Precursor frequency analysis of bryostatin activated lymphocytes

Martin Fleming, Sandra K. Barrett, Harry D. Bear

Research output: Contribution to journalArticle

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Abstract

Bryostatin 1 (Bryo) is a potent activator of protein kinase C. When T cells are stimulated with Bryo and the calcium ionophore ionomycin (Io), they expand rapidly in low-dose IL-2 (20 U/ml). We have shown that Bryo/Io- activated T-cells from murine tumor-draining lymph nodes have striking antigen-specific antitumor efficacy in vivo. To account for the specificity despite using a nonspecific T cell activation method, it was postulated that the Bryo/Io combination might preferentially activate antigen-sensitized T cells. To test this hypothesis, an allogeneic response model was used. C57BL/6 mice were primed by intraperitoneal (ip) injection with DBA/2 mouse splenocytes. After 9 days, the C57BL/6 spleens were harvested and H-2d- specific cytolytic T lymphocyte (CTL) precursor frequency (PF) was determined by limiting dilution analysis (LDA). A portion of the same spleen cells was treated for 18 hr with Bryo/Io and expanded for 7 days in IL-2 (20 U/ml); the LDA was then repeated to analyze PF after expansion. The entire experiment was also done with responder and stimulator strains reversed (DBA/2 mice immunized with C57BL/6 cells). Normal spleen cells treated with Bryo/Io exhibited <10% release vs the same targets at any effector:target ratio, ruling out nonspecific cytolysis after Bryo/Io. T lymphocytes or CD8+ T cells were selected from primed splenocytes and the PF values before and after Bryo/Io were analyzed. These data showed that the increase in PF after expansion could not be attributed to T cell or CD8+ cell enrichment alone. Therefore, Bryo/Io treatment of lymphocytes from antigen sensitized mice apparently causes preferential activation and expansion of sensitized or memory T cells.

Original languageEnglish (US)
Pages (from-to)74-79
Number of pages6
JournalJournal of Surgical Research
Volume57
Issue number1
DOIs
StatePublished - Jan 1 1994

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Bryostatins
Ionomycin
Lymphocytes
T-Lymphocytes
Inbred DBA Mouse
Spleen
Antigens
Interleukin-2
bryostatin 1
Calcium Ionophores
Intraperitoneal Injections
Inbred C57BL Mouse
Protein Kinase C
Lymph Nodes

All Science Journal Classification (ASJC) codes

  • Surgery

Cite this

Precursor frequency analysis of bryostatin activated lymphocytes. / Fleming, Martin; Barrett, Sandra K.; Bear, Harry D.

In: Journal of Surgical Research, Vol. 57, No. 1, 01.01.1994, p. 74-79.

Research output: Contribution to journalArticle

Fleming, Martin ; Barrett, Sandra K. ; Bear, Harry D. / Precursor frequency analysis of bryostatin activated lymphocytes. In: Journal of Surgical Research. 1994 ; Vol. 57, No. 1. pp. 74-79.
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