locomotion. Purified T lymphocytes also underwent transendothelial chemotaxis to 
      PHA supernatant. Using monoclonal antibodies to several human adhesion receptors, 
      we found that the interaction between LFA-1 and ICAM-1/ICAM-2 was more important 
      for transendothelial lymphocyte chemotaxis than the interaction between VLA-4 and 
      VCAM-1. A monoclonal antibody to the beta 1 integrin subunit inhibited chemotaxis 
      more than antibodies to the VLA alpha 2, alpha 3, alpha 4, or alpha 5 subunits. 
      The transendothelial assay was used to guide purification of the lymphocyte 
      chemoattractant activity, which we reported previously to be monocyte 
      chemoattractant protein-1 (MCP-1) (Carr et al., Proc. Natl. Acad. Sci. USA (1994) 
      91, 3652). The adhesion molecules required for chemotaxis to MCP-1 were similar 
      to those with PHA supernatant. The use of HUVEC in the assay enhances the 
      signal-to-background ratio of chemotaxis and provides a model that is 
      physiologically relevant to lymphocyte emigration from the bloodstream into sites 
      of inflammation.
FAU - Roth, S J
AU  - Roth SJ
AD  - Department of Cardiology, Children's Hospital, Boston, MA, USA.
FAU - Carr, M W
AU  - Carr MW
FAU - Rose, S S
AU  - Rose SS
FAU - Springer, T A
AU  - Springer TA
LA  - eng
PT  - Journal Article
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Cell Adhesion Molecules)
RN  - 0 (Immunosuppressive Agents)
RN  - 0 (Phytohemagglutinins)
SB  - IM
MH  - Antibodies, Monoclonal/pharmacology
MH  - Cell Adhesion Molecules/immunology
MH  - Cell-Free System/physiology
MH  - Cells, Cultured
MH  - Chemotaxis, Leukocyte/*immunology
MH  - Endothelium, Vascular/*immunology
MH  - Humans
MH  - Immunosuppressive Agents/pharmacology
MH  - Lymphocyte Culture Test, Mixed
MH  - Phytohemagglutinins/pharmacology
MH  - T-Lymphocytes/drug effects/*physiology
MH  - Umbilical Veins
EDAT- 1995/12/15 00:00
MHDA- 1995/12/15 00:01
CRDT- 1995/12/15 00:00
PHST- 1995/12/15 00:00 [pubmed]
PHST- 1995/12/15 00:01 [medline]
PHST- 1995/12/15 00:00 [entrez]
AID - 0022175995002081 [pii]
AID - 10.1016/0022-1759(95)00208-1 [doi]
PST - ppublish
SO  - J Immunol Methods. 1995 Dec 15;188(1):97-116. doi: 10.1016/0022-1759(95)00208-1.