Measurements of Cytokine mRNA Expression by Quantitative Polymerase Chain Reaction in Studies of Celiac Disease

2003 ◽  
pp. 185-202
Author(s):  
Ellen M. Nilsen ◽  
Finn-Eirik Johansen ◽  
Per Brandtzaeg
Keyword(s):  
Polymerase Chain Reaction ◽  
Celiac Disease ◽  
Mrna Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Cytokine Mrna ◽  
Cytokine Mrna Expression ◽  
Polymerase Chain

Compartmentalization of the acute cytokine response in humans after intravenous endotoxin administration

1993 ◽  
Vol 74 (6) ◽  
pp. 3027-3033 ◽  
Author(s):  
A. J. Boujoukos ◽  
G. D. Martich ◽  
E. Supinski ◽  
A. F. Suffredini
Keyword(s):  
Polymerase Chain Reaction ◽  
Mrna Expression ◽  
Human Subjects ◽  
Peak Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Cytokine Mrna ◽  
Cytokine Mrna Expression ◽  
Normal Human ◽  
Polymerase Chain

Lung cytokine production was examined after the intravenous administration of endotoxin to 23 normal human subjects. Bronchoalveolar lavage (BAL) was performed 7 days before and 1.5 or 5 h after endotoxin (4 ng/kg). Cytokine mRNA was evaluated in cell pellets (> 98% macrophages) by use of reverse transcription and the polymerase chain reaction. Immunoreactivity was measured by enzyme-linked immunosorbent assay of 20- to 40-fold concentrated BAL. Interleukin- (IL) 8 was detected in BAL (4–130 pg/ml) but not in the serum at baseline. Few neutrophils were found in BAL (< 1%) despite this IL-8 gradient. Peak serum IL-8 levels occurred 2 h after endotoxin (3,930 +/- 241 pg/ml), but BAL neutrophils and IL-8 did not increase. Peak serum tumor necrosis factor (TNF) levels occurred 1.5 h after endotoxin (1,844 +/- 210 pg/ml), but TNF was detected in only 1 of 20 BAL samples. TNF and IL-8 mRNA were detected by polymerase chain reaction in > 70% of the BAL samples before endotoxin, whereas IL-1 alpha, IL-1 beta, and IL-6 were detected in < 25% of the BAL samples. After endotoxin, no change was detected in cytokine mRNA expression. Actinomycin D treatment of the BAL did not alter the pattern of cytokine mRNA expression. These data suggest that mechanisms exist to insulate the alveolar space from the stimulatory effects of endotoxin and high circulating levels of cytokines. Additional factors appear to control the chemotactic effects of IL-8 on neutrophils in the air spaces during acute systemic inflammation.

Analysis of cytokine mRNA levels in interleukin-4-transgenic mice by quantitative polymerase chain reaction

1992 ◽  
Vol 22 (5) ◽  
pp. 1179-1184 ◽  
Author(s):  
Cornelia Platzer ◽  
Günther Richter ◽  
Klaus Überla ◽  
Werner Müller ◽  
Helmut Blöcker ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Transgenic Mice ◽  
Quantitative Polymerase Chain Reaction ◽  
Interleukin 4 ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Cytokine Mrna ◽  
Polymerase Chain

scholarly journals Expression of the Multidrug Resistance-Associated Protein Gene in Refractory Lymphoma: Quantitation by a Validated Polymerase Chain Reaction Assay

Blood ◽  
1997 ◽  
Vol 89 (10) ◽  
pp. 3795-3800 ◽  
Author(s):  
Zhirong Zhan ◽  
Victor A. Sandor ◽  
Erick Gamelin ◽  
Joanna Regis ◽  
Bruce Dickstein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multidrug Resistance ◽  
Mrna Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Newly Diagnosed ◽  
Chain Reaction ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Refractory Lymphoma ◽  
Mdr 1

Abstract Previous work investigating the role of MDR-1 overexpression in relapsed and refractory lymphoma led us to investigate a possible role for multidrug resistance-associated protein (MRP) as a cause of resistance in patients who did not overexpress MDR-1. A quantitative polymerase chain reaction (PCR) method for measuring MRP expression was validated. Immunoblot analysis suggested that no major discrepancy was present between mRNA expression and protein levels. MRP levels were found to be independent of sample tumor content by immunophenotyping, suggesting that the presence of normal cells had no significant impact on measurements of MRP expression. We evaluated MRP in 55 biopsy samples from 40 patients with refractory lymphoma enrolled on a trial of infusional chemotherapy (EPOCH). Pre- and post-EPOCH samples were available from 15 patients. MRP levels were also evaluated in 16 newly diagnosed, untreated lymphoma patient samples. No significant difference in MRP mRNA expression was noted between pre- and post-EPOCH groups. Also, MRP levels in the newly diagnosed patient samples were not significantly different from either pre- or post-EPOCH groups. Two of 15 paired pre- and post-EPOCH patient samples exhibited overexpression of MRP after EPOCH chemotherapy, with measured increases of 10-fold and 18-fold. We conclude that MRP overexpression is not responsible for non–P-glycoprotein (Pgp)–mediated drug resistance in the majority of these patients, although it may be important in a subset of patients. Defining this subset prospectively could aid in the development of clinical trials of MRP modulation in drug-resistant lymphoma.

Expression of the Multidrug Resistance-Associated Protein Gene in Refractory Lymphoma: Quantitation by a Validated Polymerase Chain Reaction Assay

Blood ◽  
1997 ◽  
Vol 89 (10) ◽  
pp. 3795-3800 ◽  
Author(s):  
Zhirong Zhan ◽  
Victor A. Sandor ◽  
Erick Gamelin ◽  
Joanna Regis ◽  
Bruce Dickstein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multidrug Resistance ◽  
Mrna Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Newly Diagnosed ◽  
Chain Reaction ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Refractory Lymphoma ◽  
Mdr 1

Previous work investigating the role of MDR-1 overexpression in relapsed and refractory lymphoma led us to investigate a possible role for multidrug resistance-associated protein (MRP) as a cause of resistance in patients who did not overexpress MDR-1. A quantitative polymerase chain reaction (PCR) method for measuring MRP expression was validated. Immunoblot analysis suggested that no major discrepancy was present between mRNA expression and protein levels. MRP levels were found to be independent of sample tumor content by immunophenotyping, suggesting that the presence of normal cells had no significant impact on measurements of MRP expression. We evaluated MRP in 55 biopsy samples from 40 patients with refractory lymphoma enrolled on a trial of infusional chemotherapy (EPOCH). Pre- and post-EPOCH samples were available from 15 patients. MRP levels were also evaluated in 16 newly diagnosed, untreated lymphoma patient samples. No significant difference in MRP mRNA expression was noted between pre- and post-EPOCH groups. Also, MRP levels in the newly diagnosed patient samples were not significantly different from either pre- or post-EPOCH groups. Two of 15 paired pre- and post-EPOCH patient samples exhibited overexpression of MRP after EPOCH chemotherapy, with measured increases of 10-fold and 18-fold. We conclude that MRP overexpression is not responsible for non–P-glycoprotein (Pgp)–mediated drug resistance in the majority of these patients, although it may be important in a subset of patients. Defining this subset prospectively could aid in the development of clinical trials of MRP modulation in drug-resistant lymphoma.

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