RESUMEN
CD16 and natural killer (NK) cells appear to play a central role in mediating the anti-tumor effects of monoclonal antibody (mAb) therapy, yet little is known about changes in NK cells that result from interaction of the NK cells with mAb-coated tumor cells under physiologic conditions. We developed a system using peripheral blood mononuclear cells (PBMCs) and either transformed B cells or breast cancer cells to assess how mAbs impact on NK cell phenotype. Rituximab, apolizumab and trastuzumab induced modulation of CD16 and upregulation of CD54 on NK cells when the appropriate target cells were present. Higher concentrations of mAb were needed to induce these changes on NK cells from subjects with the lower affinity CD16 polymorphism. Phenotypic changes were greater in NK cells from subjects with the higher affinity polymorphism even when saturating concentrations of mAb were used, demonstrating increased concentration of mAb can overcome some, but not all, of the influence CD16 polymorphisms have on NK activation. These studies provide a straightforward and easily reproducible technique to measure the ability of mAb-coated tumor cells to activate NK cells in vitro which should be particularly useful as mAbs with varying affinity for both target antigen and Fc receptor (FcR) are developed.
Asunto(s)
Anticuerpos Monoclonales/farmacología , Antígenos CD/genética , Pruebas Inmunológicas de Citotoxicidad , Células Asesinas Naturales/inmunología , Activación de Linfocitos/inmunología , Receptores de IgG/genética , Antígenos CD/biosíntesis , Línea Celular Transformada , Línea Celular Tumoral , Técnicas de Cocultivo , Pruebas Inmunológicas de Citotoxicidad/métodos , Regulación hacia Abajo/genética , Regulación hacia Abajo/inmunología , Citometría de Flujo/métodos , Proteínas Ligadas a GPI , Regulación Neoplásica de la Expresión Génica/inmunología , Humanos , Molécula 1 de Adhesión Intercelular/biosíntesis , Molécula 1 de Adhesión Intercelular/genética , Interferón gamma/biosíntesis , Células Asesinas Naturales/metabolismo , Activación de Linfocitos/genética , Receptores de IgG/antagonistas & inhibidores , Receptores de IgG/biosíntesis , Regulación hacia Arriba/genética , Regulación hacia Arriba/inmunologíaRESUMEN
Despite years of efforts to quantify cation distribution as a function of composition in the magnetite-ulvöspinel solid solution, important uncertainties remain about the dependence of cation ordering on temperature and cooling rate. Here we demonstrate that Curie temperature in a set of natural titanomagnetites (with some Mg and Al substitution) is strongly influenced by prior thermal history at temperatures just above or below Curie temperature. Annealing for 10(-1) to 10(3) h at 350-400 °C produces large and reversible changes in Curie temperature (up to 150 °C). By ruling out oxidation/reduction and compositional unmixing, we infer that the variation in Curie temperature arises from cation reordering, and Mössbauer spectroscopy supports this interpretation. Curie temperature is therefore an inaccurate proxy for composition in many natural titanomagnetites, but the cation reordering process may provide a means of constraining thermal histories of titanomagnetite-bearing rocks. Further, our theoretical understanding of thermoremanence requires fundamental revision when Curie temperature is itself a function of thermal history.
RESUMEN
Growing evidence indicates that the affinity of monoclonal antibodies (mAbs) for CD16 (FcgammaRIII) plays a central role in the ability of the mAb to mediate antitumor activity. We evaluated how CD16 polymorphisms, and mAb with modified affinity for target antigen and CD16, affect natural killer (NK) cell phenotype when CD20(+) malignant B cells were also present. The mAb consisted of rituximab (R), anti-CD20 with enhanced affinity for CD20 (AME-B), and anti-CD20 with enhanced affinity for both CD20 and CD16 (AME-D). Higher concentrations of mAb were needed to induce CD16 modulation, CD54 up-regulation, and antibody-dependent cellular cytotoxicity (ADCC) on NK cells from subjects with the lower affinity CD16 polymorphism. The dose of mAb needed to induce NK activation was lower with AME-D irrespective of CD16 polymorphism. At saturating mAb concentrations, peak NK activation was greater for AME-D. Similar results were found with measurement of CD16 modulation, CD54 up-regulation, and ADCC. These data demonstrate that cells coated with mAb with enhanced affinity for CD16 are more effective at activating NK cells at both low and saturating mAb concentrations irrespective of CD16 polymorphism, and they provide further evidence for the clinical development of such mAbs with the goal of improving clinical response to mAb.