Variations in MALT1 Gene Disruptions Detected by FISH in 109 MALT Lymphomas Occurring in Different Primary Sites.

Mucosa-associated lymphoid tissue (MALT) lymphoma is a type of B-cell non-Hodgkin lymphoma that can originate in the gastrointestinal (GI) tract, thyroid, breasts, lungs, and skin. The most common genetic abnormality occurring in MALT lymphomas involves t(11;18)(q21;q21) in the gene MALT1. This translocation results in a chimeric fusion product between the genes ATI2 and MALT1, which generates a survival advantage in the lymphoma cells. The MALT1 disruption is more common in some MALT lymphomas, distinguished by site, than others. If identified, this variation in frequency could affect treatment courses and outcomes for each type of MALT lymphoma. The study included 109 MALT lymphoma sample specimens. The sample paraffin-embedded slides were pretreated, hybridized for FISH using a MALT1 break-apart probe, post-washed, and viewed using a fluorescent microscope. On each slide, 100 individual cells were counted by two independent readers, totaling 200 cells per case, and the percentage of cells containing a translocation within each sample was recorded. A conservative threshold of 8% was used to make a positive call. There were a total of 18 positive results in the 109 samples tested. The tissue specimens tested that yielded positive results include the colon (62.5%), lung (57.14%), skin (25%), eyelid/lacrimal gland (16.67%), stomach (6.45%), kidney (50%), and thyroid (100%). The sites that yielded only negative results (0%) include the breast, salivary gland, salivary gland/parotid, soft tissue/skin, conjunctiva/orbital, small intestine, nasal, and epidural mass. As hypothesized, a variation in the MALT1 disruption was found. This is the first study to examine MALT1 disruption in the soft tissue, nasal, and epidural mass. The positive results of this study, specifically the results for the colon and lung, and the negative breast and salivary gland results are consistent with previous studies examining the genetic aberrations in MALT lymphomas. These results indicate that MALT lymphoma at different locations differentially display MALT1 disruption, and these disruptions may be responsible for the variance in patient response to therapy. Surgery, radiation therapy, and/or administration of cladribine (2CdA) result in the best outcomes in treating MALT lymphomas with MALT1 disruption.

[The place of dendritic cells in the microenviroment in Hodgkin's lymphoma].

Dendritic cells (DCs) are the least studied microenvironment elements in Hodgkin's lymphoma (HL). Ninety-five lymph nodes were morphologically and immunohistochemically studied in patients with classical HL. The authors found CDla- and langerin-positive DCs in the presence of T lymphocytes, which were localized in the close proximity to neoplastic Reed-Sternberg cells (RSCs). There were also CD23-positive follicular DCs, which were typically located at the periphery of tumor foci, in the areas of residual follicular structures, were surrounded by B lymphocytes and were rarely associated with RSCs. There were two morphologically distinct groups among the CD 1a- and langerin-positive DCs; one had developed cytoplasmic processes, which formed a network around the RSCs (probably a more mature/functionally active form); the other had round cells with short cytoplasmic processes, loosely distributed around the neoplastic cells (a less mature/active form). The function and role of CD1a- and langerin-positive DCs in HL call for further investigation. The topographic closeness of these DCs to RSCs suggests that these cells play a considerable role in the pathogenesis of HL.

Granulocytic sarcoma involving the uterus and right fallopian tube with negative endometrial biopsy.

Granulocytic sarcoma is an extramedullary tumor associated with acute myelogenous leukemia (AML) and it is rarely seen in the female genital tract. We report an unusual case of granulocytic sarcoma of the uterus and fallopian tube in an AML patient who presented with vaginal bleeding and persistent abdominal pain. She was under chemotherapy. Biopsy did not reveal the diagnosis. After laparoscopic examination, hysterectomy with bilateral salpingo-oophorectomy was performed. Pathology showed atypical myeloid cells infiltrating the muscle bundles which was consistent with granulocytic sarcoma involving the uterus and right fallopian tube. Immunohistochemistry confirmed the diagnosis. The patient is in complete remission for AML and being followed-up for granulocytic sarcoma. Granulocytic sarcoma of the uterus and fallopian tube is very rare, and in AML patients with abnormal uterine bleeding but negative endometrial biopsy it should still be considered in the differential diagnosis.

High-dose chemotherapy and autologous peripheral blood stem cell transplantation for primary breast cancer refractory to neoadjuvant chemotherapy.

The role of high-dose chemotherapy (HDCT) in patients with refractory breast cancer is not well established. Forty-two female patients (median age of 46 years) with breast cancer refractory to neoadjuvant chemotherapy received HDCT (cyclophosphamide, carmustine and thiotepa) supported by an autologous peripheral blood stem cells transplant. Their disease had been refractory (defined as less than partial response) to one (18 patients) or two (24 patients) regimens of neoadjuvant chemotherapy. Twenty-nine patients had surgery before HDCT. The best response after surgery, HDCT, and radiation therapy was assessed 60 days after transplantation. Thirty patients had complete remission, eight had a PR, one had a minor response, and three had progressive disease. In seven of 13 patients whose disease was inoperable before HDCT, it became operable. After a median follow-up of 42 months, 21 patients were alive, and 15 remained disease free. Five-year overall survival (OS) was 57% (CI, 50-64%), and the estimated 5-year progression-free survival was 40% (CI, 32-48%). Both OS and PFS were better in patients whose disease became operable after chemotherapy than in those whose disease remained inoperable. A randomized study is warranted in this patient population.

Cytokeratin immunohistochemistry: a procedure for exclusion of pregnancy in chorionic villi-negative specimen.

The confirmation or exclusion of the presence of products of conception (POC) in biological material may be crucial. This analysis has been undertaken to check how efficient the H&E examination of the putative POC specimen is in practice and to prove the hypothesis that the use of cytokeratin immunostaining can increase the diagnostic accuracy. Of 1078 POC specimens, 94 cases which were signed out as 'No chorionic villi identified' or 'Negative for POC' and in which all submitted material was histologically examined, were re-reviewed histologically and examined immunohistologically using the low molecular weight cytokeratin antibody CAM5.2. In six cases, the originally missed trophoblastic cells were found on re-review of the H&E slides, which was confirmed by CAM5.2 immunostaining. Of seven further cases which were suspicious for trophoblastic cells on re-review, three cases were positive for trophoblastic cells by CAM5.2 and four were negative. Of the remaining 81 cases negative for trophoblastic cells on H&E re-review, CAM5.2 was positive for the trophoblast in 13 (16 per cent). There were no false positive cases on CAM5.2 immunostaining of the endometrial curettings in 22 ectopic pregnancies and 22 cases with dysfunctional endometrial bleeding clinically and secretory endometrium histologically. Therefore, although the overall efficiency of the original diagnosis could have been as high as 98 per cent (1029 of 1051), the false negative rate was 23 per cent (22 of 94) in the chorionic villi-negative material. Even though the morphology of trophoblastic cells on conventional H&E staining is well known, it may be practically difficult to exclude pregnancy by the examination of uterine contents using conventional H&E staining only. The use of the cytokeratin immunostaining can decrease the rate significantly and should be considered in all chorionic villi-negative specimens without obvious extravillous trophoblasts.

Stromal cells prevent apoptosis of AML cells by up-regulation of anti-apoptotic proteins.

The aim of this study was to study interactions between stromal bone marrow microenvironment and leukemic cells. We tested the hypothesis that stromal cells prevent apoptosis of AML cells by up-regulating anti-apoptotic proteins in leukemic blasts. In HL-60 and NB-4 cells, serum deprivation- and ara-C-induced apoptosis was diminished when cells were cocultured with murine MS-5 stromal cells (P < 0.02). This effect was reproduced with conditioned medium from MS-5 cells. Cocultivation with stromal cells induced Bcl-2 expression levels, both by PCR analysis and flow cytometry. In primary AML (n = 14), ara-C-induced apoptosis was significantly lower in cells cocultured with MS-5 cells than in controls (P < 0.001). This effect was partially preserved when leukemic cells were separated from stromal cells by a microporous insert (in 5/9 samples, P = 0.04). In addition, Bcl-2 levels were significantly higher in stroma-supported than in control CD34(+) AML cells (P < 0.01). Bcl-X(L) levels were higher in 5/7 samples grown on stromal layers. Of note, in AML patients resistant to induction chemotherapy (n = 6), Bcl-2 increased significantly after cultivation with stromal cells, but no such increase was noted in cells from chemotherapy-sensitive patients. In conclusion, MS-5 stromal cells prevented apoptosis in HL-60 cells and in primary AML blasts via modulation of Bcl-2 family proteins. The observed association of high Bcl-2 expression in stroma-supported AML blasts in vitro with resistance to chemotherapy in vivo suggests that the same mechanisms may be operational in vivo.

HER-2/neu overexpression as a poor prognostic factor for patients with metastatic breast cancer undergoing high-dose chemotherapy with autologous stem cell transplantation.

PURPOSE: High-dose chemotherapy with autologous stem cell transplantation (HDCT) produces a high tumor response rate for patients with metastatic breast cancer and have 20% long-term progression-free survival. Overexpression of HER-2/neu oncoprotein predicts outcome in patients with breast cancer given standard-dose chemotherapy. Therefore, we evaluated whether the HER-2/neu overexpression in the primary tumor predicts clinical outcome in patients with metastatic breast cancer given HDCT. EXPERIMENTAL DESIGN: A total of 236 patients were given standard-dose induction chemotherapy followed by stem cell collection; high-dose chemotherapy with cyclophosphamide, thiotepa, and carmustine; and stem cell infusion. HER-2/neu expression was assessed by immunostaining with anti-HER-2/neu e2-4001 monoclonal antibody in 63 patients. RESULTS: Clinical characteristics and survival were similar for patients with known and unknown HER-2/neu status. HER-2/neu was overexpressed in 22 of 63 tumors (35%). There was some tendency for HER-2/neu overexpression to be associated with the absence of estrogen or progesterone receptors. In considering the association of HER-2/neu expression with patient outcomes, HER-2/neu overexpression was associated with generally shorter overall survival (P = 0.02) and progression-free survival (P < 0.01), and this association persisted to a lesser extent after adjustment for differences in important prognostic factors between the two groups. CONCLUSION: We conclude that HER-2/neu overexpression may represent an additional prognostic factor for patients with metastatic breast cancer who undergo HDCT.

Cytomegalovirus pneumonia in adult autologous blood and marrow transplant recipients.

CMV pneumonia is a major cause of morbidity and mortality among allogeneic BMT recipients. To assess the frequency, timing, risk factors and response to therapy of CMV pneumonia among autologous BMT recipients, we reviewed our experience with 795 patients. Sixteen (2%) patients were diagnosed with CMV pneumonia. The frequency was higher among patients who were seropositive than those who were seronegative (3.3% vs 0%, P = 0.008). Among seropositive patients, the frequency was higher among patients with hematological malignancies than patients with solid tumors (5.0 % vs 1.0%, P = 0.019). Eleven cases occurred <30 days, and five cases occurred >100 days post transplant. The overall CMV pneumonia-related mortality rate was 31%. Seven (78%) of nine patients treated with ganciclovir and IVIG prior to respiratory failure survived; neither of two patients treated after respiratory failure survived. Four of five (80%) untreated patients survived. In conclusion, CMV is a not infrequent cause of pneumonia among autologous BMT recipients. Risk factors include CMV seropositivity and an underlying hematological malignancy. A favorable response hinges on the prompt initiation of therapy. The survival of 25% of the patients without antiviral therapy suggests that the isolation of CMV from a BAL specimen occasionally reflects oropharyngeal contamination or that CMV pneumonia may sometimes be self-limited in more immunocompetent autologous BMT recipients.

Cytomegalovirus pneumonia in adults with leukemia: an emerging problem.

Cytomegalovirus (CMV) pneumonia is reportedly unusual among adults with leukemia who have not undergone transplantation. To assess the frequency of CMV pneumonia and its outcome during the present time, we reviewed the experience of 2136 hospitalized adults with leukemia. Sixty-one patients (2.9%) had CMV pneumonia diagnosed. The frequency doubled from 1.4% in 1992--1994 to 2.8% in 1995--1997 (P<.05). Fifty-four patients (89%) had received treatment with an immunosuppressive chemotherapeutic regimen that contained fludarabine (n=37), high-dose cytoxan (n=17), or both (n=10), and 15 patients (25%) had received granulocyte transfusions that were stimulated with hematopoietic growth factors from unscreened donors. The overall CMV pneumonia--associated mortality rate was 57%. Among autopsied patients who had leukemia, the frequency of CMV pneumonia increased from 0%, 2.3%, and 0% in 1992, 1993, and 1994, respectively, to 4.6%, 6.5%, and 16% in 1995, 1996, and 1997, respectively (P<.05). CMV has emerged as an important cause of life-threatening pneumonia in adults with leukemia who have received potent immunosuppressive therapies and stimulated granulocyte transfusions from unscreened donors.

[The effect of the murine stromal cell line MS-5 on the proliferation and apoptosis of human erythromyeloid leukemic cell line K562]. / Vliianie myshinoi stromal'noi linii MS-5 na proliferatsiiu i apoptoz éritroleikemicheskoi linii k562.

Effects of stromal environment on human leukemic cell apoptosis and proliferation have been investigated. The MS-5 cell line (kindly provided by Dr.Itoh) capable of supporting human hemopoietic cells and the K562 human erythromyeloid leukemic cell line were used. The total increment of cultured leukemic cells was lower than in control due to their co-cultivation with stromal cells, 96% of cells remaining viable. Colony formation by K562 cells appeared to be significantly lower, too (7.2 vs. 22 colonies per 10(3) cells). Inhibition by stromal cells was reproduced when stromal and leukemic cells were separated by a 0.5% agar layer as well as when leukemic cells were cultured on an agar layer which contained a 50% condition medium. Morphological signs of apoptosis in 78.75 +/- 7.64% of leukemic cells, as a result of serum deprivation, were in evidence after 72 hr. while only 14.33 +/- 3.69% of cells co-cultured with MS-5 died. Only 4% of adherent cells showed morphological signs of programmed death. Since protein bcl-2 was not detected in K562 cells it may be suggested that apoptosis of leukemic cells co-cultured with stromal ones does not use the bcl-2 gene pathway. Stromal cell-mediated inhibition of proliferation may be effected through humoral mechanisms.