PC84045 human lung carcinoma cells (PC84045 HLCCs) were mixed with the BM cells activated by anti-CD3 McAb and IL-2 and then CD3-ALTO4 bi-specific McAbs was put into the cell suspension. The mixed cells were cultured for 3 days in presence of IL-2 and IL-3. Tumor cell colony formation method in half-solid culture system was used to assess the purging efficiency (PE) of the activated T cells to PC84045 HLCCs. The ratio of T cells to PC84045 HLCCs was 8 to 1, the PE was 4 Logs. The rario of T cells to PC84045 HLCCs was 16 tol, the PE was 5 Logs. When the purged BM cells were injected into the NC nude mice, none of subcutaneous node was found. The number of the CFU-GM, BFU-E of the purged BM cells was more than 85 % compared with that of the fresh BM cells (P

Objective To study the feasibility of HLA non-identical allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBHSCT) for the treatment of patients with severe aplastic anemia (SAA). Methods Four patients with SAA received non-identical allo-PBHSCT from HLA one or three loci mismatched related donors. Donors were given G-CSF 5?g/(kg?d) for 5 days, then PBSC were harvested on days 4 and 5. Prime regimen consisted of fludarbine, CTX, ATG and TBI. Combination of CsA, MTX, MMF and Simulect (CD25 monoclonal antibody) were used for GVHD prophylaxis. Results Engraft was achieved in all patients. The median days of neutrophil exceeding 0.5?109/L and platelet exceeding 20?109/L were 15.3 days (range 14~16 days) and 17.5 days (range 16~19 days), respectively. In the 14-month median follow-up duration, only one of four patients developed grade Ⅱ aGVHD, and one patient died of systemic candidiasis, the other three were alive in disease-free condition. Conclusion HLA non-identical allo-PBHSCT was well tolerated by patients with SAA, and it provided rapid and sustained engraftment without increase in incidence of GVHD.

0.05). The survival time of the second control group and experimental groups was much longer than that of the first control group (P

Objective:To investigate the effects of TJU103 on T lymphocytes proliferation and cytokine production in vitro.Methods:MTT and ELISA assays were used respectively to detect the proliferation and the cytokine production of donor T lymphocytes blocked by TJU103.Results:There are significant difference in the inhibition of T-cell proliferation and the cytokine production between groups of with TJU103 or without.Conclusion:TJU103 could significantly inhibit the T-cell proliferation and the cytokine production.

Objective To observe the effect of combination of Tju103 and CTLA4-Ig on engraftment, graft versus host disease (GVHD), graft versus leukemia (GVL) and anti-infection post major histocompatibility complex (MHC) haploidentical bone marrow transplantation in mice in order to seek an effective access to transplantation with less GVHD and more potential GVL and anti-infection.Methods In the presence of the recipient's antigen (normal CB6F1, H-2 bd) as a stimulus for induction of specific immune tolerance, T cells from the MHC haploidentical donors (C57BL/6, H-2 b) were first in vitro cultured with Tju103 and CTLA4-Ig, then were transfused with the donors' bone marrow cells into the preconditioned recipients. At last, the effect of combination of Tju103 and CTLA4-Ig on hematopoietic rebuilding, GVHD, GVL and anti-infection was observed in compared with CsA, Tju103 and CTLA4-Ig as controls.Results The only irradiated group (group A): All the mice (10 mice) died of failure of hematopoiesis within 11 days post irradiation, of which most (8 mice) died within 4～7 days post transplantation. The CTX-treated leukemia group (group B): All the mice (10 mice) died of leukemia within 16～23 days post leukemia cells infusion (11～18 days post BMT). CTX treatment prolonged the survival time. The only transplanted group (group C): The mice began to die from day 16 post transplantation, and all (10 mice) died of GVHD within 3 weeks. The CsA prophylaxis group (group D): 4 mice died within 8～22 days after transplantation, of which one died of leukemia, two died of infection and one died of GVHD, and the remaining 6 survived over 30 days post transplantation. The Tju103 treated group (group E): 4 mice died within 9～26 days post transplantation, of which one died of leukemia, one died of infection and two died of GVHD, and the remaining 6 mice survived over 30 days post transplantation. The CTLA4-Ig treated group (group F): 3 mice died within 14～23 days after transplantation, of which one died of infection and two died of GVHD, and the remaining 7 survived over 30 days post transplantation. The Tju103/CTLA4-Ig treated group (group G): one died of GVHD on day 19 after transplantation, and the remaining 9 mice survived over 30 days post transplantation. Conclusions CsA, Tju103 or CTLA4-Ig alone could prolong survival time and reduce incidence and degree of GVHD severity. But CTLA4-Ig could spare much ability of GVL and anti-infection while Tju103, just like CsA, couldn't. Combination of both was the most favorable way for transplantation with the most remarkable efficiency on prolongation of survival time and reduction of GVHD.

Objective To investigate the effect of donors' activating killer cell immunoglobulin-like receptor (aKIR) on recipients prognosis post haemopoietic stem cell transplantation (HSCT).Methods Fifty-nine cases of related HLA full matched HSCT were studied. Sequence specific primer polymerase chain reaction (SSP-PCR) was used to type donors' KIR. Virus,bacterium,and fungi infection as well as bleeding,relapse,survival were observed in the recipients post transplantation. Relationship between aKIR and clinical indexes mentioned above were analyzed.Results Donors' aKIR showed no beneficial effects on recipients' virus and bacterium infection. Furthermore,the incidence of fungi infection was increased in HSCT if donors expressed KIR3DS1 (?2= 4.804 ,P= 0.028 ). No significant difference was found in survival curve between donor aKIR positive HSCT or negative one (KIR2DS1+/-: P= 0.651 ; KIR2DS2+/-: P= 0.847 ; KIR3DS1+/-: P= 0.341 ). No effect of donor's aKIR on relapse was observed,too.Conclusion In Guangdong Han,so far as related HLA full matched HSCT is concerned,no improvement in recipient prognosis due to donor's aKIR is verified.

Objective To investigate the photodynamic therapy(PDT)of human erythroleukemia cell line K562 and drug-resistance cell line K562/ADM using hexyl 5-aminolevulinate(He-ALA)in vitro.Methods Four groups were set:PDT group(photosensitizer with light irradiation),laser group(light irradiation only),dark-cytoxocity group(photosensitizer only)and normal control group(neither photosensitizer nor light irradiation).K562 cells and K562/ADM cells cultured were incubated with various concentrations of He-ALA(0.025,0.1,0.4 and 1.6mmol/L)for 4 hours,then illuminated with different light doses(4.5,9,18 and 36 J/cm2)using a laser with a wave length of 630nm.After 12 hours incubation,Wright's staining method was used to observe the cell's morphological changes,the survival rates of cells were analyzed by CCK8 assay,and the changes in colony-forming abilities of cells were analyzed by methylcellulose colony-forming assay.Results He-ALA or light irradiation alone had no evident cytotoxicity,while the application of He-ALA plus light irradiation effectively killed the leukemia cells.With the increase of the concentrations of He-ALA and the dosages of irradiation,the viability of cells decreased.Under the same conditions,the survival rate of K562 was significantly lower than that of K562/ADM(P

BACKGROUND:Bone marrow mesenchymal stem cels have low immunogenicity and can induce immune tolerance. At present, the mechanism of immune regulation of bone marrow mesenchymal stem cels is not completely understood. It has been rarely reported whether the bone marrow mesenchymal stem cels can migrate to the thymus after transplantation.
OBJECTIVE:To observe the distribution and survival of bone marrow mesenchymal stem cels in the thymus of aging rats after transplantation.
METHODS: Bone marrow mesenchymal stem cels cultured in vitrowere transfected by adenovirus vectors expressing green fluorescent protein. Transfected bone marrow mesenchymal stem cels were injected into the portal vein of aging rats. At days 3, 7, 14, 21 after transplantation, the survival of bone marrow mesenchymal stem cels homing to the thymus was observed under fluorescence microscope. At day 3 after transplantation, thymus tissues were taken and stained with hematoxylin-eosin for pathological observation.
RESULTS AND CONCLUSION:Green fluorescent protein-labeled bone marrow mesenchymal stem cels had a strong green fluorescence at days 3 and 7 after transplantation, and the cel contour was clear. There was no significant difference in the mean absorbance values at days 3 and 7 (P> 0.05). Expression of green fluorescent protein was weakened significantly at days 14 and 21 compared with that at day 3 (P < 0.05). At 3 days after transplantation, the transplanted bone marrow mesenchymal stem cels were clearly visible in the thymus, and acute rejection was not observed. The results show that bone marrow mesenchymal stem cels can migrate to the damaged thymus tissue through the blood circulation, and can survive at least 1 week.

Objective To observe the clinical efficacy of staphylococcal protein A immunoadsorption plus nonmyeloablative chemotherapy with CD34+ autologous peripheral blood stem cell transplantation in the treatment of refractory systemic lupus erythematosus (SLE). Methods Three patients with active SLE were enrolled into this study. All patients were diagnosed with lupus nephritis by renal biopsy and poorly responded to routine therapy. Before transplantation, patients were given 6 sessions of immunoadsorption apheresis using columns of staphylococcal protein A-silica with an interval of 3 days; each session processed 3 L plasma and a total of 18 L plasma was processed over the 6 treatments. Three days following the immunoadsorption apheresis, the mobilization of stem cells was realized by intravenous cyclophosphamide at a dose of 2 g per square meter of body surface area and subcutaneous recombinant human granulocyte colony-stimulating factor (G-CSF) at a dose of 5 g per kilogram of body weight per day for 5 days. Then, peripheral blood raonoclonal cells were obtained by CS-3000 Cell Separator, and passed through the Clini Macs CD34+ cell selection device, with the final concentration of CD34+ cells being 2.6×106, 2.1×106 and 2.4×106 per kilogram of body weight respectively, and that of CD3+ cells being 3×105, 2.1×105, and 2.0×105 per kilogram of body weight, respectively, in these three patients. The conditioning regimen consisted of oral fludarabine of 50 mg/d for 5 days plus intravenous pig anti-human thymocyte immunoglobulin (ATG) at a daily dose of 90 mg/kg for 5 days. After 72-hour treatment with ATG, the frozen stem cells were infused back to the patients. Clinical manifestations and lupus-correlated immune parameters were compared in patients at baseline and after transplantation. Results Following immunoadsorption apheresis, an obvious decrease was observed in the level of serum anti-dsDNA, antinuclear antibody and IgG antibodies, while an increase in the level of serum complement 3. All patients achieved the reconstruction of hemopoiesis 2-3 days after the transplantation. Also, an apparent clinical remission was achieved with the SLEDAI score being less than 3. Six months after the transplantation, serum anti-dsDNA and antinuclear antibodies as well as urine protein were undetectable, the level of complement 3 reached the normal range, and renal function was restored. Conclusions Staphylococcal protein A immunoadsorption plus nonmyeloablative CD34+ autologous peripheral blood stem cell transplantation are effective and safe for refractory SLE, but the long-term effect remains to be connfirmed by further studies.

BACKGROUND:Bone marrow mesenchymal stem cells (BMSCs) can differentiate into renal paranchymal cells including renal intercapillary cells.BMSCs can repair kidney structure and function after damage.OBJECTIVE:To investigate renal histology and function changes following BMSC transplantation in a rat model of radiation-inducad damage.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Laboratory of the Department of Hematology,Zhujiang Hospital from January to October 2009.MATERIALS:A total of 35 clean male Sprague Dawley rats were selected.Of them,20 were used to prepare BMSCs.The remaining was randomly assigned to a normal control,model and cell transplantation groups,with 5 in each group.METHODS:Rat BMSCs were incubated by the whole bone marrow method.When 90% cells were confluent,BMSCs were digested in trypsin for subculture.In the model and cell transplantation groups,rats were used to establish radiation-induced models,and then underwent X-ray general irradiation,at a dose of 500 cGy/min,100 cm from the target,6 Gy each,once per week,for consecutively 3 weeks.24 hours following irradiation,BMSCs of 3 passage were collected at logarithmic phase.In the cell transplantation group,1 mL cell suspension was infused into the rat caudal vein,containing 3×10~6 cells,totally three times.In the normal control and model groups,rat caudal vein received an equal volume of saline.MAIN OUTCOME MEASURES:The following parameters were measured:results of hematoxylin-eosin staining;serum and kidney malondialdehyde (MDA) content and superoxide dismutase (SOD) activity;serum creatinine and urea nitrogen levels.RESULTS:Kidney histopathology demonstrated thin renal cortex,increased number of mesenchyme,uneven renal corpuscle,and disordered structure of renal glomerulus,narrow renal glomerular vessel,partial disappeared capsular space,and degeneration and sclerosis of some glomerulus in the model group.In the cell transplantation group,renal cortex became thick,with clear structure;interstitial hyperemia and edema was significantly relieved;many complete renal corpuscles were observed;partial renal glomerulus presented degeneration and sclerosis;significant capsular space could be seen.Oxygen free radical examination results showed that compared with the model group,SOD activity was significantly higher (P ＜ 0.05),MDA levels were significantly lower (P ＜0.05) in the cell transplantation group.Renal function examination results demonstrated that compared with the model group,serum creatinine and urea nitrogen levels were significantly reduced in the cell transplantation group (P ＜ 0.05).CONCLUSION:BMSC transplantation can effectively treat renal radiation injury and improve renal function.