title: Limiting-dilution analysis of T cells extracted from solid human lung tissue: comparison of precursor frequencies for proliferative responses and lymphokine production between lung and blood T cells from individual donors.
abstract: This study evaluates the frequency and functions of immunocompetent T cells at the clonal level in solid human lung tissue versus peripheral blood. Enzymatic digestion of slices of histologically normal human lung yielded 18-42 x 10(6) viable mononuclear cells per gram wet weight tissue, of which 60-72% were lymphocytes; based upon these recoveries and the known weight of adult lung, the (median) lung parenchymal lymphocyte population can be estimated as 6 x 10(9), being of the same order as the blood pool and 15-30-fold that recoverable by broncho-alveolar lavage. Flow cytometric analysis indicated that the bulk of these lymphocytes was OKT3+/T11+ (CD3/CD2) T cells. Purified blood and lung T cells from each subject were cultured at limiting dilution in the presence of PHA, irradiated feeder cells and recombinant human IL-2. The mean frequency estimates for PHA-responsive T cells in these populations were 1 in 1.23 (81%) and 1 in 3.22 (31%) for blood and lung, respectively. This difference was seen for T cells from each donor and was highly significant by paired t-test (P less than 0.002). Analysis of surface phenotypes and functions of individual blood and T-cell clones indicated comparable frequencies for OKT4 (CD4) and OKT8 (CD8) expression, TNF production and mitogen-induced cytotoxicity. However, a striking inverse relationship was observed between the overall frequency of IL-2-producing clones (79% for blood versus 47% for lung) and interferon-gamma (IFN-gamma)-producing clones (46% versus 87%). These differences were found for each subject, and both were highly significant (P less than 0.001) by paired t-test. The available literature suggests that the majority of these lung T cells represent transient immigrants derived from the blood. Accordingly, the functional differences we have observed suggest either selective trapping within the lung vascular bed of peripheral blood T cells of certain functional phenotypes or alternatively selection/modulation of T cells by lung-derived factors during their transit through the tissue.
authors.affiliation: Clinical Immunology Research Unit, Princess Margaret Hospital, Subiaco, West Australia.
authors.forename: P G
authors.initials: PG
authors.lastname: Holt
authors.forename: U R
authors.initials: UR
authors.lastname: Kees
authors.forename: M A
authors.initials: MA
authors.lastname: Shon-Hegrad
authors.forename: A
authors.initials: A
authors.lastname: Rose
authors.forename: J
authors.initials: J
authors.lastname: Ford
authors.forename: N
authors.initials: N
authors.lastname: Bilyk
authors.forename: R
authors.initials: R
authors.lastname: Bowman
authors.forename: B W
authors.initials: BW
authors.lastname: Robinson
mesh_terms: D000328:Adult; D000368:Aged; D000954:Antigens, Surface / Q000032:analysis; D002455:Cell Division; D002469:Cell Separation; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D006801:Humans; D007958:Leukocyte Count; D008168:Lung / Q000276:immunology*; D008222:Lymphokines / Q000096:biosynthesis*; D008875:Middle Aged; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
chemical_list: D000954:Antigens, Surface; D008222:Lymphokines