Membrane expression of HLA-Cw4 free chains in activated T cells of transgenic mice.
Transgenic mice were produced in which human HLA-Cw4 is stably integrated, behaves as a single Mendelian trait, and, being under the transcriptional control of human CD2, is selectively and efficiently expressed in T lymphocytes. These mice were used as a model system to determine whether HLA-type C molecules can be exposed on the surface of activated lymphocytes as free heavy chains, non-associated with beta2-microglobulin (beta2m). In our transgenic mice we could identify HLA-Cw4 molecules either as free chains or as beta2m-associated molecules by the use of monoclonal antibodies specific for either conformation of HLA class I and nonreactive to mouse H2 molecules. Resting mouse lymphocytes were shown by western transfer analysis to contain sizeable amounts of HLA-Cw4 free chains, but they exposed on their surface HLA-Cw4 only in association with beta2m, as indicated by flow cytometric measurements. Conversely, where the content of total HLA-Cw4 was increased, lectin-activated mouse lymphocytes exposed on their outer cell membrane HLA-Cw4 molecules in both conformations, namely, also as free heavy chains. Isoelectrofocusing analysis confirmed the presence of both HLA-Cw4 molecular conformations in activated T cells and indicated that HLA-Cw4 heavy chains can bind to mouse beta2m with the same low affinity displayed for human beta2m. The results of our experiments led us to conclude that (1) association with beta2m is not necessary for the exposure of HLA-C on the surface of activated T lymphocytes and (2) cell activation affects the balance between the two conformational forms of HLA-C.
Aiuti
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Dip. Biopatologia Umana, Sez. Biologia Cellulare, Universita di Roma "La Sapienza", Viale Regina Elena, 324, 00161 Roma, Italia.
Forte
P
P
Simeoni
L
L
Lino
M
M
Pozzi
L
L
Fattorossi
A
A
Giacomini
P
P
Ginelli
E
E
Beretta
A
A
Siccardi
A
A
D000818:Animals; D002462:Cell Membrane / Q000276:immunology; D015870:Gene Expression; D015236:HLA-C Antigens / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007526:Isoelectric Point; D008213:Lymphocyte Activation; D046911:Macromolecular Substances; D051379:Mice; D008822:Mice, Transgenic; D012333:RNA, Messenger / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology*; D019076:Transgenes; D001613:beta 2-Microglobulin / Q000378:metabolism
D015236:HLA-C Antigens; C039966:HLA-C*04 antigen; D046911:Macromolecular Substances; D012333:RNA, Messenger; D001613:beta 2-Microglobulin