RESUMO
OBJECTIVE: In our laboratory, we have developed an immunorosette technique for the depletion of T cells from bone marrow transplants. Tetrameric complexes of monoclonal antibodies are able to form very stable immunorosettes, which are efficiently depleted with the aid of a blood cell separator. Major improvements over the original sheep red blood cell depletion are the use of human (patient or donor derived) erythrocytes instead of sheep-derived cells, and the possibility of using a closed system for separation in a cell separator. In contrast to bone marrow, mobilized haematopoietic stem cell transplants obtained after leucocytapheresis contain higher numbers of T cells. Therefore, a different approach is necessary. METHOD: We have used two CD34 selection systems (Isolex 300SA and the Clinimacs) to perform T-cell depletions from peripheral blood stem cell (PBSC) transplants. RESULTS: Immunorosette T-cell depletion, with CD2/CD3 tetrameric complexes, of bone marrow transplants resulted in a mean 2.5 log depletion of T cells with a yield of 50% of the CD34+ cell population. Stem cell selection of PBSC transplants using one of the CD34 selection procedures resulted in a 4.5 log depiction of T cells for both systems, but with different results for the recovery of CD34+ cells. An increased yield of CD34+ cells was obtained with the Clinimacs procedure (57.9+/-9.0%) in comparison to the Isolex procedure (40.1+/-12.5%). CONCLUSION: Our own immunorosette depletion technique and the two tested CD34 selection methods for stem cell transplants both resulted in a very efficient T-cell depletion with the recovery of 40-60% of the CD34 haematopoietic stem cells present in the transplant.
Assuntos
Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/citologia , Depleção Linfocítica/métodos , Linfócitos T/citologia , Condicionamento Pré-Transplante/métodos , Antígenos CD34/metabolismo , Artrite Juvenil/terapia , Células da Medula Óssea/citologia , Antígenos CD2/metabolismo , Complexo CD3/metabolismo , Criança , Humanos , Linfócitos T/metabolismoAssuntos
Células Sanguíneas/citologia , Preservação de Sangue/métodos , Criopreservação/métodos , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/citologia , Ensaio de Unidades Formadoras de Colônias , Humanos , Leucaférese , Leucemia/sangue , Leucemia/terapia , Linfoma/sangue , Linfoma/terapia , Albumina Sérica , Transplante AutólogoAssuntos
Células da Medula Óssea , Neuroblastoma/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Anticorpos Monoclonais/uso terapêutico , Medula Óssea/imunologia , Criança , Proteínas do Sistema Complemento/uso terapêutico , Humanos , Formação de Roseta , Células Tumorais CultivadasRESUMO
In this article a technique is described for the cryopreservation of human lymphocytes employing a programmed cooling device. A preset temperature curve was used which compensated for the heat exchange during crystallization. The influence of this method of lymphocyte preservation on a variety of the in vitro functions of these cells was evaluated by means such as the lymphocyte transformation test, CML and investigations for lymphocyte membrane markers. All the lymphocyte functions that were tested were found to be retained in full.
