Transfected murine cells expressing HLA class II can be used to generate alloreactive human T cell clones.
Alloreactive human T cells are conventionally generated in vitro using peripheral blood mononuclear cells (PBMCs). The disadvantage of such an approach is that PBMCs express multiple HLA class II molecules and, as a consequence, it is difficult to generate T cells specific for an individual HLA alloantigen. This paper describes a technique in which T cell clones can be generated using stimulators which do express only one alloantigen. This has permitted the generation of HLA-DR-specific T cell clones and will be applied to produce T cell clones specific for other isotypes which cannot easily be obtained using other techniques. Murine DAP.3 cells were transfected with cDNAs encoding human class II molecules and used to stimulate primary alloresponses by purified human CD4+ T cells. The cloning of these T cells provided a good yield of cells allospecific for the class II molecule expressed by the transfected cells. A large percentage of the T cell clones were able to recognise human cells, suggesting that specificity for DR-bound peptides of mouse origin does not limit the applicability of this approach. Despite having been raised against mouse stimulators cells, the responses of the T cell clones to alloantigen-expressing human B cell lines were profoundly inhibited by anti-human LFA-3 monoclonal antibody. The possible mechanisms responsible for these results are discussed.
Warrens
A N
AN
Department of Immunology, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
Heaton
T
T
Sidhu
S
S
Lombardi
G
G
Lechler
R I
RI
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002999:Clone Cells; D005434:Flow Cytometry; D015870:Gene Expression; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007519:Isoantigens / Q000276:immunology*; D007739:L Cells; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D014162:Transfection
D000954:Antigens, Surface; D006684:HLA-DR Antigens; D007519:Isoantigens