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1.
Article in Chinese | MEDLINE | ID: mdl-17429540

ABSTRACT

OBJECTIVE: To observe the therapeutic effect of autologous cytokine-induced killer cells (CIK) on HBV DNA positive patients with liver cirrhosis. METHODS: HBV DNA positive 33 patients with cirrhosis were treated with CIK. Before and after cultured in vitro and post-treatment, CD3+, CD3+CD4+, CD3+CD8+, CD3+CD56+ cells, mDC and pDC were detected by flow cytometry. The indexes of virus and liver function were compared between pre- and post-treatment. RESULTS: CD3+, CD3+CD8+ cells and CD3+CD56+ cells were higher after cultured in vitro and after transfused back than those before culture (91.5 +/- 10.3, 74.4 +/- 9.9 vs. 67.9 +/- 12.8; 60.9 +/- 15.5, 37.3 +/- 15.1 vs. 27.9 +/- 10.9; 18.4 +/- 11.7, 14.5 +/- 7.5 vs. 10.6 +/- 7.1). The percentages of mDC and pDC also increased after-treatment vs. pre-treatment (0.54 +/- 0.18 vs. 0.70 +/- 0.29; 0.26 +/- 0.13 vs. 0.41 +/- 0.25). HBV DNA became undetectable in 12 patients and decrease exceeded 100 times in 4 patients after treatment. HBeAg became undetectable in 10 of 14 patients who were HBeAg positive pretreatment patients, among them 2 patients had HBeAb sero conversion. The liver function was improved after treatment. All patients tolerated the treatment. CONCLUSION: CIK treatment can increase immune effector cells and has some antiviral effect and is safe.


Subject(s)
Adoptive Transfer/methods , Cytokine-Induced Killer Cells/transplantation , Hepatitis B/complications , Liver Cirrhosis/therapy , Adoptive Transfer/adverse effects , Adult , Aged , Cells, Cultured , Cytokine-Induced Killer Cells/cytology , Cytokine-Induced Killer Cells/immunology , Fatigue/etiology , Female , Headache/etiology , Hepatitis B/virology , Humans , Liver Cirrhosis/etiology , Liver Cirrhosis/immunology , Male , Middle Aged , Transplantation, Autologous , Treatment Outcome
2.
Zhonghua Zhong Liu Za Zhi ; 26(8): 465-8, 2004 Aug.
Article in Chinese | MEDLINE | ID: mdl-15555334

ABSTRACT

OBJECTIVE: To compare the inhibitory effects of cytokine-induced killer (CIK) cells alone, chemotherapeutic drug alone, and CIK cells combined with chemotherapeutic drug on the growth of hepatocellular carcinoma (HCC) cells transplanted in nude mice. METHODS: Peripheral blood mononuclear cells (PBMC) collected from five healthy donors by blood cell separator were incubated in vitro to induce CIK cells in the presence of interferon-gamma (IFN-gamma), IL-2 and anti-CD3 monoclonal antibody (mAb). The phenotype of CIK cells was characterized by flow cytometric analysis. BEL-7402 HCC cells were inoculated subcutaneously to nude mice. On day 5, at the inoculation site were injected normal saline (group 1), CIK cells (3 x 10(7) and 6 x 10(7), group 2 and 3), mitomycin-C (MMC 80 microg in 0.2 ml, group 4), and CIK cells combined with MMC (group 5), respectively. RESULTS: The percentage of CD3(+), CD3(+)CD8(+), CD3(+)CD56(+), CD25(+) cells increased from 64.0%, 28.0%, 7.8%, and 9.1% to 94.7%, 67.7%, 61.3%, and 84.0% respectively after cytokine induction. The percentage of CD3(+) and CD3(+)CD8(+) cells remained at high levels during incubation period, but that of CD25(+) and CD3(+)CD56(+) cells peaked respectively on day 7 and 13 and then declined. During the 90-day observation, the tumor formation rates were 100%, 70.0%, 80.0%, 70.0% and 66.7%; and the mouse survival rates were 10.0%, 60.0%, 40.0%, 50.0% and 75.0%, respectively from group 1 to group 5. Compared to the other groups, in the combined therapy group of mice, not only the tumor grew slowly and but also showed more marked tissue necrosis. CONCLUSION: The growth inhibitory effect on human HCC transplanted in nude mice of combined CIK cells and MMC treatment is more potent than that of CIK cells or MMC alone.


Subject(s)
Antibiotics, Antineoplastic/therapeutic use , Carcinoma, Hepatocellular/therapy , Immunotherapy, Adoptive , Killer Cells, Natural/transplantation , Liver Neoplasms/therapy , Mitomycin/therapeutic use , Animals , Carcinoma, Hepatocellular/immunology , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cells, Cultured , Combined Modality Therapy , Cytokines/metabolism , Cytokines/pharmacology , Female , Humans , Liver Neoplasms/immunology , Liver Neoplasms/pathology , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation
3.
World J Gastroenterol ; 10(8): 1146-51, 2004 Apr 15.
Article in English | MEDLINE | ID: mdl-15069715

ABSTRACT

AIM: To investigate the influence of autologous cytokine-induced killer (CIK) cells on the phenotypes of CIK effector cells, peripheral T lymphocyte subsets and dendritic cell subsets in patients with primary hepatocellular carcinoma (HCC). METHODS: Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 13 patients with HCC, then expanded by priming them with interferon-gamma (IFN-gamma) followed by monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2) the next day. The phenotypic patterns of CIK cells were characterized by flow cytometry on d 0, 4, 7, 10, 13 and 15 of incubation, respectively. Then, 5 mL of venous blood was obtained from HCC patients before or 8-10 d after CIK cells were transfused into patients to assess the influence of CIK cells on the percentages of effector cells, and proportions of DC1 or DC2 in peripheral blood by flow cytometry. RESULTS: After two weeks of in vitro incubation, the percentages of CD3(+)CD8(+), CD3(+)CD56(+), and CD25(+) cells increased significantly from 33.5+/-10.1%, 7.7+/-2.8%, and 12.3+/-4.5% to 36.6+/-9.0% (P<0.05), 18.9+/-6.9% (P<0.01), and 16.4+/-5.9% (P<0.05), respectively. However, the percentages of CD3(+)CD4(+) and NK cells had no significant difference. The percentages of CD3(+) and CD3(+)CD8(+) cells were kept at high levels during the whole incubation period, but those of CD25(+), and CD3(+)CD56(+) cells began to decrease on d 7 and 13, respectively. The proportions of type I dendritic cell (DC1) and type II dendritic cell (DC2) subsets increased from 0.59+/-0.23% and 0.26+/-0.12% before CIK cell therapy to 0.85+/-0.27% and 0.43+/-0.19% (all P<0.01) after CIK cell transfusion, respectively. The symptoms and characteristics of HCC patients were relieved without major side effects. CONCLUSION: Our results indicated that autologous CIK cells can efficiently improve the immunological status in HCC patients, and may provide a potent approach for HCC patients as the adoptive immunotherapy.


Subject(s)
Adoptive Transfer/methods , Carcinoma, Hepatocellular/therapy , Killer Cells, Natural/transplantation , Liver Neoplasms/therapy , Adult , Antibodies, Monoclonal/pharmacology , Antineoplastic Agents/pharmacology , CD3 Complex/immunology , Carcinoma, Hepatocellular/immunology , Female , Hepatitis B, Chronic/immunology , Hepatitis B, Chronic/therapy , Hepatitis B, Chronic/virology , Humans , Immunophenotyping , Interferon-gamma/pharmacology , Interleukin-2/pharmacology , Killer Cells, Natural/drug effects , Liver Neoplasms/immunology , Male , Middle Aged , T-Lymphocyte Subsets/immunology , Viral Load
4.
Zhonghua Liu Xing Bing Xue Za Zhi ; 24(7): 595-8, 2003 Jul.
Article in Chinese | MEDLINE | ID: mdl-12975017

ABSTRACT

OBJECTIVE: To investigate the V249I and T280M allelic polymorphisms of human immunodeficiency virus (HIV) coreceptor CX3CR1 in HIV-1 infected and uninfected population of Chinese indigenous Han and Uygur people and to probe the association between I249-M280 haplotype and HIV-1 susceptibility as well as AIDS progression. METHODS: Genomic DNA of 223 Uygur subjects and 316 Han subjects were purified from PBMC. I249 and M280 allelic frequencies were identified by polymerase chain reaction (PCR)/nest polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. All data were tested by chi(2) or u statistics analysis. RESULTS: Allelic frequencies of I249 and M280 were 16.1% and 13.3% in Uygur people, and 3.3% and 2.4% in Han people. No obvious difference existed between three groups of either ethnic group. However the allelic frequencies of HIV infected population were higher than those of general population, and those of general population higher than those of HIV-1 high-risk group. There was a strong linkage between I249 and M280 (P almost zero). CONCLUSIONS: I249 mutation was the sine qua non of M280 mutation, and most I249 alleles were accompanied by M280. The frequency of I249-M280 haplotype in Uygur population (13.3%) was adjacent to Caucasian people (15.8%), and that of I249-T280 haplotype (2.8%) was obviously lower than Caucasian people (12.5%); while both of them in Han people were much lower (0.9% and 2.4%). I249-M280 haplotype could accelerate AIDS progression according to Faure et al, while might be associated with HIV-1 susceptibility.


Subject(s)
HIV Infections/genetics , HIV-1/genetics , Membrane Proteins/genetics , Polymorphism, Restriction Fragment Length , Receptors, Chemokine/genetics , Alleles , Asian People/genetics , CX3C Chemokine Receptor 1 , China/epidemiology , China/ethnology , Chromosomes, Human, Pair 3 , Ethnicity , HIV Infections/epidemiology , HIV Infections/virology , Haplotypes , Humans , Membrane Proteins/metabolism , Point Mutation , Receptors, Chemokine/metabolism , Receptors, HIV/deficiency , Receptors, HIV/genetics , Receptors, HIV/physiology , Risk Factors
5.
Article in Chinese | MEDLINE | ID: mdl-12870001

ABSTRACT

OBJECTIVE: To study the polymorphism of human immunodeficiency virus (HIV)-1 coreceptor CXCR4 in Chinese Han ethnic group for AIDS prevention and treatment. METHODS: Totally 48 individuals were enrolled into the study. CXCR4 (cDNA No-AF147204) was cloned by PCR amplification using 2 pairs of primers, then sequenced using sequencing primers. The results of the same sequencing primers were analyzed by DNAstar software to find and identify single nucleotide polymorphism (SNP) sites. RESULTS: Totally 7 SNPs were found in the coding region of CXCR4, among them 3 were synonymous mutation (C-->T at loci 129, 426 and 968), 3 were missense mutation (C-->T at locus 38, A-->T at locus 90, and A-->C at locus 712) and 1 was stop mutation (C-->T at 106, which converted the codon for glutamic acid into stop codon). CONCLUSIONS: The polymorphism of CXCR4 coding region in Chinese Han is probably different from that of the other ethnic groups. Six of the 7 SNPs were discovered for the first time. Their influences on AIDS progression are worthy of studying.


Subject(s)
HIV-1/genetics , Point Mutation , Receptors, CXCR4/genetics , Adult , Asian People , Base Sequence , China/ethnology , Female , Gene Frequency , Humans , Male , Middle Aged , Molecular Sequence Data , Polymorphism, Single Nucleotide
6.
J Acquir Immune Defic Syndr ; 32(2): 124-30, 2003 Feb 01.
Article in English | MEDLINE | ID: mdl-12571520

ABSTRACT

The aim of this study is to determine in indigenous Chinese ethnic groups the frequencies of the chemokine (SDF1 3'A) and chemokine receptors (CCR5 delta32, CCR5 m303, and CCR2b 64I) HIV-1/AIDS restriction alleles. The study includes two cohorts; the first comprised 3165 indigenous healthy subjects representing eight ethnic groups: Han (n = 1406), Uygur (n = 316), Mongolia (n = 134), Hui (n = 386), Tibetan (n = 330), Zhuang (n = 378), Dai (n = 101), and Jingbo (n =114). The second cohort consisted of 330 HIV-1-infected (86 subjects infected by sexual transmission and 198 subjects infected by HIV-1-contaminated blood or by sharing injection equipment; the remaining 46 subjects said nothing about HIV-1 transmission) and 474 HIV-1-uninfected Han Chinese belonging to one of two HIV-1 high-risk groups: intravenous drug users (n = 215) and individuals with sexually transmitted diseases (n = 259). Genotypes for the four genes were obtained using PCR (CCR5 delta32) or PCR-restriction fragment length polymorphism. Randomly selected amplified PCR products were further confirmed by direct DNA sequencing. The variant allele frequencies were determined to be 0% to 3.48% for CCR5 delta32, 0% for CCR5 m303, 16.23% to 28.79% for CCR2b 64I, and 17.70% to 27.76% for SDF1 3'A in Chinese healthy individuals from eight ethnic groups. These findings show that allele frequencies differ among the eight Chinese ethnic groups for CCR5 delta32, CCR2b 64I, and SDF1 3'A and that the CCR5 m303 and CCR5 delta32 mutant alleles were absent or infrequent in Chinese, which may be helpful for studies of specific anti-HIV-1 vaccine trials and coreceptor inhibitor drug targets in Chinese populations. Furthermore, we observed no significant differences in allele or genotypic frequencies between HIV-1-infected and HIV-1-uninfected groups from the Han ethnic group. Our finding is the first reporting that there is likely no effect of the examined polymorphisms in our study on HIV-1 transmission in the Chinese Han population, However, the genetic effects of these and other AIDS-modifying polymorphisms on the pathogenesis and clinical outcome of HIV-1/AIDS diseases is under investigation in Chinese populations.


Subject(s)
Chemokines, CXC/genetics , Gene Frequency , HIV Infections/immunology , HIV-1 , Population Surveillance , Receptors, CCR5/genetics , Receptors, Chemokine/genetics , Adult , Chemokine CCL2 , Chemokine CXCL12 , China/ethnology , Cohort Studies , Disease Transmission, Infectious , Ethnicity , Genetic Testing , Genotype , HIV Infections/blood , HIV Infections/transmission , Humans , Middle Aged , Mutation , Receptors, CCR2 , Risk Factors , Substance Abuse, Intravenous/blood , Substance Abuse, Intravenous/immunology
7.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 20(1): 72-4, 2003 Feb.
Article in Chinese | MEDLINE | ID: mdl-12579509

ABSTRACT

OBJECTIVE: To investigate the single nucleotide polymorphism(SNP) loci of HIV-1 coreceptor CCR5 gene in Chinese Han people. METHODS: The coding region of CCR5 was amplified using 2 pairs of primers and the PCR products of all 42 healthy subjects were sequenced by 4 different primers. The results of sequencing were analyzed by DNAstar in search of SNP loci. RESULTS: Six SNP loci were discovered in the coding region of CCR5, among them four SNPs, i.e. 184A-->G, 503G-->T, 688G-->A and 999G-->T, cause amino acids changes and two SNPs are nonsense mutations. One cytosine deletion at the 894nt results in frame shift mutation and prematured termination. 184A-->G, 503G-->T and 999G-->T were found in Chinese Han people for the first time. The allelic frequencies of mutant 184G, 503T and 999T alleles were 1.1%, 21.1% and 10.0% in healthy Hans, respectively. The population distribution of G503T markedly deviated from Hardy-Weinberg equilibrium. CONCLUSION: The SNP loci in the coding region of CCR5 in Chinese Han people has its own characteristics, which is not consistent with those of Japanese and obviously different from those of Caucasian and African.


Subject(s)
Polymorphism, Single Nucleotide/genetics , Receptors, CCR5/genetics , Adult , Base Sequence , China , DNA/chemistry , DNA/genetics , DNA Mutational Analysis , Female , Gene Frequency , HIV-1/metabolism , Humans , Male , Middle Aged , Point Mutation , Receptors, CCR5/metabolism , Sequence Deletion
8.
Zhonghua Yi Xue Za Zhi ; 83(23): 2049-53, 2003 Dec 10.
Article in Chinese | MEDLINE | ID: mdl-14703414

ABSTRACT

OBJECTIVE: To investigate the alteration of the cellular profiles of T lymphocyte subsets and dendritic cell subsets in peripheral blood of primary hepatocellular carcinoma (HCC) patients after being transfused with autologous cytokine-induced killer cells (CIK) in patients, then to evaluate the clinical efficacy of the immune therapeutic strategy. METHODS: Peripheral blood mononuclear cells (PBMCs) from 13 patients with primary were collected using blood cell separator, and expanded in the fresh AIM-V medium in the presence of cytokine cocktail including interferon-gamma (IFN-gamma), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). The phenotypic patterns of CIK cells were longitudinally characterized by flow cytometry on day 0, 4, 7, 10,13 and 15 during the incubation period. PBMCs obtained from HCC patients before or after CIK cells transfusion into bodies to assay the changes of proportion of DC1 or DC2 in peripheral blood. RESULTS: After in vitro incubation for 14 or 15 days, a large of CD3(+)CD56(+) cells were produced from their progenitors and the percentages of CD3(+)CD8(+), CD3(+)CD56(+), CD25(+) cells significantly increased from 33.5% +/- 10.1%, 7.7% +/- 2.8%, and 12.3% +/- 4.5% at the beginning to 36.6% +/- 9.0% (P < 0.05), 18.9% +/- 6.9% (P < 0.01), and 16.4% +/- 5.9% (P < 0.05) at the day 15, respectively. In contrast, the percentages of CD3(+)CD4(+) and NK cells displayed no significant difference. The percentages of CD3(+), CD3(+)CD8(+) cells was held at a higher level during the whole incubation period, however those of the CD25(+), and CD3(+)CD56(+) cells began decreasing on day 7 and day 13, respectively. The proportion of type I of dendritic cells (DC1) and type II of dendritic cells (DC2) subsets increased from 0.59% +/- 0.23% and 0.26% +/- 0.12% before CIK cell transfusion to 0.85% +/- 0.27% and 0.43% +/- 0.20% (all P < 0.01) after CIK cell transfusion. The symptom of HCC patients receiving the CIK cell therapy was markedly ameliorated, and not side effect was seen in the treatment. CONCLUSION: Our results indicated that autologous CIK cells is able to boost the cellular immunological function in HCC patients, which probably provide a potent immune therapeutic strategy for HCC patients.


Subject(s)
Carcinoma, Hepatocellular/immunology , Cytokines/pharmacology , Killer Cells, Natural/immunology , Liver Neoplasms/immunology , Adult , Aged , Carcinoma, Hepatocellular/therapy , Dendritic Cells/immunology , Female , Humans , Liver Neoplasms/therapy , Male , Middle Aged , T-Lymphocyte Subsets/immunology
9.
World J Gastroenterol ; 8(3): 464-8, 2002 Jun.
Article in English | MEDLINE | ID: mdl-12046071

ABSTRACT

AIM: To characterize the anticancer function of cytokine-induced killer cells (CIK) and develop an adoptive immunotherapy for the patients with primary hepatocellular carcinoma (HCC), we evaluated the proliferation rate, phenotype and the antitumor activity of human CIK cells from healthy donors and HCC patients in vitro and in vivo. METHODS: Peripheral blood mononuclear cells (PBMC) from healthy donors and patients with primary HCC were incubated in vitro and induced into CIK cells in the presence of various cytokines such as interferon-gamma (IFN-gamma), interleukin-1 (IL-1), IL-2 and monoclonal antibody (mAb) against CD3. The phenotype and characterization of CIK cells were identified by flow cytometric analysis. The cytotoxicity of CIK cells was determined by (51)Cr release assay. RESULTS: The CIK cells were shown to be a heterogeneous population with different cellular phenotypes. The percentage of CD3+/CD56+ positive cells, the dominant effector cells, in total CIK cells from healthy donors and HCC patients, significantly increased from 0.1-0.13% at day 0 to 19.0-20.5% at day 21 incubation, which suggested that the CD3+ CD56+ positive cells proliferated faster than other cell populations of CIK cells in the protocol used in this study. After 28 day in vitro incubation, the CIK cells from patients with HCC and healthy donors increased by more than 300-fold and 500-fold in proliferation cell number, respectively. CIK cells originated from HCC patients possessed a higher in vitro antitumor cytotoxic activity on autologous HCC cells than the autologous lymphokine-activated killer (LAK) cells and PBMC cells. In in vivo animal experiment, CIK cells had stronger effects on the inhibition of tumor growth in Balb/c nude mice bearing BEL-7402-producing tumor than LAK cells (mean inhibitory rate, 84.7% vs 52.8%, P<0.05) or PBMC (mean inhibitory rate, 84.7% vs 37.1%, P<0.01). CONCLUSION: Autologous CIK cells are of highly efficient cytotoxic effector cells against primary hepatocellular carcinoma cells and might serve as an alternative adoptive therapeutic strategy for HCC patients.


Subject(s)
Carcinoma, Hepatocellular/therapy , Killer Cells, Natural/immunology , Liver Neoplasms/therapy , Animals , Carcinoma, Hepatocellular/immunology , Cell Division , Cytokines/pharmacology , Cytotoxicity, Immunologic , Humans , Immunophenotyping , Immunotherapy, Adoptive , Killer Cells, Natural/pathology , Liver Neoplasms/immunology , Mice , Mice, Nude , Neoplasm Transplantation , Transplantation, Heterologous , Tumor Cells, Cultured
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