Establishment of Acute GVHD Mouse Model of Haplo identical Hematopoietic Stem Cell Transplantation and Its Pathogenesis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the acute graft-versus-host disease(GVHD) mouse model based on haplo identical hematopoietic stem cell transplantation (HSCT) and to further investigate the pathogenesis of GVHD.</p><p><b>METHODS</b>Recipient mice ([C57BL/6×CBA/Ca]F1(H-2)) received lethal irradiation (11 Gy) of γ ray (Cs), followed by transplantation with donor-derived [C57BL/6(H-2)] T cell-depleted bone marrow cells (TD-BM) with or without donor-derived T cells. Recipients were randomly divided into 4 groups, including irradiation group, TD-BM group, TD-BM+T cell group 1 and TD-BM+T cell group 2. Survival rate and weight change were detected after HSCT. Pathological scoring were performed on the collected organs from recipients on day 14. Moreover, bone marrow chimerism was detected on days 14 and 18 after HSCT. Additionally, differentiation of donor-derived T cells towards Th1 cells in vivo was analyzed by flow cytometry. Furthermore, proliferation of donor-derived T cells in vivo was tested using CFSE.</p><p><b>RESULTS</b>The average survival in TD-BM, irradiation, TD-BM+T cell group 1 and TD-BM+T cell groups 2 were 60, 13.29±5.50, 33±2.3 and 29.14±1.77 days, respectively. On day 14 after transplantation, recipients of TD-BM + T cell group 1 displayed significantly more severe pathology in liver, colon, lung and skin. On days 14 and 28 after HSCT, bone marrow chimerism in recipients of both TD-BM group and TD-BM+ T cell groups 1 was over 94%. On day 14 after HSCT, serum cytokines IFN-γ, TNF-α and IL-1 levels in TD-BM+T cell group 1 were significantly higher than those in TD-BM group. Percentage of H-2bH-2kCD4IFN-γin spleen cells of recipients in TD-BM+T cell group 1 was higher than that of TD-BM group, with (0.5240±0.08447)% vs (7.912±0.6087)% (P<0.05). On days 14 after HSCT, donor-derived T cells displayed obvious proliferation in vivo.</p><p><b>CONCLUSION</b>C57BL/6(H-2)→［C57BL/6×CBA/Ca］F1(H-2) HSCT model can be used as a aGVHD model of haploidentical HSCT. Additionally, Th1 may play an important role in the pathogenesis of aGVHD resulting from haplo-identical HSCT.</p>

Bioinformatic analysis and identification for a novel antigen MLAA-22 in acute monocytic leukemia / 中国实验血液学杂志

This study was aimed to investigate a novel MLAA-22 antigen derived from a U937 cDNA library by the SEREX approach and search for gene expression in various samples. Bioinformatic analysis was performed to forecast MLAA-22 information mined from databases and experimental datasets. CTL epitope was predictied and the specific antibody for MLAA-22 was elicited by using peptide-microspheres and adjuvants. Furthermore, SYBR Green real-time PCR and immunoblotting method were used to evaluate the specificity of gene expression. The results showed that the full length cDNA of MLAA-22 located on chromosome 17q11.2 was 2.0 kb in size, and a putative protein was approximately 72.4 kD for 631 amino acids. The MLAA-22 encoded a cancer/testis antigen in human, which is nonsecreting type, plasmosin, labile protein, hydrophilia, thermostability, and without signal peptide. Many motifs might related to growth, proliferation, differentiation, and apoptosis. Antigenic peptides was synthesized as the antigen with Fmoc/PyBOP method. Rabbits were immunized by injecting the synthetic peptide-KLH to obtain antibody and the immune sera analyzed with ELISA were 1:8000. SYBR Green real-time PCR and Western blot showed that MLAA-22 presented with a higher number of copy messages in M(5), lower in CML, but not in gastric carcinoma, renal carcinoma, LNCaP cell lines and normal adult tissues, etc. It is concluded that mlaa-22 is a novel acute monocytic leukemia-associated antigen gene and be extended to further discovery.

Immunological screening for multiple myeloma-associated antigens and their bioinformatics analysis / 中国实验血液学杂志

This study was aimed to screen the cell cDNA expression library of multiple myeloma HMy2 (MM HMy2) by using "serological analysis of cDNA expression library (SEREX)" technique. The obtained 30 positive clones were all sequenced, and analyzed by BLAST (basic local alignment search tool). The results indicated that 6 known genes and 12 new MM-associated genes were obtained, part of which sequences were spliced by EST (expressed sequence tag) splicing. 6 known genes such as for ring finger protein 167, KLF10, TPT1 protein, p02 protein, cDNA FLJ46859 fis, DNMT1 methyltrasferase etc. have been demonstrated a certain relationship with other tumor's formation, progress and prognosis. The structures and functions of the new genes preliminarily analyzed and predicted by means of bioinformatics showed that MMSA-3, MMSA-8 and MMSA-11 encoding 215, 160 and 122 amino acid residues respectively had the full open reading frames (ORF). All the new genes might be located at euchromosomes but MMSA-1 at sex chromosome. MMSA-4 was highly similar to the protein controlling the transcription of tumor antigen, MMSA-5 might take part in cell phagocytosis, MMSA-7 might inactivated NF-kappaB, and MMSA-12 might be a lymphocytic cytoplasmic protein. The specificity of new genes such as MMSA-3 and MMSA-7 were higher, by a preliminary analysis using CrELISA. It is concluded that tumor antigens screened by this study can be used for early immunological diagnosis, surveillance of minor residual foci, assessment of prognosis, and preparation of tumor vaccine and so on.

Relationship between oxidative stress and depression in patients with acute leukemia / 中国实验血液学杂志

This study was aimed to investigate the relationships between oxidative stress and depression in patients with acute leukemia. Ninety two cases of acute leukemia were randomly enrolled in the study. Depressive disorder was assessed by self-rating depression scales (SDS) and multiple items questionaires. The total anti-oxidation capability (T-AOC), reactive oxygen species (ROS) and superoxide dismutase (SOD) activities, as well as malondialdehyde (MDA) and nitric oxide (NO) levels were measured in pre-treatment periods. Meanwhile, the steady state level of human 8-hydroxyguanine glycosylase (hOGG1) mRNA transcript was monitored by quantitative real-time PCR. The results showed that the defence of antioxidant system was impaired in patients with acute leukemia. The incidence of depression was 47.83% in 92 cases. T-AOC and SOD activities were significantly decreased in patients with depression, while ROS, NO, MDA levels and hOGG1 mRNA expression were reverse of the former. It revealed that depression positively correlated with course of disease and hOGG1, and negatively correlated with T-AOC. It is concluded that oxidative damage occurs in patients with acute leukemia, moreover, lower antioxidant defences exist in depressive patients. These results underscore the notion that oxidative stress may promote the development of depression.

Retrospective observation of curative effects on MDS refractory anemia with combination of all-trans retinoic acid, 1, 25-dihydroxyvitamin D3 and androgen / 中国实验血液学杂志

This study was aimed to examine whether a combination of all-trans retinoic acid (ATRA), 1, 25-dihydroxyvitamin D(3) and androgen possesses the therapeutic value for the MDS-refractory anemia (MDS-RA), and to analyze the mechanisms in detail. 62 cases receiving a scheme of combination of ATRA, 1, 25-dihydroxyvitamin D(3) and androgen (group A) were monitored. The remaining 33 cases (group B) were provided with vitamin supplementation, chalybeate drugs, and one or two of the combination. Bone marrow aspiration and biopsy were performed for collecting the specimens at the baseline and afterwards. The conditions of the patients were monitored by means of weekly complete blood counts and the monthly examination, including toxicity test, physical examination, electrocardiography, and biochemistry panel. The results showed that after treating for 8 weeks in group A, 4 out of 62 patients showed complete remission and 12 patients showed partial remission according to the defined response criteria, and 43 patients (69.35%) showed hematological improvement (HI). The further treatment for 16 out of 62 patients (25.81%), 13 failures (10 deaths, 2 RAEB and 1 RAEB-T) and 3 transformations (M(2), M(3), M(5)) with a median survival interval of 26.25 months, were observed and interrupted for some reasons. However, partial remission was observed only in 3 patients in group B, and HI amounted to 51.51%. Furthermore, the disease progression was observed in 12 out of 33 patients (36.36%) with a median survival interval of 16 months, 9 failures (including 6 deaths, 2 RAEB and 1 RAEB-T) and 3 transformations (M(2), M(3), M(4)). The overall ratios of survival for 3 and 5 years in group A, which received the combination, reached to 69.24% and 53.72% respectively, in comparison with 52.23% and 31.34% in the patients of group B (log-rank, P = 0.016). The following requirements, if were met, would be significant for prognosis: the combination regiment, no transformation, children, no complication, female, 90-120 g/L of hemoglobin concentration, normal cellular bone marrow and uni-cytopenias (P < 0.05). Moreover, Cox regression showed that therapy, transformation and age are all the independent factors (P < 0.05). It is concluded that the combination of above mentioned 3 drugs may be effective and safe treatment for the patients with MDS-RA. Its relevant mechanisms can be involved in the combination, that elicits a wide range of pharmacological effects, such as differentiation, anti-tumor-promotion, anti-apoptosis, anti-angiogenesis, anti-cachexia and immunoregulation.