Abstract
Quiescent human peripheral blood lymphocytes have been shown to maintain a relatively constant intracellular pH of 7.0-7.2 over an extracellular pH range of 6.9-7.4. Two methods of measuring intracellular pH were used in these studies, 19F nuclear magnetic resonance and [14C]5,5-dimethyloxazolidine-2,4-dione (DMO) equilibrium distributions. When ATP levels were decreased in these cells, actively maintained pH regulation was abolished and cells exhibited a constant pH gradient of 0.2 pH unit (acid inside relative to outside). Possible mechanisms for pH regulation are discussed. The effects of the Na+ and K+ composition of the medium on pH regulation showed no correlation with their effects on mitogen-induced proliferative response, which we have previously determined (Deutsch, C., and M. Price, 1982, J. Cell. Physiol., 111:73-79). In low-Na+ mannitol medium, pH regulation was similar to that observed for lymphocytes in normal medium, whereas mitogen-induced proliferation was severely inhibited in low-Na+ mannitol. In contrast, high-K+, low Na+ medium caused loss of pH homeostasis, whereas it restored the proliferative response. Loss of pH homeostasis was also observed on prolonged exposure of lymphocytes to mitogen (greater than 6 h in culture). However, mitogen stimulation led to little or no change in intracellular pH in the first few hours of cell culture. Therefore, a shift in intracellular pH is not a necessary or general event in mitogen-stimulated proliferation of lymphocytes.
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- Chang H., Saccomani G., Rabon E., Schackmann R., Sachs G. Proton transport by gastric membrane vesicles. Biochim Biophys Acta. 1977 Jan 21;464(2):313–327. doi: 10.1016/0005-2736(77)90006-2. [DOI] [PubMed] [Google Scholar]
- Deamer D. W. Proton permeability in biological and model membranes. Kroc Found Ser. 1981;15:173–187. [PubMed] [Google Scholar]
- Deutsch C., Price M. A. Cell calcium in human peripheral blood lymphocytes and the effect of mitogen. Biochim Biophys Acta. 1982 May 7;687(2):211–218. doi: 10.1016/0005-2736(82)90548-x. [DOI] [PubMed] [Google Scholar]
- Deutsch C., Price M. A., Johansson C. A sodium requirement for mitogen-induced proliferation in human peripheral blood lymphocytes. Exp Cell Res. 1981 Dec;136(2):359–369. doi: 10.1016/0014-4827(81)90015-x. [DOI] [PubMed] [Google Scholar]
- Deutsch C., Price M. Role of extracellular Na and K in lymphocyte activation. J Cell Physiol. 1982 Oct;113(1):73–79. doi: 10.1002/jcp.1041130113. [DOI] [PubMed] [Google Scholar]
- Deutsch C., Slater L., Goldstein P. Volume regulation of human peripheral blood lymphocytes and stimulated proliferation of volume-adapted cells. Biochim Biophys Acta. 1982 Nov 17;721(3):262–267. doi: 10.1016/0167-4889(82)90078-7. [DOI] [PubMed] [Google Scholar]
- Deutsch C., Taylor J. S., Wilson D. F. Regulation of intracellular pH by human peripheral blood lymphocytes as measured by 19F NMR. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7944–7948. doi: 10.1073/pnas.79.24.7944. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Felber S. M., Brand M. D. Concanavalin A causes an increase in sodium permeability and intracellular sodium content of pig lymphocytes. Biochem J. 1983 Mar 15;210(3):893–897. doi: 10.1042/bj2100893. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Felber S. M., Brand M. D. Early plasma-membrane-potential changes during stimulation of lymphocytes by concanavalin A. Biochem J. 1983 Mar 15;210(3):885–891. doi: 10.1042/bj2100885. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerson D. F., Kiefer H., Eufe W. Intracellular pH of mitogen-stimulated lymphocytes. Science. 1982 May 28;216(4549):1009–1010. doi: 10.1126/science.6281887. [DOI] [PubMed] [Google Scholar]
- Gerson D. F., Kiefer H. High intracellular pH accompanies mitotic activity in murine lymphocytes. J Cell Physiol. 1982 Jul;112(1):1–4. doi: 10.1002/jcp.1041120102. [DOI] [PubMed] [Google Scholar]
- Gillies R. J., Deamer D. W. Intracellular pH changes during the cell cycle in Tetrahymena. J Cell Physiol. 1979 Jul;100(1):23–31. doi: 10.1002/jcp.1041000103. [DOI] [PubMed] [Google Scholar]
- Gillies R. J., Ogino T., Shulman R. G., Ward D. C. 31P nuclear magnetic resonance evidence for the regulation of intracellular pH by Ehrlich ascites tumor cells. J Cell Biol. 1982 Oct;95(1):24–28. doi: 10.1083/jcb.95.1.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gonzalez-Mendez R., Wemmer D., Hahn G., Wade-Jardetzky N., Jardetzky O. Continuous-flow NMR culture system for mammalian cells. Biochim Biophys Acta. 1982 Jun 8;720(3):274–280. doi: 10.1016/0167-4889(82)90051-9. [DOI] [PubMed] [Google Scholar]
- Heinz A., Sachs G., Schafer J. A. Evidence for activation of an active electrogenic proton pump in Ehrlich ascites tumor cells during glycolysis. J Membr Biol. 1981;61(3):143–153. doi: 10.1007/BF01870520. [DOI] [PubMed] [Google Scholar]
- Holian A., Deutsch C. J., Holian S. K., Daniele R. P., Wilson D. F. Lymphocyte response to phytohemagglutinin: intracellular volume and intracellular [K+]. J Cell Physiol. 1979 Jan;98(1):137–144. doi: 10.1002/jcp.1040980115. [DOI] [PubMed] [Google Scholar]
- Kiefer H., Blume A. J., Kaback H. R. Membrane potential changes during mitogenic stimulation of mouse spleen lymphocytes. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2200–2204. doi: 10.1073/pnas.77.4.2200. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee J., Simpson G., Scholes P. An ATPase from dog gastric mucosa: changes of outer pH in suspensions of membrane vesicles accompanying ATP hydrolysis. Biochem Biophys Res Commun. 1974 Sep 23;60(2):825–832. doi: 10.1016/0006-291x(74)90315-5. [DOI] [PubMed] [Google Scholar]
- Levin G. E., Collinson P., Baron D. N. The intracellular pH of human leucocytes in response to acid-base changes in vitro. Clin Sci Mol Med. 1976 Apr;50(4):293–299. doi: 10.1042/cs0500293. [DOI] [PubMed] [Google Scholar]
- Negendank W., Shaller C. The effect of metabolic inhibition on ion contents and sodium exchange in human lymphocytes. J Cell Physiol. 1982 Mar;110(3):291–299. doi: 10.1002/jcp.1041100312. [DOI] [PubMed] [Google Scholar]
- Rink T. J., Montecucco C., Hesketh T. R., Tsien R. Y. Lymphocyte membrane potential assessed with fluorescent probes. Biochim Biophys Acta. 1980;595(1):15–30. doi: 10.1016/0005-2736(80)90243-6. [DOI] [PubMed] [Google Scholar]
- Rink T. J., Tsien R. Y., Pozzan T. Cytoplasmic pH and free Mg2+ in lymphocytes. J Cell Biol. 1982 Oct;95(1):189–196. doi: 10.1083/jcb.95.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roos A., Boron W. F. Intracellular pH. Physiol Rev. 1981 Apr;61(2):296–434. doi: 10.1152/physrev.1981.61.2.296. [DOI] [PubMed] [Google Scholar]
- Sachs G., Chang H. H., Rabon E., Schackman R., Lewin M., Saccomani G. A nonelectrogenic H+ pump in plasma membranes of hog stomach. J Biol Chem. 1976 Dec 10;251(23):7690–7698. [PubMed] [Google Scholar]
- Schackmann R., Schwartz A., Saccomani G., Sachs G. Cation transport by gastric H+:K+ ATPase. J Membr Biol. 1977 Apr 22;32(3-4):361–381. doi: 10.1007/BF01905228. [DOI] [PubMed] [Google Scholar]
- Segel G. B., Simon W., Lichtman M. A. Regulation of sodium and potassium transport in phytohemagglutinin-stimulated human blood lymphocytes. J Clin Invest. 1979 Sep;64(3):834–841. doi: 10.1172/JCI109531. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shapiro H. M., Natale P. J., Kamentsky L. A. Estimation of membrane potentials of individual lymphocytes by flow cytometry. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5728–5730. doi: 10.1073/pnas.76.11.5728. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slonczewski J. L., Rosen B. P., Alger J. R., Macnab R. M. pH homeostasis in Escherichia coli: measurement by 31P nuclear magnetic resonance of methylphosphonate and phosphate. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6271–6275. doi: 10.1073/pnas.78.10.6271. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor J. S., Deutsch C. Fluorinated alpha-methylamino acids as 19F NMR indicators of intracellular pH. Biophys J. 1983 Sep;43(3):261–267. doi: 10.1016/S0006-3495(83)84349-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsien R. Y., Pozzan T., Rink T. J. T-cell mitogens cause early changes in cytoplasmic free Ca2+ and membrane potential in lymphocytes. Nature. 1982 Jan 7;295(5844):68–71. doi: 10.1038/295068a0. [DOI] [PubMed] [Google Scholar]
- Zieve P. D., Haghshenass M., Krevans J. R. Intracellular pH of the human lymphocyte. Am J Physiol. 1967 May;212(5):1099–1102. doi: 10.1152/ajplegacy.1967.212.5.1099. [DOI] [PubMed] [Google Scholar]
