HER-2/neu-derived peptide epitopes are also recognized by cytotoxic CD3(+)CD56(+) (natural killer T) lymphocytes

The human HER-2/neu gene encodes a 185 kDa transmembrane glycoprotein recognized by MHC class I-restricted CTLs. Here, we report that HER-2/neu peptide CTL epitopes can also be recognized by cytotoxic NK-T lymphocytes. Unfractionated peptides derived from HLA-A2(+), HER-2/neu(+) tumor cells acid cell extract (ACE), collected from patients with metastatic ovarian cancer, were used as antigen to generate in vitro cytotoxic effectors. ACE was able to elicit from cancer patients' PBMCs both alphabetaTCR(+)CD3(+)CD56(-) and alphaTCR(+)CD3(+)CD56(+) (NK-T) CTLs that lysed ACE-sensitized T2 cells in an HLA-A2-restricted manner. The same CTL lines also recognized T2 cells pulsed with HER-2/neu-derived CTL peptide epitopes, a HER-2/neu-transfected HLA-A2(+) cell line and autologous tumor cells. alphaTCR(+)CD3(+)CD56(+) CTL lines also exhibited NK-like cytotoxicity against autologous tumor cells. CTL clones were isolated from alphaTCR(+)CD3(+)CD56(+) bulk cultures displaying both MHC- and non-MHC-restricted cytotoxicity, thus confirming the dual cytolytic function of such cells. Our data demonstrate that ACE from metastatic ovarian tumors can be used as multiepitope vaccines for generating in vitro, besides classical CTLs, NK-T cells exerting efficient MHC- and non-MHC-restricted cytotoxicity against autologous tumor targets. Such NK-T cells expressing dual cytotoxic activity may prove advantageous in cancer immunotherapy.     

Recurrent Bacterial Vaginosis in a Virgin Adolescent: A New Method of Treatment

Bacterial vaginosis (BV) is a polymicrobial infection of the vagina and should not be considered an exclusively sexually transmitted disease. We describe the case of a 17-year-old female virgin adolescent with recurrent malodorous vaginal discharge for 6 months. Before referral to us she had been treated unsuccessfully with conservative treatment options. Our investigation revealed Gardnerella vaginalis as the responsible factor for the vaginal infection. Because metronidazole treatment had failed as monotherapy, a new method was applied. Repeated vaginal washings with 3% H₂O₂, 15% NaCl and 10% providone iodine were initiated. At the end of each washing, vaginal walls were thoroughly cleaned up with a small gauze. After 10 days of treatment the odor and the vaginal discharge had ceased and 12 months later no relapse had occurred. It seems to be reasonnable to use this kind of treatment in recurrent BV. Received: January 3, 2001 · Revision accepted: April 23, 2002 E. G. Papanikolaou (corresponding author)     

Increased signal intensity on fat-suppressed three-dimensional T1-weighted pulse sequences in patellar tendon: magic angle effect?

Objective. To assess the frequency of increased signal intensity in the patellar tendon using three-dimensional T1-weighted MRI pulse sequences. Design and patients. Sixty patients were examined with a 1.0 T scanner (15mT/m gradient strength) using a quadrature coil. Three pulse sequences were applied in the sagittal plane: PD turbo spin echo (PD-TSE), 3D T1-weighted gradient echo with fat suppression (3D-T1-FFE-FS) and 3D T1-weighted echo planar imaging with fat suppression (3D-T1-EPI-FS). The high signal intensity areas were measured in their maximum length. The angle of the patellar tendon relative to the main field position was measured in the same slice. In eight patients with anterior knee pain, and in 11 with no anterior knee pain, a fourth T2-weighted TSE pulse sequence (T2-TSE) was obtained to rule out patellar tendinitis. Results. The correlation of the high signal intensity areas with the relative position of the tendon was found to be significant with the 3D sequences (P=0.03 for 3D-T1-FFE-FS and P=0.003 for 3D-T1-EPI-FS). The length of the high signal intensity area in the tendon was 5.4 mm with 3D-T1-FFE-FS, 4.9 mm with 3D-T1-EPI-FS and 3.1 mm with PD-TSE images. No patellar tendinitis was demonstrated on the T2-TSE images. Conclusion. The magic angle effect is commonly observed in the 3D based T1-weighted pulse sequences with fat suppression. The presence of the above sign must be recognized by radiologists, so that misdiagnosis of patellar tendinitis is avoided. Received: 31 March 2000 Revision requested: 11 July 2000 Revision received: 2 August 2000 Accepted: 26 October 2000     

Pathogenic BRCA1 mutations may be necessary but not sufficient for tissue genomic heterogeneity: Deep sequencing data from ovarian cancer patients

Background

Tissue genomic heterogeneity (t-HET) in patients with epithelial ovarian cancer (OVCA) is related to tissue plasticity, i.e., flexibility to adapt to adverse molecular environments. Here, we interrogated the presence and clinical relevance of OVCA t-HET.

Assessment of proliferating-cell nuclear antigen immunostaining in soft-tissue tumours: Relationship to histological grade and mitotic activity

The proliferative activity in 70 cases of soft-tissue tumours was estimated immunohistochemically using the monoclonal antibody PC-10, which recognizes proliferating-cell nuclear antigen (PCNA) in paraffin sections. The PCNA index (i.e. the percentage of positive neoplastic nuclei) and to a lesser degree the PCNA count (i.e. the number of positive neoplastic nuclei per ten high-power fields) positively correlated with the malignancy grade (PCNA index:P<0.001; PCNA count:P<0.01). However, the range of values of PCNA index and PCNA count was similar between benign and grade I tumours. A statistically significant positive correlation was also established between PCNA index and PCNA count on the one hand and mitotic count on the other, but the correlation coefficient was low (r=0.351,P<0.01, andr=0.290,P<0.05 respectively). These results indicate that PCNA immunostaining may successfully be used as an adjunct to the conventional histopathological parameters in assessing the malignancy grade in soft-tissue tumours although it is of limited value in distinguishing between benign and grade I tumours.Abbreviations PCNA proliferating-cell nuclear antigen - HPF high-power field     

Sensitivity to vecuronium in myasthenia gravis: a dose-response study

A cumulative dose plus infusion technique and integrated EMG monitoring of the first dorsal interosseous muscle were used to determine the potency of vecuronium in 20 normal patients and in ten patients with myasthenia gravis under thiamylal, N2O, O2, fentanyl anaesthesia. The mean (+/- SEM) values for ED50, ED90, and ED95 in the normal patients were 19 +/- 1, 31 +/- 1 and 36 +/- 2 micrograms.kg-1, respectively. Myasthenic patients showed increased sensitivity to vecuronium, the mean values for ED50, ED90, and ED95 were 10 +/- 2, 17 +/- 2 and 20 +/- 3 micrograms.kg-1, being 50, 55 and 56 per cent of normal, respectively. We did not demonstrate a difference in sensitivity to vecuronium between those myasthenic patients who received pyridostigmine preoperatively and those who did not, nor among those chronically treated with corticosteroids, compared with those who were not.     

Hypermethylation of CpG islands in the promoter region of the p15INK4B gene in childhood acute leukaemia

It has been reported that the cyclin-dependent kinase inhibitor (CDKI) gene p15INK4B is frequently inactivated by genetic alterations and may be responsible for various malignant tumours. Another way of inactivation of this CDKI is by hypermethylation of 5'CpG islands in the promoter region of the p15INK4B gene and this inactivation seems to be a frequent event in various haematological malignancies. In the present study, we investigated the methylation status of the p151NK4B gene to clarify its role in the pathogenesis of childhood acute myeloid (AML) and acute lymphoblastic leukaemia (ALL). The study included 23 cases of B-cell origin ALL, 13 cases of T-cell origin ALL, 32 cases of AML, and 10 apparently healthy controls. Hypermethylation was studied by methylation-specific polymerase chain reaction. Hypermethylation of the p15INK4B gene was more frequent in cases with T-cell origin ALL (46.2%), but similar among children with B-cell origin ALL (13.0%) and AML (18.8%). Hypermethylation of p15INK4B may be involved in the pathogenesis of T-cell origin ALL, but not in that of AML or B-cell origin ALL.