TY - JOUR
T1 - Membrane depolarization of human B cells follows stimulation by either anti-μ or B-cell growth factor, but only anti-μ causes cell volume changes
AU - Yokoyama, Wayne M.
AU - Chien, Millie M.
AU - Engardt, Susan E.
AU - Aguiar, Susan W.
AU - Ashman, Robert F.
N1 - Funding Information:
ACKNOWLEDGMENTS The work was supported by NIH Grant GM36261. The authors thank Dr. Ronald Levy for monoclonal antibody 4H9, Dr. John Cambier for a starting supply of cyanine dye, and Dr. Shashikant Mehta for helpful discussions regarding ~he properties of BCGF12. We are also grateful to Mrs. Deanna OUendick for manuscript preparation and Mr. Donald Mower for technical assistance.
PY - 1988
Y1 - 1988
N2 - Human peripheral blood B cells were separated from monocytes and T cells, depleted of null cells by an anti-Leu 9 rosetting technique, and fractionated on discontinuous Percoll gradients to yield a highly purified, small dense B-cell population. These cells responded F(ab′)2 goat anti-μ at 10 and 100 μ/ml with membrane depolarization (measured by immunofluorescence with 3,3′-dipentyloxacarbocyanine dye) at 1 h, cell volume enlargement by 48 h, and modest thymidine incorporation by 72 h. They also responded to the 12-kd human B-cell growth factor of Maizel with membrane depolarization, but not with cell volume increase. F(ab′)2 anti-μ and B-cell growth factor together induced greater depolarization than was seen with either alone, but there was no synergy. The cell volume increase seen with F(ab′)2 anti-μ was not increased by B-cell growth factor. Comparison of data analysis methods showed that mean fluorescence intensity most readily detected significant depolarization. We conclude that in human B cells: (1) depolarization may be a "general response" to a variety of membrane stimuli, because F(ab′)2 anti-μ and B-cell growth factor acting through different receptors both induce it, and (2) depolarization does not inevitably lead to cell volume increase.
AB - Human peripheral blood B cells were separated from monocytes and T cells, depleted of null cells by an anti-Leu 9 rosetting technique, and fractionated on discontinuous Percoll gradients to yield a highly purified, small dense B-cell population. These cells responded F(ab′)2 goat anti-μ at 10 and 100 μ/ml with membrane depolarization (measured by immunofluorescence with 3,3′-dipentyloxacarbocyanine dye) at 1 h, cell volume enlargement by 48 h, and modest thymidine incorporation by 72 h. They also responded to the 12-kd human B-cell growth factor of Maizel with membrane depolarization, but not with cell volume increase. F(ab′)2 anti-μ and B-cell growth factor together induced greater depolarization than was seen with either alone, but there was no synergy. The cell volume increase seen with F(ab′)2 anti-μ was not increased by B-cell growth factor. Comparison of data analysis methods showed that mean fluorescence intensity most readily detected significant depolarization. We conclude that in human B cells: (1) depolarization may be a "general response" to a variety of membrane stimuli, because F(ab′)2 anti-μ and B-cell growth factor acting through different receptors both induce it, and (2) depolarization does not inevitably lead to cell volume increase.
UR - http://www.scopus.com/inward/record.url?scp=0023883763&partnerID=8YFLogxK
U2 - 10.1016/0198-8859(88)90067-5
DO - 10.1016/0198-8859(88)90067-5
M3 - Article
C2 - 3131276
AN - SCOPUS:0023883763
SN - 0198-8859
VL - 21
SP - 155
EP - 164
JO - Human Immunology
JF - Human Immunology
IS - 3
ER -