Short-term curative efficacy of autologous cytokine induced killer cells combined with low-dose IL-2 regimen containing immune enhancement by thymic peptide in elderly patients with B-cell chronic lymphocytic leukemia / 中国实验血液学杂志

This study was purposed to evaluate the safety and curative effect of autologous cytokine induced killer cells (CIK) combined with low-dose IL-2 regimen containing immune enhancement of thymic peptide on elderly patients with B-cell chronic lymphocytic leukemia (B-CLL). Thymic peptide α1 was subcutaneously given as the immunoenhancement agent at a dose of 1.6 mg/d, 14 days as one cycle. Peripheral blood mononuclear cells (PBMNC) from 5 patients with B-CLL were isolated once a week to induce ex vivo CIK cells through culture in the context of interferon (IFN)-γ, interleukin (IL)-2 and anti-CD3 monoclonal antibody. The PBMNC were separated from patients before and after 14 days as one cycle of thymic peptide α1 administration. Parameters of amplification ability, effector cells quantity, lymphocyte subgroups percentage and antitumor cytotoxicity were compared before and after thymic peptide administration. The 5 patients were treated with CIK cells combined with low-dose IL-2 regimen immediately after injection of thymic peptide α1. The CIK cells plus low-dose IL-2 regimen containing thymic peptide enhancement was defined as: thymic peptide α1 1.6 mg/d was subcutaneously administered once every other day; (4 - 6) ×10(9) of CIK cells were transfused followed by IL-2 subcutaneous administration of 1 mU/d on days 1-10, 28 days as one cycle. Clinical evaluation parameters including cellular immunity function, CLL related biomarkers, disease state and infectious frequency and degree were investigated before and after CIK cells infusion puls IL-2. The results showed that the amount of amplified CIK cells, the percentage and amplification times of effector cells and antitumor cytotoxicity more significantly increased after thymic peptide α1 treatment than before its use (P < 0.05). The total 46 cycles of CIK cells infusion plus IL-2 were completed in the 5 CLL patients. No adverse reaction was observed. After treatment of CIK cells plus IL-2, the general conditions of 5 CLL patients were to different extent improved. Simultaneously, percentages of CD3(+), CD3(+)CD8(+), and CD3(+)CD56(+) cells in peripheral blood remarked by raised (P < 0.05), the serum level of β2 microglobulin was significantly declined (P < 0.05), and the frequency and degree of infection was also decreased (P < 0.05). Following CIK cells plus IL-2 therapy, the transformation of disease state from partial remission (PR) to complete remission was seen in 3 patients, from stable disease (SD) to PR in 1 patient, and from progress of disease to SD in 1 patient. It is concluded that the regimen of autologous CIK cells combined with low-dose IL-2 containing immune enhancement of thymic peptide is safety and effective for the treatment of elderly patients with B-CLL.

Research progress on immunocyte senescence - review / 中国实验血液学杂志

The function of immune system degenerates in an aging-dependent manner and this results in immunosenescence. Human immune system includes two parts: genetic/innate immunity and adaptive immunity. The former is involved in monocytes, nature killer cells, and dendritic cells, the later is involved in acquired B and T lymphocytes. During the aging of immunity system, the both parts of immunity are damaged to some degree. Generally, innate immunity seems well-retained and the acquired immunity is degenerative seriously with aging. Immunocyte senescence is closely related to the elderly decreased ability to control infectious disease, cancer and to their generally poor response to vaccination. This review summarized the research progress on immunosenescence characteristics in aged phase.

Prediction and bioinformatics analysis of human gene expression profiling regulated by amifostine / 中国实验血液学杂志

Objective of this study was to perform bioinformatics analysis of the characteristics of gene expression profiling regulated by amifostine and predict its novel potential biological function to provide a direction for further exploring pharmacological actions of amifostine and study methods. Amifostine was used as a key word to search internet-based free gene expression database including GEO, affymetrix gene chip database, GenBank, SAGE, GeneCard, InterPro, ProtoNet, UniProt and BLOCKS and the sifted amifostine-regulated gene expression profiling data was subjected to validity testing, gene expression difference analysis and functional clustering and gene annotation. The results showed that only one data of gene expression profiling regulated by amifostine was sifted from GEO database (accession: GSE3212). Through validity testing and gene expression difference analysis, significant difference (p < 0.01) was only found in 2.14% of the whole genome (460/192000). Gene annotation analysis showed that 139 out of 460 genes were known genes, in which 77 genes were up-regulated and 62 genes were down-regulated. 13 out of 139 genes were newly expressed following amifostine treatment of K562 cells, however expression of 5 genes was completely inhibited. Functional clustering displayed that 139 genes were divided into 11 categories and their biological function was involved in hematopoietic and immunologic regulation, apoptosis and cell cycle. It is concluded that bioinformatics method can be applied to analysis of gene expression profiling regulated by amifostine. Amifostine has a regulatory effect on human gene expression profiling and this action is mainly presented in biological processes including hematopoiesis, immunologic regulation, apoptosis and cell cycle and so on. The effect of amifostine on human gene expression need to be further testified in experimental condition.

Inhibitory effect of dendritic cells combined with cytokine-induced killer cells on the growth and liver metastasis of human primary malignant melanoma of the small intestine / 中华消化外科杂志

Objective To investigate the inhibitory effect of dendritic cells (DCs) and homologous cytokine-induced killer cells (CIKs) on the growth and liver metastasis of human primary malignant melanoma of the small intestine. Methods A histologically intact liver metastasis fragment derived from a surgical specimen of patients with primary malignant melanoma of the small intestine was implanted in the mucous layer of small intestine in nude mice to construct the liver metastasis model of the malignant melanoma. Peripheral blood mononuclear cells from healthy donors and patients with malignant melanoma of the small intestine were isolated. DCs and CIKs were separately produced according to corresponding culture method, and the DC-CIKs were harvested after coculorthotopically implanted in 56 nude mice, and 72 hours later, all the mice were equally divided into normal saline group (0.3 ml), mitomycin treatment group (400 μg/0.3 ml), healthy donor CIK group (6 × 107/0.3 ml),healthy donor DC-CIK group (3 × 107/0.3 ml), autologous CIK treatment group (6 × 107/0.3 ml), autologous DC-CIK group (6 × 107/0. 3 ml) and DC-CIK (6 × 107/0.3 ml) + mitomycin (400 μg/0. 3 ml) treatment group. Mice in the experimental groups were administered the agents once daily by vena caudalis injection for 28 days. Mice were sacrificed 72 hours after treatment, and the orthotopic xenografts were removed for weighing,and the liver metastases were simultaneously evaluated by macroscopy and microscopy. All data were analyzed using the analysis of variance, LSD test and chi-square test. Results A liver metastatic model of human primary malignant melanoma of the small intestine ( HSIM-0603 ) was established. Of the control group, healthy donor CIK group, healthy donor DC-CIK group, autologous CIK treatment group, autologous DC-CIK group and DC-CIK +mitomycin group, the tumor weights were ( 1.18 ± 0. 17), (0.71 ± 0.06), (0.68 ± 0. 15 ), (0. 43 ± 0.08 ),(0.67 ± 0.07 ), (0.37 ± 0.08 ) and (0.20 ± 0.05 ) g, respectively; the tumor inhibition rates were 0, 39.82%,42.37%, 65.56%, 43.22%, 68.64% and 83.05%, respectively; liver metastases were detected in 8, 8, 5,4, 4, 3, 2 rats, respectively, in the above mentioned groups. There were significant differences in the tumor weight, tumor inhibition rate and liver metastasis rate among the seven groups (F = 152. 300, x2= 200. 538,94. 325, P ＜ 0.05 ). Conclusions The liver metastatic model could be applied to the studies on the pathogenesis,invasion, metastasis and treatment of human primary malignant melanoma of the small intestine. DC-CIK has the potential in inhibiting the growth and liver metastasis of the malignant melanoma of the small intestine.

A liver-metastatic model of human primary gastric lymphoma in nude mice orthotopically constructed by using histologically intact patient specimens / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>In recent years, incidence and mortality of lymphoma are markedly increasing worldwide. However, the pathogenesis and mechanism of invasion and metastasis for lymphoma are not yet fully clarified. It is mainly due to the lack of ideal animal models, which can precisely simulate the invasion and metastasis of lymphoma in the human body. So, it is very necessary to establish a highly metastatic nude mouse model of human lymphoma. This study developed a liver-metastatic model of primary gastric lymphoma in nude mice by using orthotopic surgical implantation of histologically intact patient specimens into the corresponding organs of the recipient small animals.</p><p><b>METHODS</b>A histologically intact fragment of liver metastasis derived from a surgical specimen of a patient with primary gastric lymphoma was implanted into the submucosa of the stomach in nude mice. Tumorigenicity, invasion, metastasis, morphologic characteristics (via light microscopy, electron microscopy, and immunohistochemistry), karyotype analysis, and DNA content of the orthotopically transplanted tumors were studied.</p><p><b>RESULTS</b>An orthotopic liver metastatic model of human primary gastric lymphoma in nude mice (termed HGBL-0304) was successfully established. The histopathology of the transplanted tumors showed primary gastric diffuse large B-cell lymphoma. CD19, CD20, CD22, and CD79alpha were positive, but CD3 and CD7 were negative. The serum level of lactate dehydrogenase (LDH) was elevated [(1010.56+/-200.85) U/L]. The number of chromosomes ranged from 75 to 89. The DNA index (DI) was 1.45+/-0.25 (that is, heteroploid). So far, the HGBL-0304 model has been passed on for 45 generations of nude mice. A total of 263 nude mice were used for the transplantation. Both the growth and resuscitation rates of liquid nitrogen cryopreservation of the transplanted tumors were 100%. The transplanted tumors autonomically invasively grew and damaged a whole layer in the stomach of nude mice. The metastasis rates of liver, spleen, lymph node, and peritoneal seeding were 100%, 94.3%, 62.6%, and 43.5%, respectively.</p><p><b>CONCLUSIONS</b>The study successfully establishes an orthotopic liver metastatic model of human primary gastric lymphoma in nude mice. The HGBL-0304 model can completely simulate the natural clinical process of primary gastric lymphoma and provides an ideal animal model for the research on the biology of metastasis and antimetastatic experimental therapies of primary gastric lymphoma.</p>

Establishment of a nude mouse model of highly metastatic gastric lymphoma constructed with orthotopic transplantation of surgical specimen / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To construct a mouse model of highly metastatic gastric lymphoma with orthotopic transplantation of human primary gastric lymphoma specimen.</p><p><b>METHODS</b>A fresh surgical specimen of primary gastric lymphoma was obtained intraoperatively and implanted into the submucosa of stomach in nude mice. Tumor formation, invasion, metastasis, morphological characteristics under light microscopy and electron microscopy, immunohistochemistry,and the karyotype of orthotopically transplanted tumor cells were studied.</p><p><b>RESULTS</b>An orthotopic highly metastatic model of human primary gastric lymphoma in nude mice(HGBL-0305) was successfully established. Histopathology of transplanted tumors showed primary gastric diffuse large B cell lymphoma. CD19, CD20, CD22 and CD79alpha were positive, while CD3 and CD7 were negative. The number of chromosome ranged from 56 to 69. DNA index(DI) was 1.47+/-0.12(i.e. heteroploid). Until now, HGBL-0305 model has been maintained for 45 generations by orthotopic passage for almost 4 years in nude mice. A total of 156 nude mice were used for transplantation. The growth rate and resuscitation rate of liquid nitrogen cryopreservation of transplanted tumor cells were both 100%. The autonomic growth of the transplanted tumor cells invaded and destructed all the layers of the nude mice stomach. The metastasis rates of liver, spleen, lymph node, and peritoneal seeding were 69.5%, 55.6%, 45.7%, and 30.5%, respectively.</p><p><b>CONCLUSIONS</b>An orthotopic highly metastatic model of human primary gastric lymphoma in nude mice is successfully established. HGBL-0305 model may simulate the natural course of primary gastric lymphoma in human and provides an ideal animal model for studies on pathogenesis, metastasis biology and anti-metastatic therapies of primary gastric lymphoma.</p>

Establishment of lung metastasis model of human primary malignant melanoma in the small intestine in nude mice / 中华消化外科杂志

Objective To provide an ideal animal model for exploring the pathogenesis and experimental treatment of malignant melanoma in the small intestine.Methods Fresh tissue of lung metastatic lesions from patients with malignant melanoma of the smallintestine were transplanted into mucosa of the small intestine in nude mice.After 4 times of screening.the tissue of the lung metastatic lesions from the nude mice were transplanted into the small intestine of additionat nude mice.Tumorgenecity and metastasis of transplanted tumors were observed,and were analyzed by morphology,karyotype and flow cytometry.Results A lung metastatic model of human primary malignant melanoma of the small intestine in nude mice was successfully constructed and named HSIM-0601.Massive melanin granules and melanin complex were seen in cytoplasm of tumor cells.Immunohistochemical straining of S-100 and HMB-45 were positive.The number of chromosome was between 57 and 59.DNA index was 1.49.HSIM-0601 was passed for 26 generations.A total of 173 nude mice were used for tumor transplantation.The growth rate of the transplanted tumors and resuscitation rate of liquid nitrogen cryopreservation were both 100%.In HSIM-0601.lung metastasis rate was 100%(173/173)and lymph node metastasis rate was 61.3%(106/173).Conclusions The HSIM-0601 successfully mimics the natural clinicopathologic course of patients with primary small intestinal melanoma,and provides an ideal animal model for research on pathogenesis,metastasis and experimentM therapy of malignant lymphoma in the small intestine.

Establishment of nude mice liver metastatic model of human primary malignant melanoma of the small intestine / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To provide ideal animal models for exploring pathogenesis and experimental therapy of primary malignant melanoma of the small intestine.</p><p><b>METHODS</b>The histologically intact primary and liver metastatic fragments derived from surgical specimens of one patient with metastatic malignant melanoma of the small intestine were orthotopically implanted in the small intestinal mucous layer of nude mice. The take rate, invasion and liver metastasis were observed. Morphology (light microscopy, electron microscopy), immunophenotype analysis, flow cytometry and karyotype analysis were applied for the original human tumors and the transplanted tumors.</p><p><b>RESULTS</b>The primary and liver metastatic fragments of malignant melanoma of the small intestine were successfully implanted in nude mice. After continuous passages in nude mice,an orthotopic model of human primary malignant melanoma of the small intestine(from the primary focus)in nude mice (termed HSIM-0501) and a liver metastatic model of human primary malignant melanoma of the small intestine (from the liver metastatic focus) in nude mice (termed HSIM-0502) were established. Histological examination of transplanted tumors revealed high-grade melanoma. S-100 protein and HMB45 were positive. Massive melanin granules and melanin complex were seen in cytoplasm of tumor cells.Chromosomal modal number was between 55 and 59. DNA index (DI) was 1.49-1.61, representing heteroploid. HSIM-0501 and HSIM-0502 were maintained for 25 and 27 passages in nude mice respectively. Three hundred and seventeen nude mice were used for transplantation. Both the take rate after transplantation and resuscitation rate of liquid nitrogen cryopreservation were 100%. HSIM-0501 exhibited 46.2% liver metastasis and 36.7% lymph node metastases. In HSIM-0502, both liver and lymph node metastases were 100%.The transplanted tumors autonomically and invasively grew in the small intestines of nude mice and hematogenous metastasis, lymph node metastasis and celiac planting metastasis occurred.</p><p><b>CONCLUSION</b>Two nude mice liver metastatic models of human primary malignant melanoma of the small intestine are successfully established, which provide ideal animal models for the research of pathogenesis,metastasis biology and anti-metastatic experimental therapy of primary malignant melanoma of the small intestine.</p>

Reblication of high metastasis model of malignant lymphoma of human small intestine in nude mice / 解放军医学杂志

Objective To replicate an ideal animal model for exploring pathogenesis and experimental treatment of malignant lymphoma of small intestine. Methods Fresh lymphoma tissues derived from primary lesion and liver metastasis of malignant lymphoma of human small intestine obtained during operation were respectively transplanted into mucosa of small intestine and subcutaneous space in the interscapular region in nude mice. Tumorigenicity, invasion and metastasis of transplanted tumors were observed. Morphology (light microscopy, electron microscopy and immunohistochemistry), karyotype analysis, DNA quantitative assay (FCM) were also studied. Results From five patients of malignant lymphoma of small intestine, tumor tissue from 3 patients was successfully transplanted. According to the new classification of the World Health Organization, a strain of high metastatic model of orthotopically transplanted malignant lymphoma in nude mice (HSIL-0101) and another of subcutaneously transplanted highly metastatic model were reproduced from the same human neoplasm (non-Hodgkin B cell type); orthotopically Pathological study showed that the tumor was high-grade large B-cell lymphoma. Histochemitry showed that it was CD19, CD20, CD22 and CD45 positive, and CD3 and CD7 negative. The number of chromosome is ranged from 55 to 59; DI (DNA Index) was 1.47～1.61 (ie, heteroploid). HSIL-0101 and HSIL-0102 had been passaged for 32 and 38 generations in nude mice separately. 357 nude mice were transplanted. Rate of growth of neoplasm transplantation and resuscitation rate of liquid nitrogen cryopreservation were both 100%. In HSIL-0101, metastasis rate of liver and lymph node was 100%. In HSIL-0102, metastasis rate of liver was 63.5% and metastasis rate of lymph node was 62.7%. Transplanted tumors invasively grew in small intestine and subcutaneous region of nude mice. Blood metastasis was found (liver and spleen metastases). There were also lymph metastasis and seeding metastasis in the peritoneal cavity. Conclusions The study first successfully established spontaneous high metastasis models of malignant lymphoma of human small intestine in nude mice by orthotopic and subcutaneous transplantation. HSIL-0101 and HSIL-0102 could be used to carry out the research on pathogenesis, invasion, metastasis, and experimental therapy of malignant lymphoma of small intestine.