Cloning of T lymphocytes from bronchoalveolar lavage fluid.
We have prepared T-cell clones from bronchoalveolar lavage fluid (BALF) from four healthy, nonsmoking persons and from four patients with allergic asthma. T cells were cloned by direct limiting dilution and with the use of a fluorescent activated cell sorter with an automated cell deposition unit. T-cell clones from the blood (PB) were prepared as well. The cloning efficiencies of T cells from BALF ranged from 3 to 40% and were lower than those obtained from PB T cells (18 to 72%). The cloning conditions generated CD4+ as well as CD8+ clones. The very late antigen-4, VLA-4, was more frequently expressed on CD4+ T-cell clones from BALF than from the blood (P < 0.05). CD8+ clones from BALF were more frequently VLA-1+ than those from blood (P < < 0.01). Mitogen- and monoclonal antibody-driven proliferation of CD4+ clones showed that BALF clones were well responsive to proliferation stimuli similar to those from the blood. Analysis of interleukin-4 production by 10 BALF and 10 PB clones showed large variations between individual CD4+ clones (BALF: range, < 100 to 700 pg/ml; PB: range, < 100 to 1,100 pg/ml), indicating the generation of different types of clones, which was also clear from analysis of interferon-gamma production. The analysis of properties of BALF T-cell clones and their regulation will improve insight into immunologic reactions in the lungs.
Hol
B E
BE
Department of Pulmonology, Academic Medical Centre, Amsterdam, The Netherlands.
Krouwels
F H
FH
Bruinier
B
B
Reijneke
R M
RM
Mengelers
H J
HJ
Koenderman
L
L
Jansen
H M
HM
Out
T A
TA
D000328:Adult; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology*; D002999:Clone Cells; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D015847:Interleukin-4 / Q000096:biosynthesis; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism
D015847:Interleukin-4; D007371:Interferon-gamma