Acceleration of Immune Reconstitution after Bone Marrow Transplantation in Mice by Bone Marrow Stromal Cell Line Transfected with IL-6 Gene

After bone marrow transplantation, dysfunction of immunesystem is a critical problem to be resolved in clinical treat ment. Efficient bone marrow transplantation needs com pleted immune reconstitution, which is associated to manyfactors. Among them, Chemo/radiotherapy after and/orbefore BMT can cause great damage to hematopoietic mi croenvironment, which decrease the ability of proliferationand differentiation of stem cell[1]. To observe the effectof bone marrow stromal cell on the imm…     

IL-3、IL-6联合转基因的基质细胞促进异基因小鼠骨髓移植后的免疫重建

探讨转染IL 3和IL 6基因的骨髓基质细胞系QXMSC1对异基因骨髓移植 (BMT )小鼠免疫功能重建的促进作用。用逆转录病毒载体将小鼠IL 3和IL 6基因转染到骨髓基质细胞系QXMSC1(H 2 d) ,分别建立基质细胞系QXMSC1IL 3、QXMSC1IL 6及QXMSC1IL 3/IL 6 ,供体BALB/c(H 2 d)小鼠骨髓去除T细胞 ,致死量照射 (9 0Gy)受体小鼠C5 7BL/ 6 (H 2 b)后 ,输入供体骨髓细胞 (1× 10 5/只 )的同时输入基质细胞QXMSC1IL 3/IL 6 (5× 10 5/只 )。在骨髓移植后 30d、 6 0d ,检测BMT小鼠脾细胞对LPS、ConA的反应强度、脾细胞中辅助性T淋巴细胞前体频数 (HTLp )、杀伤性T淋巴细胞前体频数 (CTLp )、抗体生成细胞 (PFC )的能力及对迟发型超敏反应 (DTH )的能力来评价BMT后免疫功能的恢复情况。异基因BMT并输入基质细胞系QXMSC1IL 3/IL 6可明显增强BMT后淋巴细胞对LPS、ConA反应强度 ,小鼠对异基因小鼠脾细胞DTH反应增强 ,脾淋巴细胞HTLp、CTLp及PFC数明显增加。基质细胞QXMSC1可作为有效的基因载体明显促进异基因骨髓移植小鼠免疫功能重建。联合转入IL 3和IL 6对BMT后免疫功能的恢复有协同作用     

Immune Reconstitution after Haploidentical Hematopoietic Stem Cell Transplantation

Haploidentical hematopoietic stem cell transplantation (HSCT) offers the benefits of rapid and nearly universal donor availability and has been accepted worldwide as an alternative treatment for patients with hematologic malignancies who do not have a completely HLA-matched sibling or who require urgent transplantation. Unfortunately, serious infections and leukemia relapse resulting from slow immune reconstitution remain the 2 most frequent causes of mortality in patients undergoing haploidentical HSCT, particularly in those receiving extensively T cell–depleted megadose CD34⁺ allografts. This review summarizes advances in immune recovery after haploidentical HSCT, focusing on the immune subsets likely to have the greatest impact on clinical outcomes. The progress made in accelerating immune reconstitution using different strategies after haploidentical HSCT is also discussed. It is our belief that a predictive immune subset–guided strategy to improve immune recovery might represent a future clinical direction.     

Immunodeficiency after Allogeneic Bone Marrow Transplantation in Man

This study was undertaken to clarify the mechanism behind the severely decreased lymphocyte proliferative response upon stimulation with mitogens and antigens seen after allogeneic bone marrow transplantation (BMT) in man. We investigated eight BMT patients and eight controls and found that the proliferative response of patient cells was reduced both when the cells were stimulated with phytohaemagglutinin (PHA) and when they were stimulated with a combination of phorbol myristate acetate (PMA), which is an activator of protein kinase C (PKC), and the calcium ionophore A23187, which irreversibly opens for calcium transport into the cell (median relative responses were 41 and 37%, respectively). However, the PHA-induced increase in the concentration of intracellular free calcium in post-BMT cells was not significantly different from the values found in control cells and the expression of interleukin 2 (IL-2) receptors (CD25) was only slightly decreased. However, the production of IL-2 was severely decreased in patient cells after stimulation with A23187/PMA (median 3541 units), although it was higher than in PHA-stimulated control cells (median 354 units). These results show that a direct activation of PKC by PMA combined with an increase in intracellular free calcium by A23187 cannot overcome the lymphocyte proliferation deficiency in cells from patients after allogeneic BMT. The data suggests that the defect is affecting the diacylglycerol pathway considerably more than the inositol triphosphate pathway.     

Use of Multivariate Immune Reconstitution Patterns to Describe Immune Reconstitution after Allogeneic Stem Cell Transplantation in Children

Immune reconstitution after hematopoietic stem cell transplantation (HSCT) is a complex process. Impacts of the reconstitution of different immune cells over time are complex and difficult to understand. New mathematical models are needed to better understand this process. In this study, we used principal component analysis to better analyze the process of immune reconstitution after HSCT. Forty-six consecutive patients receiving HSCT for malignant and nonmalignant disorders were included in the study. All patients were followed for at least 24 months after transplantation with regular blood sampling for analysis of lymphocyte subset numbers and function. Exponentially transformed lymphocyte subset counts and lymphocyte functional markers were analyzed to identify major trends in the reconstitution process. Using our multivariate model for mapping immune reconstitution after HSCT, we showed that dysfunctional reconstitution patterns precede severe complications, such as chronic graft-versus-host disease, relapse, and death.     

Involvement of MHC-Linked Hemopoietic-Histocompatibility Genes in Allogeneic Bone Marrow Transplantation in Mice

Genes controlling resistance of irradiated mice of allogeneic hemopoietic cells were mapped within, or closely linked to the D region of MHC and were designated Hemopoietic-histocompatibility genes (Hha). Hhp genes responsible for resistance to parenteral hemopoietic cells had also previously been detected on the D-end of MHC. Hh genes are regarded as determinants of cell surface antigens (Hh antigens) phenotypically expressed, in contrast to Histocompatibility antigens (H antigens) only on blood-forming and leukemic cells. The inheritance of Hh genes is not codominant, unlike thformed on peripheral blood lymphocytes and platelets. Sixteen Tunisian families were typed with 37 patients and 108 relatives. Genetic transmission of the disease of the HLA system seemed to be independent in this study. Comparison of HLA gene frequencies (unrelated) parents of patients and a control population showed no difference, proving that there is no clear association in population between deleterious XP genes and a particular HLA gene. However, an excess of identical HLA among pairs of diseased siblings would suggest that the disease is polymorphic and a form of the XP could be linked to HLA.     

Immune Reconstitution in Pediatric Aplastic Anemia after Allogeneic Hematopoietic Stem-cell Transplantation

Background: Previous studies had revealed that immune reconstitution (IR) after allogeneic hematopoietic stem-cell transplantation (allo-HSCT) affected the clinical prognosis of patients. However, few studies were based on pediatric patients and patients with aplastic anemia (AA). The purpose of this research was to analyze IR of pediatric AA after HSCT and further explore its clinical prognostic value.Methods: The whole of 61 pediatric patients with AA who underwent HSCT were enrolled. Lymphocyte subsets count in peripheral blood, CD4⁺/CD8⁺ T cell ratio, and serum concentration of immunoglobulins were detected using flow cytometry at regular intervals after HSCT.Results: Innate immunity recovered faster than adaptive immunity, T lymphocytes recovered faster than B lymphocytes. The number of transfused CD34⁺ cells and the implantation time of ANC significantly affected the early rapid IR of CD3⁺ T cells. The degree of HLA site coincidence significantly affected the early rapid IR of CD19⁺ B cells. The number of transfused MNC and CD34⁺ cells significantly affected the early rapid IR of CD56⁺ NK cells. The overall survival (OS) and failure-free survival (FFS) of CD56⁺ NK cells in early rapid IR group were higher than those in non-IR group. The CD3⁺ T cell early rapid IR group and CD8⁺ T cell early rapid IR group had higher OS than the non-IR group.Conclusion: Early rapid IR after HSCT is a good predictor of clinical prognosis in children with AA. This study provides a reasonable prediction for early rapid IR, which may improve clinical outcomes of children.     

移植耐受诱导的异基因胎肝移植小鼠的免疫功能重建

本文用门静脉注射异型脾细胞加腹腔注射环磷酰胺方法，成功地诱导了成年Ｂａｌｂ／ｃ小鼠（Ｈ－２ｄ）对Ｃ５７ＢＬ／６（Ｈ－２ｂ）小鼠的免疫耐受。致死照射的耐受Ｂａｌｂ／ｃ小鼠用Ｃ５７ＢＬ／６（Ｂ６）小鼠的胎肝细胞移植后，无移植排斥产生。嵌合状态分析的结果表明，在胎肝移植后９０ｄ和２４０ｄ，重建的Ｂａｌｂ／ｃ小鼠的脾细胞分别有７４．４％和８３．７％来自于供体Ｂ６小鼠．证明Ｂ６小鼠胎肝造血干细胞已经在致死照射的Ｂａｌｂ／ｃ小鼠体内稳定植入。免疫功能检测的结果表明，在胎肝移植后９０ｄ，照射Ｂａｌｂ／ｃ小鼠的免疫功能已经重建。     

Transplantation of Human Embryonic Myoblasts and Bone Marrow Stromal Cells into Skeletal Muscle of C57BL/10J-mdx Mice

We studied expression of dystrophin in skeletal muscles of C57BL/10J-mdx mice after transplantation of human embryonic and fetal myoblasts and bone marrow stromal cells. Dystrophin-positive areas corresponding to the location of transplanted cell were detected in muscles of all recipient mice after transplantation of different cell cultures, but the distribution of dystrophin characteristic of normal muscle fibers was detected only after transplantation of embryonic myoblasts. Dystrophin distribution in muscle fibers after transplantation of fetal myoblasts and bone marrow stromal cells was atypical.     

Counts of Stromal Precursor Cells in the Bone Marrow and Heterotopic Bone Marrow Transplants from Mice Immunized with Group A Streptococcus Antigens during Different Periods after Immunization

The counts of stromal precursor cells in bone marrow transplants obtained from animals 2 months after their immunization with killed type 5 group A streptococcus vaccine drop almost 3-fold in comparison with transplants from normal donors. Six months after donor immunization, the count of stromal precursor cells in the transplants reaches the normal level. The count of stromal precursor cells in bone marrow transplants from normal mice transplanted to recipients 6 months after their immunization with killed streptococcus vaccine also virtually did not change in comparison with the counts in bone marrow transplants from normal donors transplanted to normal recipients. The weight and size of bone capsules of 6.5-month bone marrow transplants in intact recipients after transplantation from donors immunized 2 months before with killed type 5 group A streptococcus vaccine was 3-fold lower than in bone marrow transplants collected from intact donors. The content of stromal precursor cells in the femoral bone marrow of animals immunized with killed streptococcus vaccine was 2.5 time higher in comparison with the parameter in the femoral bone marrow of normal mice even 8 months after immunization. The results indicate a significant long-acting effect of streptococcal antigens on the bone marrow stromal tissue, specifically, on its osteogenesis potential. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 147, No. 1, pp. 78-81, January, 2009     

Shortened-Duration Tacrolimus after Nonmyeloablative,HLA-Haploidentical Bone Marrow Transplantation

With post-transplantation cyclophosphamide (PTCy) as graft-versus-host disease (GVHD) prophylaxis, nonmyeloablative HLA-haploidentical (NMA haplo) and HLA-matched blood or marrow transplantation (BMT) have comparable outcomes. Early discontinuation of immunosuppression may reduce the risk of relapse and improve immune reconstitution, but may increase the risk of GVHD. We conducted a prospective trial of NMA haplo BMT for patients with hematologic malignancies (median age, 61 years), evaluating the safety of early discontinuation of tacrolimus. All patients received T cell-replete bone marrow followed by high-dose PTCy, mycophenolate mofetil, and tacrolimus. Tacrolimus was prespecified to stop without taper at day +90, +60, or +120, contingent on having ≥5% donor T cells, no relapse, and no grade II-IV acute or significant chronic GVHD. Safety stopping rules were based on ≥5% graft failure, ≥10% nonrelapse mortality (NRM), or a ≥20% combined incidence of severe acute and chronic GVHD from the tacrolimus stop date through day +180. Of the 47 patients in the day +90 arm, 23 (49%) stopped tacrolimus as planned. Of the 55 patients in the day +60 arm, 38 (69%) stopped as planned. Safety stopping criteria were not met. In both arms, at day +180, the probability of grade II-IV acute GVHD was <40%, that of grade III-IV acute GVHD was <8%, and that of NRM was <5%. The 1-year probabilities of chronic GVHD and NRM were <15% and <10%, respectively, in both arms. The 1-year GVHD-free relapse-free survival was higher in the day 60 arm. Thus, stopping tacrolimus as early as day +60 is feasible and carries acceptable risks after NMA haplo BMT with PTCy. This approach may facilitate post-transplantation strategies for relapse reduction.