Procedures for the expansion of CD14(+) precursors from acute myeloid leukemic cells to facilitate dendritic cell-based immunotherapy.

AIMS: Vaccination with acute myeloid leukemia (AML)-derived dendritic cells (DCs) is a promising immunotherapeutic approach to prevent relapse of AML. However, in clinical practice AML-derived DC culture is unfeasible in 40% of cases. Here, we demonstrate that AML cells can be expanded in vitro prior to differentiation with cocktails of cytokines with known myeloid growth-promoting effects. RESULTS: Nine out of 13 initially CD14(-) samples gain de novo CD14 (>10%) expression (69% increment; p = 0.01) after in vitro expansion. These expanded CD14(+) leukemic cells displayed a high probability (six out of six initially CD14(-) samples tested) to differentiate into DCs upon culture with GM-CSF, TNF-α and IL-4. CONCLUSION: Induction of CD14 on initially CD14(-) AML cells potentially increases the number of patients eligible for DC-based immunotherapy.

Virulence of Newcastle disease virus is determined by the cleavage site of the fusion protein and by both the stem region and globular head of the haemagglutinin-neuraminidase protein.

Virulence of Newcastle disease virus (NDV) is mainly determined by the amino acid sequence surrounding the fusion (F) protein cleavage site, since host proteases that cleave the F protein of virulent strains are present in more tissues than those that cleave the F protein of non-virulent strains. Nevertheless, comparison of NDV strains that carry exactly the same F protein cleavage site shows that significant differences in virulence still exist. For instance, virulent field strain Herts/33 with the F cleavage site 112RRQRRF117 had an intracerebral pathogenicity index of 1.88 compared with 1.28 for strain NDFLtag, which has the same cleavage site. This implies that additional factors contribute to virulence. After generating an infectious clone of Herts/33 (FL-Herts), we were able to map the location of additional virulence factors by exchanging sequences between FL-Herts and NDFLtag. The results showed that, in addition to the F protein cleavage site, the haemagglutinin-neuraminidase (HN) protein also contributed to virulence. The effect of the HN protein on virulence was most prominent after intravenous inoculation. Interestingly, both the stem region and the globular head of the HN protein seem to be involved in determining virulence.