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IL-6 mRNA synthesis by peripheral blood mononuclear cells (PBMC) in patients with chronic renal failure
1First Department of Internal Medicine, Nara Medical University, Japan.
Abstract:
We measured levels of IL-6 mRNA in PBMC obtained from patients with chronic renal failure, using polymerase chain reaction (PCR). PBMC were isolated from 45 patients on haemodialysis (HD) at the start of HD. PBMC were also isolated from 35 patients on HD at the end of HD, 23 patients on continuous ambulatory peritoneal dialysis (CAPD), 24 undialysed patients with chronic renal failure (CRF), and 19 healthy controls. Total RNA was extracted from PBMC with RNA zol and reverse transcribed into cDNA. To prepare samples containing identical amounts of beta-actin cDNA, we performed competitive PCR by co-amplifying serial dilutions of mutant templates containing a single point mutation which generated a unique Eco RI site. Next, to measure IL-6 cDNA semiquantitatively in the samples containing identical amounts of beta-actin (100 pg), we performed PCR amplification using 2 fg of the IL-6 mutant template containing a unique Eco RI site. Higher levels of IL-6 mRNA in the PBMC were observed in the HD patients than in the CAPD patients and healthy controls. The levels of IL-6 mRNA in the PBMC at the end of HD were not significantly higher than those at the start of HD. These results suggest that the dialysis session itself did not significantly affect IL-6 mRNA levels in the PBMC, but that chronic stimulation by maintenance HD may be associated with higher levels of IL-6 mRNA observed in HD patients.
Insights
Patients undergoing maintenance hemodialysis (HD) show higher interleukin-6 (IL-6) mRNA levels in peripheral blood mononuclear cells (PBMC). Dialysis sessions did not significantly alter these IL-6 mRNA levels, suggesting chronic stimulation is key.
Area of Science:
- Nephrology
- Immunology
- Molecular Biology
Background:
- Chronic renal failure (CRF) is associated with altered immune responses.
- Interleukin-6 (IL-6) is a key inflammatory cytokine implicated in various diseases.
- Understanding IL-6 mRNA expression in CRF patients is crucial for managing complications.
Purpose of the Study:
- To quantify and compare IL-6 mRNA levels in peripheral blood mononuclear cells (PBMC) of patients with chronic renal failure undergoing different dialysis modalities and healthy controls.
- To investigate the impact of a single hemodialysis (HD) session on IL-6 mRNA levels.
Main Methods:
- PBMC were isolated from patients with chronic renal failure (CRF) on hemodialysis (HD), continuous ambulatory peritoneal dialysis (CAPD), undialysed CRF patients, and healthy controls.
- Total RNA was extracted, reverse transcribed to cDNA, and IL-6 mRNA was quantified using competitive polymerase chain reaction (PCR) co-amplified with beta-actin.
- IL-6 mRNA levels were measured semiquantitatively using a specific IL-6 mutant template.
Main Results:
- HD patients exhibited significantly higher IL-6 mRNA levels in PBMC compared to CAPD patients and healthy controls.
- IL-6 mRNA levels in PBMC did not differ significantly between the start and end of an HD session.
- Undialysed CRF patients' IL-6 mRNA levels were not explicitly compared to HD or CAPD in the provided text, but the implication is they are lower than HD patients.
Conclusions:
- Maintenance hemodialysis is associated with elevated IL-6 mRNA levels in PBMC.
- The dialysis procedure itself does not appear to acutely increase IL-6 mRNA levels.
- Chronic stimulation from maintenance HD may contribute to the observed higher IL-6 mRNA levels in these patients.