PMID- 9075772
OWN - NLM
STAT- MEDLINE
DCOM- 19970411
LR  - 20190723
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 202
IP  - 1
DP  - 1997 Mar 10
TI  - Whole-blood incubation method to study neutrophil cytoskeletal dynamics.
PG  - 59-66
AB  - To reduce artifactual effects in the study of filamentous (F)-actin dynamics in 
      neutrophils, we have developed a whole-blood incubation method. Neutrophils in 
      whole blood contained significantly less basal F-actin than did separated 
      neutrophils. Although the peak relative F-actin content of neutrophils in whole 
      blood after formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation was 
      significantly higher than that of separated neutrophils at 10(-9) to 10(-6) M 
      fMLP concentrations (p < 0.05), there was no significant difference in increase 
      in mean fluorescence intensity and the EC50 (concentration of stimulant giving a 
      half-maximum response). On the other hand, the EC50 of platelet-activating factor 
      (PAF) between separated neutrophils and whole-blood-incubated neutrophils 
      differed significantly (1.6 +/- 1.1 x 10(-9) M in separated neutrophils and 2.0 
      +/- 0.7 x 10(-8) M in whole-blood-incubated neutrophils, p < 0.05). The 
      whole-blood incubation method described presently reduces the sample volume, cost 
      and time needed to separate neutrophils, prevents neutrophil activation during 
      separation, and reserves all blood components that may affect neutrophil 
      function. For these reasons, the conditions adopted in the present method are 
      thought to simulate well neutrophils circulating in vivo and the method would be 
      preferable to other neutrophil function tests performed to study actin dynamics.
FAU - Mineshita, M
AU  - Mineshita M
AD  - 2nd Department of Internal Medicine, Hiroshima University School of Medicine, 
      Japan.
FAU - Kimura, T
AU  - Kimura T
FAU - Murai, H
AU  - Murai H
FAU - Moritani, C
AU  - Moritani C
FAU - Ishioka, S
AU  - Ishioka S
FAU - Kambe, M
AU  - Kambe M
FAU - Yamakido, M
AU  - Yamakido M
LA  - eng
PT  - Journal Article
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Actins)
RN  - 0 (Platelet Activating Factor)
RN  - 59880-97-6 (N-Formylmethionine Leucyl-Phenylalanine)
SB  - IM
MH  - Actins/blood
MH  - Blood
MH  - Cell Separation
MH  - Cytoskeleton/*immunology/*metabolism
MH  - Dose-Response Relationship, Drug
MH  - Flow Cytometry/methods
MH  - Humans
MH  - N-Formylmethionine Leucyl-Phenylalanine/pharmacology
MH  - Neutrophils/*cytology/drug effects/*metabolism
MH  - Platelet Activating Factor/pharmacology
MH  - Time Factors
EDAT- 1997/03/10 00:00
MHDA- 1997/03/10 00:01
CRDT- 1997/03/10 00:00
PHST- 1997/03/10 00:00 [pubmed]
PHST- 1997/03/10 00:01 [medline]
PHST- 1997/03/10 00:00 [entrez]
AID - S0022-1759(96)00242-6 [pii]
AID - 10.1016/s0022-1759(96)00242-6 [doi]
PST - ppublish
SO  - J Immunol Methods. 1997 Mar 10;202(1):59-66. doi: 10.1016/s0022-1759(96)00242-6.