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- Title
Limiting Dilution Analysis of the Response of Murine L3T4+ Spleen Cells to Alloantigen.
- Authors
Reimann, J.; Bellan, A.
- Abstract
Cell sorter-purified small splenic L3T4+ cells from B6 mice were clonally expanded under limiting dilution (LD) conditions by coculture for 4–6 days with irradiated allogeneic stimulator cells in culture medium supplemented with various growth factor preparations. Proliferating L3T4+ cell clones were detected by [³H]lthymidine uptake; interleukin 2 (IL-2) production of restimulated L3T4+ cell clones was measured in a sensitive colorimetric assay, IL-3 (but not IL-1 or IL-4) supported clonal expansion in vitro of many L3T4+ spleen cells reactive to class II MHC alloantigen. The large majority of proliferating L3T4+ cell clones produced IL-2, The data were consistent with the hypothesis that only a single titrated precursor cell was limiting in the system, fn the response to class II (bm12) H-2 alloantigen. 1 in 40–200 L3T4+ cells was induced to clonal growth: in the response lo class I (bm1) H-2 alloantigen. a tenfold lower frequency (1 in 600–80) of inducible L3T4+ B6 cells was measured, A fraction of the generated L3T4+ cell clones showed lL-2-independent growth: anti-IL-2 receptor monoclonal antibodies (MoAb) (7D4 and PC61.5) blocked the proliferation of about 80% of the IL-2-producing L3T4+ cell clones, while about 20% of these clones seemed resistant to inhibition of proliferation by these MoAb. We have thus defined an LD system with high cloning efficiency for L3T4+ cells that does not depend on exogenous IL-2 supplements.
- Subjects
SPLEEN diseases; SPLEEN; CLONE cells; HEMATOPOIETIC system; IMMUNOLOGIC diseases; MONOCLONAL antibodies
- Publication
Scandinavian Journal of Immunology, 1988, Vol 27, Issue 4, p373
- ISSN
0300-9475
- Publication type
Article
- DOI
10.1111/j.1365-3083.1988.tb02360.x