Premium
Studies of Folate Uptake by Phytohaemagglutinin‐Stimulated Lymphocytes
Author(s) -
Das K. C.,
Hoffbrand A. V.
Publication year - 1970
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1970.tb01617.x
Subject(s) - phytohaemagglutinin , biochemistry , in vitro , chemistry , microbiology and biotechnology , metabolism , biology
S ummary Cultures of lymphocytes with and without phytohaemagglutinin (PHA) were used as an in vitro model system to study the cellular uptake of tritiated folic acid, [ 3 H]PteGlu and of 14 C labelled 5 methyltetrahydrofolic acid, 5 [methyl‐ 14 C]‐H 4 PteGlu. Total folate uptake by PHA‐stimulated transformed lymphocytes, measured both by liquid scintillation counting and by autoradiography, was much greater than by non‐stimulated mature lymphocytes. This suggests that growing and dividing cells take up folate more avidly than mature non‐dividing cells. Uptake of both folate compounds, measured over a 4 hr period, was (a) approximately 80% less at 4°C than at 37°C, (b) exhibited saturation kinetics, and (c) was inhibited by methotrexate in concentrations of 10 −3 M, 10 −4 M, but was not affected by methotrexate at a concentration of 10 −6 M. The results suggest that an active transport mechanism may be at least partly involved in cellular folate uptake. The two folate compounds, PteGlu and 5 methyl H 4 PteGlu, however, do not appear to share the same uptake pathway, since PteGlu did not inhibit the uptake of 5 [methyl‐ 14 C]H 4 PteGlu by PHA‐stimulated lymphocytes. Further, the uptake of 5 [methyl‐ 14 C]H 4 PteGlu by PHA‐stimulated lymphocytes from six patients with untreated pernicious anaemia was found to be significantly impaired, whereas the uptake of [ 3 H]PteGlu by those cells was normal. Diphenylhydantoin did not show any consistent effect on folate uptake by lymphocytes. A sequential relation was found between the peak rates of RNA synthesis, folate uptake and DNA synthesis by PHA‐stimulated lymphocytes. The peak rate of folate uptake occurred at 44–48 hr of culture, and preceded peak DNA synthesis by 24 hr and followed peak RNA synthesis by 24 hr. Folate uptake, however, did not appear to be directly coupled to either DNA or RNA synthesis, since actinomycin D inhibited both DNA and RNA synthesis by transformed lymphocytes over a 4 hr period without significantly affecting folate uptake.