BACKGROUNDWe distinguished graft-versus-host disease (GVHD) from graft-versus-leukemia (GVL) effects and to investigate the distribution of T-cell receptor (TCR) V beta gene repertoire in individuals with leukemia before and after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSPeripheral blood mononuclear cells (PBMC) were obtained from 10 normal individuals, 8 donors and 11 patients with leukemia before and after transplantation. Polymerase chain reaction (PCR) amplification of complementarity-determining region 3 (CDR3) of 24 TCR V beta genes was used to examine serial samples of PBMC. The PCR products were further analyzed by genescan to evaluate clonality of T cells.

RESULTSThe 24 TCR V beta gene repertoire displayed highly diverse and polyclonal spectratypes in all normal individuals and 4 of 8 donors. Another 4 donors expressed part of the 24 TCR V beta subfamily and 1 donor had oligoclonality. The expressions of the 24 TCR V beta subfamilies were skewed and restricted in 11 leukemia patients before and after transplantation. Some absences of 24 TCR V beta subfamily expression were quite similar between the recipients pro-transplantation and related donors. The number of subfamilies expressed increased over time post-transplantation, but the restricted expressions of the subfamily could last 6 - 30 months after transplantation. All patients with GVHD and some without GVHD exhibited T cell clonal expansion. The expansive T cell clone was distributed in V beta 2-3, 16-17, 18-19, 21 and V beta 23 in patients with GVHD and in V beta 7, 9, 16 and 19 in patients without GVHD. One patient with syngeneic-HSCT (syn-HSCT) had V beta 15 and 16 T cell expansion after transplantation. One patient displayed V beta 18 T cell expansion after donor lymphocyte infusion (DLI).

CONCLUSIONSNormal individuals express the entire 24 TCR V beta gene repertoire and have polyclonal distribution. However, the TCR V beta gene repertoire is only partially expressed in some donors. The TCR V beta gene repertoire is restrictedly expressed in a skew fashion in patients with leukemia before and after transplantation. The number of TCR V beta gene subfamilies increases over time post-transplantation. GVHD and GVL effects may induce the proliferation of T cell clones. Clinical GVL response may be distinguished from GVHD alloreactivity through the host MHC antigen.

Graft vs Host Disease ; genetics ; Graft vs Leukemia Effect ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; genetics ; therapy ; Polymerase Chain Reaction ; Receptors, Antigen, T-Cell, alpha-beta ; genetics

OBJECTIVETo explore the critical dose of T lymphocyte for preserving graft versus leukemia (GVL) while preventing GVHD in murine acute leukemia model treated with H-2 haploidentical hematopoietic stem cell transplantation (HSCT).

METHODS(C57BL/6 x 615) F1 (H-2bk) mice which was inoculated with L615 cells to develop leukemic murine model was the recipient. The healthy C57BL/6 (H-2b) mice was the donor. CD(34)(+) cells from bone marrow and CD(3)(+)cells from spleen of the donor were purified by miniMACS. The purity of CD(34)(+) cells and CD(3)(+) cells were (81.5 +/- 2.4)% and (95.4 +/- 2.9)% respectively. Sixty-nine recipient mice were divided into seven groups. Group A received no treatment, group B received TBI only, the rest groups were irradiated 9 Gy by (60)Co and transfused 10(5) CD(34)(+) cells or mixed with 10(7) (E), 10(8) (F), 1.5 x 10(8) (G) of CD(3)(+) cells respectively. The mice were raised for 60 days, The cause of death was identified by pathology.

RESULTSAll mice in group E survived more than 60 days being significantly longer than that in the rest groups (p < 0.0001). The chimerisms from donor were 100% in the mice survived > 60 days. Mice died of leukemia relapse in group D and group E were significantly less than those in group C (p < 0.001). Mice died from GVHD in group G were significantly more than those in group E and group F (p < 0.001).

CONCLUSIONSThe leukemia relapse rate was highest in mice that were transplanted with CD(34)(+) cells alone. Those mice transfused with CD(3)(+) T lymphocyte in the graft higher than 10(8) cells died from the GVHD was significantly higher. The inclusive dosage of 5 x 10(7) CD(3)(+) T lymphocyte was enough to separate the GVHD from GVL.

Animals ; Antigens, CD34 ; CD3 Complex ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Host Reaction ; Graft vs Leukemia Effect ; Hematopoietic Stem Cell Transplantation ; methods ; Leukemia, Experimental ; immunology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Recurrence, Local ; prevention & control ; T-Lymphocytes ; immunology

Transfusion-associated graft-versus-host disease (TA-GVHD) is known as one of common complications of blood transfusion. The blood irradiation is generally accepted as a proven method to prevent from this disease, for the reason that it makes lymphocytes inactivated in blood products. The blood irradiation indicator provides a guarantee of proper radiation dose, thus improving the transfusion safety. Though widely used in developed countries for decades, the blood irradiation indicator is still in the initial stage in our country. In this review, the action principle, applications and applied value of the blood irradiation indicator are summarized briefly.
Blood Transfusion ; Graft vs Host Disease ; Humans ; Lymphocytes ; Transfusion Reaction

Animals ; Disease Models, Animal ; Graft vs Host Reaction ; Liver Cirrhosis, Biliary ; Mice

OBJECTIVETo assess the value of short tandem repeat (STR) analysis with fluorescence labeling polymerase chain reaction (PCR) in evaluating the status of engraftment and predicting the occurrence of graft-versus-host disease (GVHD) following orthotopic liver transplantation.

METHODSThe method of STR analysis with fluorescence labeling PCR was established. DNA was extracted by monoclonal magnetic beads from the peripheral blood nucleated cells of 23 recipients and labeled with 4 fluorescence dyes before and at 7 days after orthotopic liver transplantation.

RESULTSIn the 23 patients studied, 5 patients showed mixed chimeras after orthotopic liver transplantation. Two of the 5 patients with mixed chimeras developed GVHD and died. The other 18 patients did not show mixed chimeras, and only one of them developed GVHD. The incidence of GVHD was significantly different between the patients with and without mixed chimeras (40% vs 5.6%, P<0.05).

CONCLUSIONPCR-STR analysis after orthotopic liver transplantation can be indicative of engraftment and help to predict the occurrence of GVHD following orthotopic liver transplantation.

Adult ; Female ; Graft vs Host Disease ; diagnosis ; Humans ; Liver Transplantation ; Male ; Microsatellite Repeats ; Middle Aged ; Polymerase Chain Reaction ; methods

BACKGROUND: The ability of non-myeloablative allogeneic stem cell transplants to eradicate host neoplastic cells is based on the accumulating evidence of a graft-versus-malignancy (GVM) effect. Stable mixed chimerism (MC) is associated to the lower risk for the development of graft-versus-host diseases (GVHD), but this possibly occurs at the expense of the GVM effect. Therefore, assessment of the chimerism status is critical to allow immune intervention to maintain a state of donor-host tolerance and to prevent loss of the graft. METHODS: Serial post-transplant peripheral blood samples were collected from 17 patients with various malignant diseases following non-myeloablative allogeneic stem cell transplantation. DNA was amplified from the T-cells, and the polymerase chain reaction (PCR) products were quantified by an automated fluorescent DNA analyzer. RESULTS: All 17 patients showed T-cell MC at post-transplant, but this varied in degree and duration, and then 3 patterns emerged. Group 1: 5 patients experienced a short interval of T-cell MC prior to conversion to complete donor chimerism (CC) (median: 25 days). Group 2: 5 patients showed a rapid increase of host cells after a brief MC at a median of 21 days. They never achieved CC, and they relapsed or showed progressive diseases. Group 3: 7 patients showed persistent T-cell MC for 40-50 days, and they subsequently gradually converted to CC after a median of 112 days. CONCLUSION: All the patients achieved T-cell MC in post-transplant, but the CC development differed in frequency and speed. GVHD preceded the onset of T-cell CC in the majority of the patients. Serial engraftment monitoring of the T-cell chimerism status during the first 100 days after non-myeloablative stem cell transplantation is important in aiding the clinical management of such patients.
Chimerism* ; DNA ; Graft vs Host Disease ; Humans ; Polymerase Chain Reaction* ; Stem Cell Transplantation* ; Stem Cells* ; T-Lymphocytes* ; Tissue Donors ; Transplants

BACKGROUND: Human herpesvirus-6 (HHV-6) is recently known as a major pathogen associated with various diseases in hematopoietic stem cell transplant (HSCT) recipients. We prospectively evaluated the frequency and clinical manifestations of HHV-6 infection in HSCT recipient in a single HSCT center in Korea. METHODS: Serum and peripheral blood mononuclear cells (PBMC) were weekly obtained from 1 week before HSCT to 4 weeks after HSCT. Three months' and six months' samples were obtained in some cases. HHV-6 was detected by nested polymerase chain reaction. RESULTS: Two hundred and seventy-eight samples from 54 HSCT recipients were collected from February to November, 1999. HHV-6 was detected in 32 out of 54 recipients (59.3%) at least once during study period in their PBMC or serum. HHV-6 DNA positive rate of PBMC and serum samples were 38.1% and 4.3 % respectively. HHV-6 DNAemia (HHV-6 DNA positive in serum) was detected and peaked at 2 weeks after HSCT and continued to 4 weeks. HHV-6 DNA in peripheral blood was not associated with unexplained fever, acute graft-versus-host disease, engraftment delay, or cytomegalovirus infection in this study. CONCLUSION: Reactivation and development of DNAemia of HHV-6 certainly occurred after HSCT, but the clinical manifestations and association with other diseases were unclear in this study. The large-scaled, nation-wide detail studies about the prevalence and characteristics of HHV-6 in general population and patients of specific disease entities must be considered.
Cytomegalovirus Infections ; DNA ; Fever ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells* ; Herpesvirus 6, Human ; Humans* ; Korea ; Polymerase Chain Reaction ; Prevalence ; Prospective Studies ; Transplantation*

OBJECTIVETo investigate the clinical difference of cytomegalovirus (CMV) infection between HLA-matched allogeneic hematopoietic stem cell transplantation (allo-HSCT) and haploidentical hematopoietic stem cell transplantation (hi-HSCT).

METHODSThe clinical data of 83 patients who had undergone allo-HSCT were retrospectively analyzed. Out of them 50 patients underwent hi-HSCT and 33 patients received grafts from HLA-matched donors. The sera of all recipients and donors were CMV-negative before transplantation. All patients accepted myeloablative regimen without total body irradiation. PCR was performed to detect CMV in the peripheral blood twice a week after neutrophil recovery. CMV-DNA>500 copies/ml was defined as CMV viremia.

RESULTS68 patients (81.9%) were diagnosed as CMV viremia before 100 days after transplantation. The incidence of CMV infection in hi-HSCT group was 90% and significantly higher than that in HLA-matched HSCT group (69.7%) (P < 0.05). All the patients responded well to anti-CMV therapy; however, 63 cases experienced CMV reactivation. The occurrence rate of CMV reactivation in hi-HSCT group (95.6%) was comparable to that in HLA-matched HSCT group (87.0%) (P > 0.05). Univariate analysis showed that the transplantation pattern, the recovery time of peripheral neutrophils and the occurrence of acute graft-versus-host disease (aGVHD) significantly related to the episode of CMV viremia, while the sex and age of the recipients, and the recovery time of platelets did not associate with the incidence. Further analysis found that the recovery time of neutropils and platelets in HLA-matched HSCT group were greatly shorter than those in hi-HSCT group (P < 0.05). The incidence of aGVHD was comparable between two groups however, incidence of severe aGVHD was significantly higher in hi-HSCT (P < 0.05).

CONCLUSIONThe hi-HSCT is more susceptible to CMV infection, which may be related to the higher incidence of severe aGVHD and the relative delay of hematopoietic reconstruction as compared with HLA-matched HSCT.

Cytomegalovirus Infections ; blood ; diagnosis ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Incidence ; Polymerase Chain Reaction ; Retrospective Studies ; Risk Factors ; Tissue Donors

OBJECTIVETo investigate the microRNA (miRNA) expression in plasma of patients with aGVHD and without aGVHD after allo-hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe miRNAs (miR-423, mirR199a-3p, miR93*, miR377) expression levels in peripheral blood plasma of 25 patients before and after allo-HSCT were detected by real-time PCR.

RESULTSmiR-423, miR199a-3p and miR-93* in aGVHD group were significantly upregulated (P<0.05); miR-377 expression was not significantly different between aGVHD and non-aGVHD (P>0.05).

CONCLUSIONThe expression of miR-423, miR-199a-3p, miR-93* are upregulated in aGVHD group, which can be used as biomarkes to monitor and to diagnose aGVHD.

Biomarkers ; blood ; Graft vs Host Disease ; blood ; pathology ; Hematopoietic Stem Cell Transplantation ; Humans ; MicroRNAs ; blood ; Real-Time Polymerase Chain Reaction ; Up-Regulation

Preterm infants are a special group, and related severe neurological, respiratory, and digestive disorders have high disability/fatality rates. Allogeneic cell transplantation may be an effective method for the prevention and treatment of these diseases. At present, animal studies have been conducted for allogeneic cell transplantation in the treatment of hypoxic-ischemic encephalopathy, bronchopulmonary dysplasia, and necrotizing enterocolitis. The main difficulty of this technique is graft-versus-host reaction (GVHR), and successful induction of immune tolerance needs to be achieved in order to solve this problem. This article reviews the research advances in immune tolerance of allogeneic cell transplantation in preterm infants.
Apoptosis ; Cell Transplantation ; adverse effects ; Cytokines ; physiology ; Graft vs Host Reaction ; Humans ; Immune Tolerance ; Infant, Newborn ; Infant, Premature ; immunology ; Transplantation, Homologous