bluuebunny/crossref_metadata_2025 huggingface dataset:
Hypersensitivity Reaction of Gemtuzumab Ozogamicin (Mylotarg™) Associated with Platelet Transfusion: Two Case Reports.
Authors: Hanbali Amr , Wollner Ira , Neme Klodiana , Ulreich Cynthia
Published: April 2020

Introduction

Gemtuzumab ozogamicin (Mylotarg™) is a monoclonal antibody developed for the treatment of myeloid leukemias. Mylotarg is composed of a recombinant humanized IgG4 kappa antibody conjugated with a cytotoxic antitumor antibiotic, calicheamicin. It binds specifically to the CD33 antigen, which is expressed on the surface of leukemic myeloblasts and immature normal cells of myelomonocytic lineage. It is approved for the treatment of CD33 positive AML in first relapse in patients >= 60 years of age and are not considered candidates for cytotoxic chemotherapy. Hypersensitivity reactions, including anaphylactoid reactions, anaphylactic shock, dyspnea, and severe hypotension, have been reported during clinical use of gemtuzumab ozogamicin. Infrequently, hypersensitivity reactions have been fatal. It is unknown if the risk of hypersensitivity reaction increases with administration of blood products in the same day of Mylotarg infusion. We are reporting 2 cases of severe hypersensitivity reaction to Mylotarg on 2 patients who received platelete transfusion in the same day.

Case reports:

Case number 1: A 75 year old male diagosed with AML with normal cytogenetics in April 2005. Relapse was noted in March 2006. He was admitted for Mylotarg infusion at a dose of 9mg/m2 which was started at 11:00 a.m. after receiving the appropriate premedications and finished at 1:00 p.m. without any signs of distress or reaction. He received 6 units of platelets at 6:40 p.m. At 1:00 a.m. the next morning he went into severe respiratory distress and died shortly after that in the ICU.

Case number 2: A 68 y/o male with the history of NHL in 1991, received CHOP chemotherapy in 1991 and 1992, now has secondary AML (diagnosed in October 2005 and underwent chemo in November 2005, and relapsed after induction and consolidation. He was admitted for Mylotarg infusion. Initially he received 6 units of platelets at 8:30a.m. Mylotarg was started at 10:15a.m. and finished at 12:10p.m.No signs of distress or reaction post infusion. At 2:30 p.m. the patient started having chills, rigors with fever, tachycardia, hypertension and hypoxia (pO2 86%). The patient was treated with Solu-medrol, Benadryl, Tylenol and Demerol. The reaction ceased with no further adverse events

Conclusion:Mylotarg is known to cause hypersensitivity reaction, but it is unknown if the risk of this reaction increases with blood products transfusion around the time of Mylotarg transfusion. We are reporting these 2 cases to draw attention to this serious observation.
Treatment with Homoharringtonine and Cytarabine in Patients with High Risk Acute Myelogenous Leukemia (AML).
Authors: Chang Naibai , Wei Jianping , Zhao Shengming , Liu Hui , Fan Yun , Li Jiangtao , Xu Xiaodong , Gu Xichun
Published: April 2020

Objective: To evaluate response of homoharringtonine in patients with high risk AML or as a second-line therapy in patients with AML refractory to anthracycline based chemotherapy.

Patients and methods: From Jan 1998-Jan 2006, there were 66 patients enrolled in this regimen. Male:female=40:26. Median age was 47 (17–69). There were 48 newly diagnosed AML, 5 relapsed AML and 13 secondary AML (secondary to MDS) in this group. Among these patients, there were 40 patients untreated previously — 20 patients had unfavourable chromasomal abmormalities, 10 patients had pgp positive, 10 patients had normal chromasome but high CD34 expression on leukemic cells. The remaining 26 patients were previously treated with daunorubincin or mitoxantrane combined with cytarabine at least two cycles and failed to achieve response. Homoharringtonine was given at dose of 4mg/m2/d(2.5mg/m2/d for age≥60 years) intraveniously for 7 days. Cytarabine was given at dose of 100mg/m2/d at same time. The therapy was repeated every 21 days.Post remission therapy was divided into two cohot —same regimen maintainance in 20 cases and cytarabine 1g/m2/d q12h for 4 days at least 4 cycles in 20 cases.

Results: 40/66 patients achieved complete remission. 3/66 achieved partial remission. In patients refactory to anthracycline based regimen, the CR rate was 57.7%(15/26). In previously untreated high risk AML patients,the CR rate was 62.5%(25/40). Median disease free survival were 4.25 months (2–30) in cohot 1 and 18 months (12–47) in cohot 2 (P<0.05). CD95(APO-1/Fas) was tested by flow cytometry during treatment. APO-1/Fas (CD95) increased from (9.56±5.58)% to (25.64±0.70)% after induction chemotherapy. Main toxicities were marrow suppression and infection. There were 7 early deaths, 5 patients died of cerebral hemorrage and 2 cardiovascular events.

Conclusions: Homoharringtonine is effective in patients with high risk AML. It is also a choice of second line therapy in patients refractory to anthracycline based chemotherapy. Intensive post remission therapy is superior to conventional dose maintainance therapy in high risk AML. One of the ways for homoharringtonine to induce leukemic cell apoptosis is probably through Fas pathway.
Salvage Therapy with Flag/Idarubicin/Mylotarg (Flag-IM) Results in a Superior CR/CRp Rate and Low Toxicity When Compared to Mitoxantrone/Etoposide/Cytarabine (MEC) in Patients with Relapsed and Refractory AML.
Authors: Van Tine Brian A. , Augustin Kristan M. , Kuperman David I. , Nervi Bruno , Westervelt Peter , DiPersio John F.
Published: April 2020

Salvage therapy for relapsed AML has been historically unsatisfactory. Response to salvage therapy for AML was investigated using a retrospective chart review of patients at the Siteman Cancer Center who failed standard induction therapy. We identified and studied 42 patients with relapsed (N=24) or refractory (N=18) AML who received either (33/42 patients) Mitoxantrone 8mg/m2, Etoposide 100 mg/m2, Cytarabine 1000 mg/m2 (days 1–5) (MEC) or (9/42 patients) Mitoxantrone 10 mg/m2, Etoposide 100 mg/m2 (days 1–5) (MV) as salvage therapy. 40/42 patients were evaluable for response by formal restaging after day +30. 13/40 entered a CR and 7/40 entered a CRp. Only 7/20 (35%) CR/CRp remained progression free with a median follow up of 5.3 months (range = 2–10 months). 6/7 who remained in CR/CRp received an allogeneic stem cell transplant and/or donor lymphocyte infusion after treatment. Median follow up for transplanted patients is 38 months with a range of 10–80 months. A similar analysis was performed on 16 patients with relapsed (N= 12) or refractory (N= 4) AML that received Fludarabine 25mg/m2, Cytarabine 2000 mg/m2, Idarubicin 12mg/m2 (days 1–5), G-CSF 5mcg/kg (day 1+), and Gemtuzumab 9 mg/m2 (day 8) (Flag-IM) for CD33+ AML. Only 15/16 patients were evaluable for response by formal restaging after day +30. Flag-IM treatment resulted in 12/15 patients achieving CR/CRp (6 CR, 6CRp). This group included 3 patients that had previously failed MEC or MV salvage therapy. 10/12 CR/CRp patients received allogeneic transplants (4 sib, 6 unrelated) and 5 (50%) are alive and disease-free with a median survival of 13.4 months (range2–27) months. In spite of a short median duration of only two months (range 1–4) from Flag-IM to allogeneic transplant, no veno-occulsive disease (0/10) was observed, and only 4/10 patients developed mild hyperbilirubinemia (range 1.3–3.8) after myeloablative conditioning for allogeneic transplant.

In conclusion, in this limited retrospective analysis, Flag-IM results in a superior CR/CRp rates when compared to MEC/MV (80% versus 50%) (p = 0.045 by 2 sided t-test) in patients with relapsed/refractory AML. Thirty day treatment related mortality after MEC/MV (2/42) and Flag-IM (1/16) were similarly low, as was VOD in patients receiving myeloablative allotransplant after Flag-IM (0/10). DFS after allogeneic transplant in those patients achieving CR/CRp after MEC/MV (6/20) and Flag-IM (4/10) (p=0.34) were similar. No patient survived greater then 10 months without allogeneic transplant. Flag-IM is an effective salvage therapy associated with acceptable toxicity in patients with relapsed refractory AML.
Differential Effects of Bexarotene on Intrinsic and Extrinsic Pathways in TRAIL-Induced Apoptosis in Myeloid Leukemia Cell Lines.
Authors: Ying Shao Xu , Li Xiao , Seal Sudeshna Seal , Abbassi Nissa , Pagel John M. , Gopal Ajay K. , Deeg H. Joachim
Published: April 2020

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an attractive candidate for cancer therapy since it induces apoptosis preferentially in tumor cells. However, not all human cancer cells are sensitive to TRAIL. Here, we show that a synthetic RXR-selective retinoid, bexarotene, overcomes TRAIL-resistance in KG1a leukemia cells (P<0.001) by down-regulating FLIPLong and activating caspase-8, thereby triggering the extrinsic apoptotic signaling pathways. Over-expression of FLIPLong all but abolished apoptosis induced by TRAIL plus bexarotene. Bexarotene and TRAIL also showed an enhanced apoptosis-inducing effect in ML1 leukemia cells (P<0.01), in this case, however, involving the intrinsic pathway with downregulation of Bcl-xL and cleavage of caspase-9. Blockade of caspase-8 almost completely prevented enhancement of TRAIL-induced apoptosis by bexarotene, while the RXR-specific antagonist HX531 failed to block apoptosis induced by TRAIL plus bexarotene in both KG1a and ML1 cells. These data indicate that the positive interactions between bexarotene and TRAIL involved the death ligand-dependent extrinsic pathway and were able to overcome TRAIL resistance. We conclude that bexarotene overcame resistance or enhanced the effects of TRAIL in both type I (KG1a) and type II (ML1) leukemic cell lines, apparently via downregulation of multiple antiapoptotic molecules, including FLIPLong, and Bcl-xL.
MMRI-23: A Potential Role in AML Antibody-Mediated Therapy.
Authors: Modra Courtney J. , Jamriska Lubomira , Rao Min , Clark Georgina J. , Hart Derek N.J.
Published: April 2020

Novel targets expressed on early AML precursor cell populations, with restricted expression on normal tissues, provide attractive options for antibody based therapeutics. We identified the CD300 family as Ig superfamily members with the capacity, through signaling motifs, to induce triggering and inhibitory signals to the leukocytes on which they are expressed. Here we report on an inhibitory member of this family, 35-L5, as a potential target for antibody-mediated therapy in AML. To assess its potential, we analyzed 35-L5 transcript and cell surface expression on normal blood cells, cell lines, and AML cells using quantitative RT-PCR and a monoclonal antibody (MMRI-23) made against a recombinant 35-L5 Ig fusion protein. RT-PCR showed its expression in normal PBMC was restricted to CD14+ monocytes, CD11c+ dendritic cells (myeloid DCs), monocyte derived dendritic cells (MoDC), with limited expression by B cells. MMRI-23 bound CD14+ monocytes, myeloid DC and MoDC, but not CD19+ B, CD4+ T, CD56+ NK cells and CD11c− (plasmacytoid) DC. Interestingly, MMRI-23 binding varied between donors and 35-L5 expression was upregulated on mature MoDC, when compared to immature MoDC from the same donor (3 from 6 donors). However, MMRI-23 binding to PBMC remained stable when cultured with activation agents PMA and ionomycin, LPS, or PHA for 24 and 48 hours. 35-L5 transcript and surface protein expression was evident in the monocytic derived U937, leukemic derived HL60, erythroleukemic derived HEL, CML derived K562, and B lymphoblast derived Daudi cell lines, but was limited in the KMH2 and HDLM2 cell lines and absent in the Jurkat, Raji, L428 and Mann cell lines. Preliminary studies in AML using quantitative RT-PCR showed that 35-L5 transcript was increased compared to control normal CD14 cell expression (n=9). Furthermore, MMRI-23 bound to 6 of 8 AML samples of the FAB M1, M2 and M5 subtypes. MMRI-23 crosslinking induced neither cell apoptosis, nor cell differentiation and change of CD14, CD33, CD64, CD16, CD11b expression. Defining the role of 35-L5 on normal and AML progenitor cells is necessary to evaluate its potential as a possible target for antibody-mediated therapy. MMRI-23 may have potential for drug conjugated antibody-mediated therapy, but 35-L5’s inhibitory function may also be exploited to develop a novel mechanism for controlling myeloid leukemias.
“Fulminant” Acute Promyelocytic Leukemia: Clinical and Biological Features.
Authors: Di Bona Eros , Ferrara Felicetto , Specchia Giorgina , Leoni Franco , Melillo Lorella , Falanga Anna , Rossi Giuseppe , Fazi Paola , Coco Francesco Lo , Mandelli Franco
Published: April 2020

Acute promyelocytic leukemia (APL) is associated with severe hemorrhagic coagulopathy, which is the main cause of early death. The introduction of ATRA ameliorated the complete remission rate and improved the overall survival compared to treatment with chemotherapy alone. However, the pioneristic large randomized trials comparing ATRA alone versus chemotherapy alone in previously untreated patients failed to demonstrate a significant reduction of early hemorrhagic deaths, notwithstanding ATRA capability to quench the ongoing coagulopathy. Nevertheless, the incidence of such very early deaths remains unsettled, leading to a bias in the evaluation of real impact of ATRA in clinical outcome of APL. We have retrospectively collected and examined a total of 31 very early deaths from APL from the database of 7 Italian Centres. Very early death was defined as death occurring within 72 hours from the first clinical observation. The incidence was 9.7% (31 of 318 APL enrolled in the same period in GIMEMA APL trials). Median age was 53 years (21–88), M/F ratio 0.9, 13 were variant forms of APL, median peripheral blood cell count was: WBC 41.2 ×109/L (0.68–181), with 94% of blast cells, platelets 24 ×109/L (6–96); BCR type 1, 2, 3 were observed in 10, 2, and 7 cases, respectively; clinical features of disseminated intravascular coagulopathy (DIC) were present in all but two patients, while median WHO grade of hemorrhage was 3. Median time from symptoms onset to diagnosis and from diagnosis to death was respectively 72 and 44 hours. One third of the patient did not receive any therapy, the others received some doses of ATRA with or without chemotherapy. The majority of these patients was not enrolled in any clinical trial. These preliminary data suggest that a relevant percentage of APL patients died before starting any effective treatment. Every clinical trial should include this slice of patients to give realistic clinical outcome of APL therapies. Seventy-two percent of patients showed WBC≥10×109/L and variant form was more frequent (42%) than expected (usually 20–25% of APL). Based on the possible confused morphological features with monocytic leukaemia, diagnosis required rapidly confirmative molecular methods and heavy supportive and transfusional therapies. Moreover, underlying APL should be considered as a differential diagnosis when apparently spontaneous cerebral hemorrhage occurs; in some instance, a prompt supportive and differentiating therapy could be lifesaving.
Reed-Sternberg Cells Have Shortened Telomere Length.
Authors: Ben-Ezra Jonathan M. , Jackson-Cook Colleen
Published: April 2020

As cells age, the ends of their chromosomes, the telomeres, become shortened. The chromosomal length eventually shortens to a critical point, which is thought to lead to chromosomal instability and/or cell death. Neoplastic cells have increased amounts of telomerase, an enzyme that prevents this shortening; this may be one mechanism by which tumor cells grow. Due to the scarcity of neoplastic Reed-Sternberg cells in tissues of Hodgkin Lymphoma, studies of telomere length and telomerase expression have been difficult and few. These few studies have tended to show an increase in telomerase expression in tissues from Hodgkin Lymphoma. In order to determine telomere length in Reed-Sternberg cells, we performed an immunofluorescence/FISH assay using a PNA pantelomeric probe and a CD30 antibody with which to identify the Reed-Sternberg cells (Am J Pathol 160:1259–68, 2002). Six cases were studied, with patient ages ranging from 18– 51 years. Telomere length was assessed on a semiquantitative scale. Of the six cases, five had evaluable hybridization signal. In these five cases, 53 CD30-positive cells (between 3–20 per case) were studied. In forty cells, the hybridization signal (telomere length) was less than that of surrounding cells, and in 13 it was the same; in no case was telomere length greater in the Reed-Sternberg cell than in surrounding lymphocytes (p< 0.001, Wilcoxon signed ranks test). There was no correlation between the age of the patient or the age of the sample and telomere probe signal intensity/quantity. Similar to epithelial and soft tissue tumors, it appears that the neoplastic cells in Hodgkin Lymphoma have shortened telomere length as compared to the surrounding normal cells.
NK/T Cell Versus Peripheral T Cell Lymphoma: Significant Differences in Clinical Characteristics and Survival.
Authors: Lim Soon-Thye , Gao Fei , Lim Lay-Cheng , Quek Richard , Lim Daryl , Yap Swee-Peng , Loong Susan , Sng Ivy , Tao Miriam
Published: April 2020

Background: To compare the clinico-pathologic characteristics and prognosis of Natural Killer/T cell lymphoma (NK/TL) with peripheral T cell lymphoma (PTCL).

Methods: A total of 556 resident patients (pts) with lymphoma were treated in the departments of medical oncology and hematology in an Asian institution from 2000 to 2005. Of these pts, 71 (12.8%) had NK/TL or PTCL and were included in this analysis. Pathology was centrally reviewed and classified according to the WHO classification.

Results: NK/TL and PTCL comprised of 4.7% (26/556) and 7.9% (45/556) of all cases. Of the PTCL cases, histology was PTCL-NOS in 21, anaplastic large cell in 12 (5 were ALK-1 positive) and angioimmunoblastic T cell in 8 pts. Subcutaneous panniculitis T cell and γ/δ T cell lymphoma accounted for one case each. There were no significant differences between the two groups of pts in terms of sex, performance status, extranodal involvement and LDH level at presentation. However, more patients with NK/TL presented with stage I/II disease (65% vs. 31%, p=0.003). Among pts with NK/TL, 17 (65%) received CHOP-based chemotherapy, 4 received radiation alone and 5 received palliative chemotherapy. In the PTCL group, 39 (87%) received CHOP-based chemotherapy, 2 received radiation alone and 3 received palliative treatment only. Compared to PTCL, NK/TL was associated with a significantly inferior rate of complete remission (27% vs. 58%, p=0.01) and inferior overall survival (5 vs. 28.4 mos, p=0.001). Although age > 60, ECOG ≥ 2, elevated LDH, advanced stage, IPI ≥ 2 and NK/T cell histology were each associated with decreased survival on univariate analysis, only NK/T cell histology and advanced stage were independently associated with decreased survival (see table 1).

Conclusions:

Contrary to expectation, the incidence of PTCL based on WHO classification in this Asian series is not higher than that reported in Western series. Compared to PTCL, the NK/T subtype is associated with a paricularly inferior prognosis and overrides the prognostic significance of IPI. These data suggest that NK/TL should be considered as a seperate entity and should not be considered together with other subtypes of T cell lymphoma in clinical trials.

Table 1. NK/TL vs. PTCL: Univariate and Multivariate Analyses Univariate Analysis Multivariate Analysis Median (yr) P Hazard Ratio 95% CI P Male vs. Female 1.03 vs. Not reached 0.06 0.62 0.28 to 1.40 0.25 Age<60 vs. ≥ 60 2.37 vs. 0.51 0.01 1.41 0.70 to 2.83 0.33 ECOG 0/1 vs. ≥ 2 1.99 vs. 0.36 0.002 1.52 0.63 to 3.65 0.354 LDH normal vs. High Not reached vs. 0.75 0.03 1.29 0.53 to 3.13 0.57 Stage I/II vs. III/IV 1.99 vs. 1.41 0.16 2.91 1.17 to 7.2 0.02 IPI 0/1 vs. ≥ 2 Not Reached vs. 0.42 0.002 2.22 0.82 to 5.99 0.12 PTCL vs. NK/TL 2.37 vs. 0.42 0.001 5.8 2.36 to 14.24 <0.001
JNK Pathway Is Constitutively Activated and Is Required for Survival in Human Mantle-Cell Lymphoma Fresh Cells and Cell-Lines.
Authors: Camara-Clayette Valerie , Vainchenker William , Koscielny Serge , Hermine Olivier , Ribrag Vincent
Published: April 2020

We previously reported that GST-pi, a cellular JNK effector and activator, was strongly expressed in MCL patient’s samples (Clin Cancer Res. 2004). Recently, several primary murine and human B lymphomas were found to constitutively express high levels of activated form of c-jun N-terminal kinase (JNK), a member of the MAP kinase family and that JNK was required for survival in these lymphoid cellular models. To our knowledge, JNK activity has not been assessed in Mantle cell lymphoma cell lines as well as MCL patient’s samples. In vitro kinase assays showed that JNK was constitutively activated in 2 EBV negative cell-lines (JEKO and UPN1) and in 5 MCL patient’s lymphoma cells. JNK and one of its major substrate ATF-2 were found to be constitutively phosphorylated in the same cell-lines and patients MCL samples. JNK inhibition by SP600125, an anthrapyrazolone pharmacological JNK inhibitor impaired cell viability (IC 50: 20 microMol), indicating that JNK exerts a major role in antiapoptotic signals in MCL JEKO and UPN1 cell-lines and fresh human MCL cells. In vitro combination showed that SP600125 and Bortezomib or cytarabine had additive effects in MCL cell-lines and patients fresh lymphoma cells. We also investigated SP600125 activity in a nude model transplanted with UPN1. SP600125 was given IP every 2 days at 40 mg/kg during 10 days, 7 days after UPN1 subcutaneous transplantation. SP600125 therapy significantly delay tumour growth in 6 mice compared to 6 mice who received vehicle alone (DMSO); p=0,001. Thus, JNK pathway activation plays a major role in MCL survival and can be an important target for future therapies.
Up-Regulation of Death Receptor 5 and Bax Translocation Is Necessary To Induce Apoptosis by Sanguinarine in Primary Effusion Lymphoma.
Authors: Hussain Azhar R. , Al-Jomah Naif A. , Siraj Abdul K. , Pulicat Manugaran S. , Al-Hussein Khaled A. , Al-Kuraya Khawla S. , Uddin Shahab
Published: April 2020

Primary effusion lymphoma (PEL) is an aggressive and fatal type of cancer. PEL cells produce a variety of autocrine cytokines and growth factors, which provides cyto-protection against conventional chemotherapeutic agents. In efforts to identify novel approaches to block the proliferation of PEL cells, we found that Sanguinarine, a natural compound isolated from the root plant Sanguinaria canadendid, that is being used as an anti-microbial agent, inhibited cell proliferation and induced apoptosis in a dose dependent manner in several PEL cell lines through a bax-dependent signaling pathway. Five PEL cell lines used in this study were treated with various doses of Sanguinarine ranging between 0.5–4μM inhibited cell proliferation in all the cell lines in a dose dependent manner (BC1 40–97%, BC3 46–93%, BCBL1 11–94%, BCP1 20–97% and HBL6 7–95%). Treatment with varying doses of Sanguinarine also induced apoptosis in all cell lines as determined by cell cycle analysis, annexinV/PI dual staining, TUNEL assay and DNA laddering. Sanguinarine treatment resulted in up-regulation of death receptor 5 (DR5) expression, activation of caspase-8 and Bid leading to Bax conformational changes and translocation to the mitochondrial causing loss of mitochondrial membrane potential as measured by JC1 staining and release of cytochrome c to the cytosole. Sanguinarine induced release of cytochrome c resulted in activation of caspase-3, followed by polyadenosin-5′-diphosphate-ribose polymerase (PARP) cleavage leading to inhibition of proliferation and induction of caspase-dependent apoptosis. Furthermore, pre-treatment of PEL cells with z-VAD-fmk, a universal inhibitor of caspases, abrogated caspase-3 and PARP activation and prevented cell death induced by Sanguinarine. Inhibitor of apoptosis proteins (IAPs), play an important role in protecting cells against apoptosis through their direct action on caspases-9 and -3. Treatment of PEL cells with Sanguinarine down-regulated the expression of IAPs; XIAP, cIAP1 and cIAP2. Taken altogether, our findings suggest that Sanguinarine induces apoptosis via up-regulation of DR5, activation of Bax in a caspase-dependent pathway and down-regulation of IAPs. These results provide the molecular basis and preliminary data for new treatment strategies that may incorporate Sanguinarine in regimens for primary effusion lymphoma treatment.
Chlorambucil Induced Angioedema of the Tongue in a Patient with Lymphoma.
Authors: Shojania A. Majid
Published: April 2020

A Caucasian female, born in 1941, was first seen in November 2002 because of tiredness, weight loss and splenomegaly. She was taking glyburide, procylidine and venlafaxine for diabetes, parkinsonism and depression. Physical examination, revealed marked splenomegaly but no lymphadenopathy. Flowcytometry of peripheral blood showed a monoclonal B cell lymphoproliferative disorder consistent with marginal zone lymphoma. She was treated with chlorambucil 6 mg daily and prednisone 25 mg daily. She responded well to therapy. Chemotherapy was discontinued in December 2003. In April 2005 she was admitted to the hospital with pneumonia. She had pancytopenia and marked splenomegaly. Bone marrow apirate showed infiltration with marginal zone lymphoma cells. She was prescribed Chlorambucil 6 mg daily for management of her recurrent lymphoma. She took her first 6 mg of Chlorambucil at 8:00 PM of May 18, 2005 and woke up around 5:30 AM with grossly swollen tongue. She was brough to Emergency ward at 06:45 hr with grossly swollen, tender tongue protrudinng form her widely opened mouth. Angioedema of the tongue was diagnosed and she was given epinephrin, IV Benadryl, Solumedrol and morphine. By 10:00 hr, she was in moderate respiratory distress. She was taken to OR at 10:20 for urgent intubation or tracheotomy. Endoscopy revealed ++ oropharyngeal and supraglotic edema. A nasotracheal tube was inserted at 11:00 hr. The picture included was taken one hour after intubation.(16 hrs after taking chlorambucil). She was given Solumedrol 125 mg IV Q 12 hr. By 24 hours after intubation her tongue had receded into her mouth and she was extubated. Her serum C1 inhibitor was 0.13 g/L (normal 0.15–0.35) and functional C1 esterase inhibitor was 87% (normal >67%). Immunoglobulines, C3 & C4 were normal. Her lymphoma was treated with 6 courses of R-CVP regimen. She has not received any more chlorambucil and she has not had any more angioedema or anaphylactic reaction of any kind. In May 2005 her C1 inhibitor was 0.38 g/L and functional C1 esterase inhibitor was 96%.

Discussion: on the day before experiencing angioedema of the tongue, she had not taken any unusual food or any other new medication. The time sequence and subsequent course of this case strongly suggest that the angioedema of the tongue was due to Chlorambucil. The literature does not show any report of chlorambucil induced angioedema. However, angioedema is listed as one of the adverse reactions of Chlorambuci both in AHFS Drug Information and in Compendium of Pharmaceuticals and Specialities. Following communication with GlaskoSmithKline, the manufacturer of Chlorambucil, I was told that, they are not aware of any published report, but they have reports of angioedema induced by Chlorambucil in their file. I was told that they are not in position to reveal the number of such reactions.

Figure Figure
Cutaneous Lymphomas: Single Institution Experience in Lima-Peru.
Authors: Beltrán-Gárate Brady E. , Malaga José , Portugal Karen , Morales Domingo , Riva Luis , de Mendoza Fernando Hurtado , Castillo-Aguirre Jorge , Quiñones Pilar , Carrasco-Yalan Antonio A.
Published: April 2020

Background: The clinicopathologic characteristics of malignant lymphomas may vary according to geography. We previously described Adult T -cell leukaemia/lymphoma (ATLL) cases associated with human T-cell lymphotropic virus type-I (HTLV-I) in their different clinical presentation: acute, lymphomatous, chronic and smoldering and the recently primary cutaneous subtype in Peru (EHA2001: abstract 129). The aim of this study is to determine the relative frequency of cutaneous lymphomas and evaluate the clinical relevance of the new WHO/EORTC classification in a General Hospital in Lima-Peru

Methods: We conducted a clinicopathologic retrospective study of primary cutaneous lymphomas diagnosed from 1997 to 2004 in our General Hospital. Clinical records, haematoxylin & eosin-stained slides and immunohistochemical stains from 78 patients were reviewed. HTLV-1 serology was made using ELISA and Western Blot method. The statistical method was descriptive and survival was calculated using the Kaplan-Meier method.

Results: The mean age at time of presentation was 62 years and the female/male ratio 1,5:1. T-cell lymphomas were 88.6% and 11.4% were B-cell lymphomas. Eight-six percent (67/78) were primary cutaneous lymphomas and fourteen percent (11/78) were secondary cutaneous lymphomas. The most frequent primary cutaneous lymphomas was mycosis fungoides (MF): 44.7% (30/67); cutaneous / smoldering ATLL sutypes included 13/67 (19.4%) patients; unspecified peripheral T-cell lymphoma 4/67 (6%), lymphomatoid papulosis 2/67 (3%), leg-type diffuse large B-cell lymphoma 2/67 (3%), diffuse large B-cell lymphoma 2/67 (3%), subcutaneous panniculitis-like T-cell lymphoma 2/67 (3%), one case of the following lymphomas: anaplastic large cell, Sézary syndrome, nasal type extranodal NK/T-cell lymphoma, marginal zone B-cell lymphoma, follicle center lymphoma and intravascular lymphoma; finally unclassifiable lymphomas 5/67 (7.4%). Most frequent secondary cutaneous lymphomas were acute and lymphomatous subtypes of ATLL with 72% of the cases. Five-years overall survival for MF was 77%. The 5-years overall survival for primary cutaneous ATLL lymphomas was 18% and 0% for the secondary cutaneous ATLL group.

Conclusions: In this retrospective analysis, both ATLL and MF are the most frequent cutaneous lymphomas in our General Hospital. ATLL has a poor overall survival.
BCL6 Rearrangement Detected by FISH Analysis Is a Poor Prognostic Factor in the “Non-Germinal Center Phenotype” of Diffuse Large B-Cell Lymphoma.
Authors: Takizawa Jun , Aoki Sadao , Momoi Akihito , Kitajima Toshiki , Higashimura Masutaka , Nakamura Naoya , Aizawa Yoshifusa
Published: April 2020

Background: Diffuse large B-cell lymphoma (DLBCL), which is the most common type of adult non-Hodgkin lymphoma, is considered to be heterogeneous in cytogenetics, immunophenotype and clinical feature. As the results of gene expression profiling, DLBCL can be divided into prognostically significant 3 subgroups of germinal center B-like (GCB), activated B-like and type 3. Chromosomal translocations affecting the BCL6 locus at the 3q27 locus are common in DLBCL, however, the prognostic significance of BCL6 rearrangement is still controversial.

Methods: Twenty-six cases of DLBCL were examined with interphase fluorescence in situ hybridization (FISH) on touch preparations of lymph nodes using LSI BCL6 dual color probes (Vysis) for the incidence of BCL6 rearrangement and immunohistochemistry on paraffin section using CD10, BCL6 and MUM1 for subclassfying “GCB phenotype” and “non-GCB phenotype”. The correlation of BCL6 rearrangement with survival was investigated in two subgroups of DLBCL.

Results: Of the 26 DLBCL cases, 6 cases (23%) were considered GCB phenotype and 20 cases (77%) non-GCB phenotype. BCL6 rearrangements were detected in 2 of 6 cases (33%) with GCB phenotype and 9 of 20 (45%) with non-GCB phenotype (total 11/26, 42%). ALL 6 cases with the GCB phenotype achieved sustained complete remission after chemotherapy and are alive. On the other hand, complete remission rate was 22% for the cases with BCL6 rearrangement but 73% for the cases without BCL6 rearrangement in the non-GCB phenotype (p=0.069). BCL6 rearrangement had a significant adverse effect on progression free survival within the non-GCB phenotype (P=0.016), but there was no significant correlation between BCL6 rearrangement and overall survival.

Conclusion: FISH-based technique of the BCL6 rearrangements using touch preparations of lymph nodes could be developed for the retrospective analysis on survival. BCL6 rearrangement showed a poor prognostic effect particular in the non-GCB subgroup of DLBCL.

Overall survival Overall survival Progression free survival Progression free survival
Incidence, Clinical Characteristics and Survival of Malignant Lymphomas in the Province of Modena (Italy); a Population-Based Study.
Authors: Luminari Stefano , Cesaretti Marina , Rashid Ivan , Pennese Elsa , Mammi Caterina , Montanini Antonella , Barbolini Elisa , Bellei Monica , Marcheselli Luigi , Partesotti Giovanni , Bari Alessia , Maiorana Antonio , Artusi Tullio , Federico Massimo
Published: April 2020

The incidence of lymphomas has increased in many parts of the world in recent decades. We conducted a population-based study of peripheral lymphoma (PL) in the province of Modena (Italy). According to the international classification for lymphomas (WHO/ICD-O-3) we evaluated incidence patterns and time trends of 1,582 cases of PLs diagnosed between 1997–2003, also providing details of clinical characteristics, treatment and outcome of different entities. World Age Standardized Rates (ASR) varied substantially among lymphoid neoplasm subtypes. Overall, ASR (per 100,000) was 13.4 for B-cell lymphomas (16.5 and 10.6 for males and females respectively), 2.2 for T-cell lymphomas (3.1 and 1.5) and 3.4 for Hodgkin lymphoma. Among B-NHL the highest ASR was observed for Diffuse large B-cell lymphoma (DLBCL, 4.8) and Small lymphocytic lymphoma (CLL/SLL, 3.3). Among T-cell lymphomas, ASR was 1.4 for Mycosis fungoides/Sezary syndrome (MF/SS) and 0.4 for both Peripheral T-cell lymphoma (PTCL) and Anaplastic large cell lymphoma (ALCL). The overall annual percent change (APC) of the World ASR was 1.62 (C.I.95%: −4.32; 7.93). Extranodal (EN) involvement was observed in 49% of cases; a diagnosis of Primary EN lymphoma (PENL) was confirmed in 27.8% of cases. In addition to MF/SS, the most frequently reported PENL were marginal zone lymphomas (MZL) and DLBCL. The documented five years Overall Survival (OS) for the whole series was 61.9%, being 61.7%, 55.3% and 83% for B-NHL, T-NHL and HL, respectively. These data confirm an overall improvement when compared with available historical controls for the main European countries. In addition, using period analysis we could estimate more up-to-date survival data within our studied series (all patients vs those diagnosed after 2002): survival of patients with MZL, DLBCL and HL strongly improved during the study period. Among different lymphoma subtypes, patients with Hairy cell leukemia (HCL), HL and MF/SS had the best survival with 5-year OS of 94%, 83% and 82%, respectively. In particular, for HCL and MF/SS the analysis of the Relative Survival suggested that there was no substantial difference in survival compared with healthy population. Our study provides a comprehensive description of both epidemiological and clinical features of PL in the province of Modena, recognizing in a population-based approach, major advances in the curability of some histological subtypes.
BCL2 Expression Is a Prognostic Marker for the Activated B-Cell-Like Type of Diffuse Large B-Cell Lymphoma.
Authors: Chiattone Carlos , Carvalho Marineide P. , Paes Roberto P. , Ribeiro Karina C.B. , Soares Fernando
Published: April 2020

Diffuse large B-cell lymphoma (DLBCL) is heterogeneous both clinically and morphologically reflecting a mixture of underlying biologic or genetic differences. Therefore, it is important to identify at diagnosis biological markers which permit determination of subgroups with favorable or unfavorable evolution. The goal of this study was to evaluate the impact of BCL-2 tumor expression on overall survival (OS) in germinal center B-cell (GCB) and non-GCB subgroup.

Patients and Methods. Seventy-four untreated pts (median age: 58 yrs: 36M/34F) with DLBCL de novo diagnosed in a single institution, treated with CHOP-like regimens. Tissue microarrays (TMA) blocks were created from paraffin-embedded, formalin-fixed block and stained with antibodies to CD20 (clone L26, Dako), CD10 (clone 56C6; Novocastra; NCL-CD10–270), BCL-6 (clone GI 191E/A8; Cell Mark; CMC 798) and MUM1 (clone MUM1p; Dako, CA; M7259).

Results. Tumor expression of BCL-2 (cut off 10%), by TMA, was seen in 47.3% (35/74) and was associated with a worse OS (44.7%) (p=0.038). There was no significant correlation between BCL-2 protein expression and OS (p=0.587) OS (p=0.587), whereas for the group non-GCB was associated with a worse OS (p = 0.017).

Conclusion: The immunohistochemical expression of BCL- 2 are able to identify for the GCB and non-GCB subtypes of DLBCL outcome.
The Successful Use of Archived Formalin-Fixed Paraffin-Embedded (FFPE) Material for Gene Expression Studies.
Authors: Lilakos Konstantinos , Angelopoulou Maria K. , Georgiou George , Salpeas Vassilios , Korkolopoulou Penelope , Panayiotidis Panayiotis , Pangalis Gerassimos A.
Published: April 2020

BACKGROUND: FFPE tissues are widely used as a source of genomic DNA (gDNA), as well as for immunohistochemistry. RNA amplification and its usage for gene expression studies from archived FFPE material has been hampered due to RNA degradation.

AIM: The description of a protocol for RNA extraction from archived FFPE tissues followed by reverse transcription (RT) and real time PCR and its implementation in the study of reference genes.

METHODS: RNA was extracted from single 15μm sections from 18 samples 30 months old and 6 samples 10 years old from patients with diffuse large B-cell lymphoma (DLBCL). The FFPE RNA Extraction kit (Roche) was used, followed by DNA digestion and run on an Agilent2100 Bioanalyzer. RT was performed using the Transcriptor System (Roche) with modified temperatures followed by real time PCR for the c-abl and Survivin genes using the Universal Library probes and primers (Roche). In addition for the 10-year old samples, beta2 microglobulin (b2m) and G6PDH were also amplified.

RESULTS: RNA that was yielded, was around 250 bp. c-abl was successfully amplified in all 18 samples up to 30 months old with a median cycle (Ct) number of 33.74. All 3 control genes were successfully amplified from the 10-year old samples with a median Ct number of 30.3 for G6PDH, 23.4 for b2m and 28.6 for c-abl. To further evaluate the amplification potency we successfully amplified Survivin transcripts with a median Ct number of 35.2. Survivin/abl ratio was calculated at a median of 0.5 for all DLBCL samples. gDNA interferenence was excluded, since no amplification signal was observed when gDNA was used as a template. The special RNA extraction methods, elevated temperatures for RT, as well as the use of the Universal Library probes, which are specifically designed to be short (8–9bp) and hybridize to a short amplicon (72bp for c-abl and 97bp for Survivin) were the major novelties of this method.

Conclusions: Specialized RNA extraction methods and the use of Universal Library Probes can succesfully lead to several target gene amplification from archived FFPE material and potentially provide templates for gene expression studies.
CCL2 Chemokine Gene Expression in Lymph Nodes May Have Prognostic Value in Non-Hodgkin’s Lymphoma.
Authors: Mazur Grzegorz , Kryczek Ilona , Wrobel Tomasz , Dlubek Dorota , Klimczak Aleksandra , Jaskula Emilia , Jelen Michal , Lange Andrzej , Kuliczkowski Kazimierz
Published: April 2020

Background: Non-Hodgkin’s lymphomas (nHL) constitute complex group of lymphoproliferative disorders in which clinical outcome is difficult to predict at the moment of diagnosis. Formation of International Prognostic Index has provided criteria dividing patients into groups of risk, but still new prognostic factors are being searched. There are several reports that some chemokines including CCL2 play a role in progression of solid tumours (bladder, ovarian, non-small cell lung cancer) and increased level of CCL2 has been found in myeloma multiple. CCL2 is the chemokine produced by tumour cells and some stromal cells, such as: fibroblasts, endothelial cells and monocytes. CCL2 is chemoattractant for monocytes, T, NK and dendritic cells and basophlis. CCL2 has also angiogenic activity and is necessary for tumour growth and metastatic process.

Aim: The purpose of this study was to evaluate gene expression of CCL2 chemokine in lymphoma and reactive lymph nodes.

Material and methods: CCL2 gene expression was determined in 37 lymph nodes of lymphoma patients (26 B-cell lymphoma: 12 females and 14 males aged 26–73 years; 4 T-cell lymphoma: males aged 41–81 years; 7 Hodgkin’s lymphoma: 4 females and 3 males aged 21–58 years) and 25 reactive lymph nodes (15 females, 10 males, aged 18–59 years, median age 32 years). Gene expression was determined by the reverse transcription (RT)-polymerase chain reaction method. Scale of expression was 0–3 AU. Statistical analysis was performed using Kruskall-Wallis and Mann-Witney tests (p<0,05).

Results: In lymphoma lymph nodes CCL2 expression was significantly higher (2–3 AU) than in reactive lymph nodes (p=0,0008). Increased CCL2 expression was detected in 19/26 (73%) B-cell lymphoma lymph nodes and all (4/4, 100%) T-cell lymphomas. In reactive lymph nodes 2 AU expression was observed in 7/25 cases (28%). Particularly high expression characterized diffuse large B-cell lymphomas and Burkitt lymphomas. Patients with high CCL2 expression had significantly shorter survival than those with low expression (p=0,004). There was positive correlation between CCL2 expression and Ki-67 proliferation marker (p<0,05; coefficient 0,39).

Conclusions: CCL2 expression is higher in B-cell lymphoma lymph nodes than in reactive lymph nodes. Multivariate analysis has proved CCL2 as independent prognostic factor in nHL.
B-Chronic Lymphoproliferative Disorders (BCLD) Presenting with Splenomegaly: Differential Diagnosis and Outcome.
Authors: Angelopoulou Maria K. , Papakonstantinou Maria , Kontopidou Flora N. , Dimopoulou Maria N. , Siakantaris Marina P. , Vassilakopoulos Theodoros P. , Kalpadakis Christina , Kokoris Styliani , Dimitriadou Evangelia M. , Galani Zacharoula , Sachanas Sotirios , Tsirkinidis Padelis , Kyrtsonis Marie-Christine , Korkolopoulou Penelope , Panayiotidis Panayiotis , Pangalis Gerassimos A.
Published: April 2020

BACKGROUND: The differential diagnosis of BCLD in patients (pts) presenting with splenomegaly as the sole clinical finding is extremely difficult, due to the lack of lymph node histology. The diagnosis is based on morphology and immunophenotype (IF) from the blood and/or bone marrow. However, the reproducibility of morphology is limited and there is considerable overlap in IF findings. AIMS: The comparative analysis of pts with BCLD presenting with sole splenomegaly.

METHODS: All pts presented to our Unit with a probable BCLD and splenomegaly as the sole clinical finding, between 1980 and 2005 were retrospectively analyzed. Pts were classified into 4 groups according to their IF: Group A: CD5+/CD23+ [B-CLL phenotype], Group B: CD5+/CD23− [Mantle cell lymphoma phenotype], Group C: CD5−/CD23+, and Group D: CD5−/CD23− [Groups C and D: Splenic marginal zone lymphoma phenotype]. Hairy cell leukemia cases were excluded from the analysis. Pts’ characteristics, first line treatment, disease specific survival (DSS), freedom from 1st treatment (FF1T)and freedom from 2nd treatment (FF2T) were analyzed.

RESULTS: 71 pts (41 males) with a median age of 63 years (range: 39–83 y) were recorded. The 4 groups differed significantly in all their main characteristics, except of age, platelet count and frequency of lymphocytosis as shown in the table. 6 pts have not received any treatment, 27 were splenectomized, 16 received chlorambucil, 11 Rituximab and the remaining 11 other first-line treatments. The median follow up of the currently alive pts is 44.7 months (3–303). 34 pts have not required 2nd therapeutic intervention. The 10-year DSS of all 71 pts was 71±7%. The 10-year DSS and 5-year FF2T did not differ significantly between the 4 groups. However group B had a worse DSS (p=0.05) and FF2T (p=0.0036) compared to all other groups together. Cases who were CD38+ (p=0.055), CD11c− (p=0.001) and had Hb ≤12g/dL (p=0.03) had a worse prognosis.

CONCLUSIONS: Pts with BCLD and splenomegaly can be classified into different clinicopathologic entities according to basic IF characteristics. CD5+/CD23− splenomegalic BCLDS seem to have a worse outcome. CD38+, CD11c− and anemia are poor prognostic features. The best therapeutic strategy remains to be established.

Characteristics of the 4 Splenomegalic Groups Group A Group B Group C Group D p N 19 16 20 16 Spleen (cm) 14 (6–20) 16 (9–34) 10 (5–30) 10 (4–20) 0.03 Hb (g/dL) 12.0 11.0 10.0 12.4 0.02 WBC (×10 ^9/L) 30 10.8 10.1 10.8 0.02 CD38+ 28% 75% 5% 6% <0.001 CD25+ 63% 38% 19% 0% 0.002 CD11c+ 56% 27% 47% 80% 0.03
Mutational Analysis of NOTCH1 Gene in Mature T-Cell Leukemia/Lymphoma.
Authors: Shimizu Daisuke , Taki Tomohiko , Utsunomiya Atae , Nakagawa Hitoshi , Nomura Kenichi , Matsumoto Yosuke , Nishida Kazuhiro , Horiike Shigeo , Taniwaki Masafumi
Published: April 2020

A somatic mutation of the NOTCH1 gene is frequently observed in T-cell acute lymphoblastic leukemia (T-ALL) of immature T-cell leukemia, but its prevalence in mature T-cell leukemia/lymphoma remains unknown. T-cell non-Hodgkin lymphomas (T-NHL) are relatively uncommon malignancies that represent approximately 12% of all lymphomas. The current WHO/EORTC classification recognizes 9 distinct clinicopathologic peripheral T-NHLs. The identification of genetic abnormalities and chromosomal rearrangements has been helpful in recognizing and defining lymphoma entities according to the WHO classification. However, the molecular changes that could play a role in leukemogenesis and progression remain unknown. NOTCH1 gene mutation was analyzed in 53 mature T-cell leukemia/lymphoma in adults, comprising 21 with adult T-cell leukemia (ATL), 25 with T-cell non-Hodgkin lymphoma (T-NHL), seven with T-cell prolymphocytic leukemia (T-PLL). We analyzed the hotspots of the NOTCH1 activating mutation in T-ALL cases located in exons 26 and 27, which encodes the N-terminal and C-terminal region of the heterodimerization (HD-N, HD-C) domain, and in exon 34, which encodes the PEST domain and terminal activation domain (TAD). Mutation detection in these two locations was performed by means of PCR-based denaturing high-performance liquid chromatography (dHPLC) using a WAVE DNA fragment analysis system followed by sequencing. We detected a nonsense mutation, C7249T, resulting in Q2417X, in an ATL patient, and a missense mutation, A6955G, resulting in M2319V, as well as a 3-bp deletion between 7234 and 7236 that resulted in deletion of the proline at codon 2412 in the same allele in a T-NHL, peripheral T-cell lymphoma, unspecified (PTCL-u), patient. NOTCH1 mutation was infrequent in mature T-cell leukemia/lymphoma, but NOTCH1 may be involved in leukemogenesis associated with a variety of T-cell leukemia/lymphoma rather than only with T-ALL.

Figure 1. (A) ATL case No.9, (B) T-NHL case No.38, (C) T-NHL case No.38 Figure 1. (A) ATL case No.9, (B) T-NHL case No.38, (C) T-NHL case No.38