Abstract
This study investigated the requirements for lymphokines derived by recombinant (rec.) DNA technology for the induction of growth and maturation in highly purified lectin reactive T cell subsets. Nylon purified C57BL/6 lymph node T cells were treated with monoclonal anti-Lyt-2.2 or anti-L3T4 antibodies and fluorescence labeled (FITC) anti- immunoglobulin antibodies and were positively selected into Lyt-2+ (L3T4-) and Lyt-2- (L3T4+) lymphocyte subsets using a fluorescence-activated cell sorter. Sorted T lymphocytes, which were devoid of accessory cells were incubated either in bulk culture (2 x 102 - 3 x 104 cells/microculture) or under limiting dilution conditions (2.5–1000 cells/well) with lectin (Concanavalin A, Leukoagglutinin) and rec. human Interleukin 2 (rec. hIL-2) and/or rec. mouse Interferon Y (rec. mIFN-γ). The data show that Lyt-2+ lymphocytes respond to lectin and rec. hIL-2 with growth and development of cytolytic activity in the absence of other exogenous factor(s) or accessory cells. The presence of monoclonal antibodies to the Interleukin 2 receptor during the sensitization phase ablated the induction of Con A reactive precursor cells of cytolytic lymphocytes (CTL-P) by either rec. hIL-2 or conventional IL-2 containing lymphokine sources, indicating the essential role of IL-2 during activation of Lyt-2+ T lymphocytes. In contrast, Lyt-2- lymphocytes could not be induced by lectin and rec. hIL-2 alone for proliferation and always required the presence of accessory cells for significant growth. Exogenous rec. m IFNy was unable to induce growth and cytolytic activity in Con A reactive Lyt-2+ cells and did not significantly effect their response to rec. hIL-2. Limiting dilution experiments revealed that 10–16 % of the Lyt-2+ lymphocytes responded to Con A and rec.
Original language | English |
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Pages (from-to) | 274-301 |
Number of pages | 28 |
Journal | Immunobiology |
Volume | 171 |
Issue number | 3 |
DOIs | |
State | Published - 1986 |
Keywords
- AC
- B6
- C57BL/6 mouse strain
- CTL
- CTL-P
- Con A
- ConASN
- Concanavalin A
- FCF
- FCS
- FITC
- GTL-P
- HYBSN
- IFN-γ
- IL-2
- IL-2R
- Interferon gamma
- Interleukin 2
- LA
- LD
- LN
- Leukoagglutinin
- MLC
- MLCSN
- PBS
- PHA
- PTL-P
- Phytohemagglutinin
- SC
- SD
- SN
- T cell deprived, irradiated spleen cells
- T,-P
- [H]dThd
- cytotoxic T lymphocytes
- fetal calf serum
- flow cytofluorometry
- fluorescein isothiocyanate
- irradiated whole spleen cells
- limiting dilution
- lymph node
- mixed lymphocyte culture
- phosphate-buffered saline
- precursor cell of CTL
- precursor cell of IFN-,γ-secreting T cell
- precursor cell of growing T cell
- precursor cell of proliferating T cell
- rec. hIL-2
- rec. mIFNγ
- receptor for Interleukin 2
- recombinant human Interleukin 2
- recombinant mouse Interferon gamma
- standard deviation
- supernatant
- supernatant from Con A-stimulated rat spleen cell
- supernatant from hybridoma
- supernatant from secondary mixed lymphocyte culture
- tritiated thymidine