B Cell Lymphomagenesis

Lymphoid neoplasms are a heterogeneous group of malignancies whose diagnosis depends on a very good analysis of hematopathology and morphology, immunophenotype, cytogenetic, molecular, and clinical characteristics. B-cell lymphomas begin from different developmental stages of B cells in germinal centers of secondary lymphoid tissue. The evolution of B-cell lymphomagenesis depends on different numbers of signal pathways. Proteins that play key point of signaling networks are changed by aberrant chromosomal expression, translocation, and/or accumulation, and those events determine the fate of the affected B cells. Many chemokines and cytokines have been implicated in providing the line for the cellular surviving and interaction in lymphoid organogenesis. Specific chromosomal alterations were associated with significant changes in gene-expression signatures that reflect various aspects of lymphoma cell biology as well as the host response to the lymphoma. The goal of this study was to find out a correlation between tumor markers and survival in patients with subgroups of DLBCL. The goal is to find out chronic autoimmune or pathogen-induced immune reactions resulting in lymphoid neogenesis. So we address (i) chemokines and adhesion molecules involved in lymphoid neogenesis, (ii) the autoimmune diseases and pathogens which are associated with the development of B-cell lymphomas, and (iii) the molecular mechanisms involved in the initiation and progression of DLBCL

Genetic, immunohistochemical and morphological features of subgroups in diffuse large B-cell lymphoma

Difuzni B velikostanični limfom (engl. diffuse large B-cell lymphoma, DLBCL) je najučestaliji tip limfoma u bolesnika odrasle životne dobi. Zbog tako visoke zastupljenosti mi smo odlučili kod naših bolesnika ispitati prisutnost VEGFR2 i NF-ĸB, te ocijeniti njihov utjecaj na ishod kod bolesnika s DLBCL. Mi smo kod 99 bolesnika ispitali prisutnost tih tumorskih biljega, te abnormalnosti gena BCL2, BCL6 i c-MYC kod iste grupe bolesnika. Rezultati su pokazali da je kod imunohistokemijski određenih tumorskih biljega VEGFR2 i NF-κB koji se nalaze u citoplazmi i jezgri nađeno da samo NF-κB, koji je prisutan u citoplazmi ukazuje na značajno lošije preživljenje (p=0,037). Metodom FISH analize BCL2, BCL6 i c-MYC nije nađena značajna razlika u preživljenju među podgrupama DLBCL (p=0,714).Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma in patient's elderly population. With such a high representation we decided our patients asses the pressence of VEGFR2 and NF-ĸB and assess their impact on outcome in patients with DLBCL. We investigated in 99 patients the presence of tumor markers and abnormalities of genes BCL2, BCL6 and c-MYC in the same group of patients. The results showed that in immunochistochemical markers of tumor VEGFR2 and NF-ĸB which are located in the cytoplasm and in the nucleus was found that only NF-ĸB, which is present in the cytoplasm indicates a singnificantly worse survival (p=0.037). FISH analaysis of BCL2, BCL6 and c-MYC was not found significant differences in survival among subgroups of DLBCL (p=0.714)

Treatment of severe acquired haemophilia A with an immunosuppressive agent

Acquired haemophilia A (AHA) is a rare hemorrhagic disease caused by an autoantibody against coagulation factor VIII. Nonhaemophiliac patients develop autoantibodies (inhibitors) directed against the factor VIII circulating coagulation protein. Disease is associated with an increased morbidity and mortality. Inhibitors against FVIII induce acute and life-threatening hemorrhagic diathesis because of abnormal blood clotting. FVIII inhibitors demonstrate bleeding disorders and prolonged activated partial thromboplastin time and a normal prothrombin time. AHA should be considered in the differential diagnosis particularly in postpartum women and in the elderly patients with bleeding tendency. Treatment of acute hemorrhage is focused in the control of the acute bleeding episode and the long term suppression of the autoantibody. In congenital hemophilia A with inhibitors, in which using repetitive infusions of high dose FVIII concentrates is effective for inhibitor eradication. This report presents one patient treated with immunosuppressive regimens. The most effective first-line treatment for the eradication of factor VIII autoantibodies is the combination of steroides and cyclophosphamide

PROGNOSTIC ROLE OF NF-κB EXPRESSION IN DIFFUSE LARGE B-CELL LYMPHOMA SUBGROUPS

Difuzni B-velikostanični limfom (DLBCL) s fenotipom germinalnog centra B stanica (GCB) ima bolju prognozu nego limfom aktiviranih zrelih B-stanica (ABC) fenotip ili tip 3 podskupina. Prethodne studije su navodile da nuklearni faktor-κB (NF- κB) ima značajnu ulogu kod ABC i tip 3 fenotipa, dok GCB fenotip karakteriziraju česte REL amplifi kacije. Upotrebom imunohistokemijske metode analizirali smo kod 99 bolesnika kojima je postavljena dijagnoza DLBCL prisutnost DC10, BCL6 i MUM1, da bismo mogli bolesnike podijeliti u GCB, ABC i tip 3 podskupine, te smo zatim analizirali prisutnu ekspresiju NF- κB u jezgri i u citoplazmi. Kod 22 (22 %) slučajeva prisutna je ekspresija NF-κB u jezgri kod svih podskupina DLBCL. NF-κB ekspresija u citoplazmi prisutna je u svim podskupinama DLBCL i nalazi se kod 77 (77%) bolesnika, i rezultat je značajan. Analizirali smo prisutnost CD10, Bcl6 i MUM1 i prisutnost NF-κB u jezgri i rezultati nisu značajni. Analiza nuklearne akumulacije NF-κB nije povezana ni s jednim kliničkim parametrom uključujući dob, spol, stadij bolesti, primijenjenu terapiju i MPI. Bolesnici s prisutnim NF κB u citoplazmi bez obzira na podskupine prema Hansu i sur. imaju značajno lošije preživljenje nego bolesnici koji nemaju prisutan NF-κB i rezultati su značajni. Bolesnici s GCB fenotipom i negativnom ekspresijom NF-κB u jezgri imaju bolje preživljenje nego bolesnici s GCB fenotipom i pozitivnom ekspresijom NF-κB u jezgri i rezultati su značajni. Bolesnici koji su primali kemoterapiju po shemi R-CHOP i imaju pozitivan NF-κB u jezgri imaju bolje preživljenje, ali rezultati nisu značajni. Ovi rezultati ukazuju na to da NF-κB ekspresija u jezgri i citoplazmi može biti prognostički čimbenik kod DLBCL i to može pojasniti slojevitost rizika kod bolesnika u kombinaciji s GCB / ne-GCB fenotipom. Naša analiza pokazuje da je NF-κB aktivnost kod svih podskupina DLBCL ključna i na taj način može predstavljati obećavajući molekularni cilj za buduće terapije.Objective: Diffuse large B-cell lymphoma (DLBCL) with germinal center B-cell (GCB) phenotype has better prognosis than activated mature B-cell (ABC) phenotype or type 3 subgroup. Previous studies have reported on a major role of the nuclear factor-κB (NF-κB) in ABC and type 3 phenotypes, whereas GCB phenotype is characterized by frequent REL amplifi cations. Methods: In 99 patients diagnosed with DLBCL, the presence of CD10, BCL6 and MUM1 was analyzed by immunohistochemical method to divide them into the GCB, ABC and type 3 subgroups. Then, NF-κB expression was analyzed in the nucleus and cytoplasm. Nuclear NF-κB expression was detected in 22 (22%) cases from all DLBCL subgroups. NF-κB expression in the cytoplasm was recorded in 77 (77%) cases from all DLBCL subgroups and this fi nding was signifi cant. Results: Results on the presence of CD10, BCL6 and MUM1 and the presence of NF-κB in the nucleus were not signifi cant. Analysis of nuclear NF-κB accumulation is not associated with any of the clinical parameters including age, sex, stage of disease, therapy administered and IPP. According to Hans et al., patients with NF-κB expressed in the cytoplasm have signifi cantly poorer survival irrespective of the subgroup than patients without cytoplasmic NF-κB, and these results are signifi cant. Patients with GCB phenotype and negative nuclear NF-κB expression have better survival than those with GCB phenotype and positive nuclear NF-κB expression, and these results are also signifi cant. Patients having received chemotherapy according to the R-CHOP schedule and with positive nuclear NF-κB expression have better survival; however, these results are not signifi cant. Discussion and Conclusion: These data suggest that nuclear and cytoplasmic NF-κB expression may be a prognostic factor in DLBCL, thus explaining the stratifi ed risk observed in patients in combination with GCB/non-GCB phenotype. Our study showed the NF-κB activity to be crucial in all DLBCL subgroups, thus potentially representing a promising molecular target for future therapies

CLINICAL FEATURES IN DLBCL AND TRANSLOCATION BCL2/c-MYC “DOUBLE HIT” LYMPHOMA

Difuzni B velikostanični limfom (DLBCL) je klasificiran kao limfom različitog entiteta i pomoću genske ekspresije proteina klasificiran je u tri podskupine. Cilj ovog rada je da pojasni kliničke, biološke, imunofenotipske i citogenetske značajke DLBCL s translokacijom t(14;18) i 8q24/c-MYC. Jedanaest bolesnika s DLBCL s dvostrukom translokacijom praćeno je u razdoblju od 2000. do 2009. god. Obilježja tih bolesnika uključuju morfološke, imunohistokemijske i citogenetske analize. Svi bolesnici imaju agresivna obilježja, prisutnost B simptoma (64 %), opće stanje bolesnika prema ECOG ljestvici ≥2 (55 %), povišenu aktivnost serumske laktat dehidrogenaze (73 %), klinički stadij III i IV (82 %), ekstranodalnu zahvaćenost bolesti (73 %), IPI ≥2 (73 %). Parcijalna remisija je postignuta kod 73 %, svi su bolesnici (73 %) umrli u kratkom roku. Bolesnici su liječeni CHOP i sličnim protokolima (COP, CVP, CNOP) uz dodatak Mabthere. Učinjena je imunofenotipizacija te određena ekspresija biljega CD20, CD3, CD10, Bcl6 i MUM1. u svih je učinjena citogenetska analiza ⌠fluorescentna hibridizacija in situ - FISH)⌡i nađene su kompleksne kariotipske promjene. Tako smo analizirali prisutnost gena BCL2, BCL6 i c-MYC, osam bolesnika je imalo translokacije gena BCL2 i c-MYC, dok je troje imalo translokaciju gena BCL6 i c-MYC. Usprkos provedenoj adekvatnoj terapiji, prognoza bolesnika je loša. Medijan preživljenja tih bolesnika je 1,85 godina. Zaključujemo da je DLBCL s BCL2 i c-MYC preuređenjem podskupina limfoma s vrlo lošim preživljenjem. Prisutnost tih dviju translokacija ima agresivan klinički tijek.Diffuse large B-cell lymphoma (DLBCL) is classified as lymphoma and various entities using the gene expression of proteins are classified into three groups. The aim of this study was to clarify the clinical, biological, immunophenotypic and cytogenetic features of DLBCL with translocation t (14; 18) and 8q24/c-MYC. Eleven DLBCL patients with dual translation were monitored during the 2000-2009 period. The characteristics of these patients included morphological, immunohistochemical and cytogenetic analysis. Study results showed that all patients had aggressive characteristics, presence of B symptoms (64%), general patient condition according to ECOG scale ≥2 (55%), elevated serum lactate dehydrogenase activity (73%), clinical stage III and IV (82%), extranodal involvement of the disease (73%), and IPI ≥2 (73%). Partial remission was achieved in 73% of all patients and all patients (73%) died within a short time. Patients were treated with CHOP and similar protocols (COP, CVP, CNOP), with the addition of MabThera. Immunophenotyping was performed and determined expression of the CD20, CD3, CD10, BCL6 and MUM1 markers. The cytogenetic analysis/fluorescence in situ hybridization revealed complex karyotype changes. Thus, we analyzed the presence of BCL2, BCL6 and c-MYC genes and found eight patients to have BCL2 and c-MYC translocation genes, while three had translocation of the BCL6 and c-MYC genes. Despite appropriate therapy, the patient prognosis is poor. The median survival in these patients was 1.85 years. DLBCL with BCL2 and c-MYC rearrangement of the subgroups of lymphoma is associated with very poor survival. The presence of these two translocations has an aggressive clinical course

CLINICAL FEATURES IN DLBCL AND TRANSLOCATION BCL2/c-MYC “DOUBLE HIT” LYMPHOMA

Difuzni B velikostanični limfom (DLBCL) je klasificiran kao limfom različitog entiteta i pomoću genske ekspresije proteina klasificiran je u tri podskupine. Cilj ovog rada je da pojasni kliničke, biološke, imunofenotipske i citogenetske značajke DLBCL s translokacijom t(14;18) i 8q24/c-MYC. Jedanaest bolesnika s DLBCL s dvostrukom translokacijom praćeno je u razdoblju od 2000. do 2009. god. Obilježja tih bolesnika uključuju morfološke, imunohistokemijske i citogenetske analize. Svi bolesnici imaju agresivna obilježja, prisutnost B simptoma (64 %), opće stanje bolesnika prema ECOG ljestvici ≥2 (55 %), povišenu aktivnost serumske laktat dehidrogenaze (73 %), klinički stadij III i IV (82 %), ekstranodalnu zahvaćenost bolesti (73 %), IPI ≥2 (73 %). Parcijalna remisija je postignuta kod 73 %, svi su bolesnici (73 %) umrli u kratkom roku. Bolesnici su liječeni CHOP i sličnim protokolima (COP, CVP, CNOP) uz dodatak Mabthere. Učinjena je imunofenotipizacija te određena ekspresija biljega CD20, CD3, CD10, Bcl6 i MUM1. u svih je učinjena citogenetska analiza ⌠fluorescentna hibridizacija in situ - FISH)⌡i nađene su kompleksne kariotipske promjene. Tako smo analizirali prisutnost gena BCL2, BCL6 i c-MYC, osam bolesnika je imalo translokacije gena BCL2 i c-MYC, dok je troje imalo translokaciju gena BCL6 i c-MYC. Usprkos provedenoj adekvatnoj terapiji, prognoza bolesnika je loša. Medijan preživljenja tih bolesnika je 1,85 godina. Zaključujemo da je DLBCL s BCL2 i c-MYC preuređenjem podskupina limfoma s vrlo lošim preživljenjem. Prisutnost tih dviju translokacija ima agresivan klinički tijek.Diffuse large B-cell lymphoma (DLBCL) is classified as lymphoma and various entities using the gene expression of proteins are classified into three groups. The aim of this study was to clarify the clinical, biological, immunophenotypic and cytogenetic features of DLBCL with translocation t (14; 18) and 8q24/c-MYC. Eleven DLBCL patients with dual translation were monitored during the 2000-2009 period. The characteristics of these patients included morphological, immunohistochemical and cytogenetic analysis. Study results showed that all patients had aggressive characteristics, presence of B symptoms (64%), general patient condition according to ECOG scale ≥2 (55%), elevated serum lactate dehydrogenase activity (73%), clinical stage III and IV (82%), extranodal involvement of the disease (73%), and IPI ≥2 (73%). Partial remission was achieved in 73% of all patients and all patients (73%) died within a short time. Patients were treated with CHOP and similar protocols (COP, CVP, CNOP), with the addition of MabThera. Immunophenotyping was performed and determined expression of the CD20, CD3, CD10, BCL6 and MUM1 markers. The cytogenetic analysis/fluorescence in situ hybridization revealed complex karyotype changes. Thus, we analyzed the presence of BCL2, BCL6 and c-MYC genes and found eight patients to have BCL2 and c-MYC translocation genes, while three had translocation of the BCL6 and c-MYC genes. Despite appropriate therapy, the patient prognosis is poor. The median survival in these patients was 1.85 years. DLBCL with BCL2 and c-MYC rearrangement of the subgroups of lymphoma is associated with very poor survival. The presence of these two translocations has an aggressive clinical course

Genetic, immunohistochemical and morphological features of subgroups in diffuse large B-cell lymphoma

Difuzni B velikostanični limfom (engl. diffuse large B-cell lymphoma, DLBCL) je najučestaliji tip limfoma u bolesnika odrasle životne dobi. Zbog tako visoke zastupljenosti mi smo odlučili kod naših bolesnika ispitati prisutnost VEGFR2 i NF-ĸB, te ocijeniti njihov utjecaj na ishod kod bolesnika s DLBCL. Mi smo kod 99 bolesnika ispitali prisutnost tih tumorskih biljega, te abnormalnosti gena BCL2, BCL6 i c-MYC kod iste grupe bolesnika. Rezultati su pokazali da je kod imunohistokemijski određenih tumorskih biljega VEGFR2 i NF-κB koji se nalaze u citoplazmi i jezgri nađeno da samo NF-κB, koji je prisutan u citoplazmi ukazuje na značajno lošije preživljenje (p=0,037). Metodom FISH analize BCL2, BCL6 i c-MYC nije nađena značajna razlika u preživljenju među podgrupama DLBCL (p=0,714).Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma in patient's elderly population. With such a high representation we decided our patients asses the pressence of VEGFR2 and NF-ĸB and assess their impact on outcome in patients with DLBCL. We investigated in 99 patients the presence of tumor markers and abnormalities of genes BCL2, BCL6 and c-MYC in the same group of patients. The results showed that in immunochistochemical markers of tumor VEGFR2 and NF-ĸB which are located in the cytoplasm and in the nucleus was found that only NF-ĸB, which is present in the cytoplasm indicates a singnificantly worse survival (p=0.037). FISH analaysis of BCL2, BCL6 and c-MYC was not found significant differences in survival among subgroups of DLBCL (p=0.714)

PROGNOSTIC ROLE OF NF-κB EXPRESSION IN DIFFUSE LARGE B-CELL LYMPHOMA SUBGROUPS

Difuzni B-velikostanični limfom (DLBCL) s fenotipom germinalnog centra B stanica (GCB) ima bolju prognozu nego limfom aktiviranih zrelih B-stanica (ABC) fenotip ili tip 3 podskupina. Prethodne studije su navodile da nuklearni faktor-κB (NF- κB) ima značajnu ulogu kod ABC i tip 3 fenotipa, dok GCB fenotip karakteriziraju česte REL amplifi kacije. Upotrebom imunohistokemijske metode analizirali smo kod 99 bolesnika kojima je postavljena dijagnoza DLBCL prisutnost DC10, BCL6 i MUM1, da bismo mogli bolesnike podijeliti u GCB, ABC i tip 3 podskupine, te smo zatim analizirali prisutnu ekspresiju NF- κB u jezgri i u citoplazmi. Kod 22 (22 %) slučajeva prisutna je ekspresija NF-κB u jezgri kod svih podskupina DLBCL. NF-κB ekspresija u citoplazmi prisutna je u svim podskupinama DLBCL i nalazi se kod 77 (77%) bolesnika, i rezultat je značajan. Analizirali smo prisutnost CD10, Bcl6 i MUM1 i prisutnost NF-κB u jezgri i rezultati nisu značajni. Analiza nuklearne akumulacije NF-κB nije povezana ni s jednim kliničkim parametrom uključujući dob, spol, stadij bolesti, primijenjenu terapiju i MPI. Bolesnici s prisutnim NF κB u citoplazmi bez obzira na podskupine prema Hansu i sur. imaju značajno lošije preživljenje nego bolesnici koji nemaju prisutan NF-κB i rezultati su značajni. Bolesnici s GCB fenotipom i negativnom ekspresijom NF-κB u jezgri imaju bolje preživljenje nego bolesnici s GCB fenotipom i pozitivnom ekspresijom NF-κB u jezgri i rezultati su značajni. Bolesnici koji su primali kemoterapiju po shemi R-CHOP i imaju pozitivan NF-κB u jezgri imaju bolje preživljenje, ali rezultati nisu značajni. Ovi rezultati ukazuju na to da NF-κB ekspresija u jezgri i citoplazmi može biti prognostički čimbenik kod DLBCL i to može pojasniti slojevitost rizika kod bolesnika u kombinaciji s GCB / ne-GCB fenotipom. Naša analiza pokazuje da je NF-κB aktivnost kod svih podskupina DLBCL ključna i na taj način može predstavljati obećavajući molekularni cilj za buduće terapije.Objective: Diffuse large B-cell lymphoma (DLBCL) with germinal center B-cell (GCB) phenotype has better prognosis than activated mature B-cell (ABC) phenotype or type 3 subgroup. Previous studies have reported on a major role of the nuclear factor-κB (NF-κB) in ABC and type 3 phenotypes, whereas GCB phenotype is characterized by frequent REL amplifi cations. Methods: In 99 patients diagnosed with DLBCL, the presence of CD10, BCL6 and MUM1 was analyzed by immunohistochemical method to divide them into the GCB, ABC and type 3 subgroups. Then, NF-κB expression was analyzed in the nucleus and cytoplasm. Nuclear NF-κB expression was detected in 22 (22%) cases from all DLBCL subgroups. NF-κB expression in the cytoplasm was recorded in 77 (77%) cases from all DLBCL subgroups and this fi nding was signifi cant. Results: Results on the presence of CD10, BCL6 and MUM1 and the presence of NF-κB in the nucleus were not signifi cant. Analysis of nuclear NF-κB accumulation is not associated with any of the clinical parameters including age, sex, stage of disease, therapy administered and IPP. According to Hans et al., patients with NF-κB expressed in the cytoplasm have signifi cantly poorer survival irrespective of the subgroup than patients without cytoplasmic NF-κB, and these results are signifi cant. Patients with GCB phenotype and negative nuclear NF-κB expression have better survival than those with GCB phenotype and positive nuclear NF-κB expression, and these results are also signifi cant. Patients having received chemotherapy according to the R-CHOP schedule and with positive nuclear NF-κB expression have better survival; however, these results are not signifi cant. Discussion and Conclusion: These data suggest that nuclear and cytoplasmic NF-κB expression may be a prognostic factor in DLBCL, thus explaining the stratifi ed risk observed in patients in combination with GCB/non-GCB phenotype. Our study showed the NF-κB activity to be crucial in all DLBCL subgroups, thus potentially representing a promising molecular target for future therapies

Treatment of severe acquired haemophilia A with an immunosuppressive agent

Acquired haemophilia A (AHA) is a rare hemorrhagic disease caused by an autoantibody against coagulation factor VIII. Nonhaemophiliac patients develop autoantibodies (inhibitors) directed against the factor VIII circulating coagulation protein. Disease is associated with an increased morbidity and mortality. Inhibitors against FVIII induce acute and life-threatening hemorrhagic diathesis because of abnormal blood clotting. FVIII inhibitors demonstrate bleeding disorders and prolonged activated partial thromboplastin time and a normal prothrombin time. AHA should be considered in the differential diagnosis particularly in postpartum women and in the elderly patients with bleeding tendency. Treatment of acute hemorrhage is focused in the control of the acute bleeding episode and the long term suppression of the autoantibody. In congenital hemophilia A with inhibitors, in which using repetitive infusions of high dose FVIII concentrates is effective for inhibitor eradication. This report presents one patient treated with immunosuppressive regimens. The most effective first-line treatment for the eradication of factor VIII autoantibodies is the combination of steroides and cyclophosphamide