Residual disease in B-cell chronic lymphocytic leukemia patients and prognostic value.

Twenty-two B-cell chronic lymphocytic leukemia (CLL) patients were investigated to evaluate residual disease in clinico-hematological remission. Residual disease was determined by monotypy of surface light-chain expression and by dual-color staining with CD5 and CD19 markers. Samples were analyzed on flow cytometer. Total CD19+ cells above 25%, the CD5+CD19+/total CD19+ cells ratio above 0.25, clonal excess above 0.4 were considered positive for residual disease. According to these immunological criteria, only four cases achieved phenotypic remission. Our data confirm that dual marker analysis is more sensitive than clonal excess and may predict an early relapse. Ig gene rearrangements were studied by Southern blot analysis using IGHJ and IGKC probes in fifteen cases. All 12 cases that retained a detectable rearrangement displayed a phenotypic residual disease. Conversely, in two cases, DNA analysis failed to detect the residual disease characterized by flow cytometry. In conclusion, this study suggests that in B-CLL, dual marker analysis is sensitive in predicting an early relapse in sequential evaluations of residual disease, whereas rearranged bands are undetectable when the proportion of malignant cells is low.

Expression of the hyaluronan-binding glycoprotein hyaluronectin in leukemias.

Hyaluronectin (HN), a hyaluronan (hyaluronic acid, HA)-binding glycoprotein is normally expressed in the nervous system, found in the desmoplasia of tumours, and is also produced in vitro by peripheral blood mononuclear cells. We have therefore investigated the expression and the production of HN by leukemic cells, with the hypothesis that HN would be expressed in leukemias of the myeloid lineage. Fresh and frozen leukemic cells were studied from 70 patients of whom 53 had acute myeloblastic leukemia (AML). HN was strongly expressed (> 80% blood cells) in two out of 13 M4 AMLs and four out of four M5B AMLs. One further M4 AML displayed 25% positive cells and two 20% cell positivity cases were seen, in one case of M4 AML and in one case of chronic myelomonocytic leukemia (CMML). The rest of the cases of AML as well as all cases of acute lymphoblastic leukemia (ALL) showed almost no positivity (< 1%). The residual positive cells appeared to be normal blood promonocytes. Taken together > or = 20% positive cells was seen in eight out of 56 (14%) examined myeloid leukemias. The HN production was significantly higher (p < 0.0001) in cell culture media of M4 and M5 AML cells than in other AML or ALL cell culture media. A significant correlation was found (p < 0.0001) between the number of HN-positive leukemic cells and the number of cells with a monocytic morphology, suggesting that HN is a marker for the promonocyte.

Rearrangements of immunoglobulin light and heavy chain genes and correlation with phenotypic markers in B-cell chronic lymphocytic leukemia.

Twenty-five patients with B-cell chronic lymphocytic leukemia (B-CLL) were investigated to correlate the immunological phenotype with the description of the Ig gene rearrangements of the B-cell clone. All patients were positive for the CD19 antigen and one pan B-antigen, markers of late cells (CD20, CD37 or Y2955). Twenty-four of the 25 patients tested expressed monoclonal cell surface immunoglobulin (SIg). The CD5 antigen was present in 21 of the 25 tested patients. Immunoglobulin gene rearrangements were detected by hybridization of the BamHI, EcoRI, BgIII, and HindIII digested genomic DNAs to the IGHJ, IGKC, IGLC, and IGLJ2 probes. Twenty-four of 25 patients had two rearranged IGH loci. The IGKC rearrangements were observed in 20 patients. In four patients, the IGK loci were deleted on both chromosomes. One patient without SIg displayed a germline pattern. All six patients with lambda producing B-CLL showed a lambda gene rearranged band, although the use of IGL polymorphism to investigate IGL rearrangements must be noted. These clonal rearrangements of IGL genes, together with the detection of either the kappa or lambda light chain of SIg, confirm that patients with B-CLL meet the developmental scheme of ordered light chain gene rearrangements.

Daily evaluation of circulating granulocyte-monocyte progenitors during bone marrow recovery from induction therapy in acute leukemia.

The notable increase of the circulating granulocyte-monocyte progenitors (PB CFU-GM) during bone marrow recovery following chemotherapy is a well known phenomenon. It has led to consider harvesting a large number of autologous stem cells by cytapheresis. Daily assessments have been conducted on the PB CFU-GM level in 9 patients with acute leukemia at the time of bone marrow regeneration after the first induction course in order to identify the circumstances of this rise. The PB CFU-GM maximum peak, of an average of 2142/ml, occurs between day 17 and day 23 after the chemotherapy has ended. A ten-fold increase of the PB CFU-GM level above normal values is maintained between 2 and 10 days. The PB CFU-GM peak coincides with that of the circulating immature myeloid cells and monocytes. The platelet rise above 100 X 10(9)/1 always occurs 2-7 days before the PB CFU-GM peak.

Del(14)(q22) in diffuse B-cell lymphocytic lymphoma.

Some recurrent chromosomal abnormalities have recently been found to be associated with distinctive histologic subtypes of non-Hodgkin's lymphoma (NHL). In a study of 62 patients with NHL whose karyotypes was determined at diagnosis, 3 patients were found to have a deletion of the long arm of chromosomes 14 at band 22 (del[14][q22]). All had a diffuse lymphoma with generalized lymphadenopathy and bone marrow involvement. All three lymphomas were of B-cell origin, as shown by the presence of surface immunoglobulin and monoclonal antibody phenotyping. For each patient, a trisomy 12 was associated with del(14)(q22) in a clone. These data suggest that del(14)(q22), perhaps in association with trisomy 12, could identify a subtype of NHL and that band 22 of chromosome 14 may be implicated in the B-cell ontogeny.

New evaluations of circulating granulocyte and macrophage stem cells in healthy adults using conditioned media and recombinant human growth factors.

Peripheral human blood contains granulo-monocyte (CFU-GM) and eosinophil (CFU-Eo) progenitors. In vitro, the number of colony forming units is thought to range from 0.1-14 per 2 x 10(5) plated cells. We show that the number of CFU-GM, Eo depends on culture methods. By modifying the usual assay method (using human umbilical cord plasma and the association of 2 stimulating conditioned media: activated lymphocyte conditioned medium and bone marrow fibroblast conditioned medium), we found different circulating CFU-GM, Eo numbers. The mean number of circulating CFU-GM, Eo in 107 healthy adults was 22.4 per 2 x 10(5) plated cells (range: 1-84). There was a slight difference between males (mean number: 23.6) and females (mean number: 20.4). The mean number of CFU-GM, Eo harvested on Percoll gradient was 123/ml of peripheral blood (range: 7-513). These results are far over those commonly reported in literature. This suggests that these latter results were probably underestimated. The use of recombinant human interleukin 3 and recombinant human GM (granulocyte-monocyte) colony-stimulating factors shows that CFU-GM, Eo numbers are found to be comparatively increased compared to that obtained with our modified method (using rhIL-3 alone), or that the size of those colonies is notably increased (using rhIL-3 + rhGM-CSF).

[Splenectomy in chronic idiopathic thrombopenic purpura in adults. Apropos of 49 cases]. / Splénectomie pour purpura thrombopénique idiopathique chronique de l'adulte. A propos de quarante-neuf cas.

The authors reviewed the case files of 49 adult patients undergoing splenectomy for chronic idiopathic thrombocytopenic purpura at the Centre Henri Becquerel between 1970 and 1987. Although the postoperative course was straightforward in 83.7% of cases, one reoperation for subphrenic abscess was necessary and there was one postoperative death. Remission from thrombocytopenia was obtained in 87.5% of the patients, but only transiently in 8.5% of them. No preoperative predictive factors could be demonstrated. An early postoperative rise in the platelet count to more than 500 G/litre appears to ensure a good subsequent result. Secondary infectious complications are not exceptional and can be fatal (one death in our series); they require prophylaxis by anti-pneumococcal vaccination. The place of prophylactic antibiotic therapy has yet to be defined.

[First-line treatment of febrile episodes in leukemia in adults. Randomized, multicenter study of ceftazidime in single antibiotic therapy versus a cefotaxime-amikacin combination]. / Traitement de première intention des épisodes fébriles des leucémies de l'adulte. Etude randomisée, multicentrique de la ceftazidime en monoantibiothérapie versus l'association céfotaxime-amikacine.

A prospective study was conducted in 10 haematology departments of university hospitals on 174 leukaemic patients with prolonged bone marrow aplasia and presenting with a febrile episode. The patients were allocated at random to either ceftazidime or the cefotaxime-amikacin combination. The two treatment group were similar as regards age, sex, underlying blood disease, duration of neutropenia, presence of a venous catheter, type of digestive tract contamination, clinical and bacteriological findings. Results were assessed mainly on the course of the fever at 48 hours and on the clinical and bacteriological changes observed until the patients came out of aplasia. Documented infections were specifically analyzed. There was no significant difference in terms of success or failure between the two treatment groups. Ceftazidime administered as monotherapy proved as effective as the cefotaxime-amikacin combination in the empirical first-line treatment of febrile episodes in leukaemic patients with neutropenia.

[Preventive treatment of infection caused by gram-negative bacteria using anti-lipopolysaccharide antibodies. Evaluation in patients with aplasia]. / Traitement préventif des infections à bacille à Gram négatif par des anticorps anti-lipopolysaccharides. Evaluation chez l'aplasique.

The prophylaxis of severe Gram-negative infections with human antiserum to lipopolysaccharide (LPS) was evaluated in a randomised study of 60 patients with therapeutic aplasia for leukaemia. The antiserum was found to be ineffective in preventing Gram-negative infections. The levels of anti-LPS antibodies showed that passive immunization was obtained in only one half of the patients. These disappointing results warrant further investigations to evaluate the effectiveness of this prophylactic treatment.