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T‐Cell Activation. III. Attempts to Activate MHC Class I‐Negative and Class I‐Transfected EL4 T‐Lymphoma Cells by Immobilized Anti‐CD3 Antibody
Author(s) -
CLAESSON M. H.,
DISSING S.,
PLESNER T.,
STURMHÖFEL K.,
BRAMS P.
Publication year - 1990
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1990.tb02888.x
Subject(s) - transfection , ionomycin , microbiology and biotechnology , biology , clone (java method) , cd3 , antibody , mhc class i , mhc class ii , t cell , major histocompatibility complex , antigen , cd8 , cell culture , immunology , intracellular , gene , immune system , biochemistry , genetics
The aim of this study was to examine whether the unresponsiveness of MHC class I‐negative subclones of the EL‐4 thymoma lo CD3 cross‐linking can be restored by transfection of class I genes into the H‐2‐negative cells. Cell activation experiments with selected MHC class I‐negative subclones and H‐2 b ‐and H‐2L d ‐positive transfectants showed that these cells are equally capable of secreting interleukin 2 (IL‐2) after exposure to the phorbol ester phorbol 12‐mvristalc 13‐acetale (PMA) and ionomycin. In contrast, only the parental H‐2‐positive EL4 cells are capable of responding to treatment with immobilized anti‐CD3 antibody with IL‐2 secretion and IL‐2 receptor expression. Measurements of intracellular free Ca 2+ (Ca 2+ i ) following anti‐CD3 antibody‐induced cross‐linking of parental EL4 cells and H‐2‐negativeand H‐2 b gene‐transfected subclones showed that the parental cells and two of the class I transfectants, one H‐2‐positiveand one H‐2‐negative. responded with a slow rise in Ca 2+ ,. whereas one H‐2‐positive transfected cell clone was completely refractory to CD3 cross‐linking. Modulation experiments using parental EL 4 cells, H‐2‐negative subclones and H‐2‐positive transfectants demonstrated that the CD3 and class I molecules of these different cells are modulated to the same extent after exposure lo specific antibodies. The present findings thus indicate that the unresponsiveness of H‐2‐negative EL4 subclone cells not CD3 cross‐linking is not functionally associated with a lack of class I surface expression.