scholarly journals The Application of Inosine 5′-Monophosphate Dehydrogenase Activity Determination in Peripheral Blood Mononuclear Cells for Monitoring Mycophenolate Mofetil Therapy in Children with Nephrotic Syndrome

2020 ◽  
Vol 13 (8) ◽  
pp. 200
Author(s):  
Joanna Sobiak ◽  
Alicja Jóźwiak ◽  
Honorata Wziętek ◽  
Jacek Zachwieja ◽  
Danuta Ostalska-Nowicka
Keyword(s):  
Nephrotic Syndrome ◽  
Mycophenolate Mofetil ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Transplant Recipients ◽  
Peripheral Blood Mononuclear ◽  
Impdh Activity ◽  
Modified Method ◽  
Blood Mononuclear Cells

In pediatric nephrotic syndrome, recommended mycophenolic acid (MPA) pharmacokinetics are higher than those for transplant recipients. In MPA therapeutic monitoring, inosine-5′-monophosphate dehydrogenase (IMPDH) activity may be useful. We modified the method established for renal transplant recipients and determined IMPDH activity in peripheral blood mononuclear cells (PBMCs) from healthy volunteers and children (4–16 years) with nephrotic syndrome treated with mycophenolate mofetil (MMF). From children, four blood samples were collected, and MPA concentrations were also determined. IMPDH activity was calculated using xanthosine monophosphate (XMP) normalized with adenosine monophosphate (AMP), both determined with the HPLC-UV method. The modified method was accurate, precise, and linear for AMP and XMP within 0.50–50.0 μmoL/L. Mean IMPDH activity in volunteers was 45.97 ± 6.24 µmoL·s−1·moL−1 AMP, whereas for children, the values were variable and amounted to 39.23 ± 27.40 µmoL·s−1·moL−1 AMP and 17.97 ± 15.24 µmoL·s−1·moL−1 AMP before the next MMF dose and 1 h afterward, respectively. The modified method may be applied to IMPDH activity determination in children with nephrotic syndrome treated with MMF. IMPDH activity should be determined after one thawing of PBMCs due to the change in AMP and XMP concentrations after subsequent thawing. For children, the lowest IMPDH activity was observed concomitantly with the highest MPA concentration.

scholarly journals Impact of Immunoglobulin M-Type Donor-Specific Human Leukocyte Antigen–Antibody Levels in Supernatants from Cultured Peripheral Blood Mononuclear Cells as Predictors of Antibody-Mediated Rejection

Pathogens ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 733 ◽  
Author(s):  
Ryoichi Imamura ◽  
Yoshiko Matsuda ◽  
Koichi Tsutahara ◽  
Norio Nonomura ◽  
Shiro Takahara
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Immunosuppressive Agents ◽  
Immunoglobulin M ◽  
Transplant Recipients ◽  
Peripheral Blood Mononuclear ◽  
Antibody Mediated Rejection ◽  
Single Antigen ◽  
Blood Mononuclear Cells

Background: Antibody-mediated rejection (AMR) is a crucial barrier in the long-term prognosis of transplant recipients. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from kidney allograft recipients (N = 41) and cultured in vitro for 1 week. Furthermore, the supernatants of the cultured PBMCs were analyzed by Luminex single-antigen beads. Results: Analyses using Luminex single-antigen beads revealed the presence of immunoglobulin (Ig) G donor-specific anti-HLA antibodies (DSAs) was detected in the supernatants of cultured PBMCs collected more frequently than IgM in de novo DSA-sensitized patients with AMR, and IgM were detectable in patients with stable graft function mainly and several IgM DSAs were detectable in the supernatants of the cultured PBMCs before detecting the IgG levels in sera. We also found that the DSA-specific IgM-secreting memory B cells (mBCs) were more sensitive to the chronic use of immunosuppressive agents than to the IgG-secreting mBCs. Conclusions: In the transplant recipients, the assessment of supernatants of cultured PBMCs provide more details of immune reactions than the commonly used method that directly measures IgG DSA levels in patient sera and some IgM DSA detection may be a better predictor of IgG DSAs production, which may cause AMR and enable early intervention, in initial stages of AMR development.

Transcriptional and Post-Transcriptional Alterations of I κBα in Active Minimal-Change Nephrotic Syndrome

2001 ◽  
Vol 12 (8) ◽  
pp. 1648-1658 ◽  
Author(s):  
DJILLALI SAHALI ◽  
ANDRÉ PAWLAK ◽  
SABINE LE GOUVELLO ◽  
PHILIPPE LANG ◽  
ASTA VALANCIUTÉ ◽  
...  
Keyword(s):  
Nephrotic Syndrome ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Minimal Change Nephrotic Syndrome ◽  
Binding Activity ◽  
Minimal Change ◽  
Peripheral Blood Mononuclear ◽  
Nuclear Extracts ◽  
Blood Mononuclear Cells

Abstract. Minimal-change nephrotic syndrome (MCNS) is a renal disease characterized by heavy glomerular proteinuria and increased production of cytokines by immune cells. Because of the central role of nuclear factor-κB (NF-κB) in the regulation of cytokine expression, its activity during the relapse and remission phases of steroid-sensitive MCNS was analyzed. During relapse, nuclear extracts from peripheral blood mononuclear cells displayed high levels of NF-κB DNA-binding activity, consisting primarily of p50/RelA (p65) complexes. NF-κB p65 and IκBα proteins were barely detected or not detected in cytosolic fractions during relapse, in contrast to remission. The lack of expression of IκBα protein was associated with downregulation of IκBα mRNA and increases in the levels of the mRNA encoding the proteasome α2 subunit proteolytic pathway. In addition, inhibition of proteasome activity induced cytosolic accumulation of phosphorylated IκBα and significant reductions in the NF-κB binding activity in nuclear extracts from peripheral blood mononuclear cells from patients experiencing relapses. These results suggest that alterations in the NF-κB/IκBα regulatory feedback loop may contribute to the immunologic abnormalities that occur in steroidsensitive MCNS.

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