Chronic natural killer lymphoproliferative disorders: characteristics of an international cohort of 70 patients.

BACKGROUND: The 2008 World Health Organization (WHO) classification distinguishes three entities among the large granular lymphocytic leukemia (LGL leukemia): T-cell LGL leukemia (T-LGL leukemia), aggressive natural killer (NK) cell leukemia, and chronic NK lymphoproliferative disorders (LPD), the later considered as a provisional entity. Only a few and small cohorts of chronic NK LPD have been published. PATIENTS AND METHODS: We report here clinicobiological features collected retrospectively from 70 cases of chronic NK LPD, and compared with those of T-LGL leukemia. RESULTS: There were no statistical differences between chronic NK LPD and T-LGL leukemia concerning median age [61 years (range 23-82 years)], organomegaly (26%), associated autoimmune diseases (24%), and associated hematological malignancies (11%). Patients with chronic NK LPD were significantly less symptomatic (49% versus 18%, P < 0.001) and the association with rheumatoid arthritis was more rarely observed (7% versus 17%, P = 0.03). The neutropenia (<0.5 × 10(9)/l) was less severe in chronic NK LPD (33% versus 61%, P < 0.001) without difference in the rate of recurrent infections. STAT3 mutation was detected in 12% of the cohort, which is lower than the frequency observed in T-LGL leukemia. Thirty-seven percent of the patients required specific therapy. Good results were obtained with cyclophosphamide. Overall and complete response rates were, respectively, 69% and 56%. Overall survival was 94% at 5 years. CONCLUSION: This study suggests very high similarities between chronic NK LPD and T-LGL leukemias. Since chronic NK LPD is still a provisional entity, our findings should be helpful when considering further revisions of the WHO classification.

Clinical spectrum of gammadelta+ T cell LGL leukemia: analysis of 20 cases.

We report on the clinico-biological characteristics of 20 cases of gammadelta T cell large granular lymphocyte (LGL) leukemia. All the data were compared to that of 196 cases with alphabeta T cell subtype, which represents the majority of T cell LGL leukemias. Clinical findings were quite similar in the two groups regarding age, sex ratio, recurrent infections, and association with auto-immune diseases especially rheumatoid arthritis. Gammadelta LGL predominantly expressed a CD3+/CD4-/CD8+/CD16+/CD57+ phenotype, in 50% of cases. Clinical outcome was favorable for these patients with overall survival of 85% at 3 years. Fifty percent of gammadelta patients required treatment and the response to therapy was estimated at 55%. gammadelta and alphabeta T cell LGL leukemia harbor a very similar clinico-biological behavior and represent part of an antigen-driven T cell lymphoproliferation.

[Hemophagocytic syndrome and toxoplasmic primo-infection]. / Syndrome d'activation macrophagique et primo-infection toxoplasmique.

Hemophagocytic syndrome (HPS) is a clinical entity that combines the clinical, biological and histological symptoms. The physiopathological mechanism involves the interaction between T lymphocytes/NK cells and macrophages, at the origin of an uncontrolled activation of the macrophages. The consequence is a hemophagocytosis extending to numerous organs, preferentially bone marrow. Clinical symptoms include cytopenia, fever unresponsive to antibiotics and multiple organ dysfunctions. Infections, lymphoproliferative disorders, cancers, systemic diseases are the most prevalent triggers or etiologies of HPS. Because of its high risk of mortality, HPS constitutes a diagnostic and therapeutic urgency. The search for an aetiology, in particular by serological testing, is essential because it conditions the treatment and thus the evolution of the disease. We report here the case of a 12 years-old boy presenting a HPS secondary to a toxoplasmic primo-infection. The objective of this work is to present the step of the biological diagnosis of HPS. Moreover, this observation allows the study of a very rare clinical presentation of toxoplasmic primo-infection, in an immunocompetant patient.

Minimal residual disease detection in Tunisian B-acute lymphoblastic leukemia based on immunoglobulin gene rearrangements.

IGH gene rearrangement and IGK-Kde gene deletion can be used as molecular markers for the assessment of B lineage acute lymphoblastic leukemia (B-ALL). Minimal residual disease detected based on those markers is currently the most reliable prognosis factor in B-ALL. The aim of this study was to use clonal IGH/IGK-Kde gene rearrangements to confirm B-ALL diagnosis and to evaluate the treatment outcome of Tunisian leukemic patients by monitoring the minimal residual disease (MRD) after induction chemotherapy. Seventeen consecutive newly diagnosed B-ALL patients were investigated by multiplex PCR assay and real time quantitative PCR according to BIOMED 2 conditions. The vast majority of clonal VH-JH rearrangements included VH3 gene. For IGK deletion, clonal VK1f/6-Kde recombinations were mainly identified. These rearrangements were quantified to follow-up seven B-ALL after induction using patient-specific ASO. Four patients had an undetectable level of MRD with a sensitivity of up to 10-5. This molecular approach allowed identification of prognosis risk group and adequate therapeutic decision. The IGK-Kde and IGH gene rearrangements might be used for diagnosis and MRD monitoring of B-ALL, introduced for the first time in Tunisian laboratories.

c-myc box II mutations in Burkitt's lymphoma-derived alleles reduce cell-transformation activity and lower response to broad apoptotic stimuli.

In addition to c-myc rearrangement, over 50% of Burkitt's lymphoma cases present clustered mutations in exon 2, where many of the functional activities of c-Myc protein are based. This report describes the functional consequences induced by tumour-derived c-myc mutations located in c-myc box II. Two mutated alleles were studied, focusing on the P138C mutation, and compared to wild-type c-myc. The c-Myc transformation, transactivation and apoptosis activities were explored based on cells over-expressing c-Myc. While the transcriptional activation activity was not affected, our experiments exploring the anchorage-independent growth capacity of c-Myc-transfected Rat1a cells showed that c-Myc box II mutants were less potent than wild-type c-Myc in promoting cell transformation. Considering the possibility that these mutations could be interfering with the ability of c-Myc to promote apoptosis, we tested c-Myc-transfected Rat1a fibroblasts under several conditions: serum deprivation-, staurosporine- and TNFalpha-induced cell death. Interestingly, the mutated alleles were characterized by an overall decrease in ability to mediate apoptosis. Our study indicates that point mutations located in c-Myc box II can decrease the ability of the protein to promote both transformation and apoptosis without modifying its transactivating activity.

Staurosporine-induced apoptosis of HPV positive and negative human cervical cancer cells from different points in the cell cycle.

In the present study, we compare the sensitivity of CaSki and HeLa cells (HPV positive, wild-type p53) and C33A cells (HPV negative, mutated p53) to a protein kinase inhibitor, the staurosporine (ST). We show that ST can reversibly arrest the three cervical-derived cell lines, either in G1 or in G2/M. Beyond certain ST concentrations or/and over 24 h exposure, the cells underwent apoptosis. This process took place in G1 and G2/M for C33A and CaSki plus HeLa cell lines, respectively. By using an in vitro cell-free system, we demonstrated that cytoplasmic extracts from apoptotic cells were sufficient to induce hallmarks of programmed cell death on isolated nuclei. Moreover, we found that only G2/M cytoplasmic extracts from viable CaSki and HeLa cells supplemented with ST, triggered apoptosis while exclusively G1 cytoplasmic fractions from C33A cells were efficient. Our study describes a possible involvement of the HPV infection or/and p53 status in this different ST-induced apoptosis susceptibility.

Relationship between expression of genes involved in cell cycle control and apoptosis in diffuse large B cell lymphoma: a preferential survivin-cyclin B link.

Accumulating evidence suggests that lack of balance between proliferation and apoptosis may lead to clonal expansion and cancer emergence. In diffuse large B cell lymphoma (DLBCL), survivin expression by tumor cells has been recently described as a poor prognostic marker. We assessed the relationship between survivin gene up-regulation and several other factors involved in either cell cycle or apoptosis control. The expression of 34 genes from 27 cases of DLBCL with typical IPI factor-related poor prognostic outcome was analyzed by RNase protection assay. Using non-neoplastic tissues and low grade lymphomas as control, survivin expression was high in 80% of the cases without significant relation to patient overall survival (P = 0.64). However, the expression of several genes encoding for cell cycle inhibitors, cyclins, Bcl-2 or IAP family factors was significantly associated with the survivin up-regulation. Gene expression profiling showed that both survivin and cyclin B expression can define two subgroups of DLBCL: the previously described germinal center-like and activated B-like lymphomas, determined by protein expression analysis. We also identified a preferential survivin-cyclin B relationship (P = 0.017), suggesting that cyclin B over-expression, when linked to survivin over-expression in aggressive forms of lymphoma, might demonstrate a specific G2/M transition promotion.

Effect of granulocyte colony-stimulating factor mobilization on phenotypical and functional properties of immune cells.

Some phenotypic and functional properties of lymphocytes from bone marrow or peripheral blood stem cell donors were compared in a randomized study. Lymphocyte subsets were analyzed by immunocytometry in blood harvested from bone marrow donors (n = 27) and from peripheral blood stem cell donors before and after granulocyte colony-stimulating factor mobilization (n = 23) and in bone marrow and peripheral blood stem cell grafts. Granulocyte colony-stimulating factor mobilization increased the blood T and B, but not NK, lymphocyte counts. All lymphocyte counts were approximately 10-fold higher in peripheral blood stem cell grafts than in bone marrow grafts. Analysis of CD25, CD95, HLA-DR, and CD45RA expression shows that T-cell activation level was lower after granulocyte colony-stimulating factor mobilization. Similarly, granulocyte colony-stimulating factor reduced by twofold to threefold the percentage of interferon-gamma, interleukin-2, and tumor necrosis factor-alpha-secreting cells within the NK, NK-T, and T-cell subsets and severely impaired the potential for interferon-gamma production at the single-cell level. mRNA levels of both type 1 (interferon-gamma, interleukin-2) and type 2 (interleukin-4, interleukin-13) cytokines were approximately 10-fold lower in peripheral blood stem cell grafts than in bone marrow grafts. This reduced potential of cytokine production was not associated with a preferential mobilization of so-called "suppressive" cells (CD3+CD4-CD8-, CD3+CD8+CD56+, or CD3+TCRVA24+CD161+), nor with a modulation of killer cell receptors CD161, NKB1, and CD94 expression by NK, NK-T, or T cells. Our data demonstrate in a randomized setting that quantitative as well as qualitative differences exist between a bone marrow and a peripheral blood stem cell graft, whose ability to produce type 1 and type 2 cytokines is impaired.

Epstein-Barr virus (EBV)-associated smooth-muscle tumor arising in a post-transplant patient treated successfully for two PT-EBV-associated large-cell lymphomas. Case report.

The association of Epstein-Barr virus (EBV) with smooth-muscle tumors was recently reported in the setting of acquired immunodeficiency syndrome (AIDS) and post-transplantation. We report a case of an EBV-associated smooth-muscle tumor arising in a post-transplant (PT) patient who previously was treated successfully for two EBV-associated PT large-cell lymphomas. A 4-year-old girl required cardiac transplantation for dilated cardiomyopathy when she was aged 23 months. Her PT regimen included cyclosporine, azothiaprine, and diltiazem. At 16 months PT, she presented with anemia, guaiac-positive stools, and an abdominal mass diagnosed as diffuse large-cell lymphoma of B-cell phenotype. Immunosuppressive therapy was reduced, and interferon and i.v. immunoglobulin were initiated. She rapidly developed signs of rejection, and a cardiac biopsy was performed, revealing grade IIIB rejection. Subsequently, immunosuppressive therapy increased. At 23 months PT, a biopsy was done of a large pelvic mass that was diagnosed as immunoblastic large-cell lymphoma. After treatment with chemotherapy and retinoic acid, the size of the mass markedly decreased. Follow-up computed tomography scan revealed multiple liver nodules. A needle biopsy of the liver showed a smooth-muscle tumor of indeterminate grade. Both the lymphomas and the smooth-muscle tumor contained EBV within > 95% of tumor cells by Epstein-Barr (EBER1) in situ hybridization, were of strain type A by Epstein-Barr nuclear antigen-2 (EBNA-2) polymerase chain reaction (PCR) and contained an identical 30 base-pair deletion (amino acids 346-355) of the latent membrane protein (LMP)-1 oncogene by PCR analysis. Notably, the initial large-cell lymphoma and the subsequent immunoblastic lymphoma each contained a unique p53 mutation, suggesting that they were distinct. These data suggest that the same virus contributed to the pathogenesis of both the malignant lymphomas and the smooth-muscle tumor.

Hepatitis B virus infection on a tropical island: sociodemographic and geographic risk factors in Guadeloupe.

A seroepidemiologic study on a cross-sectional sample of blood donors was carried out in Guadeloupe, a French West Indies island, to estimate the prevalence of hepatitis B virus (HBV) markers, and to investigate the influence of age, socioeconomic, and geographic factors on prior HBV infection. Blood specimens and sociodemographic data were collected in 1989 from 2,339 blood donors residing on the island. A total of 73 (3.1%) of 2,339 were found to be HBV surface antigen carriers, and 518 (22.1%) were positive for antibody to HBV core antigen. Among them, 61 were positive for both markers and consequently 530 persons (22.7%) were considered to have evidence of prior HBV infection. Multivariate logistic regression analyses identified age and low socioprofessional status as being related to HBV infection, as in many endemic areas. In addition, some major geographic risk factors were highlighted, reflecting a strongly hyperendemic situation in specific areas and the potential influence of horizontal transmission or unknown environmental factors on these particular populations.

Comparative preclinical study of three bone marrow purging methods using PCR evaluation of residual t(14;18) lymphoma cells.

The t(14;18) chromosomal translocation occurring in most follicular lymphomas can be exploited by a Bcl2/JH polymerase chain reaction (PCR) to detect residual disease and to monitor the effectiveness of ex-vivo tumor cell immunological purging. We first demonstrated the 10(-5) Bcl2/JH PCR sensitivity with serial dilutions of OCY-LY8 lymphoma cell lines in normal mononuclear cells; and then the specificity and reproductibility of this technique by analysing follicular and non follicular lymphoma samples. With the Bcl2/JH PCR, we tested the efficiency of three marrow purging protocols with an experimentally contaminated bone marrow either treated by three anti-B cell monoclonal antibodies (mAb) followed by three rounds of rabbit complement or two rounds of immunomagnetics beads. Samples obtained after each purging were amplified by Bcl2/JH PCR and hybridized with PFL3 probe. We were able to produce a 2 to 3 log tumor cell reduction after three rounds of complement and a 4 to 5 log reduction after two rounds of beads. This study showed that it is feasible to use the Bcl2/JH PCR technique for residual cell lymphoma detection in patients undergoing intensive chemotherapy or BM transplantation. These results indicate that ex-vivo immunomagnetic BM purging is probably superior to complement mediated lysis for the eradication of B lymphoma cells from the marrow of patients undergoing autologous transplantation.

[Prevention of post-transfusion hepatitis in Guadeloupe: results after application of new measures for detecting blood donors potentially at risk]. / Prévention des hépatites post-transfusionnelles en Guadeloupe: résultats obtenus après application des nouvelles mesures visant à détecter les donneurs de sang potentiellement à risque.

One of the main concerns of blood transfusion centers is viral hepatitis as a direct result of blood transfusion. Ninety-five percent of these cases are non-A, non-B hepatitis. In order to prevent this disease, blood collections were screened for antibody anti-HBc as well as the level of activity of the alanine aminotransferase in 3,051 blood donors in Guadeloupe. this revealed a particular epidemiological situation, which caused this French country to be rated among moderate endemic zones for hepatitis B virus. As a result of this new screening procedure, 25 percent of the blood collected had to be discarded and was classified with prevalence rates of 21.8 percent HBc antibody, 2.9 percent HBs antigen, and 2.6 percent alanine aminotransferase (45 IU/l). Differences were noted according to sex, age, social-economical level and geographical origin of the blood donors. These data raised many significant questions regarding the vertical transmission of hepatitis B virus, the epidemiological situation of hepatitis B virus in the Guadeloupe population as well as in the rest of the French West-Indies, and also the type of action which must be taken against non-A, non-B hepatitis in a moderate endemic zone for HBV.

A unique seroepidemiologic situation for hepatitis B markers in Guadeloupe. Results of a prospective study in blood donors.

Screening for HBsAg, anti-HBc, anti-HCV and ALAT levels is now performed on donated blood to prevent post-transfusion hepatitis. A prospective study of 2,368 blood donors was performed in Guadeloupe (French West Indies) to determine risk factors associated with serologic abnormalities: 571 donations (24%) were positive for at least 1 of the 4 analyzed markers with 3.2% positive for HBsAg, 22% for anti-HBc, 0.8% for anti-HCV and 1.4% with ALAT > or = 45 IU/L. The anti-HCV prevalence was significantly different according to ALAT levels (P < 10(-4)). Transfusion history and work status (worker or serviceman) were found to be risk factors, with an odds ratio of 1.94 for serviceman population. Other unexpected risk factors were: number of years residency in Guadeloupe (progressively increased risk with the number of years), birthplace and residence in southern part of the island as well as the existence of gastrointestinal discomfort unrelated to viral hepatitis (odds ratio = 2.91). The results of this study show a unique epidemiologic situation for hepatitis B virus in Guadeloupe necessitating careful selection of blood donors.

[Angioimmunoblastic lymphadenopathy: a pathogenetic intersection between dysimmune, viral and lymphomatous diseases]. / La lymphadénopathie angio-immunoblastique: carrefour pathogénique entre maladies dysimmunitaires, virales et lymphomateuses.

Angioimmunoblastic lymphadenopathy (AIL) still is a clinico-pathological syndrome with little known physiopathology. The advent of molecular biology has improved our understanding of this syndrome by characterization of the clonal cell. With this technique, combined with cytogenetics and immunohistochemistry, three pathological states have been individualized: 1) true AIL without evidence of monoclonal proliferation; 2) transformed AIL, and 3) AIL-like T-cell lymphoma. This clinical complex can be integrated in an evolutive continuum, starting with simple lymphoid hyperplasia and ending with frank malignant T-cell lymphoma.

[Stauffer's syndrome disclosing kidney cancer: another cause of inflammatory syndrome with anicteric cholestasis]. / Syndrome de Stauffer révélateur d'un cancer du rein: une autre cause de syndrome inflammatoire avec cholestase anicterique.

Stauffer's syndrome is characterized by a cholestasis without biliary obstruction or hepatic metastasis and a renal tumor. We report a case of Stauffer's syndrome in a 73 year-old woman. Cholestasis and inflammatory syndrome regressed with corticosteroid. Giant cell arteritis with negative temporal artery biopsy was wrongly suspected.

[Osteochonroplastic tracheopathy associated with dermatomyositis: apropos of a case]. / Trachéopathie ostéochondroplastique associée à une dermatomyosite: à propos d'un cas.

We report a case of a 35 year-old-man with dermatomyositis associated with tracheopathia osteoplastica. The swallowing perturbation secondary to myositis and airway involvement by tracheopathia induced fatal outcome. Tracheopathia osteoplastica is a rare disease and occurs exclusively in men over the age of 50. The association of two rarest disease is not a fortuitous event. The common pathogenic factor may be, in this case, the occupational exposure to silicon.

[Systemic amyloidosis in the elderly: diagnostic value of the test of subcutaneous abdominal fat and the labial salivary glands. Prospective study in 100 aged patients]. / L'amylose systémique du sujet âgé: valeur diagnostique de l'examen de la graisse sous-cutanée abdominale et des glandes salivaires accessoires. Etude prospective chez 100 patients âgés.

Primary and secondary amyloidosis are not uncommon in aging but the diagnosis is rarely made on account of the risk of bleeding in the site of biopsies and the difficulty to distinguish senile from systemic amyloidosis on the biopsy samples. We have studied the frequency of amyloid deposition in the abdominal fat aspirate (AFA), the labial salivary gland (LSG), the temporal arteries (four cases), bone marrow (two cases), digestive tract (four cases) in 100 elderly patients (aged 80 or greater). AFA was positive in 15 percent of the patients and LSG in 5%; both samples were positive in 4%. Four cases of systemic amyloidosis were found (two of the AL and two of the AA type). Sensitivity of AFA was 75%, specificity was 87% and the positive predictive value was 20%. The values were respectively 100%, 99%, 100% for LSG. In 11 patients whose AFA biopsies samples were singly positive, amyloid deposits were found in temporal arteries in four of four cases. We conclude that AFA is too sensitive for the diagnosis of systemic amyloidosis in aging. The responsibility of senile amyloid deposition on AFA should require further investigations. LSG biopsies seem to be a more reliable test for the diagnosis of primary and secondary amyloidosis in elderly.

[Mesenteric lymph node cavitations in Whipple's disease. Apropos of a case]. / Cavitations ganglionnaires mésentériques dans la maladie de Whipple. A propos d'un cas.

We report a computed tomographic and echographic description of lymphadenopathy during a Whipple's disease. The computed tomography find low density of involved lymph nodes because of the high fatty charge, ultrasound revealed diffusely echogenic aspect. This cavitation was highly suggestive of Whipple's disease. Moreover, the computed tomography allows the follow-up of the disease.

[Pleuropericarditis and Pyoderma gangrenosum during remission of a hemorrhagic rectocolitis]. / Pleuro-péricardite et Pyoderma gangrenosum au décours d'une rectocolite hémorragique en rémission.

We report the case of a 17 year-old woman who had a pleuro-pericarditis after remission of an ulcerative colitis. The patient had a rapid and complete recovery with steroids. This extra-intestinal complication can develop during all the stages of the disease and its diagnosis can be very difficult.

[Vascular and/or cardiac manifestations of type IV Ehlers-Danlos syndrome. 9 cases]. / Manifestations vasculaires et/ou cardiaques du syndrome d'Ehlers-Danlos de type IV. 9 observations.

Type IV Ehlers-Danlos syndrome, a rare disease caused by abnormal synthesis of type III collagen, often leads to vascular fragility. We report 9 cases (6 men and 3 women, mean age 35 years). For 7 of the patients, the inaugural signs were arterial complications including haemoperitoneum in 2 patients with multiple aneurysmal dystrophy of the abdominal arteries, one case of ruptured subclavian artery, two dissections of the renal artery, one case with rupture of a cerebral aneurysm, one rupture of the mesenteric artery and a haematoma after arterial puncture. Other vascular manifestations were acrosyndrome (n = 4), varicose veins (n = 3), and prolapsus of the mitral valve (n = 2). In addition, 8 of the 9 patients presented extravascular signs. There was a history of familial disease in 5 cases. Pregnancy was completed to term in three patients: a cesarean section was required in one case and intra-uterine growth retardation was seen in 2. Morbidity was important with hemiparesia, blindness and paraparesis sequellae. One patient died from haemorrhage. This series of patients with type IV Ehlers-Danlos syndrome illustrates the severity of this disease whose prevalence is often underestimated. The disease is transmitted by autosomal dominant inheritance, underscoring the importance of familial testing for early diagnosis. Clinicians should be aware of the vascular manifestations and avoid invasive punctures or operations except in exceptional indications.