Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones.
A limiting dilution microculture system, supplemented with a source of interleukin-2 (IL-2), was employed to evaluate the frequency of Moloney-murine leukaemia/sarcoma virus (M-MuLV/M-MSV)-specific cytotoxic T-lymphocyte precursors (CTL-p) which also exhibited NK-like activity. Spleen cells, obtaine...
| Main Authors: | , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
1985
|
| _version_ | 1826273837241597952 |
|---|---|
| author | Ronchese, F Collavo, D Zanovello, P Cerundolo, V Biasi, G |
| author_facet | Ronchese, F Collavo, D Zanovello, P Cerundolo, V Biasi, G |
| author_sort | Ronchese, F |
| collection | OXFORD |
| description | A limiting dilution microculture system, supplemented with a source of interleukin-2 (IL-2), was employed to evaluate the frequency of Moloney-murine leukaemia/sarcoma virus (M-MuLV/M-MSV)-specific cytotoxic T-lymphocyte precursors (CTL-p) which also exhibited NK-like activity. Spleen cells, obtained from M-MuLV/M-MSV regressor mice, were restimulated in bulk secondary mixed leucocyte-tumour cell cultures (MLTC), and subsequently plated in a culture medium supplemented with two different supernatants (SN) produced following PMA-stimulation of the same EL-4 thymoma cell line. SN 20, obtained from the cell line maintained in vitro, contained IL-2 and only negligible amounts (less than 3 U/ml) of interferon (IFN), while SN 19, obtained after passage of the ascitic form of EL-4 thymoma in syngeneic mice, contained both IL-2 and IFN in high titres. The frequency of CTL-p specific for MBL-2 lymphoma cells was high and comparable in cultures supplemented with both SN (1/2 X 84 cells and 1/2 X 40 cells, respectively), while the frequency of CTL-p directed against NK-susceptible YAC-1 target cells was low in SN 20 (1/90 cells) and high in SN 19 (1/5 X 40 cells). An analysis of individual microcultures established at low cell dose (1 cell/well) indicated that specific and NK-like activity could be ascribed to the same precursor cells. Furthermore, using different long-term CTL clones, we observed that, after passage in SN 20, double-reactive clones gradually lose the capacity to lyse NK-susceptible targets, while most of MBL-2 specific clones acquired NK-like activity following a few passages in SN 19. Therefore, the induction of NK-like activity is reversible and may be modulated by soluble factors present in supernatant in which CTL clones are maintained. Double-reactive clones were unable to lyse NK-resistant allogeneic tumour cells or normal syngeneic blast cells. A few clones cross-reacting with H-2d alloantigens also exhibited NK-like activity when maintained in SN 19. The different pattern of CTL clone activity was associated with a morphological change in the clones themselves: the acquisition of double activity was accompanied by an increase in cell size and the appearance of numerous cytoplasmic granules. All CTL clones were phenotypically Thy-1+ and Lyt-2+ on indirect immunofluorescence and complement-dependent cytotoxicity investigation.(ABSTRACT TRUNCATED AT 400 WORDS) |
| first_indexed | 2024-03-06T22:34:17Z |
| format | Journal article |
| id | oxford-uuid:59598137-9bb7-4da5-b77a-c85931b4e619 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:34:17Z |
| publishDate | 1985 |
| record_format | dspace |
| spelling | oxford-uuid:59598137-9bb7-4da5-b77a-c85931b4e6192022-03-26T17:09:16ZReversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:59598137-9bb7-4da5-b77a-c85931b4e619EnglishSymplectic Elements at Oxford1985Ronchese, FCollavo, DZanovello, PCerundolo, VBiasi, GA limiting dilution microculture system, supplemented with a source of interleukin-2 (IL-2), was employed to evaluate the frequency of Moloney-murine leukaemia/sarcoma virus (M-MuLV/M-MSV)-specific cytotoxic T-lymphocyte precursors (CTL-p) which also exhibited NK-like activity. Spleen cells, obtained from M-MuLV/M-MSV regressor mice, were restimulated in bulk secondary mixed leucocyte-tumour cell cultures (MLTC), and subsequently plated in a culture medium supplemented with two different supernatants (SN) produced following PMA-stimulation of the same EL-4 thymoma cell line. SN 20, obtained from the cell line maintained in vitro, contained IL-2 and only negligible amounts (less than 3 U/ml) of interferon (IFN), while SN 19, obtained after passage of the ascitic form of EL-4 thymoma in syngeneic mice, contained both IL-2 and IFN in high titres. The frequency of CTL-p specific for MBL-2 lymphoma cells was high and comparable in cultures supplemented with both SN (1/2 X 84 cells and 1/2 X 40 cells, respectively), while the frequency of CTL-p directed against NK-susceptible YAC-1 target cells was low in SN 20 (1/90 cells) and high in SN 19 (1/5 X 40 cells). An analysis of individual microcultures established at low cell dose (1 cell/well) indicated that specific and NK-like activity could be ascribed to the same precursor cells. Furthermore, using different long-term CTL clones, we observed that, after passage in SN 20, double-reactive clones gradually lose the capacity to lyse NK-susceptible targets, while most of MBL-2 specific clones acquired NK-like activity following a few passages in SN 19. Therefore, the induction of NK-like activity is reversible and may be modulated by soluble factors present in supernatant in which CTL clones are maintained. Double-reactive clones were unable to lyse NK-resistant allogeneic tumour cells or normal syngeneic blast cells. A few clones cross-reacting with H-2d alloantigens also exhibited NK-like activity when maintained in SN 19. The different pattern of CTL clone activity was associated with a morphological change in the clones themselves: the acquisition of double activity was accompanied by an increase in cell size and the appearance of numerous cytoplasmic granules. All CTL clones were phenotypically Thy-1+ and Lyt-2+ on indirect immunofluorescence and complement-dependent cytotoxicity investigation.(ABSTRACT TRUNCATED AT 400 WORDS) |
| spellingShingle | Ronchese, F Collavo, D Zanovello, P Cerundolo, V Biasi, G Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones. |
| title | Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones. |
| title_full | Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones. |
| title_fullStr | Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones. |
| title_full_unstemmed | Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones. |
| title_short | Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones. |
| title_sort | reversibility of lymphokine induced nk like activity in virus specific cytotoxic t lymphocyte clones |
| work_keys_str_mv | AT ronchesef reversibilityoflymphokineinducednklikeactivityinvirusspecificcytotoxictlymphocyteclones AT collavod reversibilityoflymphokineinducednklikeactivityinvirusspecificcytotoxictlymphocyteclones AT zanovellop reversibilityoflymphokineinducednklikeactivityinvirusspecificcytotoxictlymphocyteclones AT cerundolov reversibilityoflymphokineinducednklikeactivityinvirusspecificcytotoxictlymphocyteclones AT biasig reversibilityoflymphokineinducednklikeactivityinvirusspecificcytotoxictlymphocyteclones |