Max Bittrich, Katharina Kriegsmann, Carola Tietze-Stolley, Kamram Movassaghi, Matthias Grube, Vladan Vucinic, Daniela Wehler, Andreas Burchert, Martin Schmidt-Hieber, Andreas Rank, Heinz A. Dürk, Bernd Metzner, Christoph Kimmich, Marcus Hentrich, Christian Kunz, Frank Hartmann, Cyrus Khandanpour, Maike de Wit, Udo Holtick, Michael Kiehl, Andrea Stoltefuß, Alexander Kiani, Ralph Naumann, Christian W. Scholz, Hans-Joachim Tischler, Martin Görner, Franziska Brand, Martin Ehmer, Nicolaus Kröger
Introduction: In patients with a clinical indication for autologous hematopoietic stem cell transplantation (ASCT), sufficient mobilization of CD34+ precursor cells into peripheral blood is essential to ensure adequate hematopoietic stem cell (HSC) collection prior to intensive therapy. However, with standard granulocyte-colony stimulating factor (G-CSF)-based mobilization schemes, an important minority of patients fail to mobilize sufficient (e.g., >10/µL) CD34+ cell counts into the peripheral blood and are considered as poor mobilizers (PM). Because failure to achieve sufficient CD34+ cell mobilization can negatively affect important clinical treatment endpoints, the use of plerixafor (PLX) was approved to increase CD34+ mobilization in PM patients. Methods: The German non-interventional, multicenter, open-label, prospective OPTIMOB study evaluated HSC mobilization strategies prior to planned ASCT in adult patients with hematologic malignancies (lymphomas or multiple myeloma [MM]) focusing on PM patients. PM patients were defined as follows: (1) never achieving ≥20 CD34+ cells/µL before 1st apheresis, (2) receiving PLX at any timepoint of mobilization, (3) their initially planned stem cell yield had to be reduced, or (4) they had not received apheresis due to low CD34+ count in peripheral blood. Results: 168 of 475 MM patients (35%) participating in the OPTIMOB study were classified as PM, and 155 of them (92%) received PLX (PM+PLX) during the study. PM patients were 40–78 years old, slightly more often male (n = 97, 58%), mostly newly diagnosed (n = 146, 87%) and received highly individualized previous treatments. Ninety-four of the PMs underwent chemotherapy mobilization (65%), and 51 patients (35%) received steady-state mobilization with G-CSF only during 1st mobilization attempt. 92% of the total PM population (n = 155) underwent apheresis, 78% of them (n = 117) achieved >2.0 × 106 CD34+ cells/kg body weight on the 1st day of apheresis. PM+PLX had a higher median total collection result than those PM patients without PLX support (7.2 vs. 5.7 × 106 CD34+ cells/kg body weight). In total, ASCT was performed in 136 PM+PLX (88%) versus 8 PM−PLX patients (62%). Conclusion: The OPTIMOB study showed that a considerable proportion of adult MM patients in Germany are PMs. Even though most of PMs were supported with PLX in the OPTIMOB study, PM-PLX also successfully mobilized HSCs, allowing ASCT in majority of all PMs. However, further analyses are required for treatment optimization in PMs.
{"title":"A German-Wide Systematic Study on Mobilization and Collection of Hematopoietic Stem Cells in Poor Mobilizer Patients with Multiple Myeloma prior to Autologous Stem Cell Transplantation","authors":"Max Bittrich, Katharina Kriegsmann, Carola Tietze-Stolley, Kamram Movassaghi, Matthias Grube, Vladan Vucinic, Daniela Wehler, Andreas Burchert, Martin Schmidt-Hieber, Andreas Rank, Heinz A. Dürk, Bernd Metzner, Christoph Kimmich, Marcus Hentrich, Christian Kunz, Frank Hartmann, Cyrus Khandanpour, Maike de Wit, Udo Holtick, Michael Kiehl, Andrea Stoltefuß, Alexander Kiani, Ralph Naumann, Christian W. Scholz, Hans-Joachim Tischler, Martin Görner, Franziska Brand, Martin Ehmer, Nicolaus Kröger","doi":"10.1159/000531935","DOIUrl":"https://doi.org/10.1159/000531935","url":null,"abstract":"<b><i>Introduction:</i></b> In patients with a clinical indication for autologous hematopoietic stem cell transplantation (ASCT), sufficient mobilization of CD34<sup>+</sup> precursor cells into peripheral blood is essential to ensure adequate hematopoietic stem cell (HSC) collection prior to intensive therapy. However, with standard granulocyte-colony stimulating factor (G-CSF)-based mobilization schemes, an important minority of patients fail to mobilize sufficient (e.g., &gt;10/µL) CD34<sup>+</sup> cell counts into the peripheral blood and are considered as poor mobilizers (PM). Because failure to achieve sufficient CD34<sup>+</sup> cell mobilization can negatively affect important clinical treatment endpoints, the use of plerixafor (PLX) was approved to increase CD34<sup>+</sup> mobilization in PM patients. <b><i>Methods:</i></b> The German non-interventional, multicenter, open-label, prospective OPTIMOB study evaluated HSC mobilization strategies prior to planned ASCT in adult patients with hematologic malignancies (lymphomas or multiple myeloma [MM]) focusing on PM patients. PM patients were defined as follows: (1) never achieving ≥20 CD34<sup>+</sup> cells/µL before 1st apheresis, (2) receiving PLX at any timepoint of mobilization, (3) their initially planned stem cell yield had to be reduced, or (4) they had not received apheresis due to low CD34<sup>+</sup> count in peripheral blood. <b><i>Results:</i></b> 168 of 475 MM patients (35%) participating in the OPTIMOB study were classified as PM, and 155 of them (92%) received PLX (PM+PLX) during the study. PM patients were 40–78 years old, slightly more often male (<i>n</i> = 97, 58%), mostly newly diagnosed (<i>n</i> = 146, 87%) and received highly individualized previous treatments. Ninety-four of the PMs underwent chemotherapy mobilization (65%), and 51 patients (35%) received steady-state mobilization with G-CSF only during 1st mobilization attempt. 92% of the total PM population (<i>n</i> = 155) underwent apheresis, 78% of them (<i>n</i> = 117) achieved &gt;2.0 × 10<sup>6</sup> CD34<sup>+</sup> cells/kg body weight on the 1st day of apheresis. PM+PLX had a higher median total collection result than those PM patients without PLX support (7.2 vs. 5.7 × 10<sup>6</sup> CD34<sup>+</sup> cells/kg body weight). In total, ASCT was performed in 136 PM+PLX (88%) versus 8 PM−PLX patients (62%). <b><i>Conclusion:</i></b> The OPTIMOB study showed that a considerable proportion of adult MM patients in Germany are PMs. Even though most of PMs were supported with PLX in the OPTIMOB study, PM-PLX also successfully mobilized HSCs, allowing ASCT in majority of all PMs. However, further analyses are required for treatment optimization in PMs.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136079073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Challenges for Plasma-Derived Medicinal Products","authors":"Rainer Moog, Teija Dehmer-Laitinen, Uwe Taborski","doi":"10.1159/000533736","DOIUrl":"https://doi.org/10.1159/000533736","url":null,"abstract":"","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136062715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objectives: Autologous blood transfusion techniques are well applied in surgery, but the red blood cells (RBCs) collected during laparoscopic surgery may forfeit their ability to oxygenate. O3 is a potent oxidation gas. This study investigates whether O3 could improve the oxygen-carrying capacity of RBCs, reduce inflammatory reactions, and offer organ protection. Methods: We established a hemorrhagic shock model in rabbits, and simulated CO2 pneumoperitoneum and O3 were applied before autologous blood transfusion. Perioperative mean arterial pressure and arterial blood gas were recorded, blood gas and RBC morphology of collected blood were analyzed, plasma IL-6, ALT, AST, CRE, and lung histopathology POD0 and POD3 were tested, as well as postoperative survival quality. Results: Autologous blood that underwent simulated CO2 pneumoperitoneum had a lower pH and SaO2 and a higher PaCO2 than the control group. After O3 treatment, PaO2 and SaO2 increased significantly, with unchanged pH values and PaCO2. RBCs in autologous blood were drastically deformed after CO2 conditioning and then reversed to normal by O3 treatment. Rabbits that received CO2-conditioned autologous blood had a compromised survival quality after surgery, higher plasma IL-6 levels, higher lung injury scores on POD0, higher ALT and AST levels on POD3, and O3 treatment alleviated these adverse outcomes. Conclusion: O3 can restore RBC function, significantly improve blood oxygenation under simulated CO2 pneumoperitoneum, offer organ protection, and improve the postoperative survival quality in the rabbit hemorrhage shock model.
{"title":"Ozone Improves Oxygenation and Offers Organ Protection after Autologous Blood Transfusion in a Simulated Carbon Dioxide Pneumoperitoneal Environment in a Rabbit Hemorrhagic Shock Model","authors":"Yu Gan, Xue Tian, Han Yao, Fei Huo, Yi Feng","doi":"10.1159/000527934","DOIUrl":"https://doi.org/10.1159/000527934","url":null,"abstract":"<b><i>Objectives:</i></b> Autologous blood transfusion techniques are well applied in surgery, but the red blood cells (RBCs) collected during laparoscopic surgery may forfeit their ability to oxygenate. O<sub>3</sub> is a potent oxidation gas. This study investigates whether O<sub>3</sub> could improve the oxygen-carrying capacity of RBCs, reduce inflammatory reactions, and offer organ protection. <b><i>Methods:</i></b> We established a hemorrhagic shock model in rabbits, and simulated CO<sub>2</sub> pneumoperitoneum and O<sub>3</sub> were applied before autologous blood transfusion. Perioperative mean arterial pressure and arterial blood gas were recorded, blood gas and RBC morphology of collected blood were analyzed, plasma IL-6, ALT, AST, CRE, and lung histopathology POD0 and POD3 were tested, as well as postoperative survival quality. <b><i>Results:</i></b> Autologous blood that underwent simulated CO<sub>2</sub> pneumoperitoneum had a lower pH and SaO<sub>2</sub> and a higher PaCO<sub>2</sub> than the control group. After O<sub>3</sub> treatment, PaO<sub>2</sub> and SaO<sub>2</sub> increased significantly, with unchanged pH values and PaCO<sub>2</sub>. RBCs in autologous blood were drastically deformed after CO<sub>2</sub> conditioning and then reversed to normal by O<sub>3</sub> treatment. Rabbits that received CO<sub>2</sub>-conditioned autologous blood had a compromised survival quality after surgery, higher plasma IL-6 levels, higher lung injury scores on POD0, higher ALT and AST levels on POD3, and O<sub>3</sub> treatment alleviated these adverse outcomes. <b><i>Conclusion:</i></b> O<sub>3</sub> can restore RBC function, significantly improve blood oxygenation under simulated CO<sub>2</sub> pneumoperitoneum, offer organ protection, and improve the postoperative survival quality in the rabbit hemorrhage shock model.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135579483","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: At the beginning of the pandemic, COVID-19 convalescent plasma (CCP) containing anti-SARS-CoV-2 antibodies was suggested as a source of therapy. In the last 3 years, many trials have demonstrated the limited usefulness of CCP therapy. This led us to the hypothesis that CCP could contain other elements, along with the desired neutralizing antibodies, which could potentially prevent it from having a therapeutic effect, among them cytokines, chemokines, growth factors, clotting factors, and autoantibodies. Methods: In total, 39 cytokines were analyzed in the plasma of 190 blood donors, and further research focused on the levels of 23 different cytokines in CCP (sCD40L, eotaxin, FGF-2, FLT-3L, ractalkine, GRO-α, IFNα2, IL-1β, IL-1RA, IL-5, IL-6, IL-8, IL-12, IL-13, IL-15, IL-17E, IP-10, MCP-1, MIP-1b, PDGF-AA, TGFα, TNFα, and TRAIL). Anti-SARS-CoV-2 antibodies and neutralizing antibodies were detected in CCP. Results: We found no significant differences between CCP taken within a maximum of 180 days from the onset of the first COVID-19 symptoms and the controls. We also made a comparison of the cytokine levels between the low neutralizing antibodies (<160) group and the high neutralizing antibodies (≥160) group and found there were no differences between the groups. Our research also showed no correlation either to levels of anti-SARS-CoV-2 IgG Ab or to the levels of neutralizing antibodies. There were also no significant changes in cytokine levels based on the period after the start of COVID-19 symptoms. Conclusions: No elements which could potentially be responsible for preventing CCP from having a therapeutic effect were found.
{"title":"Cytokine, Anti-SARS-CoV-2 Antibody, and Neutralizing Antibody Levels in Conventional Blood Donors Who Have Recovered from COVID-19","authors":"Elvira Maličev, Klemen Žiberna, Katerina Jazbec, Ana Kolenc, Polonca Mali, Urška Rahne Potokar, Primož Rožman","doi":"10.1159/000531942","DOIUrl":"https://doi.org/10.1159/000531942","url":null,"abstract":"<b><i>Background:</i></b> At the beginning of the pandemic, COVID-19 convalescent plasma (CCP) containing anti-SARS-CoV-2 antibodies was suggested as a source of therapy. In the last 3 years, many trials have demonstrated the limited usefulness of CCP therapy. This led us to the hypothesis that CCP could contain other elements, along with the desired neutralizing antibodies, which could potentially prevent it from having a therapeutic effect, among them cytokines, chemokines, growth factors, clotting factors, and autoantibodies. <b><i>Methods:</i></b> In total, 39 cytokines were analyzed in the plasma of 190 blood donors, and further research focused on the levels of 23 different cytokines in CCP (sCD40L, eotaxin, FGF-2, FLT-3L, ractalkine, GRO-α, IFNα2, IL-1β, IL-1RA, IL-5, IL-6, IL-8, IL-12, IL-13, IL-15, IL-17E, IP-10, MCP-1, MIP-1b, PDGF-AA, TGFα, TNFα, and TRAIL). Anti-SARS-CoV-2 antibodies and neutralizing antibodies were detected in CCP. <b><i>Results:</i></b> We found no significant differences between CCP taken within a maximum of 180 days from the onset of the first COVID-19 symptoms and the controls. We also made a comparison of the cytokine levels between the low neutralizing antibodies (&lt;160) group and the high neutralizing antibodies (≥160) group and found there were no differences between the groups. Our research also showed no correlation either to levels of anti-SARS-CoV-2 IgG Ab or to the levels of neutralizing antibodies. There were also no significant changes in cytokine levels based on the period after the start of COVID-19 symptoms. <b><i>Conclusions:</i></b> No elements which could potentially be responsible for preventing CCP from having a therapeutic effect were found.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136100220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-09-21eCollection Date: 2023-10-01DOI: 10.1159/000531936
Katharina Kriegsmann, Max Bittrich, Sandra Sauer, Carola Tietze-Stolley, Kamran Movassaghi, Matthias Grube, Vladan Vucinic, Daniela Wehler, Andreas Burchert, Martin Schmidt-Hieber, Andreas Rank, Heinz A Dürk, Bernd Metzner, Christoph Kimmich, Marcus Hentrich, Christian Kunz, Frank Hartmann, Cyrus Khandanpour, Maike de Wit, Udo Holtick, Michael Kiehl, Andrea Stoltefuß, Alexander Kiani, Ralph Naumann, Christian W Scholz, Hans-Joachim Tischler, Martin Görner, Franziska Brand, Martin Ehmer, Nicolaus Kröger
Introduction: Successful mobilization and collection of peripheral hematopoietic stem cells (HSCs) are necessary for lymphoma patients eligible for myeloablative chemotherapy with subsequent autologous stem cell transplantation (ASCT). Albeit G-CSF alone or combined with chemotherapy is well-established methods for HSC mobilization, up to 40% of the patients fail to mobilize (poor mobilizer, PM). Plerixafor (PLX) is commonly used in PM patients resulting in increased migration of HSCs into peripheral blood and thus improves the collection outcome.
Methods: The prospective, multicenter, open-label, non-interventional OPTIMOB study assessed mobilization and collection parameter of patients with lymphoma or multiple myeloma to get deep insights in the treatment of those patients in clinical routine focusing on PM patients. PM was defined as follows: (1) no achievement of ≥20 CD34+ progenitor cells/µL before first apheresis, (2) PLX administration at any time point during the observational period, (3) reduction of the initially planned CD34+ progenitor cell yield as necessity due to failed mobilization or HSC collection, and (4) no performance of apheresis due to low CD34+ progenitor level. Primary objective of the study was to assess mobilization success by the proportion of PM patients achieving >2 × 106 CD34+ progenitor cells/kg body weight on the first day of apheresis. Here, the data of the lymphoma cohort are presented.
Results: Out of 238 patients with lymphoma documented in the study, 32% were classified as PM. 87% of them received PLX. Demographic data revealed no obvious differences between PM and good mobilizing (GM) patients. All patients were treated highly individualized prior to mobilization. Majority of all PM patients were able to undergo apheresis (95%) and reached their individual requested CD34+ progenitor cell target (72%). 57% of the PM patients achieved >2.0 × 106 CD34+ progenitor cells/kg body weight on day 1 of apheresis and nearby 70% of them underwent ASCT. Median time to engraftment was similar in PM and GM patients of the lymphoma cohort.
Conclusions: Majority of PM patients with lymphoma were successfully mobilized and underwent ASCT. Most of them received PLX during the study.
{"title":"Mobilization and Hematopoietic Stem Cell Collection in Poor Mobilizing Patients with Lymphoma: Final Results of the German OPTIMOB Study.","authors":"Katharina Kriegsmann, Max Bittrich, Sandra Sauer, Carola Tietze-Stolley, Kamran Movassaghi, Matthias Grube, Vladan Vucinic, Daniela Wehler, Andreas Burchert, Martin Schmidt-Hieber, Andreas Rank, Heinz A Dürk, Bernd Metzner, Christoph Kimmich, Marcus Hentrich, Christian Kunz, Frank Hartmann, Cyrus Khandanpour, Maike de Wit, Udo Holtick, Michael Kiehl, Andrea Stoltefuß, Alexander Kiani, Ralph Naumann, Christian W Scholz, Hans-Joachim Tischler, Martin Görner, Franziska Brand, Martin Ehmer, Nicolaus Kröger","doi":"10.1159/000531936","DOIUrl":"https://doi.org/10.1159/000531936","url":null,"abstract":"<p><strong>Introduction: </strong>Successful mobilization and collection of peripheral hematopoietic stem cells (HSCs) are necessary for lymphoma patients eligible for myeloablative chemotherapy with subsequent autologous stem cell transplantation (ASCT). Albeit G-CSF alone or combined with chemotherapy is well-established methods for HSC mobilization, up to 40% of the patients fail to mobilize (poor mobilizer, PM). Plerixafor (PLX) is commonly used in PM patients resulting in increased migration of HSCs into peripheral blood and thus improves the collection outcome.</p><p><strong>Methods: </strong>The prospective, multicenter, open-label, non-interventional OPTIMOB study assessed mobilization and collection parameter of patients with lymphoma or multiple myeloma to get deep insights in the treatment of those patients in clinical routine focusing on PM patients. PM was defined as follows: (1) no achievement of ≥20 CD34<sup>+</sup> progenitor cells/µL before first apheresis, (2) PLX administration at any time point during the observational period, (3) reduction of the initially planned CD34<sup>+</sup> progenitor cell yield as necessity due to failed mobilization or HSC collection, and (4) no performance of apheresis due to low CD34<sup>+</sup> progenitor level. Primary objective of the study was to assess mobilization success by the proportion of PM patients achieving >2 × 10<sup>6</sup> CD34<sup>+</sup> progenitor cells/kg body weight on the first day of apheresis. Here, the data of the lymphoma cohort are presented.</p><p><strong>Results: </strong>Out of 238 patients with lymphoma documented in the study, 32% were classified as PM. 87% of them received PLX. Demographic data revealed no obvious differences between PM and good mobilizing (GM) patients. All patients were treated highly individualized prior to mobilization. Majority of all PM patients were able to undergo apheresis (95%) and reached their individual requested CD34<sup>+</sup> progenitor cell target (72%). 57% of the PM patients achieved >2.0 × 10<sup>6</sup> CD34<sup>+</sup> progenitor cells/kg body weight on day 1 of apheresis and nearby 70% of them underwent ASCT. Median time to engraftment was similar in PM and GM patients of the lymphoma cohort.</p><p><strong>Conclusions: </strong>Majority of PM patients with lymphoma were successfully mobilized and underwent ASCT. Most of them received PLX during the study.</p>","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10601601/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71414020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-09-15eCollection Date: 2023-10-01DOI: 10.1159/000533624
Anabel Heuer, Svea Löwhagen, Stefanie Uhlig, Svetlana Hetjens, Sylvia Büttner, Britta Pflästerer, Anke Diehlmann, Stefan Klein, Harald Klüter, Karen Bieback, Patrick Wuchter
Introduction: Autologous stem cell transplantation is a successful routine procedure with only a small number of non-engraftment cases, although the time to hematopoietic recovery may vary considerably across patients. While CD34 has been the decisive marker for enumerating hematopoietic stem and progenitor cells (HSPCs) for more than 30 years, the impact of CD34-positive cellular subpopulations in autologous HSPC grafts on hematopoietic reconstitution remains unclear.
Methods: The two-color ISHAGE protocol represents the current gold standard for CD34+ cell enumeration but includes only the number of viable CD45+/CD34+ cells relative to the body weight of the recipient. We adapted a multicolor flow cytometry marker panel for advanced characterization of CD34 subpopulations in retained samples of autologous peripheral blood stem cell products (n = 49), which had been cryostored for a wide range from 4 to 15 years. The flow cytometric analysis included CD10, CD34, CD38, CD45, CD45RA, CD133, and viability staining with 7AAD. The findings were correlated with clinical engraftment data, including reconstitution of leukocytes, neutrophils, and platelets after transplantation (TPL).
Results: We demonstrated that the identification of autologous HSPC subpopulations by flow cytometry after cryopreservation is feasible. Regarding the distribution of HSPC subpopulations, a markedly different pattern was observed in comparison to previously published data obtained using fresh autologous material. Our data revealed the largest ratio of lympho-myeloid progenitors (LMPPs) after freezing and thawing, followed by multipotent progenitors and erythroid-myeloid progenitors. A high ratio of LMPPs, representing an immature stage of differentiation, correlated significantly with early neutrophilic granulocyte and leukocyte engraftment (p = 0.025 and p = 0.003). Conversely, a large ratio of differentiated cells correlated with late engraftment of neutrophilic granulocytes (p = 0.024). Overall, successful engraftment was documented for all patients.
Conclusion: We established an advanced flow cytometry panel to assess the differentiation ability of cryostored autologous peripheral blood stem cell grafts and correlated it with timely hematopoietic reconstitution. This approach represents a novel and comprehensive way to identify hematopoietic stem and progenitor subpopulations. It is a feasible way to indicate the engraftment capacity of stem cell products.
{"title":"Flow Cytometric Characterization of Hematopoietic Stem and Progenitor Cell Subpopulations in Autologous Peripheral Blood Stem Cell Preparations after Cryopreservation.","authors":"Anabel Heuer, Svea Löwhagen, Stefanie Uhlig, Svetlana Hetjens, Sylvia Büttner, Britta Pflästerer, Anke Diehlmann, Stefan Klein, Harald Klüter, Karen Bieback, Patrick Wuchter","doi":"10.1159/000533624","DOIUrl":"10.1159/000533624","url":null,"abstract":"<p><strong>Introduction: </strong>Autologous stem cell transplantation is a successful routine procedure with only a small number of non-engraftment cases, although the time to hematopoietic recovery may vary considerably across patients. While CD34 has been the decisive marker for enumerating hematopoietic stem and progenitor cells (HSPCs) for more than 30 years, the impact of CD34-positive cellular subpopulations in autologous HSPC grafts on hematopoietic reconstitution remains unclear.</p><p><strong>Methods: </strong>The two-color ISHAGE protocol represents the current gold standard for CD34<sup>+</sup> cell enumeration but includes only the number of viable CD45<sup>+</sup>/CD34<sup>+</sup> cells relative to the body weight of the recipient. We adapted a multicolor flow cytometry marker panel for advanced characterization of CD34 subpopulations in retained samples of autologous peripheral blood stem cell products (<i>n</i> = 49), which had been cryostored for a wide range from 4 to 15 years. The flow cytometric analysis included CD10, CD34, CD38, CD45, CD45RA, CD133, and viability staining with 7AAD. The findings were correlated with clinical engraftment data, including reconstitution of leukocytes, neutrophils, and platelets after transplantation (TPL).</p><p><strong>Results: </strong>We demonstrated that the identification of autologous HSPC subpopulations by flow cytometry after cryopreservation is feasible. Regarding the distribution of HSPC subpopulations, a markedly different pattern was observed in comparison to previously published data obtained using fresh autologous material. Our data revealed the largest ratio of lympho-myeloid progenitors (LMPPs) after freezing and thawing, followed by multipotent progenitors and erythroid-myeloid progenitors. A high ratio of LMPPs, representing an immature stage of differentiation, correlated significantly with early neutrophilic granulocyte and leukocyte engraftment (<i>p</i> = 0.025 and <i>p</i> = 0.003). Conversely, a large ratio of differentiated cells correlated with late engraftment of neutrophilic granulocytes (<i>p</i> = 0.024). Overall, successful engraftment was documented for all patients.</p><p><strong>Conclusion: </strong>We established an advanced flow cytometry panel to assess the differentiation ability of cryostored autologous peripheral blood stem cell grafts and correlated it with timely hematopoietic reconstitution. This approach represents a novel and comprehensive way to identify hematopoietic stem and progenitor subpopulations. It is a feasible way to indicate the engraftment capacity of stem cell products.</p>","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10601604/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71414018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Julia Zeller-Hahn, Anna Kobsar, Marius Bittl, Angela Koessler, Katja Weber, Markus Boeck, Juergen Koessler
Introduction: Refrigeration of platelets is considered to provide advantages in therapy of acute hemorrhage due to increased platelet responsiveness. The alleviation of inhibitory signaling caused by cold temperature (CT) has been identified as an important mechanism contributing to enhanced platelet reactivity, detectable in freshly prepared platelets within 1 h of cold storage. The aim of this study was to confirm the effects of short-term refrigeration in platelets from apheresis-derived platelet concentrates (APC). Methods: APC were stored under standardized conditions for 1 day or for 2 days at room temperature and then refrigerated for 1 h, followed by sampling of platelets for analysis. Platelet reactivity was measured by aggregation studies using threshold concentrations of different agonists and by detection of fibrinogen binding using flow cytometry. The exploration of inhibitory signaling comprised the detection of VASP phosphorylation using flow cytometry or Western blot and the measurement of cyclic nucleotide levels. Results: Aggregation responses induced with ADP, collagen, or thrombin receptor-activating peptide-6 (TRAP-6) were increased in APC after cold storage for 1 h, associated with elevated TRAP-6-induced fibrinogen binding. VASP phosphorylation levels were decreased after cold exposition, detectable in 1-day- and 2-day-stored APC with flow cytometry, and in 2-day-stored APC with Western blot technique. Induced cGMP levels were lower after storage at CT in APC on day 1 and on day 2, whereas cAMP levels were reduced on 2-day-stored APC. Conclusion: Short-term refrigeration for 1 h is sufficient to induce an attenuation of inhibitory signaling, accompanied with increased aggregation responses in APC stored for up to 2 days. The “on demand” refrigeration of PC may be a reasonable approach for the preparation of platelets with enhanced responsiveness to treat patients with hemorrhage more effectively, which should be further addressed in consecutive studies.
{"title":"Increased Responsiveness of Stored Platelets after Short-Term Refrigeration","authors":"Julia Zeller-Hahn, Anna Kobsar, Marius Bittl, Angela Koessler, Katja Weber, Markus Boeck, Juergen Koessler","doi":"10.1159/000533274","DOIUrl":"https://doi.org/10.1159/000533274","url":null,"abstract":"<b><i>Introduction:</i></b> Refrigeration of platelets is considered to provide advantages in therapy of acute hemorrhage due to increased platelet responsiveness. The alleviation of inhibitory signaling caused by cold temperature (CT) has been identified as an important mechanism contributing to enhanced platelet reactivity, detectable in freshly prepared platelets within 1 h of cold storage. The aim of this study was to confirm the effects of short-term refrigeration in platelets from apheresis-derived platelet concentrates (APC). <b><i>Methods:</i></b> APC were stored under standardized conditions for 1 day or for 2 days at room temperature and then refrigerated for 1 h, followed by sampling of platelets for analysis. Platelet reactivity was measured by aggregation studies using threshold concentrations of different agonists and by detection of fibrinogen binding using flow cytometry. The exploration of inhibitory signaling comprised the detection of VASP phosphorylation using flow cytometry or Western blot and the measurement of cyclic nucleotide levels. <b><i>Results:</i></b> Aggregation responses induced with ADP, collagen, or thrombin receptor-activating peptide-6 (TRAP-6) were increased in APC after cold storage for 1 h, associated with elevated TRAP-6-induced fibrinogen binding. VASP phosphorylation levels were decreased after cold exposition, detectable in 1-day- and 2-day-stored APC with flow cytometry, and in 2-day-stored APC with Western blot technique. Induced cGMP levels were lower after storage at CT in APC on day 1 and on day 2, whereas cAMP levels were reduced on 2-day-stored APC. <b><i>Conclusion:</i></b> Short-term refrigeration for 1 h is sufficient to induce an attenuation of inhibitory signaling, accompanied with increased aggregation responses in APC stored for up to 2 days. The “on demand” refrigeration of PC may be a reasonable approach for the preparation of platelets with enhanced responsiveness to treat patients with hemorrhage more effectively, which should be further addressed in consecutive studies.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134973091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Introduction: DEL is known to be one of the weakest D variants, which can be detected by the adsorption-elution technique or by molecular study. Currently, in Thailand, we do not routinely test for DEL variants serologically or genetically among serologic RhD-negative blood donors. Case Presentation: We reported 2 cases of alloimmunization after transfused with Rh DEL, RHD*DEL1 allele, in the Thai population. The first case was a 73-year-old male with anemia who presented with post-cardiac arrest and septic shock. The patient was group B, RhD-negative, and was transfused with RhD-negative red blood cells (RBCs). Antibody screening and identification found that the patient developed anti-D and anti-Mia during the admission course. The second case was a 38-year-old woman with pseudomyxoma peritonei who developed anti-D after receiving four units of RhD-negative RBCs during cytoreductive surgery with hyperthermic intraperitoneal chemotherapy. Both patients did not receive anti-D immunoglobulin and had no previous history of anti-D detection. We retrospectively investigated and found two units of RHD*DEL1 among the RBCs transfused to these patients. Discussion: Previous reports of several cases of anti-D alloimmunization in RhD-negative recipients transfused by RHD*DEL1, an Asian-type DEL, are limited only to East Asia. We first identified 2 patients with anti-D alloimmunization after receiving the RHD*DEL1 RBCs in the Thai population. This raises concern about Rh DEL screening among D-negative Thai blood donors and whether to remove DEL units from the D-negative inventory to improve patient safety.
{"title":"Two Cases of Anti-D Alloimmunization in D-Negative Thai Patients as a Result of the Asian-Type DEL on Transfused Red Cells","authors":"Kanyapon Suksard, Komon Luangtrakool, Thongbai Rungroung, Sutthisak Chamsai, Pradermchai Saetam, Kulvara Kittisares, Parichart Permpikul, Janejira Kittivorapart","doi":"10.1159/000533625","DOIUrl":"https://doi.org/10.1159/000533625","url":null,"abstract":"<b><i>Introduction:</i></b> DEL is known to be one of the weakest D variants, which can be detected by the adsorption-elution technique or by molecular study. Currently, in Thailand, we do not routinely test for DEL variants serologically or genetically among serologic RhD-negative blood donors. <b><i>Case Presentation:</i></b> We reported 2 cases of alloimmunization after transfused with Rh DEL, <i>RHD*DEL1</i> allele, in the Thai population. The first case was a 73-year-old male with anemia who presented with post-cardiac arrest and septic shock. The patient was group B, RhD-negative, and was transfused with RhD-negative red blood cells (RBCs). Antibody screening and identification found that the patient developed anti-D and anti-Mi<sup>a</sup> during the admission course. The second case was a 38-year-old woman with pseudomyxoma peritonei who developed anti-D after receiving four units of RhD-negative RBCs during cytoreductive surgery with hyperthermic intraperitoneal chemotherapy. Both patients did not receive anti-D immunoglobulin and had no previous history of anti-D detection. We retrospectively investigated and found two units of <i>RHD*DEL1</i> among the RBCs transfused to these patients. <b><i>Discussion:</i></b> Previous reports of several cases of anti-D alloimmunization in RhD-negative recipients transfused by <i>RHD*DEL1</i>, an Asian-type DEL, are limited only to East Asia. We first identified 2 patients with anti-D alloimmunization after receiving the <i>RHD*DEL1</i> RBCs in the Thai population. This raises concern about Rh DEL screening among D-negative Thai blood donors and whether to remove DEL units from the D-negative inventory to improve patient safety.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134973087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lutz Pichl, Katja Konietzko, Ludwig Hartmann, Bogdan Puscasu, Carlos Jiménez Klingberg
Introduction: According to the guidelines (GL) valid in Germany, persons born or raised in a malaria-endemic area or had continuously stayed in a malaria-endemic area for more than 6 months may only be admitted donating blood if, among other things, validated and quality-assured laboratory diagnostics show that there is no evidence of infectivity. In a statement of the Working Group "Blood" of the Federal Ministry of Health (WGB), a reduction of the deferral period from 4 to 3 years and an antibody test after the deferral period are recommended. Methods: In accordance with the GL, nucleic acid testing (NAT) by means of PCR is carried out at our institution after a retention period of 4 years. In addition to the validated molecular biological testing, an infection serological examination was performed. Case Presentation: In the present cases, Plasmodia genome was detected in the respective single PCR in two blood donors originating from malaria-endemic areas after the expiry of the deferral period. However, one donor tested negative for antibodies against Plasmodia. Discussion/Conclusion: This observation is discussed in the context of a recommendation of the WGB. The question is addressed whether PCR testing is dispensable or whether a combination of infection serological testing and NAT should be favored.
{"title":"Moleculargenetic and in Addition Partly Discrepant Infection Serological Malaria Testing in Two Blood Donors","authors":"Lutz Pichl, Katja Konietzko, Ludwig Hartmann, Bogdan Puscasu, Carlos Jiménez Klingberg","doi":"10.1159/000530141","DOIUrl":"https://doi.org/10.1159/000530141","url":null,"abstract":"<b><i>Introduction:</i></b> According to the guidelines (GL) valid in Germany, persons born or raised in a malaria-endemic area or had continuously stayed in a malaria-endemic area for more than 6 months may only be admitted donating blood if, among other things, validated and quality-assured laboratory diagnostics show that there is no evidence of infectivity. In a statement of the Working Group \"Blood\" of the Federal Ministry of Health (WGB), a reduction of the deferral period from 4 to 3 years and an antibody test after the deferral period are recommended. <b><i>Methods:</i></b> In accordance with the GL, nucleic acid testing (NAT) by means of PCR is carried out at our institution after a retention period of 4 years. In addition to the validated molecular biological testing, an infection serological examination was performed. <b><i>Case Presentation:</i></b> In the present cases, <i>Plasmodia</i> genome was detected in the respective single PCR in two blood donors originating from malaria-endemic areas after the expiry of the deferral period. However, one donor tested negative for antibodies against <i>Plasmodia</i>. <b><i>Discussion/Conclusion:</i></b> This observation is discussed in the context of a recommendation of the WGB. The question is addressed whether PCR testing is dispensable or whether a combination of infection serological testing and NAT should be favored.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135938063","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-09-05eCollection Date: 2023-10-01DOI: 10.1159/000533649
Katharina Kriegsmann, Patrick Wuchter
Dear Editor, Mobilizing hematopoietic stem and progenitor cells from the bone marrow into the peripheral blood, collecting them sufficiently by leukapheresis, and transplanting them in an allogeneic or autologous setting as “stem cell graft” is a routine procedure in collection facilities such as in many blood transfusion establishments and transplant centers worldwide. Patients with multiple myeloma and malignant lymphoma are the main candidates for an autologous stem cell transplantation. Today’s induction regimen in both entities have been improved significantly over the last two decades, but the application of high-dose chemotherapy followed by autologous stem cell rescue still represents an essential part of the therapeutic regimen. In this context, the collection of an optimal stem cell product is an important prerequisite. This special issue of Transfusion Medicine and Hemotherapy provides a detailed insight into state-ofthe art stem cell mobilization, collection, and processing techniques. Regarding patients with multiple myeloma, Jantunen et al. comprehensively analyzed current mobilization strategies [1]. The impact of clinical parameters and induction regimens on peripheral blood stem cell (PBSC) mobilization in a large cohort of multiple myeloma patients has been evaluated by Sauer and colleagues [2]. In a more specific approach, Sauer et al. [3] assessed in a second study the effectiveness of autologous stem cell collection after daratumumab-VTD versus VCD. For the entity of mantle cell lymphoma, Turunen et al. examined the cellular composition and the clinical outcome after autologous transplantation [4]. In general,more than 80%of patients succeed in collecting an autologous PBSC graft. However, 10–20% mobilize insufficiently and are so-called “poor mobilizer” [5], but a harmonized definition has not been found yet. Strategies to overcome this problem include the application of plerixafor, either used preemptively or as a rescue strategy [6, 7]. However, comprehensive information on poor mobilizing patients regarding incidence, current treatments, and mobilization strategies is lacking. The German prospective, multicenter, open-label, non-interventional OPTIMOB study addressed this lack of knowledge by analyzing mobilization and collection parameters in a large cohort of adult, transplant-eligible, good and poormobilizing patients with lymphoma or multiple myeloma. In this special issue, the results of this huge national study with 28 recruiting study centers are presented in two articles: Bittrich et al. [8] described the results in patients with multiple myeloma, whereas Kriegsmann et al. [9] showed the results of the cohort of lymphoma patients. A state-of-the-art collection of PBSCs includes a system of quality control and benchmarking. Approaches to quantitatively assess the effectiveness of autologous leukapheresis sessions have been developed, and some allow not only to calculate the collection efficiency (CE2) but also to m
{"title":"Stem Cell Mobilization, Collection, and Processing.","authors":"Katharina Kriegsmann, Patrick Wuchter","doi":"10.1159/000533649","DOIUrl":"https://doi.org/10.1159/000533649","url":null,"abstract":"Dear Editor, Mobilizing hematopoietic stem and progenitor cells from the bone marrow into the peripheral blood, collecting them sufficiently by leukapheresis, and transplanting them in an allogeneic or autologous setting as “stem cell graft” is a routine procedure in collection facilities such as in many blood transfusion establishments and transplant centers worldwide. Patients with multiple myeloma and malignant lymphoma are the main candidates for an autologous stem cell transplantation. Today’s induction regimen in both entities have been improved significantly over the last two decades, but the application of high-dose chemotherapy followed by autologous stem cell rescue still represents an essential part of the therapeutic regimen. In this context, the collection of an optimal stem cell product is an important prerequisite. This special issue of Transfusion Medicine and Hemotherapy provides a detailed insight into state-ofthe art stem cell mobilization, collection, and processing techniques. Regarding patients with multiple myeloma, Jantunen et al. comprehensively analyzed current mobilization strategies [1]. The impact of clinical parameters and induction regimens on peripheral blood stem cell (PBSC) mobilization in a large cohort of multiple myeloma patients has been evaluated by Sauer and colleagues [2]. In a more specific approach, Sauer et al. [3] assessed in a second study the effectiveness of autologous stem cell collection after daratumumab-VTD versus VCD. For the entity of mantle cell lymphoma, Turunen et al. examined the cellular composition and the clinical outcome after autologous transplantation [4]. In general,more than 80%of patients succeed in collecting an autologous PBSC graft. However, 10–20% mobilize insufficiently and are so-called “poor mobilizer” [5], but a harmonized definition has not been found yet. Strategies to overcome this problem include the application of plerixafor, either used preemptively or as a rescue strategy [6, 7]. However, comprehensive information on poor mobilizing patients regarding incidence, current treatments, and mobilization strategies is lacking. The German prospective, multicenter, open-label, non-interventional OPTIMOB study addressed this lack of knowledge by analyzing mobilization and collection parameters in a large cohort of adult, transplant-eligible, good and poormobilizing patients with lymphoma or multiple myeloma. In this special issue, the results of this huge national study with 28 recruiting study centers are presented in two articles: Bittrich et al. [8] described the results in patients with multiple myeloma, whereas Kriegsmann et al. [9] showed the results of the cohort of lymphoma patients. A state-of-the-art collection of PBSCs includes a system of quality control and benchmarking. Approaches to quantitatively assess the effectiveness of autologous leukapheresis sessions have been developed, and some allow not only to calculate the collection efficiency (CE2) but also to m","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10601598/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71414022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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