Technical advancement in regulatory T cell isolation and characterization using CD127 expression in patients with malignant glioma treated with autologous dendritic cell vaccination.

We have successfully treated over two hundred high-grade glioma (HGG) patients with immunotherapy consisting of vaccination with autologous dendritic cells (DCs) loaded with autologous tumour lysate. It has been documented that regulatory T cells (Treg) can counteract anti-tumour immune responses. Therefore, monitoring of Treg in these patients is essential. Up till now, Treg have been characterized based on the expression of the transcription factor Foxp3. Here, we validated IL-7 receptor alpha subunit (CD127)dim expression as a marker for human Treg within HGG patients, as a less laborious assay for routine use in tumour vaccination trials. We noted a strong positive correlation between Foxp3 expression and CD127dim expression in CD4+CD25+ and CD4+ cells. The suppressive function of CD4+CD127dim cells was assessed in an allogeneic mixed lymphocyte reaction (MLR). We conclude that CD127 staining is a fast, well-suited and reproducible Treg monitoring tool in HGG patients treated with immunotherapy.

Oesophageal perforation after anterior cervical surgery: management in four patients.

BACKGROUND: Oesophageal perforation related to anterior cervical surgery is an uncommon but well recognised and potentially life-threatening complication with an incidence of 0-3.4%. Our experience with this complication and a review of the literature are presented. METHOD: We retrospectively reviewed our clinical experience over 10 years and found four patients in whom an oesophageal perforation was recognised after anterior surgery for cervical spine trauma. In three patients the perforation was noticed in the early post-operative period and the other had a delayed presentation. In all patients, the hardware was removed, long-term intravenous antibiotics were administered and parenteral nutrition was instituted. In two patients a primary suture of the perforation was performed and in one of these an additional sternocleidomastoid myoplasty was carried out as well. One patient had conservative treatment and one died before closure of the perforation could be performed. FINDINGS: The two patients, in whom surgical repair of the perforation was performed, recovered well with residual neurological deficits as expected due to the cervical trauma. In the patient in whom conservative treatment was instituted, healing of the perforation occurred. One patient died due to systemic complications, indirectly related to the perforation. CONCLUSIONS: Although not very frequent and sometimes difficult to diagnose, oesophageal perforations after anterior cervical surgery constitute a potentially life-threatening complication. Diagnosis is made by imaging or endoscopic studies, but clinical suspicion is most important. Basic treatment consists of surgery with removal of hardware, drainage of abscesses, primary closure of the perforation if possible, parenteral nutrition and antibiotic therapy. Residual instability should be recognised in time and may be anticipated in patients in whom there has been little time for solid bony fusion. Successful management depends on early diagnosis and immediate institution of treatment.

Factors influencing the flow rate through a surgical defect in human fetal membranes.

OBJECTIVES: In order to determine factors influencing the flow rate trough a created defect in human fetal membranes, an ex vivo set-up was used with fetal membranes collected from patients undergoing Caesarean section at term. METHODS: The membranes were secured at the bottom of a plastic tube and traumatised with needles ranging from 14-26 Gauges (Ga), under a hydrostatic pressure of 10 to 20 cm H(2)O and an angle of 45 degrees or 90 degrees. The column was filled with amniotic fluid or Hartmann's solution. The duration of the puncture was 1 s or the time it takes to aspirate 10 ml through the needle. The flow rate through the defect in the fetal membranes and size of the defect were measured. RESULTS: The flow rate and defect size increased with increasing diameter of the needle. Increasing the pressure in the column resulted in a significant linear increase in the flow rate. Replacing the saline solution with amniotic fluid did not result in significant changes in the measured flow rates, except for the small needle size (24 Ga). Increasing the duration of the puncture did not result in increased flow rates, except for small needle size (24 Ga). CONCLUSION: These experiments suggest that needle diameter, angle of needle insertion, duration of the procedure, amniotic fluid pressure and composition could influence the incidence of amniotic fluid leakage following amniocentesis.

Biochemical differentiation of gestational compartments in the midgestational fetal rabbit.

OBJECTIVES: The fetal rabbit at midgestation is increasingly being used as a model in fetal diagnosis and therapy. In this study, we aimed to establish a reliable method for identification of the origin of sampled extra-embryonic fluids based on selected biochemical components. METHODS: In 6 pregnant does at 22 days of gestation, 18 gestational sacs were sampled for amniotic, allantoic and exocoelomic fluid. These fluids, as well as matching maternal and fetal blood samples, were assayed for levels of sodium, potassium, chloride, bicarbonate, total protein, alkaline phosphatase, gamma-glutamyl transferase and progesterone. RESULTS: Levels of sodium and potassium were, respectively, lower and higher in the allantoic fluid when compared to other extra-embryonic spaces. Amniotic fluid had a significantly lower total protein content and higher level of alkaline phosphatase when compared to the exocoelomic fluid. Significant levels of progesterone could only be detected in maternal blood. CONCLUSIONS: In the midgestational rabbit, a combined assay of potassium, alkaline phosphatase and progesterone can determine the gestational cavity of origin of the sampled fluid. The obtained gradients for these markers suggest compartment-specific production and/or inter-cavity transfer mechanisms.