Podoplanin (Aggrus), which is a type I transmembrane sialomucin-like glycoprotein, is highly expressed in malignant pleural mesothelioma (MPM). We previously reported the generation of a rat anti-human podoplanin Ab, NZ-1, which inhibited podoplanin-induced platelet aggregation and hematogenous metastasis. In this study, we examined the antitumor effector functions of NZ-1 and NZ-8, a novel rat-human chimeric Ab generated from NZ-1 including Ab-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity against MPM in vitro and in vivo. Immunostaining with NZ-1 showed the expression of podoplanin in 73% (11 out of 15) of MPM cell lines and 92% (33 out of 36) of malignant mesothelioma tissues. NZ-1 could induce potent ADCC against podoplanin-positive MPM cells mediated by rat NK (CD161a+) cells, but not murine splenocytes or human mononuclear cells. Treatment with NZ-1 significantly reduced the growth of s.c. established tumors of MPM cells (ACC-MESO-4 or podoplanin-transfected MSTO-211H) in SCID mice, only when NZ-1 was administered with rat NK cells. In in vivo imaging, NZ-1 efficiently accumulated to xenograft of MPM, and its accumulation continued for 3 wk after systemic administration. Furthermore, NZ-8 preferentially recognized podoplanin expressing in MPM, but not in normal tissues. NZ-8 could induce higher ADCC mediated by human NK cells and complement-dependent cytotoxicity as compared with NZ-1. Treatment with NZ-8 and human NK cells significantly inhibited the growth of MPM cells in vivo. These results strongly suggest that targeting therapy to podoplanin with therapeutic Abs (i.e., NZ-8) derived from NZ-1 might be useful as a novel immunotherapy against MPM.

No abstract

The gelation mechanism of polyethylene solutions was investigated by using light scattering and X-ray diffraction techniques in terms of the liquid−liquid phase separation. Three kinds of polyethylene, ultrahigh molecular weight polyethylene (UHMWPE), low molecular weight linear polyethylene (L-LMWPE), and low molecular weight branched polyethylene (B-LMWPE), were used as test specimens. When an incident beam of He−Ne gas laser was directed to the UHMWPE and B-LMWPE solutions quenched to a desired temperature, the logarithm of scattered intensity increased linearly in the initial stage and tended to deviate from this linear relationship in the latter stage. If the linear increase in the initial stage can be analyzed within the framework of the linear theory of spinodal decomposition proposed by Cahn, the gelation is obviously attributed to the phase separation leading to the concentration fluctuation of the solution. Furthermore, in the later stage showing the deviation from the linear relationship, light scattering from the UHMWPE solution with 0.5% concentration under Hv polarization condition showed an X-type pattern, indicating the appearance of optically anisotropic rods. On the other hand, the scattering from the B-LMWPE solution with 3% concentration showed a four-leaf clover pattern, indicating the appearance of optically anisotropic spherulites. No crystallite was confirmed by the X-ray diffraction measurements, when the rods and spherulites appeared. With further lapse of time, the Hv patterns became clearer and the corresponding X-ray diffraction intensity curves showed a very small diffraction peak from the (110) plane. In contrast, for the L-LMWPE solutions, the logarithm of scattered intensity against time showed a rapid increase and tended to level off because of rapid change from slightly transparent to whiten gels. In such a process, any quantitative analysis of the scattered light intensity was impossible. The corresponding X-ray diffraction revealed the strong reflections from the (110) and (200) planes, indicating the rapid formation of stable crystallites. Through a series of experiments for the three kinds of polyethylene solution, it turned out that gelation mechanism of polyethylene solutions is strongly affected by molecular weight and the degree of branching.

To analyze polarized light scattering patterns from gels, an approach is proposed to calculate the scattered intensity. In the proposed model system, difference between polar angles of the principal axes of the and elements, which were defined with respect to the axis along the distance between two elements, was given as a correlation of the distance between the two elements. Furthermore, the azimuthal angle, which makes a projection of the principal axis onto a plane perpendicular to the principal axis of the element, was also given as a correlation of the distance between the two elements. The theoretical calculation was carried out for the scattered intensity under Hv and Vv polarization conditions. The general equations proposed for Hv and Vv scattering were based on a statistical approach for polarized light scattering system. The calculated pattern under the Hv polarization condition showed an X-type pattern and was in good agreement with the pattern observed from polymer gels prepared by quenching their solutions to the desired temperatures.