Bryostatin 1 activates splenic lymphocytes and induces sustained depletion of splenocyte protein kinase C activity in vivo after a single intravenous administration.
Bryostatin 1 activates and subsequently down-regulates protein kinase C (PKC) in vitro and has potential use as an immunomodulator and as an anti-cancer agent. Despite extensive examination of its activities in vitro and anti-tumor effects in vivo, previous studies have failed to document that bryostatin 1 modulates total cellular PKC activity in tumor or normal tissues when administered in vivo. After a single bolus injection of bryostatin 1 (1.0 microgram) in normal C57BI/6 mice, blood was drawn at various intervals and assayed for bryostatin 1 levels. In addition, spleens from bryostatin-treated mice were harvested 10 min to 10 days after treatment, weighed and analyzed for cell numbers, PKC activity and cell surface phenotypes. Bryostatin 1 levels in plasma rose rapidly, reaching peak levels of 56.5 nM less than 1 min after injection, and then declined to undetectable levels by 1 h. A similar pattern was observed when bryostatin 1 was incubated with leukemia cells in vitro, raising the possibility that the rapid fall in plasma levels results from intracellular uptake and binding. Bryostatin 1 induced marked depletion of total splenocyte PKC activity (as much as 69% relative to control values) at 24-96 h after drug administration, but not at earlier times (i.e. 1 h). A single injection of bryostatin 1 also induced expression of the T cell activation marker CD69, leading to positivity in 53% of cells at 3-24 h versus 11% in control mice, and resulted in marked splenomegaly, associated with increased numbers of nucleated cells at 48-96 h. Together, these studies demonstrate that despite rapid disappearance of the drug from plasma, a single i.v. dose of bryostatin 1 exhibits significant and sustained effects on normal murine spleen cells, including early lymphocyte activation, prolonged depletion of PKC activity, splenocyte proliferation and splenomegaly. These findings may have implications for attempts to understand the in vivo effects of bryostatin 1 in normal host tissues.
Bear
H D
HD
Massey Cancer Center, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298, USA.
McFadden
A W
AW
Kostuchenko
P J
PJ
Lipshy
K A
KA
Hamad
G G
GG
Turner
A J
AJ
Roberts
J D
JD
Carr
M
M
Carr
S
S
Grant
S
S
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis; D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000494:pharmacology*; D054713:Bryostatins; D002455:Cell Division; D018922:HL-60 Cells / Q000378:metabolism; D006801:Humans; D007783:Lactones / Q000493:pharmacokinetics / Q000494:pharmacology*; D037181:Lectins, C-Type; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D018942:Macrolides; D051379:Mice; D008810:Mice, Inbred C57BL; D011493:Protein Kinase C / Q000032:analysis*; D013154:Spleen / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D013997:Time Factors; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D000970:Antineoplastic Agents; D054713:Bryostatins; D007783:Lactones; D037181:Lectins, C-Type; D018942:Macrolides; D011493:Protein Kinase C; D000099192:CD69 Antigens; C046785:bryostatin 1