Objective： Umbilical cord blood stem cell transplantation （CBSCT） has approached significant success in leukemia treatment, but it is associated with higher rates of delayed or failed engraftment and relapse. This ...Objective： Umbilical cord blood stem cell transplantation （CBSCT） has approached significant success in leukemia treatment, but it is associated with higher rates of delayed or failed engraftment and relapse. This may be caused by immature immune calls of umbilical cord blood. We try to expand stem/progenitor calls and T, NK, DC immunocytes from umbilical cord blood for transplantation and immunotherapy. Methods： CB MNCs were cultured and analyzed for progenitor/stem calls, immunocytes at day 0, 3, 7 and 14 by using flowcytometry. Results： The combinations of SCF, IL-3, IL-6, plus IL-2 or/and IL-4 showed significantly expanded results both for UCB MNCs and CD34^＋ cells. CD34^＋ percentage went up from fresh CB 1.6% to the highest group E （SCF＋IL-3, 6, 2, 4） 11.1%. The average expansion multiples of CD34^＋ calls at 7th culture days were from 10 to 50 （SCF＋IL-3, 6, 2, 4）. The CD3^＋ T cells was （18.7±4.3）% in fresh cord blood, and decreased sharply in the medium without cytokine, while markedly increased in groups with cytokines combination, in group B, E, G and F, their level were about 2 times of fresh control. The fresh UCB contained （3.6±1.9）% CD56^＋ calls, NK cells only were expanded in groups with IL-2. DCs markers CDla, CD80, CD83 and CD86 expressed a lower level at day 3 in all test groups, and then increased sharply in groups E, F and G with IL-4 cytokin at 7th culture days. Conclusion： T calls, NK cells and DCs as well as stem/progenitor calls could be expanded in the same medium from CB MNCs with the combinations of cytokines. The combination of SCF, IL-2, IL-3, IL-6 and IL-4 showed a balancaable expansion result of both CD34^＋ cells and immunocytes at 7th culture days.展开更多
Background:With an increasing number of patients with hematological malignancies being treated with umbilical cord blood transplantation(UCBT),the correlation between immune reconstitution(IR)after UCBT and graft-vers...Background:With an increasing number of patients with hematological malignancies being treated with umbilical cord blood transplantation(UCBT),the correlation between immune reconstitution(IR)after UCBT and graft-versus-host disease(GVHD)has been reported successively,but reports on double-negative T(DNT)cell reconstitution and its association with acute GVHD(aGVHD)after UCBT are lacking.Methods:A population-based observational study was conducted among 131 patients with hematological malignancies who underwent single-unit UCBT as their first transplant at the Department of Hematology,the First Affiliated Hospital of USTC,between August 2018 and June 2021.IR differences were compared between the patients with and without aGVHD.Results:The absolute number of DNT cells in the healthy Chinese population was 109(70-157)/μL,accounting for 5.82(3.98-8.19)%of lymphocytes.DNT cells showed delayed recovery and could not reach their normal levels even one year after transplantation.Importantly,the absolute number and percentage of DNT cells were significantly higher in UCBT patients without aGVHD than in those with aGVHD within one year(F=4.684,P=0.039 and F=5.583,P=0.026,respectively).In addition,the number of DNT cells in the first month after transplantation decreased significantly with the degree of aGVHD increased,and faster DNT cell reconstitution in the first month after UCBT was an independent protective factor for aGVHD(HR=0.46,95%confidence interval[CI]:0.23-0.93;P=0.031).Conclusions:Compared to the number of DNT cells in Chinese healthy people,the reconstitution of DNT cells in adults with hematological malignancies after UCBT was slow.In addition,the faster reconstitution of DNT cells in the early stage after transplantation was associated with a lower incidence of aGVHD.展开更多
To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α...To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α and rhFL for 10 days. Then the induced cells were characterized by DC′s morphological and phenotypic properties. In addition, they stimulated the proliferation of allogeneic T cells and possessed an efficient capacity for initiating T cell dependent antitumor immune responses in vitro. It is concluded that mature DCs could be obtained from human cord blood CD34 + cells.展开更多
基金Supported by a grant from Posteoctoral Sciences Foundation of China (No.2002031281).
文摘Objective： Umbilical cord blood stem cell transplantation （CBSCT） has approached significant success in leukemia treatment, but it is associated with higher rates of delayed or failed engraftment and relapse. This may be caused by immature immune calls of umbilical cord blood. We try to expand stem/progenitor calls and T, NK, DC immunocytes from umbilical cord blood for transplantation and immunotherapy. Methods： CB MNCs were cultured and analyzed for progenitor/stem calls, immunocytes at day 0, 3, 7 and 14 by using flowcytometry. Results： The combinations of SCF, IL-3, IL-6, plus IL-2 or/and IL-4 showed significantly expanded results both for UCB MNCs and CD34^＋ cells. CD34^＋ percentage went up from fresh CB 1.6% to the highest group E （SCF＋IL-3, 6, 2, 4） 11.1%. The average expansion multiples of CD34^＋ calls at 7th culture days were from 10 to 50 （SCF＋IL-3, 6, 2, 4）. The CD3^＋ T cells was （18.7±4.3）% in fresh cord blood, and decreased sharply in the medium without cytokine, while markedly increased in groups with cytokines combination, in group B, E, G and F, their level were about 2 times of fresh control. The fresh UCB contained （3.6±1.9）% CD56^＋ calls, NK cells only were expanded in groups with IL-2. DCs markers CDla, CD80, CD83 and CD86 expressed a lower level at day 3 in all test groups, and then increased sharply in groups E, F and G with IL-4 cytokin at 7th culture days. Conclusion： T calls, NK cells and DCs as well as stem/progenitor calls could be expanded in the same medium from CB MNCs with the combinations of cytokines. The combination of SCF, IL-2, IL-3, IL-6 and IL-4 showed a balancaable expansion result of both CD34^＋ cells and immunocytes at 7th culture days.
基金National Natural Science Foundation of China(Nos.82270223,82170209,and 82100230)Anhui Provincial Key Research and Development Project(No.2022e07020015)+1 种基金Anhui Health Research Project(No.AHWJ2022a 011)Fundamental Research Funds for the Central Universities(Nos.WK9110000204,and WK9110000168)
文摘Background:With an increasing number of patients with hematological malignancies being treated with umbilical cord blood transplantation(UCBT),the correlation between immune reconstitution(IR)after UCBT and graft-versus-host disease(GVHD)has been reported successively,but reports on double-negative T(DNT)cell reconstitution and its association with acute GVHD(aGVHD)after UCBT are lacking.Methods:A population-based observational study was conducted among 131 patients with hematological malignancies who underwent single-unit UCBT as their first transplant at the Department of Hematology,the First Affiliated Hospital of USTC,between August 2018 and June 2021.IR differences were compared between the patients with and without aGVHD.Results:The absolute number of DNT cells in the healthy Chinese population was 109(70-157)/μL,accounting for 5.82(3.98-8.19)%of lymphocytes.DNT cells showed delayed recovery and could not reach their normal levels even one year after transplantation.Importantly,the absolute number and percentage of DNT cells were significantly higher in UCBT patients without aGVHD than in those with aGVHD within one year(F=4.684,P=0.039 and F=5.583,P=0.026,respectively).In addition,the number of DNT cells in the first month after transplantation decreased significantly with the degree of aGVHD increased,and faster DNT cell reconstitution in the first month after UCBT was an independent protective factor for aGVHD(HR=0.46,95%confidence interval[CI]:0.23-0.93;P=0.031).Conclusions:Compared to the number of DNT cells in Chinese healthy people,the reconstitution of DNT cells in adults with hematological malignancies after UCBT was slow.In addition,the faster reconstitution of DNT cells in the early stage after transplantation was associated with a lower incidence of aGVHD.
文摘To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α and rhFL for 10 days. Then the induced cells were characterized by DC′s morphological and phenotypic properties. In addition, they stimulated the proliferation of allogeneic T cells and possessed an efficient capacity for initiating T cell dependent antitumor immune responses in vitro. It is concluded that mature DCs could be obtained from human cord blood CD34 + cells.
