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Effect of Factors Other Than Pathologic Status on Responsiveness of Peripheral Blood Mononuclear Cells from Patients With Chronic Periodontitis
Author(s) -
Suzuki Jon B.,
Sims Tom J.,
Page Roy C.
Publication year - 1983
Publication title -
journal of periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.036
H-Index - 156
eISSN - 1943-3670
pISSN - 0022-3492
DOI - 10.1902/jop.1983.54.7.408
Subject(s) - peripheral blood mononuclear cell , fusobacterium nucleatum , prostaglandin e2 , in vitro , prostaglandin e , prostaglandin , periodontitis , medicine , endocrinology , cell , chemistry , porphyromonas gingivalis , biochemistry
S tudies were designed to assess factors other than pathologic status of the cell donor which affect the blastogenic responsiveness in vitro of peripheral blood mononuclear cells (PBMs) from normal donors and patients with periodontitis. Cultures were established and activated using phytohemagglutinin‐P (PHA) or homogenates of Actinomyces viscosus (AVIS), a gram‐positive plaque microorganism, and Fusobacterium nucleatum (FUSO), a gram‐negative plaque microorganism. Activation was assessed by measuring the incorporation of labeled precursor into DNA. The effects of incubation time, vessel shape, cell concentration, prostaglandin E 2 and indomethacin on blastogenic responsiveness were studied. Blastogenic responsiveness became maximal after 5 to 8 days' activation with the bacterial substances, and after 3 days' activation with PHA. Radioactivity incorporated by cultures in microtest wells with flat, round and conical bottoms was 5.9, 7.8 and 10.6 × 10 3 cpm, respectively. Cultures of cells from all of the patients and normal subjects were activated by PHA, AVIS and FUSO, and cell concentration was a major determinant of the magnitude of the blastogenic response. Responsiveness of cultures from all patients and control subjects activated with AVIS and FUSO was inhibited significantly by Prostaglandin E‐2 (PGE 2 ) at a concentration of 10 μ m. Inhibition was generally 50% or greater. Indomethacin, an inhibitor of prostaglandin production, at a concentration of 0.5 μ g/ml significantly enhanced responsiveness of AVIS‐ and FUSO‐activated cultures from control donors and patients, indicating that prostaglandins are produced endogenously, and that they affect cell responsiveness. The effect of PGE 2 and indomethacin on PHA‐activated cultures was more variable and, where present, of a lesser magnitude than that observed for cultures activated with bacterial homogenates. In most cultures the effects were not statistically significant. Our data show that in studies of lymphocyte activation, the incubation time, culture‐vessel shape, cell concentration and presence of endogenous inhibitors need to be taken into account.