[Development of a new strategy for minimal residual disease monitoring in children with B-precursor acute lymphoblastic leukemia]. / Mise en place d'une stratégie de suivi de la maladie résiduelle chez les enfants atteints de leucémies aiguës lymphoblastiques préB.

Despite modern regimen of chemotherapy, one-third of children with acute lymphoblastic leukaemia relapse. Recent studies have shown that a high level of minimal residual disease (MRD) at the end of induction is associated with an increased risk of relapse. We have developed and validated a real-time PCR method (RQ-PCR) to quantify MRD. We monitored 57 patients using IgH and TCR (Vdelta2Ddelta3) genes rearrangements as PCR targets. RQ-PCR was performed with a primer and a TaqMan probe designed to consensus sequences in VH segments in combination with one allele specific oligonucleotide primer complementary to the junctionnal region. A sensitivity of 10(-4) was reached for 72% of the IgH alleles (n = 50) and for 54,5% of the Vdelta2Ddelta3 alleles (n = 22). We compared the results with those obtained by competitive PCR in 53 patients: no discordance between the two methods was observed. Seventeen patients were found positive (32%) and 27 negative (51%) with both techniques and 9 children (17%) were positive only with TaqMan technology. RQ-PCR is more sensitive and more specific than competitive PCR. We thus propose that RQ-PCR might be used in first intention for MRD analysis.

Candida bituminiphila, a novel anamorphic species of yeast.

A novel anamorphic species of yeast belonging to the genus Candida was isolated from tar in Canada. Morphological and physiological observations, as well as phylogenetic analyses, were performed. Conidiophores were produced, were usually short and had sympodial growth, numerous bud scars and a rachis-like structure. They bore one or more conidia. Pseudomycelium was scarcely produced and true mycelium was sparse. No sexual reproduction was observed on corn meal, malt, Gorodkowa, Dextrose Yeast Peptone or V8 agars. Zygoascus hellenicus was physiologically the most closely related species, but it differed from the novel species by its ability to assimilate D-galacturonate and L-rhamnose, ferment sucrose and grow at 37 degrees C. From sequence analysis of the 26S rDNA D1/D2 region, Z. hellenicus and Candida bertae var. bertae were the closest species with 54 and 56 bp substitutions, respectively. Similar results have been obtained from analysis of the 18S rDNA. All these data support the hypothesis that the yeast, named Candida bituminiphila, is a novel species closely related to Z. hellenicus. The holotype and only isolate of C. bituminiphila is strain CBS 8813T (= MUCL 41424T).

A new family of mouse genes homologous to the human MAGE genes.

The human MAGE genes are expressed in a wide variety of tumors but not in normal cells, with the exception of the male germ cells, placenta, and, possibly, cells of the developing embryo. These genes encode tumor-specific antigens recognized by cytolytic T lymphocytes. The MAGE genes are located on the X chromosome, in three clusters denoted MAGE-A, B, and C, mapping at q28, p21.3, and q26, respectively. The function of these genes remains unknown. Because mice offer many advantages for the study of genes that may be involved in embryonic development, we looked for the murine equivalents of the 12 human MAGE-A genes. Using a MAGE-A probe, we isolated 8 new murine genes that are homologous to the MAGE genes. On average, the open reading frames (ORFs) of these 8 closely related genes display a slightly higher degree of nucleotide identity with the MAGE-A ORFs than with the MAGE-B or MAGE-C ORFs. Furthermore, like MAGE-A genes, they encode acidic proteins, whereas the MAGE-B genes encode basic proteins. Accordingly, these 8 murine genes were named Mage-a1 to 8 (approved symbols Magea1 to 8). Mage-a genes were mapped in two different loci on the mouse X chromosome. Mage-a4 and Mage-a7 are located in a region that is syntenic to either Xp21 or Xq28. The 6 other genes are arranged in a cluster located in a region syntenic to Xp22. Like their human counterparts, Mage-a genes were found to be transcribed in adult testis, but not in other tissues. Expression of some Mage-a genes was also detected in tumor cell lines. Two Mage-a genes were found to be expressed in blastocysts.