T-cell responses in osteosarcoma patients vaccinated with an anti-idiotypic antibody, 105AD7, mimicking CD55.

We assessed T-cell responses in young osteosarcoma patients vaccinated with 105AD7, 1-6 months after having received chemotherapy. 105AD7 is a human anti-idiotypic antibody mimicking CD55, a glycoprotein that protects from attack by complement and which is overexpressed on osteosarcoma cells. Seven out of 21 investigated patients made a IFN-gamma T-cell response against the vaccine, 105AD7 as assessed by ELISPOT. Cytokine secretion was analysed using Luminex assays and revealed TNF-alpha and GM-CSF responses not only to the vaccine but also towards the native antigen, CD55, in 5 / 14 (36%) of investigated patients. Importantly, the Luminex assay was found to be more sensitive than the more established T-cell assays (ELISPOT and proliferation assay), since responses towards the native antigen were recorded in this assay. Clinical responses and induction of immune responses to both the anti-idiotype and the native CD55 antigen support the use of CD55 as a target in cancer treatment.

Analysis of circulating tumor cells in patients with multiple myeloma during the course of high-dose therapy with peripheral blood stem cell transplantation.

In multiple myeloma (MM) circulating CD19+ cells have been considered as myeloma precursors. As these cells are also possibly a reservoir of treatment resistant disease evaluation of the CD19+ cells during the course of high-dose therapy has to be a major concern. We determined the number of tumor cells in the CD19+ as well as CD19- fractions of PB of eight patients with disease sensitive to VA[I]D chemotherapy, of 10 patients who achieved partial or complete remission post-high-dose therapy (HDT) with peripheral blood stem cell transplantation (PBSCT) and of a further seven patients with disease progression post-transplantation. CD19+ cell fractions were obtained by preparative sequential magnetic and fluorescence activated cell sorting with a median purity of 97.1%. In addition, PB samples of seven patients post-transplantation were sorted for CD20+ cells (median purity, 98.7%). The number of tumor cells in the CD19+, the CD19- and the CD20+ fractions were determined using a quantitative CDR3 PCR assay. The number of CD19+ tumor cells in patients in remission post-HDT was similar to those of the patients post-VA[I]D (median, 1.05 vs 0.92 CD19+ tumor cells/ml PB, P = 0.72) providing evidence for the persistence of this tumor cell fraction during the course of HDT. This was in contrast to the CD19- compartment, in which the number of tumor cells was significantly reduced in those patients in remission post-transplantation (median, 53 vs 0 CD19- tumor cells/ml PB; P = 0.006). In patients with progressive disease the number of tumor cells in both cell fractions was significantly higher (CD19+: median, 1.05 vs 21 tumor cells/ml PB, P = 0.05; CD19-: 0 vs 63 tumor cells/ml PB, P = 0.008). While the absolute number of CD19+ cells was reduced in the group of patients after VA[I]D treatment, a polyclonal CD19+ reconstitution had occurred in patients responding to HDT. The tumor cell content in the CD19+ fractions could be confirmed by the results obtained analyzing the CD20+ cell fractions. In conclusion, these results indicate that disease progression after PBSCT in MM is accompanied by an expansion of tumor cells in both the CD19+ and CD19- fractions. Similar numbers of CD19+ clonotypic cells post-HDT suggest that these cells persist and thus, contribute to disease dissemination and relapse.

Decreased numbers of circulating B cells in myeloma patients with reduction after conventional chemotherapy.

Reports of high numbers of circulating monotypic B cells in patients with multiple myeloma (MM) have recently been published. These cells, which were identified by their expression of CD19, were reported to be resistant to conventional chemotherapy and to represent the source of relapse. We examined blood samples from 48 patients before and 53 patients after glucocorticoid containing chemotherapy by dual color flow cytometry. The absolute count of CD19+B cells in patients before treatment (212.6+/-24.8 x 10(6)/l) was decreased compared to normal controls (P = .038). In the post-treatment group, circulating B cells were highly significantly lower than in untreated patients (45.23+/-6.69 x 10(6)/l. P < .001). This reduction was also seen in 26 patients, that were followed during chemotherapy. The cytoplasmic kappa/lambda ratio was within normal range before and after treatment with no difference according to the light chain isotype of the paraprotein. We conclude that circulating B cells are not increased in patients with MM, that the majority of these cells are polyclonal, and that conventional chemotherapy effectively reduces circulating B cells without leading to dominance of resistant monotypic cells.

[Chronotropic competence in patients with the sick sinus syndrome wearing AAI or DDD pacemakers]. / Capacidad de taquicardización en pacientes con enfermedad del nódulo sinusal portadores de marcapasos AAI o DDD.

The prevalence of chronotropic incompetence in patients with sinus node disease (SND) is not well defined. To assess this, we evaluated 18 patients (7 men, 11 women; mean age: 64 +/- 11) with SND and permanent pacemakers (AAI/DDD) with Holter monitoring and treadmill stress test. Only 2 patients received active cardiac drugs (1, L-dopa an 1 propafenone). The treadmill tests results were compared with a control group of 15 men and 18 women (mean age: 66 +/- 5, p = NS) without organic heart disease. During ambulatory activity all pacemaker patients increased their own cardiac rate to a value higher than the programmed basic pacemaker rate. In 8 patients the maximal rate attained was over 100/min (mean 95 +/- 19/min). The maximal rate during treadmill test in pacemaker patients was 131 +/- 25 (control group 138 +/- 14, p = NS). Exercise tolerance in METs was similar in pacemaker patients (5.2 +/- 2.6) and in controls (5.8 +/- 1.2) (p = NS). Two pacemaker patients (12%) didn't reach 100/min during stress test. Most patients with SND and permanent pacemakers (AAI/DDD) are able to increase cardiac rate during exercise. Rate responsive pacing (AAIR/DDDR) should be limited to a minority of patients with true chronotropic incompetence.

Suppression of Torsades de Pointes with verapamil in patients with atrio-ventricular block.

Experimental data have suggested a relation between Torsades de Pointes and early post-depolarization (EPD). We have studied the effect of intravenous verapamil in three patients with atrioventricular block (AVB) and Torsades de Pointes (TP), to obtain indirect evidence of slow membrane channel involvement in the TP mechanism. In two cases TP were completely suppressed and in one there was marked, albeit partial, suppression. In two cases verapamil did not shorten QT, while in the third suppression verapamil was related to junctional escape acceleration and QT shortening. In one of the cases where QT was not changed, the abolition of long pauses may have played a role in TP suppression. The affect of verapamil on TP in our patients is consistent with a combination of mechanisms, including direct membrane effects (EPD inhibition) and junctional pacemaker acceleration. Verapamil might be of therapeutic value in this clinical setting.

Validation of double-spike electrograms as markers of conduction delay or block in atrial flutter.

Recent mapping studies of atrial flutter have shown that fragmented electrograms can be found in most cases from the posterior, posteroseptal and posterolateral walls of the right atrium. The fragmentation pattern most often consists of a double spike. To further assess double-spike electrograms as a possible marker of conduction delay, bipolar electrograms were continuously recorded during atrial overdrive pacing of common flutter from the right atrium (7 patients) and from the proximal coronary sinus (5). Baseline double-spike separation of 50 to 130 ms was unchanged in 1 patient and slightly increased (5 to 25 ms) in 4 by coronary sinus pacing. The electrogram sequence was unchanged and the surface morphology was similar to that of basal flutter. Right atrial pacing decreased double-spike separation by 25 to 85 ms from basal values of 45 to 175 ms (23 to 83%), suggesting fusion in the area of fragmented electrograms. These findings suggest that double-spike electrograms represent activation on both sides of a conduction delay zone. The changes induced in these electrograms by pacing from the anterior right atrium and the coronary sinus are consistent with flutter circuits rotating counterclockwise (frontal plane) in the posterior right atrial wall in common atrial flutter.