J. Demeter

P53 mutations in hairy cell leukemia

We have studied the frequency of p53 mutations in genomic DNA extracted from peripheral blood or the spleen of 61 patients with hairy cell leukemia using PCR-SSCP and automated cycle sequencing. We identified exon 5–8 mutations in 17 cases, corresponding to a frequency of 28%. In four cases, mutations were localized in exon 5; one patient with atypical HCL had a mutation in exon 6 at the 3′ boundary; five cases showed mutations in exon 7, while exon 8 was found to be mutated in seven cases. The mutations found could be divided into three major categories: structural (n = 9), inactivating (n = 6), and neutral (n = 2) mutations. None of the three transitions found occurred at CpG dinucleotides. The rate of p53 mutations found in this large cohort of HCL patients is unexpectedly high as in other non-Hodgkin lymphomas p53 mutations predict for poor treatment outcome. The character of the mutations we have found is entirely different from that described in other hematologic malignancies.

Immunological and molecular biological identification of a true case of T-hairy cell leukaemia.

A hairy cell leukaemia (HCL) patient is presented in whom the peripheral blood mononuclear cells (PBMCs) carried suppressor T‐cell markers (CD3 +, CD2 +, CD8 + /CD4‐, CD38 +). Analysis of genomic DNA of PBMNC showed the presence of a monoclonal population of T cells, the T‐cell receptor (TCR) beta‐chain gene being rearranged on both alleles (DR/DR), while the immunoglobulin (Ig) heavy chain‐genes were in germline configuration. The neoplastic cells were found to react with the monoclonal antibody RAB‐1 — originally described as belonging to the B lineage‐restricted monoclonal antibodies — and to carry RAB‐1/CD‐8 in a double marker assay. Natural killer activity of PBMNCs against K562 target cells was severely reduced, while the cells were found to exert strong antibody‐dependent cellular cytotoxicity.

Detection of Activation Antigens on Chronic Lymphocytic Leukaemia Cells.

The peripheral blood mononuclear cells of patients with chronic lymphocytic leukaemia were characterized by the presence of a variety of cell surface differentiation antigens. The cells of 20 patients were found to be of B-cell phenotype when studied with antibodies directed against CD19, CD20, HLA-DR and sIg. Furthermore, a significant percentage of the cells gave a positive reaction with the monoclonal antibody to CD5. On the other hand, the CLL-cells did not express the CD21 antigen (C3d receptor, EBV receptor). We studied in parallel the presence of various activation antigens using 19 monoclonal antibodies grouped into 7 clusters (CD25, CD30, CD40, CD69, CD70, CD39, CD71). A significantly higher percentage of the CLL cells expressed activation antigens than lymphocytes from healthy controls. The percentage of CD3/HLA + DR + cells, compared to the healthy control lymphocytes was not increased in the CLL patients, and the activated cells in CLL were found to have characteristics of B-cells. Based on these results, we suggest that the CLL cells, like the cells in Hodgkins disease and T-cell lymphoma, are not resting, but activated B-cells or the neoplastic abberrants of activated cells.

Development of chronic lymphocytic leukaemia after posttraumatic splenectomy

SummaryIn the course of a post-splenectomy follow-up study, 2 out of 102 patients who had been splenectomized after an abdominal trauma were found to have developed chronic lymphocytic leukaemia 5 and 31 years after splenectomy, respectively. The possible association between splenectomy and secondary leukaemia is discussed.

Serum dehydroepiandrosterone sulphate (DHEAS) and dehydroepiandrosterone (DHEA) levels in hairy-cell leukaemia.

To the editor: In hairy-cell leukaemia (HCL), a chronic lymphoproliferative disorder of B cells less mature than those of myeloma multiplex (MM), bone involvement is an infrequent problem. Nevertheless, lytic lesions that involve primarily the axial skeleton and the proximal femur have been described in this disorder (1). Humoral factors might contribute to the development of the lytic (2) as well as of the osteosclerotic type of bone lesions (3). To our knowledge there are no data available on the adrenal androgen production of HCL patients, though these steroids are known to profoundly influence bone mineral metabolism as well as immune responses. In this study, serum levels of the adrenal androgens DHEAS and DHEA have been determined in the peripheral blood of HCL patients in different clinical stages of the disease. 26 HCL patients (13 men and 13 women, aged 29 to 89 years: mean: 58 yr) were studied. 7 patients were 20-45 yr old and 19 were above 46 yr of age. 20 patients had been splenectomized prior to the study. All patients had been without treatment for at least 3 weeks at the time of the study. 1 patient had chronic alcoholic liver disease and 1 had moderate azotaemia no other accompanying disease was found. Only 1 of the patients had clinical signs of autoimmune disease (vitiligo) and none of the patients complained of localized bone pain. The functional criteria of Porzsolt were used for the determination of the clinical stage of the disease (4). The numbers of healthy controls were as follows: in females between 20 to 45 yr of age serum DHEAS and DHEA levels were determined in 59 and 54 subjects, respectively in the age group above 46 yr the corresponding numbers were 15 and 32 subjects. In males between 20 to 50 yr of age serum DHEAS and DHEA levels were determined in 49 and 70 subjects, respectively in the age group above 5 1 yr the corresponding figures were 15 and 16. DHEAS was determined in serum by a direct radioimmunobiological method elaborated in our laboratory utilizing 3H-tracer and a polyclonal antibody raised by ourselves in rabbit against DHEA-3haemisuccinate-B SA. The serum DHEA level was measured by our RIA technique without pre-assay extraction and by using 3H-tracer and a monospecific polyclonal antibody obtained in rabbit against DHEA-7-carboxymethyloxime-BSA. The inter-assay variation coefficients for either method were below 13.3 and those for intra-assay variance were below 8.2%, as described previously (5) . DHEAS levels were found to be decreased in 11 cases (as shown in Fig. l), while DHEA levels were decreased in the serum of 16 patients (Fig. 2) as compared with ageand sex-matched control values. No relationship between clinical stage and hormone levels was found. There was no statistically significant difference in the hormone levels of splenectomized and unsplenectomized patients. Serum levels of those 6 patients who have been studied in the leukaemic phase of the disease showed all kinds of variations. The DHEAS and DHEA levels of the patient with chronic alcoholic liver disease, as well as of the patient with azotaemia, were normal as were hormone levels of the only patient with HCL of T-cell origin (6). Our data concerning decreased adrenal androgen production of HCL patients might be of interest both from the biologic as well as from the therapeutic point of view. DHEAS and DHEA levels have been described to be decreased in HIV-infected patients (7) and in women with breast tumours (8). On the other hand, the serum DHEAS levels were found to be decreased in postmenopausal osteoporosis and this finding could partially explain the effectiveness of androgen therapy in this disease (9,lO). The effectiveness of androgen (oxymetholon) therapy in patients with progressive HCL might be of special interest in this respect (11, 12). In conclusion, the adrenal androgen production of patients with HCL was found to be impaired. As DHEA up-regulates host immune response (13), the decreased adrenal androgen production might contribute to the immunological impairment in HCL.

Functional Criteria for Staging and Treatment of Hairy Cell Leukemia

The determination of clinical stage in hairy cell leukemia is hampered by the lack of common criteria. Furthermore, clinical stages in which the disease should be treated have not yet been defined. In this paper we propose common criteria for the clinical staging of all hairy cell leukemia patients (treated or untreated), as well as for the assessment of responses to therapy. These criteria are absence or presence of disease-related symptoms and disease progression. Based on these criteria we propose that hairy cell leukemia patients in the most favorable stage of the functional system, non-symptomatic Stable Disease (nsSD), do not require treatment. Only in patients with disease-related symptoms and/or signs of disease progression is treatment justified and necessary. No symmetry was observed between the functional stages and Jansens stages. For evaluation of responses to treatment, our stages were more sensitive than the Leeds criteria. Experimental evidence enables the differentiation of patients with non-symptomatic stable disease from those with symptomatic and/or progressive disease.