Abstract
Parechymal cells (PC) were prepared by digestion of perfused, lavaged rat lung in a mixture of collagenase and DNase, and harvested on a discontinuous percoll gradient. The process yielded on average 1.0 X 10(8) viable cells/gram tissue. PC were pulsed with soluble antigen, and tested for their capacity to trigger antigen-specific activation of immune T-cells in vitro, or to replace adherent accessory cells necessary for Concanavalin A (Con A)-induced T-cell proliferation. Unfractionated PC exhibited only minor antigen-presenting cell (APC) activity. However, removal of adherent or FcR-positive cells unmasked substantial APC activity. Subsequent experiments indicated that the majority of the APC banded at the top of the percoll gradient (density less than 1.048 g/ml). The same cell preparations substituted for adherent accessory cells in Con A- activation of T cells, suggesting capacity to secrete soluble factors such as interleukin-1 (IL-1) as well to present antigen. The PC preparation also contained T-cells, which were refractory to Con A stimulation unless endogenous adherent cells were first removed. Collectively, these data suggest the presence of non-adherent, FcR-negative, low density accessory cells in the lung parenchyma, capable of both APC activity and soluble factor production. Their T-cell-activation functions appear to be down regulated by endogenous adherent, FcR-positive cells. It is speculated that the accessory cells in these lung preparations may be dendritic cells, the activity of which is subject to inhibition by macrophages.
MeSH Terms
Animals
Antigen-Presenting Cells/immunology
Cell Separation
Centrifugation, Density Gradient
Concanavalin A/pharmacology
Female
Immune Tolerance
Lung/cytology
Lymphocyte Activation
Macrophages/immunology
Rats
Rats, Inbred Strains
T-Lymphocytes/immunology
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Holt P G
Degebrodt A
O'Leary C
Krska K
Plozza T
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