- Email: [email protected]
Posters: Bone Marrow and Lymphoreticular System
Bone Marrow and Lymphoreticular System . 299
Posters: Bone Marrow and lymphoreticular System 89 Significant hybridization differences in comparative genomic hybridization (CGH) may be due to nucleotides used for DNA labelling as well as on the reference DNA chosen L. Wilkens, J. Tchinda, D. Burkhardt, D. Benten, R.v. Wasielewski, M. Nolte Institut fOrPathologie, Medizinische Hochschule Hannover CGH has been reported to be influenced by hybridization differences probably based on different DNA labelling systems. To define such differences in more detail we analysed 30 bone marrow samples with normal karyotype in conventional cytogenetics (CC) using two different DNA labelling systems and 7 different reference DNAs. In addition, fluorescence in situ hybridization (FISH) was performed for aberration detected in CGH. Signal deviations reaching the diagnostic threshold of 0.75 or 1.25 were found in 11/30 BMC samples when using fluorochrome conjugated nucleotides for DNA labelling. Deviations affected chromosomes 1(n=10), 2(n=4), 4(n=5), 5(n=3), 6(n=3), 12(n=2), 13(n=5), 16(n=6), 17(n=3), 19(n=11), 20(n=2), and/or 22(n=7). None of the imbalances were confirmed by (FISH). Using Digoxigenin (Dig) and Biotin (Bio) conjugated nucleotides in exemplary cases (n=3) led to the disappearance of these deviations. Repeated CGH experiments for a particular case with 7 different reference DNAs led to variations of the deviations from not detectable to obvious. Deviations not reaching the threshold of 0.75 or 1.25 but indicated by passing values of 0.87 or 1.12 were seen for hybridization of a fluorochrome labelled reference DNA with itself. These deviations disappeared using Dig/Bio for DNA labelling. The results demonstrate that the occurence of hybridization differences depends on both the haptens and fluorochrome conjugated nucleotides taken for DNA labelling and the reference DNA chosen for co-hybridization. Therefore a close control of CGH is mandatory using additional techniques such as FISH to confirm the results obtained by CGH.
91 BONE MARROW BIOPSIES IN AIDS PATIENTS: AIDS-COMPLICATION OR THERAPY-INDUCED TOXICI1Y? A Amirmaki', W. Heise2, K. Arasteh', V. Savvas', G. Grosse' ' Institut fur Pathologie, 2Innere Abteilung, Augusta-ViktoriaKrankenhaus, ' Bioptisches Institut; Berlin Aims: The different histological preperation of bone marrow in AIDSpatients raises the following question: Are pathological changes due to immunodeficiency or do they result from therapy-induced toxicity in patients with prolonged survival? Methods: We examined I1iac-Crest-Biopsies of 110 patients during the years 1996- 1998 . A greater part of the slide-preparations were not decalcified, but embadded in Methacrylat. These preparations were examined with H&E and Giemsa stains, Gomori-Silverplate, PAS and Esterase-Reaction. The rest were EDTA-decalcified; in paraffin embedded material, immunhistologicallymphoma marker were used. Results: In 71 cases we noticed quantitative deviations in the bone ~ structure presenting variously expressed atrophy also correlating with the reduction in peripheral blood cells. A granulomatous myelitis following miliar tuberculosis was found as rare infectious complication. Bone marrow involvement in atypical mycobacteriosis was diagnosed only in some cases. In the group of neoplastic complications mainly infiltration of highly malignant Non Hodgkin-Lymphoma occured. In focal lesions caused by the manifestation of Hodgkin's lymphoma, diagnosis can cause problems both in primary lymphoma diagnosis and in the differential diagnosis between neoplastic manifestation and uncharacteristic concomitant reactions. Conclusions: In a large series of 110 histologically examined bone marrow of AIDS-patients we mainly observed uncharacteristic forms of atrophy of different degree, Since an increase of these findings were found in patients with prolonged survival under intensive antiretroviral therapy, the influence of HIV-therapy needs to be discussed. Focal lesions of opportunistic or neoplastic complications were single findings in this series of biopsies.
92
Problems in the Diagnosis of Hodgkin's Disease in Needle Biospies from AIDS-Patients
90 COMBINED AUTOMATED PCR-FRAGMENT ANALYSIS OF TCELL RECEPTOR ~AND Y -CHAIN GENE REARRANGEMENT, A SENSITIVE METHOD TO DETECT CLONALlTY IN PARAFFIN-EMBEDDED TISSUE Erik Biirthel* , H .Kuhn, * lEdelmann** , M .Mittag** *, W. Ponisch* *** , T . Friedrich", *Institut fur Pathologie, **Institut fur Rechtsmedizin, *** K1inik fur Hautkrankheiten, **** K1inik II des Zentrums fur Innere Med izin der Universitat Leipzig Aims: We tried to establish a reliable and stable method to detect clonality ofTCR gene rearrangements in paraffin-embedded tissue. Methods: DNA samples from morphologically and immunologically unquestionable T-ceIl lymphomas (n=35), and reactive controls (n=I5) were extracted. We used a multiplex PCR of the rearranged TCRP. and of the TCRy-gene with fluorescence marked primers. The amplicons were analyzed by routine agarosegel and polyacrylarnid electrophoresis and by automated PCR-fragment analysis (GeneScan). Dilution studies with the JURKAT ceIl line were done . Results: 70% of the analyzed lymphomas were detected as monoclonal after TCRy-gene, 85% after additional use of TCR~ gene analysis. Some typ ical AlLD lymphomas were oligoclonal. Monoclonality could distinguished from polyclonality much easier after GeneScan than after routine gel electrophoresis. The limit of detection was 0,6% JURKAT celIs in a polyclonal solution. Conclusions: The combined TCRP. and TCRy-gene rearrangement analysis with automated PCR fragment analysis is recommended.
A Droste (a.G.)!, K. Arasteh(a.GY, W. Heise(a.G.)2, G. Grosse t, H. Stein' Institut fur Pathologie' und II. Innere Abteilung', Auguste -ViktoriaKrankenhaus Berlin und Universitatsklinikurn Benjamin Franklin Berlin' Aims:We want to show the problems in diagnosis and differential diagnostic considerations of Hodgkin's lymphoma especially in small needle biopsies from AIDS-patients, where lymphoma was clinically suspected. Extranodal manifestations of Hodgkin's disease were of special interest. ~:Biopsies from liver, bone marrow , lymph nodes and some other organs from AIDS-patients with late stage disease presenting with fever, swelling of lymph nodes or foci of unknown origin were investigated. In some patients bioptic diagnosis could be compared with autoptic findings. Conventional methods, special stains for infectious diseases and immunohistological investigations for pathogens and lymphomas were performed. Results : In Hodgkin's disease, there was an increase of extranodal manifestations with unusual locations and pattern of spread which could be confirmed by autopsy. We often found infiltrates of special types with only few neoplastic cells. Sometimes lymphoma closely imitated infectious disease. In findings without pathogens except for latent membrane protein (LMP) ofEBV, repeated biopsy and the positive reaction with CD 30 could confirm the diagnosis oflymphoma. Conclusions: Organic focuses of uncertain etiology in AIDS-pat ients especially with long-time positivity for HfV, poor immunological parameters and lack of response towards antibiotic therapy should be investigated for such a case of EBV-associated M. Hodgkin even without typical neoplastic cells if no other pathogen is found since this is important for rapid therapy.
300 . Posters
93
95
Prevalence, presentation and prognosis of Lymphocyte-Rich classical Hodgkin's Disease Rvon Wasielewski, Sabine Seth, J.Franklin, R.Fischer, K.Hiibner, M.L.Hansmann, V.Diehl, A.Georgii Medical School Hannover, University Cologne,University Frankfurt aM Aims: The REAL classification includes the provisional entity of the Lymphocyte-Rich classical Hodgkin's Disease (LRcHD), but no clinical or prognostic data is available in unselected cases with modem treatment. Design: 1.520 cases of the German Hodgkin Study Group were reviewed. Cases showing typical Hodgkin/Sternberg Reed cells and more than 90% of surrounding lymphocytes in the absence of sclerosis were regarded as LRcHD if immunohistochemistry ruled out a nodular paragranuloma. Results: Among 1.339 cases with complete follow up, 4.1% were classified as LRcHD (LP 5.2%; NS 63.3%; MC 19.6%; LD 0.9%; unclassifiable HD: 6.9%). Age distribution was similar to MC patients, but >70% presented at early stages of disease comparable with LP. Significant differences were detected for the erythrocyte sedimentation rate (vs LP, NS, MC), in B-symptoms, extended lymph node involvement, stage (vs. NS, MC), spleen tumor, hemoglobin, great mediastinal mass and sex (only vs. NS). Survival and Freedom from treatment failure were in-between LP and classical HD, but not significantly different. A comparison of the original diagnoses by local pathologists showed that LRcHD has been formerly hidden among all subtypes ofHD, i.e. LP, NS and MC. Conclusion: LRcHD is a small group previously submerged in all other categories ofHD. Even in this large study its rarity allows no final answer as to whether LRcHD cases do really worse in clinical behavior compared to the favorable course of nodular LP under modem therapy. The different clinical presentation compared to NS/MC.HD suggests that LRcHD is more closely related to LP, in spite of its clear distinction by the immunophenotype.
Tissue eosinophilia is a prognostic factor in Hodgkin's Disease R.von Wasielewski R, Sabine Seth, J.Franklin, RFischer, K.Hiibner, M.L.Hansmann, V.Diehl, A.Georgii Medical School Hannover, University Cologne,University Frankfurt aM Aims: Identification of prognostic factors in HD remains important to identify either low-risk groups that could benefit from the use of less aggressive therapies or high risk patients where an adapted therapy could reduce the rate of relapses. Contrasted with age, stage and serum factors, the prognostic differences between histologic subtypes have been mainly narrowed by improvements of modem treatment. Exceptions are nodular sclerosis grade-2 (NS-2) histology and lack ofCDI5 positivity in classical HD that are both associated with a worse clinical outcome. Tissue eosinophilia has been shown in some carcinomas to be a favorable prognostic marker, but studies investigating its influence on survival in HD failed to show any significance. Aim was the evaluation of the prognostic relevance of eosinophila and correlation of other clinical features in cases treated by standardized modem therapy. Design: 1.529 cases of HD were reviewed and the amount of eosinophils was scored either as few/low or increasedlhigh. For most of the cases, complete clinical data and follow-up was analyzable. Results: In 39% of cases there was a prominent increase of tissue eosinophils. This was never observed in nodular LP and rarely in lymphocyte rich classical HD (7%), but in 44% of nodular sclerosis and mixed cellularity. Among NS-2 it reached 56%. Eosinophilia was correlated to the erythrocyte sedimentation rate (p=0.001), but not to stage or age. There was a worse freedom from treatment failure (p=0.0002) and survival (p=0.0001) for cases with eosinophila. Conclusion: Eosinophilia is an independent unfavorable prognostic factor in HD that is not equalized by modem therapy. This observation indicates that the eosinophils in HD are more than innocent bystanders. A possible explanation might be an interaction via cytokines/chemokines pathway.
94
96
Histomorphological parameters as prognostic factors in Nodular Sclerosing Hodgkin's Disease Sabine Seth, Rvon Wasielewski, J. Franklin. RFischer, K.Hlibner, M.L.Hansmann, V. Diehl, A.Georgii Medical School Hannover, University Cologne,University Frankfurt aM Aims: Grading of nodular sclerosing Hodgkin's disease is based on the number of lymphocytes, cohesive sheets of or atypical Hodgkin/Sternberg Reed (H/SR) cells. Recent results of the German Hodgkin Study Group have ascertained that NS-2 histology is an independent unfavorable prognostic factor. However, the predictive value of the respective morphological features is unknown in patients where a modem treatment may have substantially changed the outcome in comparison to former reports. Therefore, an analysis of the prognostic relevance of several histomorphological features contributing to the grading of NS in a large series of cases from a prospective clinical trial under standardized treatment protocols was performed. Design: Number of lymphocytes, cohesive sheets of H/RS cells, atypical H/RS cells, necrosis, granulocytes and degree of sclerosis were determined in 843 cases and correlated to clinical outcome. Results: A significantly worse survival was associated with decreased lymphocytes (p=0.0029; 9% ofNS cases) as well as necrosis (p=O.OI7; 20%), which was more often seen in NS-1. Cases with atypical H1RScells showed a negative trend, but not significantly. All other factors were not significant (p>O.2) Conclusion: Surprisingly, neither sheets of nor atypical HIRS cells were of prognostic value, both being decisive discriminants for diagnosing NS2. The prognostic significance between NS-I and NS-2 cannot be contributed to one single histologic parameter investigated here.
Treatment of Large Cell Anaplastic Lymphoma with immunostimulatory CpG-oligodeoxinucleotides in a mouse model C. Jenetzky*, H. Merz*, C. Bittner*, R. Pietrzik*, G.J. Wiedemann°, A.C. Feller* * Institute of Pathology, Medical University of LUbeck o Department of Hematology and Oncology, Medical University of LUbeck Aims: Immunostimulatory synthetic CpG-Oligodeoxynucleotides CODN) elicit cellular and humoral immune reponses. They have been tested successfully as adjuvants in trials of immunotherapy for malignant tumors. We investigated the therapy of the Large Cell Anaplastic Lymphoma (LCAL) with ODNs in an animal model. Methods: There is an established murine model of IL-9transfected T-Lymphocytes which induces growth of a malignant lymphoma with the typical features of the LCAL in humans. Mice were treated at different timepoints before or after injection of the malignant cells in variable doses. Results: In relation to the amount of tumor cells injected, establishment and growth of tumors were prevented or significantly delayed by treating mice with immunostimulatory CpG-ODNs. Conclusions: Since CpG-ODNs induce ail unspecific activation of the immune response they may be used as a possible adjuvant of already established therapies of the LCAL/Hodgkin's disease, providing a support to the host's immune system and offering the possibility of an effective cure of the malignant tumor.
Posters: Bone Marrow and lymphoreticular System 89 Significant hybridization differences in comparative genomic hybridization (CGH) may be due to nucleotides used for DNA labelling as well as on the reference DNA chosen L. Wilkens, J. Tchinda, D. Burkhardt, D. Benten, R.v. Wasielewski, M. Nolte Institut fOrPathologie, Medizinische Hochschule Hannover CGH has been reported to be influenced by hybridization differences probably based on different DNA labelling systems. To define such differences in more detail we analysed 30 bone marrow samples with normal karyotype in conventional cytogenetics (CC) using two different DNA labelling systems and 7 different reference DNAs. In addition, fluorescence in situ hybridization (FISH) was performed for aberration detected in CGH. Signal deviations reaching the diagnostic threshold of 0.75 or 1.25 were found in 11/30 BMC samples when using fluorochrome conjugated nucleotides for DNA labelling. Deviations affected chromosomes 1(n=10), 2(n=4), 4(n=5), 5(n=3), 6(n=3), 12(n=2), 13(n=5), 16(n=6), 17(n=3), 19(n=11), 20(n=2), and/or 22(n=7). None of the imbalances were confirmed by (FISH). Using Digoxigenin (Dig) and Biotin (Bio) conjugated nucleotides in exemplary cases (n=3) led to the disappearance of these deviations. Repeated CGH experiments for a particular case with 7 different reference DNAs led to variations of the deviations from not detectable to obvious. Deviations not reaching the threshold of 0.75 or 1.25 but indicated by passing values of 0.87 or 1.12 were seen for hybridization of a fluorochrome labelled reference DNA with itself. These deviations disappeared using Dig/Bio for DNA labelling. The results demonstrate that the occurence of hybridization differences depends on both the haptens and fluorochrome conjugated nucleotides taken for DNA labelling and the reference DNA chosen for co-hybridization. Therefore a close control of CGH is mandatory using additional techniques such as FISH to confirm the results obtained by CGH.
91 BONE MARROW BIOPSIES IN AIDS PATIENTS: AIDS-COMPLICATION OR THERAPY-INDUCED TOXICI1Y? A Amirmaki', W. Heise2, K. Arasteh', V. Savvas', G. Grosse' ' Institut fur Pathologie, 2Innere Abteilung, Augusta-ViktoriaKrankenhaus, ' Bioptisches Institut; Berlin Aims: The different histological preperation of bone marrow in AIDSpatients raises the following question: Are pathological changes due to immunodeficiency or do they result from therapy-induced toxicity in patients with prolonged survival? Methods: We examined I1iac-Crest-Biopsies of 110 patients during the years 1996- 1998 . A greater part of the slide-preparations were not decalcified, but embadded in Methacrylat. These preparations were examined with H&E and Giemsa stains, Gomori-Silverplate, PAS and Esterase-Reaction. The rest were EDTA-decalcified; in paraffin embedded material, immunhistologicallymphoma marker were used. Results: In 71 cases we noticed quantitative deviations in the bone ~ structure presenting variously expressed atrophy also correlating with the reduction in peripheral blood cells. A granulomatous myelitis following miliar tuberculosis was found as rare infectious complication. Bone marrow involvement in atypical mycobacteriosis was diagnosed only in some cases. In the group of neoplastic complications mainly infiltration of highly malignant Non Hodgkin-Lymphoma occured. In focal lesions caused by the manifestation of Hodgkin's lymphoma, diagnosis can cause problems both in primary lymphoma diagnosis and in the differential diagnosis between neoplastic manifestation and uncharacteristic concomitant reactions. Conclusions: In a large series of 110 histologically examined bone marrow of AIDS-patients we mainly observed uncharacteristic forms of atrophy of different degree, Since an increase of these findings were found in patients with prolonged survival under intensive antiretroviral therapy, the influence of HIV-therapy needs to be discussed. Focal lesions of opportunistic or neoplastic complications were single findings in this series of biopsies.
92
Problems in the Diagnosis of Hodgkin's Disease in Needle Biospies from AIDS-Patients
90 COMBINED AUTOMATED PCR-FRAGMENT ANALYSIS OF TCELL RECEPTOR ~AND Y -CHAIN GENE REARRANGEMENT, A SENSITIVE METHOD TO DETECT CLONALlTY IN PARAFFIN-EMBEDDED TISSUE Erik Biirthel* , H .Kuhn, * lEdelmann** , M .Mittag** *, W. Ponisch* *** , T . Friedrich", *Institut fur Pathologie, **Institut fur Rechtsmedizin, *** K1inik fur Hautkrankheiten, **** K1inik II des Zentrums fur Innere Med izin der Universitat Leipzig Aims: We tried to establish a reliable and stable method to detect clonality ofTCR gene rearrangements in paraffin-embedded tissue. Methods: DNA samples from morphologically and immunologically unquestionable T-ceIl lymphomas (n=35), and reactive controls (n=I5) were extracted. We used a multiplex PCR of the rearranged TCRP. and of the TCRy-gene with fluorescence marked primers. The amplicons were analyzed by routine agarosegel and polyacrylarnid electrophoresis and by automated PCR-fragment analysis (GeneScan). Dilution studies with the JURKAT ceIl line were done . Results: 70% of the analyzed lymphomas were detected as monoclonal after TCRy-gene, 85% after additional use of TCR~ gene analysis. Some typ ical AlLD lymphomas were oligoclonal. Monoclonality could distinguished from polyclonality much easier after GeneScan than after routine gel electrophoresis. The limit of detection was 0,6% JURKAT celIs in a polyclonal solution. Conclusions: The combined TCRP. and TCRy-gene rearrangement analysis with automated PCR fragment analysis is recommended.
A Droste (a.G.)!, K. Arasteh(a.GY, W. Heise(a.G.)2, G. Grosse t, H. Stein' Institut fur Pathologie' und II. Innere Abteilung', Auguste -ViktoriaKrankenhaus Berlin und Universitatsklinikurn Benjamin Franklin Berlin' Aims:We want to show the problems in diagnosis and differential diagnostic considerations of Hodgkin's lymphoma especially in small needle biopsies from AIDS-patients, where lymphoma was clinically suspected. Extranodal manifestations of Hodgkin's disease were of special interest. ~:Biopsies from liver, bone marrow , lymph nodes and some other organs from AIDS-patients with late stage disease presenting with fever, swelling of lymph nodes or foci of unknown origin were investigated. In some patients bioptic diagnosis could be compared with autoptic findings. Conventional methods, special stains for infectious diseases and immunohistological investigations for pathogens and lymphomas were performed. Results : In Hodgkin's disease, there was an increase of extranodal manifestations with unusual locations and pattern of spread which could be confirmed by autopsy. We often found infiltrates of special types with only few neoplastic cells. Sometimes lymphoma closely imitated infectious disease. In findings without pathogens except for latent membrane protein (LMP) ofEBV, repeated biopsy and the positive reaction with CD 30 could confirm the diagnosis oflymphoma. Conclusions: Organic focuses of uncertain etiology in AIDS-pat ients especially with long-time positivity for HfV, poor immunological parameters and lack of response towards antibiotic therapy should be investigated for such a case of EBV-associated M. Hodgkin even without typical neoplastic cells if no other pathogen is found since this is important for rapid therapy.
300 . Posters
93
95
Prevalence, presentation and prognosis of Lymphocyte-Rich classical Hodgkin's Disease Rvon Wasielewski, Sabine Seth, J.Franklin, R.Fischer, K.Hiibner, M.L.Hansmann, V.Diehl, A.Georgii Medical School Hannover, University Cologne,University Frankfurt aM Aims: The REAL classification includes the provisional entity of the Lymphocyte-Rich classical Hodgkin's Disease (LRcHD), but no clinical or prognostic data is available in unselected cases with modem treatment. Design: 1.520 cases of the German Hodgkin Study Group were reviewed. Cases showing typical Hodgkin/Sternberg Reed cells and more than 90% of surrounding lymphocytes in the absence of sclerosis were regarded as LRcHD if immunohistochemistry ruled out a nodular paragranuloma. Results: Among 1.339 cases with complete follow up, 4.1% were classified as LRcHD (LP 5.2%; NS 63.3%; MC 19.6%; LD 0.9%; unclassifiable HD: 6.9%). Age distribution was similar to MC patients, but >70% presented at early stages of disease comparable with LP. Significant differences were detected for the erythrocyte sedimentation rate (vs LP, NS, MC), in B-symptoms, extended lymph node involvement, stage (vs. NS, MC), spleen tumor, hemoglobin, great mediastinal mass and sex (only vs. NS). Survival and Freedom from treatment failure were in-between LP and classical HD, but not significantly different. A comparison of the original diagnoses by local pathologists showed that LRcHD has been formerly hidden among all subtypes ofHD, i.e. LP, NS and MC. Conclusion: LRcHD is a small group previously submerged in all other categories ofHD. Even in this large study its rarity allows no final answer as to whether LRcHD cases do really worse in clinical behavior compared to the favorable course of nodular LP under modem therapy. The different clinical presentation compared to NS/MC.HD suggests that LRcHD is more closely related to LP, in spite of its clear distinction by the immunophenotype.
Tissue eosinophilia is a prognostic factor in Hodgkin's Disease R.von Wasielewski R, Sabine Seth, J.Franklin, RFischer, K.Hiibner, M.L.Hansmann, V.Diehl, A.Georgii Medical School Hannover, University Cologne,University Frankfurt aM Aims: Identification of prognostic factors in HD remains important to identify either low-risk groups that could benefit from the use of less aggressive therapies or high risk patients where an adapted therapy could reduce the rate of relapses. Contrasted with age, stage and serum factors, the prognostic differences between histologic subtypes have been mainly narrowed by improvements of modem treatment. Exceptions are nodular sclerosis grade-2 (NS-2) histology and lack ofCDI5 positivity in classical HD that are both associated with a worse clinical outcome. Tissue eosinophilia has been shown in some carcinomas to be a favorable prognostic marker, but studies investigating its influence on survival in HD failed to show any significance. Aim was the evaluation of the prognostic relevance of eosinophila and correlation of other clinical features in cases treated by standardized modem therapy. Design: 1.529 cases of HD were reviewed and the amount of eosinophils was scored either as few/low or increasedlhigh. For most of the cases, complete clinical data and follow-up was analyzable. Results: In 39% of cases there was a prominent increase of tissue eosinophils. This was never observed in nodular LP and rarely in lymphocyte rich classical HD (7%), but in 44% of nodular sclerosis and mixed cellularity. Among NS-2 it reached 56%. Eosinophilia was correlated to the erythrocyte sedimentation rate (p=0.001), but not to stage or age. There was a worse freedom from treatment failure (p=0.0002) and survival (p=0.0001) for cases with eosinophila. Conclusion: Eosinophilia is an independent unfavorable prognostic factor in HD that is not equalized by modem therapy. This observation indicates that the eosinophils in HD are more than innocent bystanders. A possible explanation might be an interaction via cytokines/chemokines pathway.
94
96
Histomorphological parameters as prognostic factors in Nodular Sclerosing Hodgkin's Disease Sabine Seth, Rvon Wasielewski, J. Franklin. RFischer, K.Hlibner, M.L.Hansmann, V. Diehl, A.Georgii Medical School Hannover, University Cologne,University Frankfurt aM Aims: Grading of nodular sclerosing Hodgkin's disease is based on the number of lymphocytes, cohesive sheets of or atypical Hodgkin/Sternberg Reed (H/SR) cells. Recent results of the German Hodgkin Study Group have ascertained that NS-2 histology is an independent unfavorable prognostic factor. However, the predictive value of the respective morphological features is unknown in patients where a modem treatment may have substantially changed the outcome in comparison to former reports. Therefore, an analysis of the prognostic relevance of several histomorphological features contributing to the grading of NS in a large series of cases from a prospective clinical trial under standardized treatment protocols was performed. Design: Number of lymphocytes, cohesive sheets of H/RS cells, atypical H/RS cells, necrosis, granulocytes and degree of sclerosis were determined in 843 cases and correlated to clinical outcome. Results: A significantly worse survival was associated with decreased lymphocytes (p=0.0029; 9% ofNS cases) as well as necrosis (p=O.OI7; 20%), which was more often seen in NS-1. Cases with atypical H1RScells showed a negative trend, but not significantly. All other factors were not significant (p>O.2) Conclusion: Surprisingly, neither sheets of nor atypical HIRS cells were of prognostic value, both being decisive discriminants for diagnosing NS2. The prognostic significance between NS-I and NS-2 cannot be contributed to one single histologic parameter investigated here.
Treatment of Large Cell Anaplastic Lymphoma with immunostimulatory CpG-oligodeoxinucleotides in a mouse model C. Jenetzky*, H. Merz*, C. Bittner*, R. Pietrzik*, G.J. Wiedemann°, A.C. Feller* * Institute of Pathology, Medical University of LUbeck o Department of Hematology and Oncology, Medical University of LUbeck Aims: Immunostimulatory synthetic CpG-Oligodeoxynucleotides CODN) elicit cellular and humoral immune reponses. They have been tested successfully as adjuvants in trials of immunotherapy for malignant tumors. We investigated the therapy of the Large Cell Anaplastic Lymphoma (LCAL) with ODNs in an animal model. Methods: There is an established murine model of IL-9transfected T-Lymphocytes which induces growth of a malignant lymphoma with the typical features of the LCAL in humans. Mice were treated at different timepoints before or after injection of the malignant cells in variable doses. Results: In relation to the amount of tumor cells injected, establishment and growth of tumors were prevented or significantly delayed by treating mice with immunostimulatory CpG-ODNs. Conclusions: Since CpG-ODNs induce ail unspecific activation of the immune response they may be used as a possible adjuvant of already established therapies of the LCAL/Hodgkin's disease, providing a support to the host's immune system and offering the possibility of an effective cure of the malignant tumor.