scholarly journals Value of gamma interferon enzyme-linked immunospot assay in the diagnosis of peritoneal dialysis-associated tuberculous peritonitis

Author(s):  
Qiuxia Fan ◽  
Xiaoyan Huang ◽  
Jieyun Zhang ◽  
Yinan Sun ◽  
Zuying Xiong ◽  
...  

Abstract Background Tuberculous peritonitis is the most common form of extrapulmonary tuberculosis infection in peritoneal dialysis patients. However, diagnosing tuberculous peritonitis quickly and early has always been a challenge for nephrologists. Mycobacterium tuberculosis antigen-specific gamma interferon enzyme-linked immunospot (IFN-γ ELISPOT) assay has been widely used in the clinical diagnosis of tuberculous pleurisy and peritonitis, but its use has not been reported for uremia. Methods This study mainly verified the feasibility of using the M. tuberculosis antigen-specific IFN-γ ELISPOT assay in the diagnosis of continuous ambulatory peritoneal dialysis (CAPD) patients with tuberculous peritonitis. Taking M. tuberculosis culture as the gold standard, the IFN-γ ELISPOT assay was used to analyze peripheral blood and peritoneal dialysis fluid of patients, and the receiver operating characteristic (ROC) curves in patients with tuberculous peritonitis (TBP) or non-tuberculous peritonitis (NTBP) were analyzed. Results The area under the receiver operating characteristic curve (AUC) was 0.927 (95% CI 0.816–1.000, P = 0.001) for the ELISPOT assay with peritoneal fluid mononuclear cells (PFMC), which was higher than that for the ELISPOT assay with peripheral blood mononuclear cells (PBMC) (0.825, 95% CI 0.6490–1.000, P = 0.011). The cutoff value for the diagnosis of TBP was 40 spot-forming cells (SFCs)/2 × 105 for the ELISPOT with PBMC, with a sensitivity of 55.6%, a specificity of 92.3%, and a diagnostic efficiency of 77.3%. The cutoff value for the diagnosis of TBP was 100 SFCs/2 × 105 for the ELISPOT on PFMC, with a sensitivity, specificity, and diagnostic efficiency 77.8%, 84.6%, and 81.8%, respectively. Parallel and serial testing algorithms appeared more accurate than single ELISPOT assays with PBMC, but ELISPOT assays with PFMC. Conclusions The IFN-γ release test can be used for the early diagnosis of CAPD-related TBP; compared with peripheral blood, peritoneal fluid may be a more effective and accurate medium to diagnose CAPD complicated with tuberculous peritonitis.

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Qiuxia Fan ◽  
Xiaoyan Huang ◽  
Zibo Xiong ◽  
Jieyun Zhang ◽  
Yan He

Abstract Background and Aims Patients with ESRD undergoing chronic dialysis are 6 to 25 times more likely to develop tuberculosis (TB) than the general population. Tuberculous peritonitis is the most common form of tuberculosis infection in peritoneal dialysis patients. However, diagnosing active TB infection can be challenging, early and rapid diagnosis of tuberculous peritonitis has always been a challenge for nephrologists. M.tuberculosis antigen-specific IFN-γ production by peripheral blood mononuclear cells (PBMC) was determined by using an in-house ELISPOT assay by Institute of Hepatology of Shenzhen Third Hospital.In this study, we evaluated diagnostic performance of an M.tuberculosis antigen-specific gamma interferon enzyme-linked immunospot (ELISPOT) assay in continuous ambulatory peritoneal dialysis (CAPD) patients with tuberculous peritonitis. Method There were nine patients (1.29%) with tuberculous peritonitis among 700 CAPD patients. M. tuberculosis accounted for 2.84 % of all peritonitis.IFN-γ response in peripheral blood and peritoneal fluid were determined in 9 patients with tuberculous peritonitis(TBP),at the same time, 13 cases diagnosed with non-tuberculous peritonitis(NTBP) were enrolled.All patients were examined for tuberculosis-specific IFN-γ ELISPOT in peripheral blood and peritoneal dialysis fluid. Results 22 patients were recruited into this study, 9 were classified with confirmed tuberculosis peritonitis, 13 patients with bacterial or fungal peritonitis were enrolled as controls during the same period, in this patients, 3 were caused by Gram positive bacteria, 3 were caused by Gram negative bacteria, 3 were caused by fungus, 4 patients were negative athogenic bacteria culture .The AUC of ROC curve was 0.927 (95%CI:0.816–1.000,P=0.001) for ELISPOT on PFMC, which was higher than that of ELISPOT on PBMC (0.825,95%CI: 0.6490–1.000,P=0.011).When the cutoff value for the diagnosis of TBP was 40.5 SFCs/2 × 105 for ELISPOT on PBMC, with a sensitivity of 55.6%, a specificity of 92.3%,a diagnostic efficiency of 77.3%,The positive predictive value was83.3%, with negative predictive value of 75%,positive likelihood ratio of 7.2, and negative likelihood ratio of 0.48.The sensitivity, specificity,diagnostic efficiency,PPV, NPV, LR+, and LR- of the ELISPOT assay for the diagnosis of active TBP when performed on the PFMC were 77.8%, 84.6%, 81.8%, 77.8%, 84.6%, 5.05 and 0.26, respectively.Receiver operating characteristic(ROC) curves were used to estimate the diagnostic values of Elispot on PFMC and PBMC in 22 patients with confirmed TBP or NTBP. Conclusion IFN-γ release test can be used for early diagnosis of CAPD-related TBP; compared with peripheral blood, peritoneal dialysate may be a more effective and accurate way to diagnose CAPD complicated with tuberculous peritonitis.


2011 ◽  
Vol 62 (6) ◽  
pp. 462-471 ◽  
Author(s):  
Oh-Hyun Cho ◽  
Ki-Ho Park ◽  
Su-Jin Park ◽  
Sun-Mi Kim ◽  
Seong Yeon Park ◽  
...  

2021 ◽  
Vol 10 (4) ◽  
pp. 875
Author(s):  
Kawaljit Kaur ◽  
Shahram Vaziri ◽  
Marcela Romero-Reyes ◽  
Avina Paranjpe ◽  
Anahid Jewett

Survival and function of immune subsets in the oral blood, peripheral blood and gingival tissues of patients with periodontal disease and healthy controls were assessed. NK and CD8 + T cells within the oral blood mononuclear cells (OBMCs) expressed significantly higher levels of CD69 in patients with periodontal disease compared to those from healthy controls. Similarly, TNF-α release was higher from oral blood of patients with periodontal disease when compared to healthy controls. Increased activation induced cell death of peripheral blood mononuclear cells (PBMCs) but not OBMCs from patients with periodontal disease was observed when compared to those from healthy individuals. Unlike those from healthy individuals, OBMC-derived supernatants from periodontitis patients exhibited decreased ability to induce secretion of IFN-γ by allogeneic healthy PBMCs treated with IL-2, while they triggered significant levels of TNF-α, IL-1β and IL-6 by untreated PBMCs. Interaction of PBMCs, or NK cells with intact or NFκB knock down oral epithelial cells in the presence of a periodontal pathogen, F. nucleatum, significantly induced a number of pro-inflammatory cytokines including IFN-γ. These studies indicated that the relative numbers of immune subsets obtained from peripheral blood may not represent the composition of the immune cells in the oral environment, and that orally-derived immune effectors may differ in survival and function from those of peripheral blood.


2002 ◽  
Vol 30 (2) ◽  
pp. 144-160 ◽  
Author(s):  
M Kodama ◽  
HK Takahashi ◽  
H Iwagaki ◽  
H Itoh ◽  
T Morichika ◽  
...  

Interleukin (IL) 18, a powerful inducer of the immunoregulatory cytokine interferon-γ (IFN-γ), presents upstream of the cytokine activation cascade in the inflammatory response. The anti-inflammatory properties of steroids permit their use in various conditions, although effects are transient and pathological states are not fully relieved by short-term steroidal use. We examined the effect of lipopolysaccharide (LPS)/IL-2 on the cytokine cascade in human peripheral blood mononuclear cells (PBMCs). We also examined the effect of steroids on LPS/IL-2-induced cytokine production in human PBMCs taken from healthy volunteers. Cell-free supernatant fractions were assayed for IL-18, IL-12, IL-2, IFN-γ and IL-10 protein, using enzyme-linked immunosorbent assays, and synergy between LPS and IL-2 in enhanced production of IL-18 was observed. Steroids suppressed the production of IL-18 and other secondary cytokines in LPS/IL-2-stimulated PBMCs, in a concentration- and time-dependent manner, although inhibition was incomplete even at high concentrations. Effects of steroid treatment on expression of membrane-bound LPS receptor antigen (mCD14) and intercellular adhesion molecule-1 (ICAM-1) in PBMCs were studied by flow cytometric analysis. Steroid treatment up-regulated mCD14 expression in a concentration-dependent manner, with no effect on ICAM-1 expression. These results suggest that the incomplete counteraction of steroids in the LPS/IL-2-initiating cytokine cascade is due, at least partly, to the up-regulation of mCD14 by steroid preparations, which increases susceptibility to bacterial endotoxins.


Author(s):  
Atsushi Satomura ◽  
Yoichi Oikawa ◽  
Akifumi Haisa ◽  
Seiya Suzuki ◽  
Shunpei Nakanishi ◽  
...  

Abstract Context Unprovoked A−β+ ketosis-prone type 2 diabetes (KPD) is characterized by the sudden onset of diabetic ketosis/ketoacidosis (DK/DKA) without precipitating factors, negative anti-islet autoantibodies (“A−”), and preservation of β-cell function (“β+”) after recovery from DKA. Although this phenotype often appears with acute hyperglycemia and DK/DKA just like acute-onset type 1 diabetes (AT1D), the involvement of anti-islet immune responses remains unknown. Objective We sought to clarify the immunological role of insulin-associated molecules in unprovoked A−β+ KPD. Methods In this cross-sectional study, blood samples from 75 participants (42 with AT1D and 33 with KPD) were evaluated for interferon (IFN)-γ-secreting peripheral blood mononuclear cells (PBMCs) reactive to four insulin B-chain amino acid 9–23-related peptides (B:9–23rPep) using an enzyme-linked immunospot (ELISpot) assay. Results Overall, 36.4% (12/33) of KPD participants showed positive IFN-γ ELISpot assay results; the positivity rate in KPD was similar to that in AT1D (38.1%; 16/42) and significantly higher than the previously reported rate in type 2 diabetes (8%; 2/25; P < 0.0167). Moreover, B:9–23rPep-specific IFN-γ-producing PBMC frequency was negatively correlated with age and ad lib serum C-peptide levels in all KPD participants and positively correlated with HbA1c level in KPD participants with positive IFN-γ ELISpot results. Conclusions These findings suggest the involvement of B:9–23rPep-specific IFN-γ-related immunoreactivity in the pathophysiology of some unprovoked A−β+ KPD. Moreover, increased immunoreactivity may reflect transiently decreased β-cell function and increased disease activity at the onset of DK/DKA, thereby playing a key role in DK/DKA development in this KPD phenotype.


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