In vitro induction of aryl hydrocarbon hydroxylase in human pulmonary alveolar macrophages by benzanthracene

1977 ◽  
Vol 2 (6) ◽  
pp. 327-333 ◽  
Author(s):  
T MCLEMORE ◽  
R MARTIN
Keyword(s):  
Alveolar Macrophages ◽  
Aryl Hydrocarbon Hydroxylase ◽  
Aryl Hydrocarbon ◽  
Pulmonary Alveolar Macrophages ◽  
In Vitro Induction

In Vitro Induction of Aryl Hydrocarbon Hydroxylase in Baboon Pulmonary Alveolar Macrophages and Peripheral Blood Lymphocytes

1985 ◽  
Vol 4 (2) ◽  
pp. 201-212 ◽  
Author(s):  
T. L. McLemore ◽  
T. J. Kuehl
Keyword(s):  
Alveolar Macrophages ◽  
Peripheral Blood ◽  
Culture Medium ◽  
Peripheral Blood Lymphocytes ◽  
Interindividual Variation ◽  
Aryl Hydrocarbon Hydroxylase ◽  
Blood Lymphocytes ◽  
Aryl Hydrocarbon ◽  
Pulmonary Alveolar Macrophages

Aryl hydrocarbon hydroxylase (AHH) activity was measured in cultured pulmonary alveolar macrophages (PAMs) (n = 12) and peripheral blood lymphocytes (n = 72) from unrelated baboons. It was determined that optional AHH induction was observed in PAMs when cells were cultured for 24 or 48 hours in the presence of a 15 μM concentration of the inducer, benzanthracene (BA). Maximal enzyme activity was observed in mitogen-activated lymphocytes when cells were cultured for 96 or 120 hours in the presence of 10 μM BA concentration. Additionally, there were no differences in PAM or lymphocyte AHH induction when cells were cultured in different types of culture medium (P > 0.10 in all instances, using paired, 2-tailed, Student's t-test). Similarly, no differences were observed between PAM AHH activity when human AB serum or autologous baboon serum was used in the culture medium (P > 0.30). However, slightly higher BA-induced AHH levels were noted in lymphocytes cultured in the presence of autologous baboon serum as compared with human AB serum (P < 0.02). Using optimal culture conditions for PAMs and lymphocytes from individual baboons a 6.5-fold interindividual variation in PAM and a 30-fold variation in lymphocyte BA-induced AHH levels were observed (n = 8 and 64, respectively). These studies document that AHH is measurable in cultured baboon PAMs and lymphocytes and that there is wide interindividual variation in BA-induced AHH levels. The distribution of AHH activity appears to be similar to that observed in the human population.

Measurement of Viability and Aryl Hydrocarbon Hydroxylase Activity in Cryopreserved Baboon Peripheral Blood Lymphocytes and Pulmonary Alveolar Macrophages

1985 ◽  
Vol 4 (2) ◽  
pp. 193-199
Author(s):  
T. L. McLemore ◽  
T. J. Kuehl
Keyword(s):  
Alveolar Macrophages ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blue Dye ◽  
Aryl Hydrocarbon Hydroxylase ◽  
Time Intervals ◽  
Blood Lymphocytes ◽  
Aryl Hydrocarbon ◽  
Aryl Hydrocarbon Hydroxylase Activity ◽  
Pulmonary Alveolar Macrophages

Pulmonary alveolar macrophages (PAMs) and peripheral blood lymphocytes obtained from healthy nonsmoking baboons were evaluated for the ability to retain viability and aryl hydrocarbon hydroxylase (AHH) activity following cryopreservation for 2, 4, 6, 8, and 12 week intervals. Viability was measured (by trypan blue dye exclusion) in lymphocytes following 2, 4, 6, 8, and 12 weeks of cryopreservation. No significant decrease in viability was observed (P > 0.05 for all comparisons; nonpaired 2-tailed, t-test; n = 22), with lymphocytes retaining 88% of their original viability following 12 weeks of cryopreservation. Similarly, a slight, but insignificant decrease in PAM viability was observed following cryopreservation for 2, 4, 6, and 8 week intervals (P > 0.05 in all instances), with PAMs retaining 93.5% of the original 0 time viability. AHH activity was also evaluated in PAMs and lymphocytes following cryopreservation. Lymphocyte noninduced and BA-induced AHH activities were slightly but not significantly decreased after 2, 4, 6, 8, and 12 week cryopreservation intervals (P > 0.05 for all comparisons). Similarly, AHH activity in cultured PAMs was not significantly decreased following cryopreservation for 2, 4, 6, and 8 week intervals (P > 0.05 for all noninduced and BA-induced AHH levels assayed at the various time intervals; n = 8). Even following cryopreservation periods, for 8 or 12 week intervals, PAMs and lymphocytes retained 91% and 90%, respectively, of the original BA-induced AHH activity. These data demonstrate that baboon PAMs and lymphocytes are capable of undergoing cryopreservation while maintaining viability and AHH activity.

scholarly journals LPS-induced expression of CD14 in the TRIF pathway is epigenetically regulated by sulforaphane in porcine pulmonary alveolar macrophages

2016 ◽  
Vol 22 (8) ◽  
pp. 682-695 ◽  
Author(s):  
Qin Yang ◽  
Maren J Pröll ◽  
Dessie Salilew-Wondim ◽  
Rui Zhang ◽  
Dawit Tesfaye ◽  
...  
Keyword(s):  
Gene Expression ◽  
Alveolar Macrophages ◽  
Methylation Status ◽  
Gene Body ◽  
Gene Body Methylation ◽  
Tnf Α ◽  
Pulmonary Alveolar Macrophages ◽  
Cluster Of Differentiation ◽  
Cd14 Gene

Pulmonary alveolar macrophages (AMs) are important in defense against bacterial lung inflammation. Cluster of differentiation 14 (CD14) is involved in recognizing bacterial lipopolysaccharide (LPS) through MyD88-dependent and TRIF pathways of innate immunity. Sulforaphane (SFN) shows anti-inflammatory activity and suppresses DNA methylation. To identify CD14 epigenetic changes by SFN in the LPS-induced TRIF pathway, an AMs model was investigated in vitro. CD14 gene expression was induced by 5 µg/ml LPS at the time point of 12 h and suppressed by 5 µM SFN. After 12 h of LPS stimulation, gene expression was significantly up-regulated, including TRIF, TRAF6, NF-κB, TRAF3, IRF7, TNF-α, IL-1β, IL-6, and IFN-β. LPS-induced TRAM, TRIF, RIPK1, TRAF3, TNF-α, IL-1β and IFN-β were suppressed by 5 µM SFN. Similarly, DNMT3a expression was increased by LPS but significantly down-regulated by 5 µM SFN. It showed positive correlation of CD14 gene body methylation with in LPS-stimulated AMs, and this methylation status was inhibited by SFN. This study suggests that SFN suppresses CD14 activation in bacterial inflammation through epigenetic regulation of CD14 gene body methylation associated with DNMT3a. The results provide insights into SFN-mediated epigenetic down-regulation of CD14 in LPS-induced TRIF pathway inflammation and may lead to new methods for controlling LPS-induced inflammation in pigs.

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