Generation of a large number of functional dendritic cells from human monocytes expanded by forced expression of cMYC plus BMI1

Anticancer vaccination therapies with monocyte-derived dendritic cells (DC) are widely conducted. A large number of primary monocytes (approximately 10⁸ cells) are needed to generate the number of DC required to achieve an effect upon vaccination, and monocytes are usually purified from peripheral blood mononuclear cells obtained by apheresis procedure, which is somehow invasive for cancer patients. As a means to facilitate the generation of DC for therapeutic use, we herein report a method to amplify human monocytes. We found that lentivirus-mediated transduction of cMYC along with BMI1 induced proliferation of CD14⁺ monocytes derived from 9 out of 12 blood donors, and we named the monocyte-derived proliferating cells CD14-ML. Their proliferation continued for 3–5 weeks in the presence of M-CSF and GM-CSF, resulting in 20–1000-fold amplification. Importantly, the expanded CD14-ML differentiated into fully functional DC (CD14-ML-DC) upon the addition of IL-4 to the culture. We successfully stimulated autologous CD8⁺ T cells with CD14-ML-DC pulsed with cytomegalovirus peptide or MART-1 peptide to generate antigen-specific CTL lines. This is the first report describing the method for in vitro expansion of human peripheral blood monocytes.     

Clinical characteristics of patients with ischemic stroke after the 2016 Kumamoto earthquake,a multi-center study

Objective

To investigate the clinical characteristics of patients with ischemic stroke following the 2016 Kumamoto earthquake.

Methods

We retrospectively studied patients with ischemic stroke admitted to 5 stroke centers for 1 year after the earthquake. We compared clinical characteristics in these patients (the post-earthquake group) to those in the patients with ischemic stroke admitted during the same period from the previous 3 years (the pre-earthquake group). Additionally, we analyzed the trend of the incidence rate of stroke before and after the earthquake.

Results

A total of 1979 patients were admitted after the earthquake; 5670 (1,890/year on average) patients were admitted before the earthquake. A first-ever ischemic stroke (71 vs. 75%) and premorbid modified Rankin Scale > 1 (26 vs. 29%) were found significantly more frequently in patients after the earthquake. National Institutes of Health Stroke Scale score ≤ 2 at discharge (60 vs. 65%) was found more frequently in patients after the earthquake, although non-discharge to home (65 vs. 70%) was more frequent in patients after the earthquake. Trend analysis revealed a decrease of small vessel occlusion and large artery atherosclerosis in the month after the earthquake.

Conclusions

The 2016 Kumamoto earthquake may have affected the characteristics of stroke during the early phase of the earthquake and increased the difficulty in returning home.

     

Lack of association of interleukin-18 gene polymorphisms with susceptibility of Japanese populations to Graves' disease or Graves' ophthalmopathy.

OBJECTIVE: To investigate whether polymorphisms of interleukin (IL)-18 gene confer susceptibility to Graves' disease (GD) and Graves' ophthalmopathy (GO). DESIGN: We performed a case control study on polymorphisms of IL-18 gene in Japanese patients with GD (n = 435), and healthy control subjects without antithyroid autoantibodies or family history of autoimmune disorders (n = 255). The C-4675G, C-607A, and G-137C polymorphisms in the promoter region and A105C (exon 5) polymorphism were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using restriction enzymes, sequence-specific PCR, and PCR-direct sequencing methods. RESULTS: None of the polymorphisms in the IL-18 gene were associated with development of Graves' disease. The CC genotype and C allele frequencies of IL-18 gene G-137C polymorphism tended to be greater in patients with ophthalmopathy than in patients without evident ophthalmopathy. However, the differences were not statistically significant. Although there were three major haplotypes, none of the haplotypes were statistically associated with susceptibility to GD or ophthalmopathy. CONCLUSIONS: These results suggest that IL-18 gene polymorphisms are not major genetic factors for susceptibility to GD in a Japanese population. Further studies with adequate sized data set in the subset analyses for GO are needed.     

Direct visualization of cortical peritubular capillary of transplanted human kidney with reperfusion injury using a magnifying endoscopy

BACKGROUND: We developed a direct imaging system of renal microcirculation by a magnifying-endoscopy that enables visualization of the movement of erythrocyte in glomerular and cortical peritubular capillary (CPC). We investigated the microcirculation of CPC in the early phase of both living- and cadaveric-donor transplant kidneys. METHODS: Erythrocyte velocity in CPC were monitored and measured in 20 renal transplants at 20, 60, 90, and 120 minutes after reperfusion. The kidney grafts came from 11 living donors and 9 non-heart-beating cadaveric donors. RESULTS: In living-donor transplants, erythrocyte velocity in CPC at 20 minutes after revascularization declined to one third of baseline value just before nephrectomy and recovered to the prenephrectomy value 120 minutes after reperfusion. In contrast, it continued to be disturbed for 90 minutes in cadaveric-donor transplants. Erythrocyte velocity in CPC more significantly deteriorated in cadaveric transplants than in those of living transplants at 20 through 60 minutes after the revascularization. In living-donor transplants, erythrocyte velocity did not correlate with donor age, both warm (WIT) and cold ischemic time (CIT), time to the initial urination, and best creatine clearance. In the cadaveric transplants, ischemic time, both WIT and CIT, did not correlate with the erythrocyte velocity. However, donor age, duration of acute tubular necrosis, and best creatine clearance after transplantation significantly correlated with the erythrocyte velocity. CONCLUSION: The measurement of erythrocyte velocity in CPC is a reliable method for predicting the recovery of renal function and reserved renal function of kidney allografts undergoing prolonged ischemia.     

A comparison of changes in cardiac preload variables during graded hypovolemia and hypervolemia in mechanically ventilated dogs

We developed an online monitoring system to measure systolic blood pressure variation (SPV) and its down (dDown) and up components, along with pulse pressure variation (dPP). Using the system, we compared different cardiac preload indicators-such as stroke volume variation (SVV) and corrected flow time (FTc)-along with central venous pressure and pulmonary artery occlusion pressure in mechanically-ventilated dogs during normovolemia, graded hypovolemia (-200 and -350 mL), and hypervolemia (+200 and +350 mL). We simultaneously measured these preload indicators along with global hemodynamic variables and investigated their validity and limitations to access preload changes. SPV increased from 4.8 +/- 1.4 mm Hg at baseline to 11.2 +/- 1.8 mm Hg during hypovolemia (-350 mL), but it did not change significantly during hypervolemia. Similar changes were observed with dDown, dPP, and SVV. FTc, conversely, increased during hypervolemia but remained unchanged during hypovolemia. The results of this study indicate that SPV, dDown, dPP, and SVV are useful indicators of hypovolemia, but not of hypervolemia. Conversely, hypovolemia could not be detected reliably by FTc, but it does reflect blood volume changes during hypervolemia. Although SPV, dDown, and dPP measurements require no additional invasion and cost beyond arterial cannulation, their limits must be kept in mind for the monitoring of blood volume status in mechanically-ventilated patients.