Tuberculosis associated haemophagocytic syndrome: two cases with a favourable outcome.

Haemophagocytic syndrome is a heterogenous disease characterized by disordered macrophage activation associated with viral, bacterial or parasitic infection. The few reports of haemophagocytosis occurring in the presence of mycobacterial infection show a high mortality rate and we present two further cases notable for their favourable issue. Rapidity of diagnosis and immediate treatment could explain the avoidance of a fatal outcome.

Cytomegalovirus expression in minor salivary glands and chronic graft-versus-host disease.

A systematic survey of human cytomegalovirus (CMV) was performed in 29 allogeneic bone marrow transplant (BMT) recipients. At day 100 a lip biopsy was performed and histological grading according to Sale's score was compared with the immunohistochemical detection of the immediate early protein IE2 of HCMV. In 10 patients without chronic graft-versus-host disease (GVHD), 3 had lip biopsy grade 1, 7 had grade 0 Sale's score and in 19 patients with chronic GVHD, 11 had grade 2, 1 had grade 1 and 7 had grade 0. On the same lip biopsies, we found IE2 protein in 8 of the 19 patients with chronic GVHD. None of the lip biopsies from patients without chronic GVHD expressed the protein, suggesting that HCMV expression is strongly associated with chronic GVHD and Sale's grade 2. To conclude, in our group of patients with a risk for HCMV infection, detection of the protein IE2 was a good predictive criterion of chronic GVHD with a sensitivity of 61% and a specificity of 100%.

High incidence of lymphoid infiltration on labial salivary gland biopsy in non-Hodgkin's lymphomas: clinical implications.

Sjögren's syndrome (SS) is characterized by an increased risk of developing a non-Hodgkin's lymphoma (NHL). We performed labial salivary gland biopsy (LSGB) in 103 patients with untreated NHL, negative for human immunodeficiency virus. Median age was 58 years (range 21-79) and M/F 1.3. Using the Working Formulation, 37 patients had low-grade NHL and 66 had intermediate- or high-grade NHL. Dense lymphocytic infiltration (positive focus score 3 or 4) was found in 28 patients. 10 (35%) of these 28 patients fulfilled criteria for possible SS. 15/28 patients had an identical monotypic infiltrate on LSGB and NHL tissue (including two of the 10 patients with criteria for SS). The significance of the lymphoid infiltrate of LSGB was unknown in the five remaining patients with positive focus score. Significant correlations were found between positive focus score and presence of two or more extranodal sites of disease (P = 0.02), impaired performance status (P = 0.004), splenomegaly (P = 0.05), increased gammaglobulin level (P = 0.03), and beta 2 microglobulin (P = 0.004). Considering intermediate- or high-grade NHL, we found significant correlation between positive focus score and unfavourable prognosis according to the two Dana Farber Cancer Institute indexes (P < 0.04), to the LNH-84 index (P = 0.05), and to the international index (P = 0.003). In conclusion, systematic evaluation of LSGB in 103 patients with NHL found lymphoid infiltration in 28% of them, but possible SS could be considered in only 10%. This lymphoid infiltration, though not correlated with any particular histological subtypes, was associated with unfavourable clinical prognostic factors, especially in intermediate- or high-grade NHL.

Non-Hodgkin's lymphoma and Sjögren's syndrome. An immunopathological study of 113 patients.

One hundred and thirteen patients with untreated non-Hodgkin's lymphoma were evaluated for Sjögren's syndrome (SS) using clinical and biological parameters, and a minor salivary gland biopsy. Concomitant immunophenotyping for T and B cells, and kappa and lambda chains was performed on both minor salivary gland and lymphomatous tissues. Patients with a positive focus score on examination of their minor salivary gland biopsy had a second biopsy, when possible, at the end of induction therapy. Fourteen of the 113 patients had SS according to the Greek criteria, and 4 of these 14 patients had an identical monotypic infiltrate in both their minor salivary gland and lymphomatous tissues. In all four cases this infiltrate disappeared from both locations when the lymphoma was in complete remission (CR). The 10 other patients with criteria for SS had a mixed infiltrate which persisted unchanged when the lymphoma was in CR. Among the 99 patients without SS, 12 had a positive focus score and an identical monotypic infiltrate in both their minor salivary gland and lymphomatous tissue. This infiltrate disappeared from both locations when the lymphoma was in CR. Among the remaining 87 patients, 82 had no minor salivary gland infiltrate, and 5 had a positive focus score with mixed lymphocytic infiltrate. Long term follow up and a large cooperative study are needed to better understand the immunopathologic lesions of patients of this latter type.

Mutations of the p53 gene in B-cell chronic lymphocytic leukemia: a report on 39 cases with cytogenetic analysis.

Mutations of exons 5 to 8 of the p53 gene were looked for in 39 cases of B-cell chronic lymphocytic leukemia (CLL) using polymerase chain reaction single-strand conformation polymorphism analysis and DNA sequencing. All patients also had cytogenetic analysis. A point mutation, leading to an amino acid change in the p53 protein was found in four cases, involving exon 7 (one case) or exon 8 (three cases). Mutations seemed to predominate in advanced clinical stages (Binet's stage C). All four patients with 17p monosomy had a mutation whereas no mutation was found in the 35 patients with cytogenetically normal 17p. These findings suggest that p53 mutations are relatively rare in B-cell CLL, and largely predominate or may even be restricted to patients with 17p monosomy (who constitute about 5% of all B-cell CLL patients in large published series). In those patients, the mutations may play a role in leukemogenesis through loss of tumor suppressive activity of normal p53 genes.

Mutations of the P53 gene in acute myeloid leukaemia.

In a previous report we found point mutations in exons 5-8 of the P53 gene in five of 46 patients with acute myeloid leukaemia (AML), with a predominance of mutations in the 10 patients with 17p monosomy. In this report we extended our findings studying such mutations in 66 unselected additional cases of AML, using polymerase chain reaction single strand conformation polymorphism (SSCP) analysis and nucleotide sequencing. Three of the 66 new cases had a point mutation, leading to a change in one encoded amino acid. Thus, eight of the 112 AML studied had P53 mutations in exons 5-8, suggesting that the incidence of P53 mutation is relatively low in AML. A predominance of mutations in exon 8 (5/8) was found. Six of the eight patients with mutations were older than 60 years of age, and all eight cases had a short survival. All seven mutated cases karyotyped showed complex cytogenetic findings, especially monosomy 5 and/or 7, thus questioning the pathogenic importance of P53 mutations in a context of multiple genetic abnormalities. However, five of them also had 17p monosomy, and in the remaining two cases SSCP and sequence analysis also suggested loss of the normal P53 allele. This supported a role for the P53 gene mutations in leukaemogenesis in the relatively small number of AML patients in whom they were found, through loss of tumour suppressive activity of both normal P53 alleles, as reported in solid tumours.

Mutations of the p53 gene in B-cell lymphoblastic acute leukemia: a report on 60 cases.

Exons 5 to 8 of the p53 gene were examined for mutations in 60 patients with B-cell acute lymphoblastic leukemia (ALL), including 50 cases of precursor-B-cell ALL, nine cases of Burkitt (L3) ALL and one case of atypical ALL with surface immunoglobulins and t(8:14) translocation but L2 morphology. Karyotype was available in all patients. DNA was analyzed by polymerase chain reaction, single strand conformation polymorphism analysis, and nucleotide sequencing. Three patients showed point mutations in exons 7 or 8, including two of the nine patients with Burkitt ALL and one of the 50 patients with precursor-B-cell ALL. These findings suggest that p53 gene mutations are rare in precursor-B-cell ALL but may be more frequent in Burkitt ALL. In the three patients with p53 mutations, however, the relevance of those mutations to the development or progression of leukemia remained uncertain.

Mutations in the p53 gene in myelodysplastic syndromes.

We have examined p53 alleles in 151 DNAs from patients with myelodysplastic syndrome using single-strand conformation polymorphism analysis of polymerase chain reaction products. We focused our study on the four highly conserved regions of the p53 gene and detected five patients with aberrantly migrating fragments. We confirmed the putative mutation in each case by direct sequencing analysis. Of these five patients, three had chromosome 17 monosomy associated with p53 mutation, one patient showed one mutated p53 allele and one wild-type allele, and the last patient demonstrated only the mutant allele, suggesting a homozygous state. Unlike many other types of human cancers, point mutations in the p53 tumor-suppressor gene appear to be a rare event in myelodysplastic syndromes.

P53 gene mutations in acute myeloid leukemia with 17p monosomy.

We looked for mutations of exons 5 to 8 of the P53 gene in 10 patients with acute myeloid leukemia (AML) and 17p monosomy, and 36 patients with AML and no cytogenetic abnormalities of 17p. DNA was analyzed by polymerase chain reaction, single-strand conformation polymorphism analysis, and nucleotide sequencing. Four of the 10 patients with 17p monosomy showed point mutation, single-nucleotide deletion, or insertion in exons 7 or 8. By contrast, only 1 of the 36 patients with AML and no cytogenetic abnormalities of 17p showed a mutation of the P53 gene in exons 5 to 8 (P less than .01). These results suggest that alterations of the P53 gene may have a role in leukemogenesis in some cases of AML. The fact that P53 gene mutations occurred more often in patients with 17p monosomy seems to support the "recessive" model of tumor suppressive activity of the P53 gene rather than the "dominant" model, in which alteration of only one allele is sufficient for the development of malignancy.

Therapy related myelodysplastic syndrome and leukemia with no "unfavourable" cytogenetic findings have a good response to intensive chemotherapy: a report on 15 cases.

We treated 15 patients with therapy related acute nonlymphocytic leukemia (tANLL) or therapy related myelodysplastic syndrome (tMDS) who had no rearrangements of chromosomes 5 and/or 7 or complex cytogenetic rearrangements by intensive chemotherapy. The median age was 43 years. Seven patients had one of the "specific" rearrangements of de novo ANLL (inv(16), t(8;21), t(15;17)or t(9;11)). Eight patients had a normal karyotype (4 cases) or single cytogenetic rearrangements not involving chromosomes 5 and 7: trisomy 8 (2 cases), t(1,2)(1 case), 20q deletion (1 case). All 7 patients with "specific" rearrangements had tANLL at presentation, without a preceding myelodysplastic phase. Seven of the 8 patients with a normal karyotype or other single cytogenetic rearrangements presented with tMDS, and the remaining patient with tANLL. Twelve patients achieved complete remission (CR), 2 had hypoplastic death and 1 had resistant disease. Median actuarial disease free interval (DFI) was 30 months. No significant prognostic factor for achieving CR was found. Significantly longer DFI was found in patients with "specific" chromosome rearrangements, compared to other karyotypes, and in patients who presented with tANLL, compared to those who presented with tMDS. Those 2 prognostic factors strongly correlated. In contrast to tANLL and tMDS with rearrangements of chromosomes 5 and/or 7 or complex karyotypes, patients with tANLL or tMDS who had other abnormal cytogenetic findings seem to achieved a high CR rate with intensive chemotherapy. tANLL with "specific" rearrangements achieved prolonged CR in many cases, whereas tMDS with other abnormal karyotypes generally had short CR, like their de novo counterparts.

Slow infusions of vinblastine in the treatment of adult idiopathic thrombocytopenic purpura: a report on 43 cases.

Forty-three adult patients with idiopathic thrombocytopenic purpura (ITP) were treated by slow intravenous infusions of vinblastine. Nineteen had ITP of recent onset (i.e. of less than 6 months duration) and had contraindication to steroids (3 patients), refractoriness to steroids (6 patients) or to steroids and high dose intravenous immunoglobulins (IVIg, 10 patients). Of the 19 patients, 10 achieved complete response (CR), 2 achieved partial response (PR), 2 had minor response (MR) and the remaining 5 patients had no response (NR). Six of the complete responders remained in CR after 12 to 48 months, whereas all other responders relapsed within 3 months, in spite of maintenance therapy. Twenty-four patients had chronic ITP (i.e. of 6 months duration or more) and had showed no or only transient response to steroids and/or splenectomy, and in many of them, to other therapeutic approaches. Four achieved CR, 4 PR, 6 MR and 10 NR. All but 3 responses were shorter than 3 months, in spite of maintenance therapy. Most responses to slow infusions of vinblastine began after the first infusion. Main side effects included leukopenia in 9 patients (but with absolute neutropenia in only one) and peripheral neuropathy in 2 patients. Interval from diagnosis was the only prognostic factor of response to treatment. We conclude that slow infusions of vinblastine may be a useful approach in ITP of recent onset, when contraindication or refractoriness to steroids and/or IVIg exists. In our experience, this treatment has limited benefit in chronic ITP. In addition, it remains to be demonstrated that slow infusions of vinca alkaloids have any superiority over intravenous bolus injections of the same drugs.

Re-evaluation of the role of azathioprine in the treatment of adult chronic idiopathic thrombocytopenic purpura: a report on 53 cases.

We treated 53 adults (mean age 54 years, range 17-89; 37 females and 16 males) with chronic idiopathic thrombocytopenic purpura (ITP) by azathioprine. All patients had received at least one form of therapy (including splenectomy in 40 patients) and had less than 50 x 10(9)/1 platelets. The duration of ITP before azathioprine was started ranged from 6 to 350 months (median 19). All patients initially received 150 mg/d of azathioprine. This was associated with a short initial course of prednisone (0.3-0.5 mg/kg d) in 10 of them, who were refractory to prednisone alone. 34 (64%) patients responded, including 24 (45%) complete remissions (CR), three (6%) partial remissions (PR) and seven (13%) minor responses (MR). Median time to achieve response was 4 months. 17 of the CR persisted after 7-182 months, 10 of them after discontinuation of azathioprine. Seven patients relapsed after 4-26 months, five of them after azathioprine was stopped or its dose was reduced. PR were short and the median duration of MR was 8 months. Overall, 21 patients (40%) had responses lasting 1 year or more and 17 (32%) lasting 2 years or more. Median duration of treatment was 18 months (range 3-84). Five patients died of bleeding while on treatment. No prognostic factors for response to azathioprine were found. Mild leucopenia was seen in seven patients and a moderate (x3) increase in transaminases in two patients. No opportunistic infections were seen and no malignancy has occurred since the onset of azathioprine. We conclude that azathioprine gives a relatively high incidence of durable responses and very limited side effects in chronic ITP, when splenectomy has failed or is contraindicated. This efficacy, in our experience, is superior to that obtained with other therapeutic approaches. As responses may be delayed, a course of azathioprine of 4 months is required before one can infer a failure to respond. In responding patients, however, the optimum duration of treatment remains to be established.

The role of danazol in the treatment of refractory idiopathic thrombocytopenic purpura. A report of 22 cases.

We treated 22 patients with refractory idiopathic autoimmune thrombocytopenic purpura (ITP) with Danazol (generic name). Median age was 57 (1 child aged 11, and 21 adults aged 18 to 77). Patients were refractory to or had contraindication to therapy by prednisone and splenectomy. Several patients had already received other treatments. All patients received Danazol at a daily dose of 600 mg/day for at least 2 months (except the child who received 400 mg/day), and other treatments for ITP were discontinued when Danazol was started. Two patients achieved complete remissions (CR), one partial remission (PR) and four had minor responses (MR). The remaining 15 patients had no response (NR). One of the complete responses lasted 34 months, but was dependent on daily maintenance doses of Danazol of 400 mg or more. One partial responder remained so after seven months, four months following the discontinuation of Danazol. All other responders relapsed within three months, while receiving Danazol 600 mg/day. The drug was generally well tolerated, with few side effects. Danazol, when used as a single drug in refractory ITP, therefore had limited benefit in our experience.