The role of apoptosis in bispecific antibody-mediated T-cell cytotoxicity.

In this report we describe the role of apoptosis in the process of tumour cell killing by bispecific monoclonal antibody (BsMAb)-redirected cytolytic T cells. The BsMAb used, BIS-1, has dual specificity for the CD3 complex on T cells and the pancarcinoma-associated 38 kDa transmembrane antigen EGP-2. BIS-1 allows activated T cells to specifically recognise and kill EGP-2-positive but not EGP-2-negative target cells. An assay was developed to quantify apoptosis in cells by separation of 3H-thymidine-labelled low-molecular, i.e. fragmented, from high-molecular, i.e. non-fragmented DNA. The presence of low molecular weight DNA was measured both within the target cells and in the cell-free supernatant. After exposure to BIS-1-redirected, -activated T cells, apoptosis was observed in EGP-2-positive target cells but not in EGP-2-negative target cells. Also no DNA fragmentation proved to be induced in the activated effector cells during assay. The degree of EGP-2-positive target DNA fragmentation depended on the concentration of BsMAb, the E/T ratio and the incubation time. Using a low E/T ratio (1/1), DNA fragmentation in and 51Cr release from target cells showed similar characteristics and kinetics. At higher E/T ratio (20/1), the 51Cr release from the target cells increased to a greater extent than the percentage fragmented target cell DNA. Inhibitors of DNA fragmentation added to the cytotoxicity assay inhibited not only DNA fragmentation, but also the release of chromium-51 from the target cells, suggesting that apoptosis and cell lysis are closely related in BsMAb-mediated cell killing

In situ infrared absorption spectroscopy of dusty plasmas

International audienceIn situ, time-resolved Fourier transform infrared spectroscopy was used to study particulate formation in rf discharges in mixtures of silane, argon, and nitrogen. The spectra were taken at a maximum rate of 20 Hz. The discharge conditions were chosen such that previous calibrations of the time evolutions of particle size and density could be used. The measurements indicate that the onset of the solid-state vibrational absorptions of the SiH and SiH 2 bands only takes place after the nucleation and coagulation phase have finished; it coincides with the previously predicted start of the deposition of amorphous hydrogenated silicon on the particles. The dissociation of the silane feed gas is found to be in the range of 30%, and its time development suggests that also the large-scale dissociation of silane only starts after the coagulation phase. This is in agreement with previously observed trends for the electron temperature. If silicon partilces are grown in the plasma, and the silane flow is stopped, the Si particles stay trapped in the glow. The infrared measurements, however, show that they almost completely oxidize: the SiH/SiH 2 vibrations disappear and a strong SiO vibration appears. If nitrogen gas is allowed into the plasma, the SiO vibration is replaced by a SiN vibration

Immunomodulatory effects of intravenous BIS-1 F(ab')2 administration in renal cell cancer patients.

We report the immunomodulatory effects of an intravenous treatment with F(ab')2 fragments of the bispecific monoclonal antibody BIS-1 during subcutaneous recombinant interleukin 2 (rIL-2) therapy of renal cell cancer (RCC) patients. BIS-1 is directed against both the CD3 antigen on T cells and the EGP-2 molecule on carcinoma cells and some normal epithelia. The amount of BIS-1 F(ab')2 bound to peripheral blood lymphocytes (PBLs) increased dose-dependently. This occupation degree was highest at the end of the 2 h infusion and rapidly decreased subsequently. During the first hour of BIS-1 F(ab')2 infusion the number of PBLs decreased slowly. This was followed by an increase in serum tumour necrosis factor alpha (TNF-alpha) concentrations and a rapid decrease in the numbers of peripheral blood lymphocytes, monocytes and eosinophils. In our view, the most likely explanation for the observed decrease in occupation degree of BIS-1 F(ab')2 and the rise in TNF-alpha levels is based on the assumption that BIS-1-carrying T cells leave the circulation. The CD3 antigens on these extravasated T cells become cross-linked by EGP-2 antigens, inducing TNF-alpha secretion. This results in an enhanced decrease in the numbers of PBLs, monocytes and eosinophils. These preliminary results suggest that BIS-1 F(ab')2 treatment during IL-2 therapy may induce local T-cell activation

Transport of argon ions in an inductively coupled high-density plasma reactor

The first direct observation of the velocity distribution of the metastable Ar1*~2G9/2! ions in the presheath of an inductively coupled plasma has been achieved by using the Doppler shifted laser induced fluorescence technique. Drift of the ions along the electric field in the presheath is observed and distribution functions of the velocity in both parallel and perpendicular directions, relative to the E field, are deduced at 5 and 40 mTorr. Present results show that in high density plasmas the velocity distribution of the metastable ions is directly related to that of the ground state argon ions. Neutral gas temperature of around 600 K is also measured from the absorption profile of a diode laser beam, set on one of the 772.4 nm argon lines

Revealing an intermediate region between the collisional radiofrequency plasma bulk and its sheath

Experimental evidence of the existence of an intermediate region between a capacitively coupled plasma and the collisional space-charge region at its borders is presented. This proof is generated by monitoring—in an airplane carrying out parabolic flights—the trajectory of plasma-confined microparticles. Based on only primary data and without the need for a sophisticated model, our analysis concludes a sharply marked transition from the sheath region into another region with a significantly lower—yet nonzero—space-charge density, i.e., a region which is often called the presheath