Biologic and Immunohistochemical Analysis of Macrophage lnterleukinâ 1α, â 1β, and Tumor Necrosis Factor Production During the Peritoneal Exudative Response

The present study examined changes in lipopolysaccharide (LPS)â induced interleukin 1 (ILâ 1) and tumor necrosis factor (TNF) production by murine peritoneal macrophages during the chronic exudative response to Freund’s complete adjuvant (CFA). Macrophages were isolated by peritoneal lavage and adherence at intervals over a 32 day period following i.p. injection of CFA. Optimal culture conditions for ILâ 1 and TNF production were predetermined, and it was found that ILâ 1 production was profoundly impaired at densities of above 150 cells/mm2, whereas TNF sythesis was more resistant to density effects. Using optimal conditions, we observed a sequential appearance of monokines. On day 0 there was minimal ILâ 1 production and no detectable TNF production. By days 4â 7, ILâ 1 production reached maximum levels with a steady decline to baseline by day 32. TNF production steadily increased after day 2, reached maximal levels by days 16â 20, and then partly declined by day 32. These findings were supported by kinetic analyses at specified days. When related to exudative events, it appeared that maximal ILâ 1 was associated with the recruitment stage of the reaction, whereas TNF production was associated with the established exudate. Immunohistochemical analysis revealed that TNF production could be related to the proportion of macrophages with cytoplasmic TNF expression. In contrast, ILâ 1α and â 1β expression was comparable among populations with 85â 100% of cells showing cytoplasmic expression 6 hr after LPS stimulus. Whereas cytoplasmic ILâ 1α persisted for the 18 hr study period, ILâ 1β disappeared from many adjuvant recruited cells. Our findings suggest that monokine production is orchestrated during macrophage recruitment and activation at sites of chronic inflammation.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141960/1/jlb0529.pd