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Am J Physiol Gastrointest Liver Physiol 274: G147-G156, 1998;
0193-1857/98 $5.00
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Vol. 274, Issue 1, G147-G156, January 1998

IL-1beta and IL-6 in mouse parotid acinar cells: characterization of synthesis, storage, and release

Naoko Tanda1,2, Hiroe Ohyama3, Midori Yamakawa1,2, Maria Ericsson4, Takanori Tsuji3, Jim McBride3, Aram Elovic3, David T. W. Wong3, and Gary R. Login1,2,3

3 Department of Oral Medicine and Diagnostic Sciences, Harvard School of Dental Medicine, 1 Department of Pathology and 2 The Charles A. Dana Research Institute, Beth Israel Deaconess Medical Center and 4 Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02215

Synthesis, storage, and secretion of the proinflammatory cytokine interleukin-1beta (IL-1beta ) and the anti-inflammatory cytokine IL-6 have not been established in normal exocrine gland secretory cells. Parotid glands and isolated acinar cells prepared from BALB/c mice were homogenized for RNA isolation and reverse transcription-polymerase chain reaction (RT-PCR). IL-1beta and IL-6 enzyme-linked immunosorbent assays (ELISAs) were done on supernatants prepared from mouse parotid acinar cell (MPAC) preparations unstimulated or stimulated between 0 and 10 min with 10-5 M norepinephrine at 37°C. MPACs were fixed in paraformaldehyde, frozen sectioned for light and electron microscopy, and labeled with antibodies to IL-1beta and IL-6. Mouse specific riboprobes to IL-1 and IL-6 were used for in situ hybridization. RT-PCR yielded the expected IL-1 (336-bp) and IL-6 (614-bp) mRNA products. By ELISA, stimulated MPACs showed a significant increase in IL-1beta (P < 0.03) and IL-6 (P < 0.01) release into supernatants by 10 min that paralleled the time course of amylase release. In situ hybridization showed the presence of transcripts for IL-1 and IL-6 in glandular epithelial cells. Gold-labeled IL-1beta and IL-6 were significantly higher (P < 0.01) in granules than in the nucleus and cytoplasm. This study shows that MPACs synthesize IL-1beta and IL-6 and release these cytokines from their granules after alpha - and beta -adrenergic stimulation.

reverse transcription-polymerase chain reaction; immunoelectron microscopy; exocytosis; secretion; parotid gland; cytokines


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