ISSN 1866-8836
Клеточная терапия и трансплантация

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Volume 1, Number 2
12/01/2008
Volume 1, Number 2
Editor-in-Chief
Afanasyev B. V. (St. Petersburg, Russia)
Co-Editors-in-Chief
Wagemaker G. (Rotterdam, Netherlands)
Zander A. (Hamburg, Germany)
Deputy Editor
Chukhlovin A. B. (St. Petersburg, Russia)
Fehse B. (Hamburg, Germany)
Novik A. (Moscow, Russia)
Managing Editor
Claudia Koltzenburg (Hamburg, Germany)
Editorial Board
Aleynikova O. (Minsk, Belarus)
Alyansky A. (St. Petersburg, Russia)
Anagnostou A. (Boston, USA)
Andreeff M. (Houston, USA)
Bacher U. (Hamburg, Germany)
Baуkov V. (St. Petersburg, Russia)
Baranov V. S. (St. Petersburg, Russia)
Barkhatov I. (St. Petersburg, Russia)
Baum C. (Hannover, Germany)
Bilko N. (Kiev, Ukraine)
Borset M. (Trondheim, Norway)
Buechner Th. (Muenster, Germany)
Bykov V. (St. Petersburg, Russia)
Dini G. (Genoa, Italy)
Drize N. (Moscow, Russia)
Egeland T. (Oslo, Norway)
Elstner E. (Berlin, Germany)
Emanuel V. (St. Petersburg, Russia)
Everaus H. (Tartu, Estonia)
Ferrara J. (Ann Arbor, USA)
Fibbe W. (Leiden, Netherlands)
Galibin O. (St. Petersburg, Russia)
Ganser A. (Hannover, Germany)
Granov D. (St. Petersburg, Russia)
Ivanov R. (Moscow, Russia)
Klimko N. (St. Petersburg, Russia)
Kolb H.-J. (Muenchen, Germany)
Konopleva M. (Houston, USA)
Koza V. (Pilsen, Czech Republic)
Kroeger N. (Hamburg, Germany)
Malikov A. (St. Petersburg, Russia)
Mikhailova N. (St. Petersburg, Russia)
Mentkevich G. (Moscow, Russia)
Nagler A. (Tel Hashomer, Israel)
Nemkov A. (St. Petersburg, Russia)
Neth R. (Hamburg, Germany)
Nevorotin A.J. (St. Petersburg, Russia)
Ostertag W. (Hamburg, Germany)
Palutke M. (Detroit, USA)
Roumiantsev A. G. (Moscow, Russia)
Savchenko V. G. (Moscow, Russia)
Smirnov A. V. (St. Petersburg, Russia)
Stamm C. (Berlin, Germany)
Tetz V. (St. Petersburg, Russia)
To B. (Adelaide, Australia)
Totolian A. A. (St. Petersburg, Russia)
Uss A.L. (Minsk, Belarus)
Vilesov A. (St. Petersburg, Russia)
Westenfelder Ch. (Salt Lake City, USA)
Wisloff F. (Oslo, Norway)
Zubarovskaya L. (St. Petersburg, Russia)
Zvartau E. (St. Petersburg, Russia)
In this Issue

Stepping ahead

Only a few months have passed since the launch issue of CTT, and the feedback from authors, reviewers, and readers has been a rewarding experience.

Apart from current articles and reviews on an interesting range of topics, this issue contains a special section on mesenchymal cell therapy, with 7 articles out of 15 dedicated to this topic.

In the same section, in addition to the MSC articles, Mariusz Z. Ratajczak et al. introduce the very small embryonic-like stem cells (VSEL) and their potential for the regeneration of tissues.

From a related field, we publish „In and out of mitosis“, Tim Hunt‘s Special Lecture given at the Wilsede Meeting 2008. This is published jointly with our sister portal www.wilsede-science-connections.com, whose multimedia resources we are starting to make available to a wider audience.

We particularly wish to encourage young medical researchers to get their cutting edge work peer-reviewed and published in CTT. From this issue onwards, the Contents page will include an icon called „FirstIA“ to highlight the authors for whom this CTT article is their first international article.

We would like to thank the following colleagues for undertaking the peer reviews for this issue of CTT: Ulrike Bacher, Alexey B. Chukhlovin, Boris Fehse, Nikolay N. Klimko, Nicolaus Kröger, Claudia Lange, Katarina Le Blanc, Nikolai N. Mamaev, and Ludmila S. Zubarovskaya.

Boris Vladimirovich Afanasyev and Axel Rolf Zander

Your Managing editor speaking

Since CTT is an open access journal, readers as well as authors can benefit enormously. How do authors benefit? You can link directly from your publication lists on the web to your CTT article. Your reader can click on a link that makes the article accessible immediately and without any cost barriers. This means that any web reader anywhere has access to your article and, without any extra effort, can cite your CTT article in their work.

CTT articles are already being indexed in CAS and DOAJ, with more visibility measures to come.

It is a great pleasure for me to run this journal on the managerial and production side together with our Editorial Office team located in both Hamburg and St. Petersburg. Our production team, too, is a meeting point for different lines of thought and it is an exceptional experience to develop a scientific journal together in managerial and technological respects.

I wish to thank the following colleagues for helping us start off so well: bitfarmers.com (with Michael Hirdes, Gina Steiner and Michael Sendke), René Hornung, Liudmila Lashkouskaya, Viktoriya Levenko, Yana Onikiychuk, Melissa Pritchard, Jutta Roebers, Anna Starikova, and Oksana Zhebel.

If you as authors and/or readers wish to contribute any questions, suggestions or comments, these are highly appreciated. Feel free to contact me directly, thank you.

Claudia Koltzenburg
managingeditor@ctt-journal.com

Keynote

Articles on mesenchymal cell therapy

Initial report on a phase I clinical trial: Prevention and treatment of post-operative Acute Kidney Injury with allogeneic Mesenchymal Stem Cells in patients who require on-pump cardiac surgery

Anna Gooch1, John Doty2, Jean Flores2, LeAnne Swenson2, Florian E Toegel1,3, George R Reiss4, Claudia Lange5, Axel R Zander5, Zhuma Hu1, Scott Poole1, Ping Zhang1 and Christof Westenfelder1,6 

A unique population of mobile very small embryonic/epiblast like (VSEL) stem cells resides in adult tissues: physiological and pathological consequences

Mariusz Z. Ratajczak1,2, Magda Kucia1, Dong-Myung Shin1, Liu Rui1, Justyna Drukala1, Wojtek Marlicz2, Janina Ratajczak1, Ewa K. Zuba-Surma1

Composition and functional properties of monolayer cell culture from human umbilical cord blood

Barkhatov I. M.1, Roumiantsev S. A.2, Vladimirskaya E. B.2, Afanasyev B. V.1

Experience of clinical mesenchymal stem cells (MSCs) usage for prophylaxis and GVHD treatment in patients undergoing allo-HSCT

Stankevich Y.1, Golovacheva A.1, Babenko E.1, Alyansky A.1, Paina O.1, Zubarovskaya L.1, Semenova E.1, Polintsev D.2,
Kruglyakov P.2, Afanasyev B.1

Autologous hematopoietic stem cell transplantation in multiple sclerosis

Yury L. Shevchenko1, Andrei A. Novik1, Alexey N. Kuznetsov1, Boris V. Afanasiev2, Igor A. Lisukov3, Oleg A. Rykavicin4, Аlexandr A. Myasnikov5, Vladimir Y. Melnichenko1, Denis A. Fedorenko1, Tatyana I. Ionova6, Roman A. Ivanov1, and Gary Gorodokin7 on behalf of the Russian Cooperative Group for Cellular Therapy

Review articles on mesenchymal cell therapy

Mesenchymal stem cells: precursor hierarchy

Irina Shipounova (Nifontova), Daria Svinareva, Josef Chertkov, Nina Drize

Platelet lysate for rapid expansion of human mesenchymal stromal cells

Claudia Lange, Figen Cakiroglu, Andrej Spiess, Heike Cappallo-Obermann, Axel R. Zander

Articles

Caspofungin as secondary prophylaxis or therapy in patients undergoing allogeneic stem cell transplantation with a prior or ongoing history of systemic or invasive fungal infections

N. Stute, T. Zabelina, N. Fehse, W. Hassenpflug, J. Panse, C. Wolschke, F. Ayuk, H. Schieder, H. Renges, A. Kratochwille, R. von Hinüber, R. Erttmann, A.R. Zander, N. Kröger

The use of Imatinib mesylate in Nigerians with chronic myeloid leukemia

Prof. Muheez A. Durosinmi1, Prof. Julius O. Faluyi2, Dr. Anthony A. Oyekunle1, Dr. Lateef Salawu1, Dr. Ismail A. Adediran1, Dr. Norah O. Akinola1, Oluwakemi O. Bamgbade3, Dr. Charles C. Okanny4, Dr. Sulaiman Akanmu4, Dr. Oche P. Ogbe5, Dr. Tambi T. Wakama5, Dr. Chijoke A. Nwauche6, Dr. Matthew E. Enosolease7, Dr. Daye N.K. Halim7, Dr. Godwin N. Bazuaye7, Dr. Chide E. Okocha8, Dr. J. A. Olaniyi9, Dr. Titi S. Akingbola9, Dr. Victor O. Mabayoje10, Dr. Ajani A. Raji10, Dr. Aisha Mamman11, Dr. Aisha Kuliya-Gwarzo12, Dr. Obike G. Ibegbulam13, Dr. Sunday Ocheni13, Dr. Yohanna Tanko14, Dr. Oladimeji P. Arewa1, Dr. Rahman A.A. Bolarinwa1, Dr. Davidson O. Kassim1, Dr. Mohammed A. Ndakotsu1, Dr. Omotilewa A. Amusu15, Prof. O. O. Akinyanju16

Clonal composition of hematopoietic tissue in reconstituted mice after repeated treatment with G-CSF

Irina Shipounova (Nifontova), Natalia Sats, Daria Svinareva, Tatiana Petrova, Nina Drize, Valeriy Savchenko

Detecting the V617F mutation of the Jak2 gene in patients with myeloproliferative disorders

Irina Y. Saburova, Yana S. Onikiychuk, Irina I. Zotova, Galina N. Sologub, Mikhail I. Zarayskiy

Review articles

Generation of regulatory T cells by T cell receptor gene transfer

Graham P. Wright, Judy Wai-Ling King and Hans J. Stauss

Advances in T cell receptor gene transfer for immunotherapy

Judy Wai-Ling King, Graham P. Wright and Hans J. Stauss

Keynote

Getting in and out of mitosis (Video Lecture)

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Tim Hunt

Cell Cycle Control Laboratory, Cancer Research UK, South Mimms, UK

Cells enter mitosis (more generally, M-phase, and much of our work has been in frog oocytes and eggs and extracts thereof) when CDK1/cyclin complexes are activated. Phosphorylation by these, and other mitotic protein kinases, is responsible for reorganizing the cell and initiating progression to metaphase.
We would like to know how many proteins needs to be phosphorylated how much to bring about this state of affairs, and have been trying to enumerate the mitotic targets for various cyclin-CDK combinations for some time. I’ll talk about our approaches, difficulties and findings.
Exit from mitosis, starting at the metaphase to anaphase transition, occurs when the anaphasepromoting factor (APC/ C) is activated and tags a small number of target proteins, including cyclins and securin, with polyubiquitin chains that signal their proteolysis by the proteasome. Chromatids part and move to opposite poles of the cell where they decondense and re-form a functional nucleus.
Cytokinesis separates the two daughter cells. Mitotic phosphoproteins revert to their interphase un- or hypo-phosphorylated state.
We recently made the accidental discovery that the activity responsible for this postmitotic dephosphorylation is almost completely inactive in M-phase cell extracts, and is reactivated when cells exit mitosis. This explains how proteins can become almost completely converted to hyperphosphorylated states: not only are kinases activated, but the counteracting phosphatase(s) are concomitantly shut down. I will present the evidence that has led us to this conclusion. It stems from studies of frog egg extracts released from cytostatic factor (CSF) arrest by added CaCl2, and the discovery that calcineurin (protein phosphatase 2B) plays a role in escaping the clutches of CSF. But the real work of restoring proteins to their interphase state of hypophosphorylation is performed by an activity we call ‘Phosphatase X’, whose identity and regulation I shall discuss.

Articles on mesenchymal cell therapy

Initial report on a phase I clinical trial: Prevention and treatment of post-operative Acute Kidney Injury with allogeneic Mesenchymal Stem Cells in patients who require on-pump cardiac surgery

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Anna Gooch1, John Doty2, Jean Flores2, LeAnne Swenson2, Florian E Toegel1,3, George R Reiss4, Claudia Lange5, Axel R Zander5, Zhuma Hu1, Scott Poole1, Ping Zhang1 and Christof Westenfelder1,6 

1Division of Nephrology, Department of Medicine, University of Utah Health Sciences Center and George E. Wahlen VA HCS, Salt Lake City, Utah, USA; 2Division of Cardiovascular Surgery, Intermountain Medical Center, Murray, Utah, USA; 3Jacobi Hospital, Albert Einstein College of Medicine affiliated Medical Center, Bronx, New York, USA; 4Division of Cardiovascular Surgery, Department of Surgery, University of Utah Health Sciences Center, and Research Service, George E. Wahlen VA HCS, Salt Lake City, Utah, USA; 5Bone Marrow Transplantation Center, University of Hamburg, Germany; 6Department of Physiology, University of Utah Health Sciences Center, Salt Lake City, Utah, USA
 
Correspondence:
Christof Westenfelder, MD, Section of Nephrology (111 N), George E. Wahlen VA Health Sciences Center, 500 Foothill Blvd., Salt Lake City, UT 84148, USA
E-mail: christof.westenfelder@spam is badhsc.utah.edu

Based on our extensive pre-clinical data that show that ischemia/reperfusion-induced Acute Kidney Injury (AKI), an essentially treatment resistant complication in patients, can be effectively treated by the administration of allogeneic Mesenchymal Stem Cells (MSC), an FDA approved, Phase I Clinical Trial (www.clinicaltrials.gov; NCT00733876) in patients who are at high risk of developing severe AKI post open heart surgery is currently being conducted. In this safety trial, patients who are undergoing on-pump coronary artery bypass surgery or cardiac valve repair, who are older than 65 years, with underlying renal disease, diabetes mellitus, hypertension, coronary artery disease, congestive heart failure and/or chronic obstructive pulmonary disease will be infused with allogeneic MSC following completion of surgery. The MSC are dosed in an escalating fashion, the initial five patients being infused via a femoral catheter that is placed into the suprarenal aorta with a defined low dose of MSC/kg body weight. This report summarizes the clinical course of the first five patients that have been treated according to this protocol. The renal function did not deteriorate post operatively in any of these patients, nor were adverse (AE) or severe adverse events (SAE) observed to date. However, one patient died suddenly 26 days after discharge from causes that both the principal investigator and the members of the Data and Safety Monitoring Board judged as being unrelated to the study drug and its route of administration. The next group of five study subjects will receive an intermediate dose of MSC/kg body weight, and if no safety concerns arise with this dose, the final five patients will be treated with a high dose of MSC/kg body weight. Preliminary efficacy of MSC therapy in the prevention and treatment of post-operative AKI in this high risk cohort of cardiac surgery patients will be assessed by comparing outcomes in study subjects (frequency, severity and duration of post-operative AKI, dialysis dependency [temporary, permanent], length of stay, and death at 30 days) to those in a large number of historical controls (data base at www.STS.org).

Articles on mesenchymal cell therapy

A unique population of mobile very small embryonic/epiblast like (VSEL) stem cells resides in adult tissues: physiological and pathological consequences

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Mariusz Z. Ratajczak1,2, Magda Kucia1, Dong-Myung Shin1, Liu Rui1, Justyna Drukala1, Wojtek Marlicz2, Janina Ratajczak1, Ewa K. Zuba-Surma1

1Stem Cell Institute at James Graham Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA; 
2Department of Physiopathology, Pomeranian Medical University, Szczecin, Poland

Accumulating evidence demonstrates that adult tissue contains a population of very primitive pluripotent stem cells (PSCs). Recently, our group identified a population of very small SCs in murine bone marrow (BM) and other adult organs that express several markers characteristic for epiblast/germ line-derived SCs. We named these rare cells “very small embryonic like stem cells (VSELs).” We hypothesized that these cells, which are deposited during early gastrulation in developing tissues/organs, play an important role in the turnover of tissue-specific/committed SCs. Based on this, we envision that germ line is not only the origin but also a “basis/skeleton” for the SC compartment in adult life forms. We noticed that VSELs could be mobilized into peripheral blood (PB) and the number of these cells circulating in PB increases during stress and tissue/organ injuries (e.g., heart infarct, stroke). Furthermore, our data indicates that VSELs are protected from uncontrolled proliferation and teratoma formation by a unique pattern of methylation of selected somatic imprinted genes. Finally, we envision that in pathological situations, VSELs could be involved in the development of some malignancies (e.g., teratomas, germinal tumors, pediatric sarcomas).

Articles on mesenchymal cell therapy

Composition and functional properties of monolayer cell culture from human umbilical cord blood

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Barkhatov I. M.1, Roumiantsev S. A.2, Vladimirskaya E. B.2, Afanasyev B. V.1

1Saint-Petersburg Pavlov State Medical University, Russia;
2Russian Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia

Summary

Objectives

It’s known that during cultivation, adherent cells of umbilical cord blood (UCB) form a monolayer reminiscent, in its composition, of the stromal monolayer of bone marrow (BM) culture. However, the presence of mesenchymal stem cells (MSCs) in UCB still remains uncertain. This study was performed to investigate the composition and some functional characteristics of MSC-like cell populations revealed in the cord blood monolayer culture.

Materials and methods

Forty-three human UCB samples were under study. All the samples were obtained during full-term deliveries. To produce monolayer cultures, mononuclear cell fractions from UCB were cultivated in a culture medium containing DMEM with 20% FCS, supplied with 1% Pen/Strep. Phenotypic patterns of UCB culture were assessed with a panel of monoclonal antibodies specific for CD34; CD117; CD45; CD14; CD3; CD19; CD31; CD90; HLA DR; and HLA ABC. To determine the functional characteristics of MSCs derived from UCB culture, their differentiation ability and stimulation of hematopoietic colony formation activity were evaluated.

Results

In most cases, the cultures of plastic-adherent cells proved to be heterogeneous. Both spindle-shaped and polygonal cells were observed. In some samples, clonal growth could be detected. However, the number of fibroblastoid cells did not increase 100 cells per colony. Large colonies were registered in three UCB samples of the 43 under study. As evidenced by immune phenotyping, the monolayer UCB cultures were rather polymorphic and dissimilar in each sample. Most of the cells present in the cultures were macrophages (CD45+). However, we also found different amounts of presumably mesenchymal cells, including cells with an endothelial phenotype (CD34+CD31+).

Specific staining showed that the cells from a UCB monolayer culture have the capacity to differentiate into adipocytes and osteoblasts. In some cultures, however, induction of differentiation lead to the detachment of a major cell fraction. Hemostimulatory ability of UCB monolayer cultures depended on the phenotype composition of the monolayer culture. CD45+ and CD14+ cells, evidently, are stimulatory for granulocyte-macrophage colony formation. Moreover, levels of non-hematopoietic subpopulations (CD90+CD31-) in UCB cultures showed a direct correlation with the numbers of CFU-GM colonies produced.

Conclusion

UCB contains a subpopulation of non-hematopoietic cells possessing phenotypic and some functional characteristics of bone marrow derived mesenchymal stem cells. However, the low content and variable numbers of such cells provide some doubts on the viability of UCB as an alternative source for MSC.

Articles on mesenchymal cell therapy

Experience of clinical mesenchymal stem cells (MSCs) usage for prophylaxis and GVHD treatment in patients undergoing allo-HSCT

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Stankevich Y.1, Golovacheva A.1, Babenko E.1, Alyansky A.1, Paina O.1, Zubarovskaya L.1, Semenova E.1, Polintsev D.2,
Kruglyakov P.2, Afanasyev B.1

1Pavlov State Medical University, St. Petersburg, Russia;
2"TransTechnology" LtD, St. Petersburg, Russia

Summary

Within bone marrow stroma, there exist subsets of nonhematopoietic cells referred to as mesenchymal stem cells (MSCs), or mesenchymal stromal cells [1]. These cells may not only improve HSC engraftment and regeneration of damaged tissues after allogeneic transplantation [7], but also modulate immune responses in vitro and in vivo [8]. Hence, co-transplantation of allogeneic HSC together with allogeneic MSC hypothetically could provide some beneficial effects, such as enhanced engraftment, acceleration of immune reconstitution [4], GVHD suppression, and it may be used for GVHD prophylaxis, like as for treatment of severe acute or chronic GVHD. This study shows that more than a half of the patients with steroid-refractory acute GVHD responded to treatment with MSCs. However, further randomized clinical trials are necessary for estimation of therapeutic effect of MSCs in allo-HSCT patients and definition of important and significant factors influenced upon MSCs infusion.

Articles on mesenchymal cell therapy

Autologous hematopoietic stem cell transplantation in multiple sclerosis

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Yury L. Shevchenko1, Andrei A. Novik1, Alexey N. Kuznetsov1, Boris V. Afanasiev2, Igor A. Lisukov3, Oleg A. Rykavicin4, Аlexandr A. Myasnikov5, Vladimir Y. Melnichenko1, Denis A. Fedorenko1, Tatyana I. Ionova6, Roman A. Ivanov1, and Gary Gorodokin7 on behalf of the Russian Cooperative Group for Cellular Therapy

1Pirogov National Medical Surgical Center, Moscow, Russia;
2Pavlov State Medical University, St. Petersburg, Russia;
3Institute of Clinical Immunology, Siberian Branch of Russian Academy of Science, Novosibirsk, Russia;
4Burdenko Central Military Hospital, Moscow, Russia;
5Republic Hospital, Petrozavodsk, Russia;
6Multinational Center of Quality of Life Research, St. Petersburg, Russia;
7New Jersey Center for Quality of Life and Health Outcome Research, NJ, USA

Although there is no effective cure for this disease, high-dose chemotherapy (HDCT), together with autologous hematopoietic stem cell transplantation (auto-HSCT) offers promising results in the treatment of multiple sclerosis (MS) patients.

Methods

In this paper we present results of a prospective clinical study of safety and efficacy of HDCT+auto-HSCT in MS patients. One hundred and nine patients with various types of MS were included in this study. The patients underwent early, conventional, or salvage/late transplantation.

Results

The transplantation procedure was well tolerated by MS patients, with no transplant-related deaths at all. The efficacy analysis was performed in 79 patients. Forty-two achieved an objective improvement of neurological symptoms (defined as a ≥0.5 point decrease in EDSS score as compared to the baseline and confirmed over 6 months), and 37 patients had disease stabilization (steady EDSS level as compared to the baseline and confirmed over 6 months). Quality of life (QoL) was assessed in 44 patients. Thirty-nine patients exhibited a QoL response 1 year after transplantation.

Conclusions

This study provides ample evidence in support of HDCT+auto-HSCT efficacy in MS patients. The results obtained show that transplantation appears to be effective in patients with various types of MS.

Review articles on mesenchymal cell therapy

Mesenchymal stem cells: precursor hierarchy

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Irina Shipounova (Nifontova), Daria Svinareva, Josef Chertkov, Nina Drize

The hierarchy of stromal precursors is the focus of this research. It has been previously shown that transplantation of the bone marrow plug under the renal capsule of the syngeneic animal leads to the formation of the foci of ectopic hematopoiesis, where a stromal microenvironment is formed by the donor's mesenchymal stem cells (MSC). In the irradiated recipients such foci are 2-3 times larger than in non-irradiated foci due to "inducible" precursors that are more differentiated than MSC. Along with the in vivo tests, the method of in vitro estimation of concentration of clonogenic stromal precursors (CFU-F) is widely used. However, the hierarchical arrangement of the described precursors is still unclear. This study describes the alterations in the number of mentioned precursors in the ectopic hematopoietic foci formed in the irradiated recipients. CFU-F was shown to be the closest MSC progeny thus far, while "inducible" precursor cells – stromal multipotent precursors – are at a lower position in the hierarchy and possibly enlarge the hematopoietic territory in the irradiated recipients directly.

Review articles on mesenchymal cell therapy

Platelet lysate for rapid expansion of human mesenchymal stromal cells

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Claudia Lange, Figen Cakiroglu, Andrej Spiess, Heike Cappallo-Obermann, Axel R. Zander

Clinic for Stem Cell Transplantation, University Hospital Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany

Correspondence:
Claudia Lange, Clinic for Stem Cell Transplantation, University Hospital Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany,
E-mail: cllange@spam is baduke.uni-hamburg.de

Human bone marrow mesenchymal stromal cells (hMSC) are promising candidates for new treatment options in transplant and regenerative medicine. However, most expansion protocols still use fetal calf serum (FCS) as growth factor supplement, which is a potential source of undesirable xenogeneic pathogens. We established an easy and reproducible expansion protocol for hMSC based on the addition of platelet lysate (PL) obtained from human thrombocyte concentrates. Both CFU-F and cumulative cell numbers were significantly increased compared to the conventional FCS-based medium. The generated cells meet all criteria for MSCs, e.g. plastic adherence, spindle-shaped morphology, surface marker expression, lack of hematopoietic markers, and differentiation capability into 3 mesenchymal lineages. Human MSC expanded with PL revealed favorable immunological properties in vitro. Gene expression profiles showed upregulation of cell cycle and DNA replication genes and downregulation of developmental, differentiation, adipogenic and MHC II genes. Thus, PL provides a safe component for accelerated and safe hMSC expansion.

Articles

Caspofungin as secondary prophylaxis or therapy in patients undergoing allogeneic stem cell transplantation with a prior or ongoing history of systemic or invasive fungal infections

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N. Stute, T. Zabelina, N. Fehse, W. Hassenpflug, J. Panse, C. Wolschke, F. Ayuk, H. Schieder, H. Renges, A. Kratochwille, R. von Hinüber, R. Erttmann, A.R. Zander, N. Kröger

Dept of Stem Cell Transplantation, University Hospital Hamburg-Eppendorf, 20246 Hamburg, Germany


Correspondence:
Prof.  Dr. med. Nicolaus Kröger, Bone Marrow Transplant Center, University Hospital Hamburg-Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany, Tel.: +49-40-42803 5864, Fax: +49-40-42803 3795, E-mail: nkroeger@spam is baduke.uni-hamburg.de

Background

Patients with a history of or ongoing invasive fungal infection (IFI) who undergo allogeneic stem cell transplantation (SCT) have a high risk of reactivation or progression. In a prospective study we evaluated the efficacy and safety of caspofungin as secondary prophylaxis or as therapy for persistent disease.

Methods

Twenty-eight adult patients were included in this study, all of whom had acute leukemia. At the time of SCT 16 patients had no signs of infection, while in 12 cases radiographic signs (CT scan) of florid fungal infections were noted. Caspofungin 50 mg intravenously was given daily from start of conditioning until stable engraftment. 

Results

No patient experienced side effects leading to the discontinuation of caspofungin. In 14 out of 16 patients (88%) without active signs of infection at start of transplantation, no fungal disease was observed after prophylaxis with caspofungin. In 10 out of 12 cases (83%) with radiographic signs of florid fungal infection pre-transplantation, complete (n=4) or partial (n=6) responses after caspofungin treatment were achieved. 

Conclusions

The use of caspofungin is safe and effective in high-risk patients with a history of IFI when undergoing allogeneic SCT.

Articles

The use of Imatinib mesylate in Nigerians with chronic myeloid leukemia

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Prof. Muheez A. Durosinmi1, Prof. Julius O. Faluyi2, Dr. Anthony A. Oyekunle1, Dr. Lateef Salawu1, Dr. Ismail A. Adediran1, Dr. Norah O. Akinola1, Oluwakemi O. Bamgbade3, Dr. Charles C. Okanny4, Dr. Sulaiman Akanmu4, Dr. Oche P. Ogbe5, Dr. Tambi T. Wakama5, Dr. Chijoke A. Nwauche6, Dr. Matthew E. Enosolease7, Dr. Daye N.K. Halim7, Dr. Godwin N. Bazuaye7, Dr. Chide E. Okocha8, Dr. J. A. Olaniyi9, Dr. Titi S. Akingbola9, Dr. Victor O. Mabayoje10, Dr. Ajani A. Raji10, Dr. Aisha Mamman11, Dr. Aisha Kuliya-Gwarzo12, Dr. Obike G. Ibegbulam13, Dr. Sunday Ocheni13, Dr. Yohanna Tanko14, Dr. Oladimeji P. Arewa1, Dr. Rahman A.A. Bolarinwa1, Dr. Davidson O. Kassim1, Dr. Mohammed A. Ndakotsu1, Dr. Omotilewa A. Amusu15, Prof. O. O. Akinyanju16

1Department of Haematology, Obafemi Awolowo University Teaching Hospital Complex, Ile-Ife, Nigeria; 2Department of Botany, Obafemi Awolowo University, Ile-Ife, Nigeria; 3Department of Pharmacy, Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria; 4Department of Haematology, University of Lagos, Nigeria; 5Department of Haematology, National Hospital, Abuja, Nigeria; 6Department of Haematology, University of Port-Harcourt, Port-Harcourt, Nigeria; 7Department of Haematology, University of Benin, Nigeria; 8Department of Haematology, Nnamdi Azikiwe University, Nnewi, Nigeria; 9Department of Haematology, University College Hospital, Ibadan, Nigeria; 10Department of Haematology, Ladoke Akintola University of Technology, Osogbo, Nigeria; 11Department of Haematology, Ahmadu Bello University, Zaria, Nigeria, 12Department of Haematology, Bayero University,  Kano, Nigeria; 13Department of Haematology, University of Nigeria, Enugu, Nigeria; 14Department of Haematology, Gwagwalada Specialist Hospital, Abuja, Nigeria; 15Department of Haematology, Army Reference Hospital, Lagos, Nigeria; 16Asaju Medical Clinic, Victoria Island, Lagos, Nigeria

Objectives

To assess response and toxicity to Imatinib mesylate (Glivec) in Nigerian Patients with chronic myeloid leukemia.

Methods

From August 2003 to August 2007, 98 consecutive, consenting patients, 56 (57%) males and 42 (43%) females, median age 36 years (range, 11-65 years) diagnosed with  CML, irrespective of disease phase received Imatinib at a dose of 300-600mg/day at the OAU Teaching Hospitals, Nigeria. Response to therapy was assessed by clinical, haematological and cytogenetic parameters. Blood counts were checked every two weeks in the first three months of therapy. Chromosome analysis was repeated sixth monthly. Overall survival (OS) and frequency of complete or major cytogenetic remission (CCR/MCR) were evaluated.  

Results

Complete haematologic remission was achieved in 64% and 83% of patients at one and three months, respectively. With a median follow-up of 25 months, the rates of CCR and MCR were 59% and 35% respectively. At 12 months of follow-up, OS and progression- free survival (PFS) were 96% and 91%, respectively. Achievement of CR at six months was associated with significantly better survival (p = 0.043).

Conclusions

Compared to treatment outcome with conventional chemotherapy and alpha interferon, as previously used in Nigeria, the results obtained with this regimen has established Imatinib as the first-line treatment strategy in patients with CML, as it is in other populations, with minimal morbidity.

Articles

Clonal composition of hematopoietic tissue in reconstituted mice after repeated treatment with G-CSF

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Irina Shipounova (Nifontova), Natalia Sats, Daria Svinareva, Tatiana Petrova, Nina Drize, Valeriy Savchenko

National Hematology Research Center, Russian Academy of Medical Science, Moscow, Russia

The clonal composition of hematopoietic tissue was studied in chimeras reconstituted with bone marrow cells expressing the human adenosine deaminase gene after repeated rounds of G-CSF treatment. Six courses
of G-CSF treatment led to an insignificant decrease of clone numbers and a considerable reduction in clone size and lifespan. The data suggest that the dissociation of the hematopoietic stem cells from their microenvironment after G-CSF treatment resulted in the exhaustion of clone size, and a decrease in the proliferative potential of the hematopoietic stem cells. Repeated G-CSF treatment adversely affects the hematopoietic system.

Articles

Detecting the V617F mutation of the Jak2 gene in patients with myeloproliferative disorders

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Irina Y. Saburova, Yana S. Onikiychuk, Irina I. Zotova, Galina N. Sologub, Mikhail I. Zarayskiy

Saint-Petersburg Pavlov State Medical University, Russia

Myeloproliferative disorders (MPD) are a heterogeneous group of hematopoietic diseases accompanied by multiple hyperplasia of bone marrow cells. They are rather difficult for diagnostics and often only revealed by excluding other conditions. One of the most valuable diagnostic criteria for MPD is the V617F mutation of the JAK2 gene. The main subject of this study was to develop a routine detection technique for the JAK2V617F mutation that will be useful for primary diagnostics. To do so, we developed two pairs of primers specific for mutated and wild-type JAK2. To ensure high sensitivity and specificity in JAK2V617F detection we first adjusted the novel PCR technique on the UKE1 cell line previously shown to be homozygous for the JAK2V617F mutation. Next we isolated genomic DNA from 58 MPD patients with different diagnoses using standard techniques. The overall mutation rate in this group was found to be 29.3%. The frequency of the JAK2V617F mutation in newly diagnosed patients with non-verified MPD was 25.7%. We conclude that the detection technique for the JAK2V617F mutation developed in our laboratory represents a useful tool as a diagnostic screening method in patients with myeloproliferative disorders.

Review articles

Generation of regulatory T cells by T cell receptor gene transfer

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Graham P. Wright, Judy Wai-Ling King and Hans J. Stauss

Department of Immunology, Royal Free Hospital, University College London, UK

Our lab and others have demonstrated T cell receptor (TCR) gene transfer as an efficient way of redirecting the specificity of a bulk T cell population to that of a known antigen. Thus far there has been considerable effort put into the use of TCR gene transfer into conventional CD8+ and CD4+ T cells in order to initiate or augment immune responses. There has, as yet, been little investigation into the potential use of TCR gene therapy at the other end of the spectrum: control of immune pathology using regulatory T cells. Here we will briefly discuss the evidence indicating that the generation of Ag-specific Tregs, potentially via TCR gene transfer, may be an efficacious treatment for various forms of immune-pathology and briefly outline the challenges towards realizing the full potential of this type of therapy.

Review articles

Advances in T cell receptor gene transfer for immunotherapy

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Judy Wai-Ling King, Graham P. Wright and Hans J. Stauss

Department of Immunology, Royal Free Hospital, University College London, UK

Adoptive T cell transfer has seen clinical success in the treatment of both malignancies and viral infections. However, one of the main limitations of this strategy has been the difficulty in producing sufficient quantities of antigen-specific T cells. In addition, donor lymphocyte infusions are commonly associated with graft-versus-host disease (GvHD), which carries with it significant morbidity and mortality. Retroviral T cell receptor (TCR) gene transfer is an attractive new strategy by which large numbers of autologous, antigen-specific T cells can be generated, since the TCR is the sole determinant of T cell specificity. The introduced TCR specificity can be targeted against viral antigens or poorly immunogenic targets such as tumor associated antigens, and recent clinical trial data has demonstrated the feasibility of this technique in melanoma patients. Furthermore, TCR gene transfer also has the potential to generate antigen-specific regulatory T cells. This review will focus on recent advances in the field of TCR gene transfer and explore the potential clinical applications of this strategy.

Review articles

Integration of leukemia research in Europe: the paradigm CML

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R. Hehlmann, S. Saußele

Medizinische Klinik, Medizinische Fakultät Mannheim der Universität Heidelberg, Wiesbadener Str. 7-11, 68305 Mannheim, Germany

Presented at the 2nd Raissa Gorbacheva Memorial Symposium in St. Petersburg, Russia, on 20 September 2008.

Leukemias are rare diseases, the investigation into which requires multicenter activities, study groups and networking. The ELN integrates leukemia research and trial groups across Europe. Progress with CML continues to promote European integration (EUTOS for CML). Median survival with CML is now expected at 25 years. The main treatment options for CML are TK inhibition and allo-SCT. Treatment optimization trials are ongoing worldwide. Looking at the speed of current progress, the prospects for a cure of CML, and possibly other forms of leukemia, are good.