Development of an immune function assay by measuring intracellular adenosine triphosphate (iATP) levels in mitogen-stimulated CD4+ T lymphocytes.

We developed an immune function assay for monitoring CD4+ T cells activity based on changes in intracellular adenosine triphosphate (iATP) levels after phytohemagglutinin (PHA) stimulation. Blood samples were obtained from 40 healthy subjects and 30 RTRs and incubated with 5 µg/mL of PHA for 15-18 hr at 37°C and 5% CO2. Afterward, the CD4+ T cells were separated by antibody-coated magnetic beads and lysed. Then, iATP content in unstimulated and stimulated conditions was measured by luciferin-luciferase reaction using a log-log standard curve. The iATP levels showed significant increase in CD4+ T cells in both healthy persons (mean: 550 ± 142 ng/mL vs. 109 ± 54 ng/mL) and RTRs (mean: 394 ± 160 ng/mL vs. 52 ± 37 ng/mL) after PHA stimulation (P < 0.001). However, the iATP production in RTRs was significantly lower than that in healthy individuals; both prior to and after stimulation with PHA (P < 0.001). No gender-specific difference in iATP production was observed between women and men subjects. This rapid and low-cost assay reflects the degree of immune cell function through assessment of CD4+ T cells activation. Thus, it can be used for evaluation of immune system status in immunodeficient individuals as well as in immunosuppressed transplant recipients who needs drug adjustment.

Monitoring cellular immune function of renal transplant recipients based on adenosine triphosphate (ATP) production by mitogen-induced CD4+ T helper cells.

Maintaining the balance between over- and under-immunosuppression has a critical role for successful immunosuppressive therapy after renal transplantation. We studied the predictive value of our functional immune assay, which works based on adenosine triphosphate (ATP) levels, in determining risk of infection and rejection among renal transplant recipients (RTRs). A total of 65 RTRs with less than 1 month (RTRL1) and 48 RTRs with more than 6 months (RTRM6) of post-transplant time, and 56 healthy individuals were included. Upon lymphocyte activation by phytohemagglutinin (PHA), CD4+ T cells were separated using magnetic beads (Dynabeads), the intracellular ATP (iATP) concentrations were measured by luciferin-luciferase reaction, and compared within and between the groups. Activated CD4+ cells iATP production directly correlated with post-transplant time (r = 0.32, P = 0.011). The iATP levels were significantly lower in both RTRL1 and RTRM6 groups compared to control (P < 0.001), and in the RTRL1 group compared to the RTRM6 (P < 0.05). The iATP concentrations were significantly lower in patients who suffered from infection versus the RTRs with stable graft function (SGF). However, the iATP levels were higher in those with allograft rejection episode (ARE). Our optimization experiments showed that best iATP levels cutoffs were 472.5 and 572.5 ng/ml for predicting risk of ARE, and 218.5 and 300.5 ng/ml for predicting risk of developing infection in RTRL1 and RTRM6 patients, respectively. iATP levels measured by immune function assay might be a promising predictive tool for identifying RTRs who are at risk of developing infection or allograft rejection.

Massive hemoptysis, a presentation of invasion of aneurysm of descending aorta to bronchopulmonary tree.

Aortobronchial (AB) fistula is a rare disease, which is presented with massive hemoptysis; lethal if not treated. It should be suspected in any patient who presents with massive hemoptysis and had previous thoracic aortic surgery, but even it may be seen in patients without any history of operation on the thoracic aorta. Although, today in many centers endovascular therapy is done for these patients, but it is not the standard approach. Surgery in urgent situations has an essential role in saving the patients. Operative management consists of double lumen intubation and one lung ventilation, followed by femoral artery and vein cannulation, posterolateral thoracotomy and achieving proximal and distal control on the aorta, applying cardiopulmonary bypass (CPB), separation the lesion, and bypass the segment of the diseased aorta by a synthetic graft.