Functions of HIV-1 Tat and its association with diseases / 中国生物制品学杂志

@#Acquired immune deficiency syndrome,or AIDS,has been a major infectious disease that troubles the public health in a global scale. Human immunodeficiency virus type 1(HIV-1)is the causative reagent responsible for AIDS development. Even though the highly active anti-retroviral therapy(HAART,or the cocktail therapy)that has been widely applied could effectively suppress the infection and replication of HIV-1,the infected people suffer from other related diseases,such as the HIV-associated neurocognitive disorder(HAND). This paper mainly focused on the function of an important regulatory protein of HIV-1,trans-activator of transcription(Tat),and its correlation with HIV-1 replication and HAND development,so as to clarify the importance of developing anti-AIDS drugs targeting Tat protein

Effects of CITED1 gene silencing on biological functions of papillary thyroid carcinoma K1 cells / 肿瘤

Objective: To investigate the effects of Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 1 (CITED1) gene silencing on the proliferation, apoptosis and migration of papillary thyroid carcinoma K1 cells. Methods: The recombinant lentivirus carrying CITED1-shRNA or the negative control (NC)-shRNA was successfully infected into thyroid cancer K1 cells. The silencing efficiency of CITED1 gene was detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The cell proliferation was detected by CCK-8 method, the cell cycle distribution and apoptosis were detected by FCM, and the cell migration ability was detected by scratch wound healing assay. Results: After the recombinant lentivirus carrying CITED1-shRNA was successfully infected into papillary thyroid carcinoma K1 cells, the expression levels of CITED1 mRNA and protein were significantly decreased (both P < 0.01), which suggested that the K1 cells with CITED1 gene silencing were successfully obtained. After silencing CITED1 gene expression, the cell proliferation was significantly inhibited (P < 0.01), the apoptosis rate was significantly increased (P = 0.001), the proportion of G0/G1-phase cells was significanlty increased (P = 0.007), the proportion of G2/M- and S-phase cells was significanlty decreased (both P < 0.05), and the cell migration abilities at 12 h and 24 h were significantly decreased (both P < 0.01). Conclusion: Silencing CITED1 gene expression can inhibit the proliferation and migration of papillary thyroid carcinoma K1 cells, and promote the apoptosis of tumor cells.

Adenovirus-mediated MHC class II transactivator mutant in gene therapy of mouse experimental autoimmune thyroiditis / 第二军医大学学报

Objective: To observe the therapeutic effect of major histocompatibility complex (MHC) class II transactivator mutant (C II TAm) for gene therapy of mouse experimental autoimmune thyroiditis (EAT) and explore the possible mechanisms. Methods: Thirty-one healthy female CBA/J mice were randomly divided into 4 groups, namely EAT model group(n= 8), C II TAm therapy group(n=9), GFP control group (n=9), and normal control group(n=5). Animals in the first 3 groups were immunized with porcine thyroglobulin (pTg) and complete or incomplete Freud's adjuvant (CFA/IFA) to establish EAT model; mice in the C II TAm therapy group and GFP control group were also treated by intravenous recombinant adenovirus Ad-CMV-C II TAm and Ad-GFP, respectively, while those in the EAT model group were injected with equal volume of normal saline. Mice in the normal control group received no special treatment. All mice were sacrificed on the 29th day after the first immunization. The thyroid pathological changes were examined using H-E staining; the expression of MHC II molecules in the thyroid was examined using immunohistochemical staining; the spleen lymphocyte proliferation and IFN-γ secretion stimulated by pTg were examined in their culture supernatant; the titer of plasma anti-pTg autoantibody was assayed by ELISA; and the expression of inducible costimulator (ICOS) on CD4+ T cells in both peripheral blood and spleen was analyzed by flow cytometry. Results: H-E staining showed that the infiltration index of thyroid lymphocyte in the C II TAm therapy group (0.3±0.5) was significantly lower than that in the EAT model group (1.4±0.4) and the GFP control group (1.5±0.2, both P<0.01). Immunohistochemical staining showed diffused expression of MHC II molecules in the thyroid of the EAT model group and GFP control group, compared to very weak expression in the C II TAm therapy group and the negative expression in the normal control group. The lymphocyte stimulation index (SI) against 80 μg/ml pTg in the C II TAm therapy group was significantly lower than that in the EAT model group and the GFP control group(P<0.05). The IFN-γ secretion in the culture supernatants showed a similar difference as SI in all the groups (P<0.01). The titer of plasma anti-pTg autoantibody in the C II TAm therapy group was significantly lower than those in the EAT model group and the GFP control group (both P<0.01). The positive rate of ICOS on CD4+ T cells in the C II TAm therapy group was significantly lower than that in the EAT model group and the GFP control group(both P<0.01). Conclusion: Ad-CMV-C II TAm recombinant adenovirus can inhibit the MHC II molecule expression in the thyroid of EAT mouse and the proliferation of self-reactive T cells, attenuate the inflammatory cells infiltration in the thyroid, and decrease the titer of plasma anti-pTg autoantibody, indicating that C II TA mutants might have therapeutic effect for EAT.

Study on the relationship between HBV infection and polymorphism of CⅡTA G-944C gene among Jinuo,Dai and Aini minority populations in Yunnan province / 中华流行病学杂志

Objective To explore the relationship between HBV infection and the genotypes and allele frequencies of CⅡTA G-944C gene polymorphism in three minority populations(Jinuo,Dai and Aini population)in Xishuangbanna district,Yunnan province.Methods Polymerase chain reaction and sequencing method were used to study the genotypes and allele frequencies distributions of CⅡTA G-944C gene polymorphism in those three populations.Relationship between the genotypes distribution and HBV infection results were also analyzed.Results The rates on HBV infection and HBsAg carrier status in Aini minority population were 89.2% and 16.3%,which were significantly higher than in Jinuo(27.9% and 3.9%,χ~2=135.196 and 10.361,P=0.000 and 0.001)and Dai population(44.9% and 6.6% χ~2=96.783 and 8.748,P=0.000 and 0.003)while among Aini population it was significantly different with the other two minority populations.The CC genotype and C allele frequencies were more distributed in Aini population than in the other two minority populations.In contrast,the GG genotype and G allele frequencies were lower than the other two minority populations,with χ~2 rates between Aini and Jinuo population were 11.841 and 12.208 and the P as 0.003 and 0.000 respectively while the χ~2 rates between Ami and Dai population were 23.902 and 20.220 with P value as 0.000 and 0.000.The genotypes frequencies of CⅡTA G-944C was significantly different in the infected individuals(IF)group and health control(HC)group in Jinuo population(χ~2=6.150 and 4.911,P=0.046 and 0.027).When compared with HBsAg+ group and HBsAg~- group,the genotypes and allele frequencies were different in Aini population and the total three minority populations(χ~2 rates in Jinuo minority were 8.650 and 5.034 with P values as 0.013 and 0.025).However,the χ~2 rates in the whole population were 13.047 and 9.416 with P values as 0.001 and 0.002,respectively.The distribution of CC genotype and C allele gene in HBsAg~+ group was increasing.Data from non-condition logistic regression analysis and adjusting for confounding factors,the HBsAg~+ group had a significantly increase of HBsAg~- group under the C allele Recessive Model(P=0.000;OR=2.964;95% CI:1.609-5.460).Conclusion The genotypes and allele frequencies distribution of CⅡTA G-944C were different in the three ethnic populations.Polymorphism of this gene was closely associated with HBsAg carrier.The CC genotype patients were more easily to become HBsAg carrier.

The enhancement of astrocytic-derived monocyte chemoattractant protein-1 induced by the interaction of opiate and HIV tat in HIV-associated dementia / 西安交通大学学报·英文版

HIV-associated dementia (HAD) is a public health problem and is particularly prevalent in drug abusers. The neuropathogenesis of human immunodeficiency virus (HIV) infection involves a complex cascade of inflammatory events, including monocyte/macrophage infiltration in the brain, glial immune activation and release of neurotoxic substances. In these events, astrocytic-derived monocyte chemoattractant protein-1 (MCP-1) plays an important role, whose release is elevated by HIV transactivator of transcription (HIV tat) and could be further elevated by opiates. This review will also consider some critical factors and events in MCP-1 enhancement induced by the interactions of opiate and HIV tat, including the mediating role of mu opioid receptor (MOR) and CCR2 as well as the possible signal transduction pathways within the cells. Finally, it will make some future perspectives on the exact pathways, new receptors and target cells, and the vulnerability to neurodegeneration with HIV and opiates.

The enhancement of astrocytic-derived monocyte chemoattractant protein-1 induced by the interaction of opiate and HIV tat in HIV-associated dementia / 药物分析学报

HIV-assodated dementia (HAD) is a public health problem and is particularly prevalent in drug abusers. The neuropathogenesis of human immunodeficiency virus (HIV) infection involves a complex cascade of inflammatory events, including monocyte/macrophage infiltration in the brain, glial immune activation and release of neurotoxic substances. In these events, astrocytic-derived monocyte chemoattractant protein-1 (MCP-1) plays an important role, whose release is elevated by HIV transactivator of transcription (HIV tat) and could be further elevated by opiates. This review will also consider some critical factors and events in MCP-1 enhancement induced by the interactions of opiate and HIV tat, including the mediating role of mu opioid receptor (MOR) and CCR2 as well as the possible signal transduction pathways within the cells. Finally, it will make some future perspectives on the exact pathways, new receptors and target cells, and the vulnerability to neurodegeneration with HIV and opiates.

Functional study of p38 mitogen-activated protein kinase based on cell-penetrating peptide delivery system / 老年心脏病学杂志（英文版）

Objective p38 Mitogen-activated protein kinase (MAPK) is a crossing center of various pathways. In this study, protein transduction system based on human immunodeficiency virus (HIV)-1 transactivator of transcription (TAT), which is an efficient delivery peptide of the foreign proteins into cells, was employed to study p38 MAPK functions in eukaryotic cells. Methods p38 And its dominant negative form, p38AF, were constructed into pET-His-TAT vector correctly to verify that the recombinant plasmids were well-founded through restriction enzyme digestion and DNA sequencing. The two proteins, His-TAT-p38 and His-TAT-p38AF, were expressed and purified in Escherichia coli by SDS-PAGE. Then they were incubated with ECV304 cells respectively and readily transduced into cells in a time-dependent and dose-dependent manner. The cells were stimulated by sorbitol. Activating transcription factor (ATF) 2 phosphorylation level was checked using Western blot to assess the activity of endogenous p38. Results Compared with controls, it was found that His-TAT-p38 increased the level ofATF2 phosphorylation in sorbitol-stimulated ECV304 cells, while His-TAT-p38AF inhibited it, indicating p38 MAPK protein delivery system based on TAT was constructed successfully. TAT-p38 and its dominant negative form possessed high biological activity after transduction into ECV304 cells by TAT protein delivery system. The results showed that p38AF fused with TAT could inhibit the transduction of endogenous p38 signal pathway in part, and other pathway might regulate p38 phosphorylation. Conclusions Our study provides a novel pathway to inhibit p38 signal pathway and establish a new method to study p38 function.

Sepsis Mortality in CIITA Deficient Mice is Associated with Excessive Release of High-mobility Group Box 1

BACKGROUND: Down regulation of major histocompatibility complex class II transactivator (CIITA) has been identified as a major factor of immunosuppression in sepsis and the level of CIITA expression inversely correlates with the degree of severity. However, it has not been fully elucidated whether the lower expression of CIITA is a cause of disease process or a just associated sign. Here we determined whether the CIITA deficiency decreased survival rate using murine sepsis model. METHODS: Major histocompatibility complex class II (MHC-II) deficient, CIITA deficient and wild type B6 mice were subjected to cecal ligation puncture (CLP) surgery. CIITA and recombination activation gene (RAG)-1 double deficient mice were generated to test the role of lymphocytes in CIITA-associated sepsis progression. RESULTS: Sepsis mortality was enhanced in CIITA deficient mice, not by impaired bacterial clearance resulted from CD4 T cell depletion, but hyper-inflammatory response such as excessive release of a pro-inflammatory cytokine, high-mobility group box 1 (HMGB1). CONCLUSION: Our results demonstrate that CIITA deficiency affects the course of sepsis via the unexpected function of CIITA, regulation of cytokine release.

Construction of the eukaryotic expression vectors containing different haplotypes DNA of human CIITA gene promoter IV / 重庆医科大学学报

Objective:To Construct the eukaryotic expression vectors containing four different haplotypes DNA of human CIITA gene promoter IV.Methods:Four haplotypes(CG,CC,TG and TC)can be constructed based on two single nucleotide polymorphism(SNPs)sites(G-944C and T-1350C)in promoter IV of human CIITA gene.The 487bp DNA fragments of CIITA promoter IV including the two SNPs were obtained by polymerase chain reaction(PCR)based on human genome DNA from the subjects with the CG/CG,CC/CC,TG/TG and TC/TC genotypes,which were TA cloned to pMD18-T Simple vectors and then were digested by restriction endonucleases Mlu I and Hind Ⅲ.After fragment recovery,those were ligated to four PGL3-Basic Vectors and four PGL3-Promoter Vectors respectively,which were digested by restriction endonucleases Mlu I and Hind Ⅲ as well.All recombinant plasmids were identified by sequencing.Results:Eight recombinant plasmids(CG-PGL3-Basic,CG-PGL3-Promoter,CC-PGL3-Basic,CC-PGL3-Promoter,TG-PGL3-Basic,TG-PGL3-Promoter,TC-PGL3-Basic,TC-PGL3-Promoter)containing four different haplotypes DNA of human CIITA promoter IV were obtained,whose sequences completely matched with the theoretical prediction demonstrated by sequencing.Conclusions:The eukaryotic expression vectors containing four different haplotypes DNA of human CIITA promoter IV are successfully constructed,and it lays the groundwork for the further study of different haplotype function.

Molecular Mechanisms of Hepatitis B Virus-associated Hepatocellular Carcinoma / 대한간학회지

Hepatocellular carcinoma (HCC) is one of the most common malignant diseases in the world. The hepatitis B virus (HBV) replicates non-cytopathically in hepatocytes, and most of the liver injury associated with this infection reflects the immune response. Epidemiological studies have clearly demonstrated that a chronic HBV infection is a major etiological factor in the development of HCC. The pathogenesis of HBV-associated HCC has been studied extensively, and the molecular changes during the malignant transformation have been identified. The main carcinogenic mechanism of HBV-associated HCC is related to the long term-inflammatory changes caused by a chronic hepatitis B infection, which might involve the integration of the HBV. Integration of the HBV DNA into the host genome occurs at the early steps of clonal tumorous expansion. The hepatitis B x protein (HBx) is a multifunctional regulatory protein that communicates directly or indirectly with a variety of host targets, and mediates many opposing cellular functions, including its function in cell cycle regulation, transcriptional regulation, signaling, encoding of the cytoskeleton and cell adhesion molecules, as well as oncogenes and tumor suppressor genes. Continued study of the mechanisms of hepatocarcinogenesis will refine our current understanding of the molecular and cellular basis for neoplastic transformations in the liver. This review summarizes the current knowledge of the mechanisms involved in HBV-associated hepatocarcinogenesis.

Molecular Pathogenesis of Hepatitis-B-virus-associated Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is one of the most frequent and malignant diseases worldwide. Epidemiological studies have clearly demonstrated that chronic hepatitis B virus (HBV) infection is a major etiological factor in the development of HCC. The pathogenesis of HBV-associated HCC has been studied extensively, and the molecular changes associated with malignant transformation have been identified. The predominant carcinogenic mechanisms of HBV-associated HCC are chronic inflammation and the effects of cytokines in the development of fibrosis and liver cell proliferation. An important role is also played by the integration of HBV DNA into host cellular DNA, which disrupts or promotes the expression of cellular genes that are important in cell growth and differentiation. Especially, HBx protein is a transactivating protein that promotes cell growth, survival, and the development of HCC. Continued investigation of the mechanisms underlying hepatocarcinogenesis will refine our current understanding of the molecular and cellular basis for neoplastic transformation in the liver. Prevention of HBV infections and effective treatments for chronic hepatitis B are still needed for the global control of HBV-associated HCC. This review summarizes the current knowledge on the mechanisms involved in HBV-associated hepatocarcinogenesis.

Overexpression of CIITA in T Cells Aggravates Th2-Mediated Colitis in Mice

The MHC class II transactivator (CIITA) is the master transcriptional regulator of genes involved in MHC class II restricted antigen presentation. Previously we suggested another role of CIITA in Th1/Th2 balance by demonstrating that forced expression of CIITA in murine T cells repressed Th1 immunity both in vitro and in vivo. However, the results were contradictory to the report that CIITA functioned to suppress the production of Th2 cytokine by CD4+T cells in CIITA deficient mice. In this study, we investigated the influence of constitutive expression of CIITA in T cells on Th2 immune response in vivo using murine experimental colitis model. In the dextran sodium sulfate-induced acute colitis, a disease involving innate immunity, CIITA transgenic mice and wild type control mice showed similar progression of the disease. However, the development of oxazolone-induced colitis, a colitis mediated by predominantly Th2 immune response, was aggravated in CIITA-transgenic mice. And, CD4+T cells from the mesenteric lymph node of CIITA-transgenic mice treated with oxazolone exhibited a high level of IL-4 secretion. Together, these data demonstrate that constitutive expression of CIITA in T cells skews immune response to Th2, resulting in aggravation of Th2-mediated colitis in vivo.

Delayed allograft rejection by the suppression of class II transactivator

We examined the effect of class II transactivator (CIITA) down-modulation on allograft rejection. To inhibit the function of CIITA, we constructed a series of CIITA mutants and found one exhibiting the dominant-negative effect on the regulation of major histocompatibility complex (MHC) class II expression. To test whether the CIITA dominant-negative mutant reduces immunogenecity, CIITA-transfected melanoma cells were injected into allogeneic host and assessed for immune evading activity against host immune cells. We demonstrated that the CIITA dominant-negative mutant allowed tumor nodules to develop earlier in the lung than control by this tumor challenge study. Furthermore, skin grafts deficient for CIITA also survived longer than wild-type in allogeneic hosts. Both the tumor challenge and skin graft studies suggest the inhibition of CIITA molecules in donor tissue would be beneficial to the control of allo-response.

Enhanced CEA-specific Immune Responses by Tat-LLO Fusion Protein

BACKGROUND: Carcinoembryonic antigen (CEA) is well-known soluble tumor marker frequently detectable in peripheral blood of carcinoma patients and considered as good target for antigen-specific immunotherapy. However, it is known that the induction of immune response to CEA is very difficult because CEA is a self-antigen expressed in fetal cells and weakly expressed in normal colorectal epithelial cells. To enhance anti-tumor immunity specific for CEA, recombinant CEA protein was modified using listeriolysin O (LLO) for endosomal lysis and transactivator of transcription (Tat) domain for transducing extracellular proteins into cytoplasm. METHODS: After immunization using dendritic cells pulsed with Tat-CEA, both Tat-CEA and LLO, and both Tat-CEA and Tat-LLO, antibody titer to CEA and LLO, cytotoxic T lymphocyte activity and the frequency of IFN-gamma producing T lymphocytes were measured. RESULTS: Immunization using DC pulsed with both Tat-CEA and Tat-LLO protein showed the increasement of production of CEA-specific antibody in serum, cytotoxic T lymphocyte activity, the frequency of IFN-gamma secreting T cells, compared with DC pulsed with both Tat-CEA and LLO. Furthermore the ratio of CD8+ T cell to CD4+ T cell among CEA-specific T cells was increased in group pulsed with both Tat-CEA and Tat-LLO. CONCLUSION: These results suggested that DC vaccine using Tat-LLO could be used for the development of effective immunotherapy for the treatment of tumor.

Hepatitis B Virus Surface Antigen: A Multifaceted Protein / 대한간학회지

Despite the small size of its genome (3.2 kb) and having only four genes that are encoded within it, the hepatitis B virus (HBV) is one of the most successful viral pathogens in human history. It is estimated that there are about 350-400 million people worldwide who are chronically infected with HBV, and even with the extensive efforts that are being done with preventive vaccination, this malady still remains a clear and present danger to the public health. How is it possible that this small double-stranded DNA virus can escape and outfox the surveillance of the complex human immune system? One explanation is that HBV gene products play multiple roles in infections and throughout the viral life cycle so that the virus can effectively survive under various hostile circumstances. Indeed, the HBV DNA polymerase, for example, exerts several functions such as reverse transcription and RNA degradation, and the HBV X protein not only acts as a transcriptional activator, but it also interferes with the host cells' DNA repair mechanism as well as inducing apoptosis and controlling signal transduction. The HBV surface protein, which is encoded in the env gene, is another intriguing example of such multifunctionality. Thus, our present article overviews and summarizes the multifaceted role of this membrane protein as shown in 1) its role as a structural protein of the virus envelope; 2) its function as the viral ligand for interacting with the viral receptors on host cells; 3) its characteristics as an energy-independent transporter molecule that can mediate the nuclear accumulation of itself and other tagged molecules; 4) its role as a viral transactivator protein that can cause hepatocellular carcinoma; 5) its hypothetical function in viral apoptotic mimicry that results in host anti-inflammatory responses; and last 6) its immunostimulatory property by providing for strong and well-defined B- and T-cell epitopes. Understanding these various functions and the versatility of this single protein will help us decipher and understand the viral- and immuno-pathogenesis of HBV itself.

Expression and significance of major histocompatibility complex classⅡgene in multiple organ dysfunction syndrome / 中华创伤杂志

Objective To investigate the expression and significance of major histocompatibility complex classⅡgene in multiple organ dysfunction syndrome.Methods Two-hit porcine model of MODS was duplicated in 18 swine that were randomly assigned into experimental group(Group M,n=9) and control group(Group C,n=9).The Group M was given compound factors including hemorrhagic shock,reperfusion injury and endotoxemia,and the Group C only underwent anesthesia and arterious/ve- nous eannula.After seven days,the animals were killed to remove splenic tissues fro extracting total RNA by Trizol method.The primer of SLA-DQA(MHC classⅡgene of swine)was designed to construct cD- NA by reverse transcription and the quantity of SLA-DQA mRNA detected with real time fluorescent quan- titative polymerase chain reaction(real time FQ-PCR).The standard curve was described by UVP com- puter image analysis system.Results The mortality of Group M was 78%(7/9),and the incidence rate of MODS was 89%(8/9).The expressing quantity of Group M was(1.376?1.006)?10~3,signifi- cantly lower than(5.330?3.053)?10~3 of Group C(P＜0.01).Conclusion Duplication of por- cine MODS model is satisfactory.Down-regulation of MHC classⅡgene may be due to control of classⅡtransactivator(CⅡTA)and release of multiple eytokine,such as TNF-?and IL-10.

Expression of TAT-Smad7-HA fusion protein and validation of its transduction activity / 第三军医大学学报

Objective To construct,express,purify,identify and label the TAT-Smad7-HA fusion protein (protein transduction domain of trans-activator,human Smad7 and hyaluronic acid) and to validate its transduction activity in the cultured human primary keloid fibroblast cells. Methods TAT-PTD,Smad7 and HA fragments were sequentially inserted into pET32a(+) to construct the pTAT-Smad7-HA prokaryotic expression vector. After the expression of target fusion protein was induced to express by IPTG,affinity purification,Western blot analysis,enterokinase cleavage,target protein capture and FITC labeling were subsequently performed by turns to obtain the FITC-TAT-Smad7-HA fusion protein and to further observe its transduction activity in the human primary KFB cells in vitro. Results The prokaryotic expression vector for the TAT-Smad7-HA fusion protein,named as pTAT-Smad7-HA was successfully constructed,and the target fusion protein was efficiently induced to express,covering more than 25% of the total bacterial proteins,successfully purified with a purity of more than 95% purity,desalted by desalting column,identified by Western blotting,thioredoxin removed and FITC labeled. Finally,a fusion protein of FITC-TAT-Smad7-HA with the approximately molecular weight of 50?103 was successfully purified and its high transduction activity in KFB cells was validated. Conclusion The highly-purified FITC-TAT-Smad7-HA fusion protein and the validation of its high transduction activity in KFB cells have provided an experimental foundation for further studies on the role the human Smad7 protein playing in the TGF-?/Smads signal transduction pathway and further elucidation of the pathogenesis of keloid formation.

Role of MHC class Ⅱ transactivator in upregulating the expression of HLA class Ⅱ antigen in HepG2 cells / 第三军医大学学报

Objective To investigate the role of MHC class Ⅱ transactivator(CⅡTA) in upregulating the expression of HLA class Ⅱ antigen in HepG2 cells.Methods Eukaryotic expression vector EBS-NPL-CⅡTA containing CⅡTA cDNA or the empty vector EBS-NPL was transfected into HepG2 cells respectively.CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were respectively detecued by RT-PCR,indirect cell immunofluorescence technique and flow cytometry in original HepG2 cells,HepG2 cells transfected with EBS-NPL-CⅡTA or the empty vector EBS-NPL.Results The expression of CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were not observed in original HepG2 cells and HepG2 cells transfected with empty vector,but in the HepG2 cells transfected with EBS-NPL-CⅡTA.Conclusion CⅡTA is a switching factor of mastering the expression of HLA class Ⅱ antigen in HepG2 cells.The lack of CⅡTA expression in HepG2 cells contributes to no expression of HLAⅡ antigen.

Association between disease susceptivity of chronic HBV infection and SNP G-944C in CⅡTA gene promoter Ⅳ / 第三军医大学学报

0.05). Conclusion The polymorphism of G-944C in CⅡTA gene promoter Ⅳ was associated with the susceptivity of chronic HBV infection, but was not associated with severity of diseases. The individuals with chronic HBV infection of CC genotype are of less possibility to develop chronic liver disease than those of other genotypes.

Comparesion of the method of CⅡTA and HLA-DR detection / 中华检验医学杂志

Objective To investigate the relation and difference of expression phase between classⅡtransactivator (CⅡTA) and HLA-DR antigens after IFN-? incubation, so as to investigate the potential effect of the class Ⅱ trasactivator (CⅡTA) in graft-versus-host disease(GVHD). Methods T lymphocyte from peripheral blood of health was incubated with IFN-? for 1 000 U/ml. RT-PCR was used to detect CⅡTA mRNA and Western blot was used to explore HLA-DR antigen in various time periods. Then Stat1? antisense oligonucleotides (AS) were given to inhibit the expression of CⅡTA, CⅡTA mRNA and HLA-DR antigen were tested again at peak point. Results CⅡTA mRNA was detectable 5 h after IFN-? treatment and peaks at 14 h; HLA-DR protein was detectable 28 h after IFN-? treatment and peaks at 52 h. The expression of CⅡTA mRNA and HLA-DR protein was lower in AS groups than that in control groups. Conclusion CⅡTA expression was positively correlated to HLA-DR expression, and earlier than the HLA-DR expression after IFN-? incubation. CⅡTA might be used as an early predicting marker of GVHD.