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HLA class-Ⅰ antigens in Gansu Chinese Han nationality with leukemia

Chinese Journal of Immunology.2004;20(10):693-695.

Objective:Host genetic factors are known to contribute to disease susceptibility.They may also be important in defining the pattern of disease presentation and progression,as well as its overall prognosis.However,no consistent HLA class-Ⅰ associations have been established in leukemia by PCR/SSOP in Gansu Chinese Han.Such studies have been reported in other counties,with conflicting results.This is the first PCR-based HLA class-Ⅰ association study in northwestern Chinese Han nationality leukemia.Methods:Compared HLA class-Ⅰ in 43 Chinese leukemia patients and 66 healthy Chinese controls as determined by polymerase chain reaction and sequence-specific olignucletide probe hybridization(PCR/SSO) DNA analysis.The present findings imply that HLA-associated genetic factors influence the risk for the development of leukemia.

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Detection of anti-Sm antibodies in SLE serum by immuno-PCR

Changqian ZENG ; Shu YAN ; Yuguo WANG

Chinese Journal of Immunology.2001;17(5):263-265.

Objective:To apply immuno-PCR for detection of anti-Sm antibodies.Methods: Immuno-PCR was established for anti-Sm antibodies in SLE sera using SaHIgG-DNA probe. The experimental conditions were optomized and methological identifieation indicated obvious in crease(×107 ) in sensitivity compared with that of ELISA. The positive rate of anti-Sm antibodies was 54.9% in SLE sera, and significantly higher than that of ELISA. Intra-and inter-CV were 2.3%～4.8% and 3.7%～6.9% respectively. Immuno-PCR was positively correlated with ELISA( p ＜0.001). Therefore, immuno-PCR for anti-Sm antibodies is specific,sensitive,reproducible and practical clinically.

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Analysis of auto-antibody profiles in 58 patients with PBC

Lifang ZHANG ; Shangen ZHENG ; Yougong YAN ; Liping FU

Chinese Journal of Immunology.1985;0(05):-.

Objective: To study the role of auto-antibody in the diagnosis of patients with PBC. Methods: ANA, SMA and AMA in serum of 58 PBC patients were tested by indirect immune fluorescence and Western blot. Such auto-antibodies as Anti-type AMAM2,anti-SLA/LP, anti-LKM-1 and anti-LC-1 were also identified. Results: Auto-antibodies existing in patients with PBC were mainly AMA(96.5%) and AMAM2(93.1%) and the titer was beyond 1∶100. 8 cases of those patients were positive with ANA and SMA simultaneously. One case had positive AMA and SLA/LP in serum and the clinical appearances were the same as those of type Ⅰ and type Ⅲ autoimmune hepatitis. 19 patients with positive AMAM2 in serum had liver-puncture and the results suggested the diagnosis of PBC in 63.7%(12/9). Conclusion: Test of auto-antibodies is clinically significant for the diagnosis of autoimmune hepatitis.

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The effect of rALR on the proliferation and IL-2 production of spleen mononuclear cells of rat in vitro

Hua XIE ; Hang SUN ; Hui GUO ; Qi LIU

Chinese Journal of Immunology.1985;0(05):-.

Objective: To study the possible role and way of ALR in the immune regulation in vitro. Methods: The proliferation of spleen mononuclear cells of rat was detected with 3H-TdR method in vitro under the following different treatments:(1)the administration of the different concentration of rALR with 5 ?g/ml ConA at same time;(2)the addition of 30 ?g/ml ALR after 5 ?g/ml ConA pretreatment in 10 h and 32 h, respectively;(3)the addition of 5 ?g/ml ConA after 30 ?g/ml ALR pretreatment until 30 h. Cyclosporin A and the supernatant from cultures of yeast were used as positive and negative controls, respectively. The IL-2 levels in the supernatants from the mononuclear cells by various treatments were detected with RIA test kit. Results: rALR inhibited the proliferation and the production of IL-2 of the mononuclear cells from rat spleen stimulated by ConA dose-dependently. The mononuclear cell proliferations were still inhibited by 30 ?g/ml rALR after stimulated by ConA for 10 h or 32 h, but the pretreatment of 30 ?g/ml rALR for 30 h had no influence on the reactivity of mononuclear cell to ConA compared with control. Conclusion: rALR could inhibit in dose-dependent way the proliferation of mononuclear cells from rat spleen stimulated by ConA in vitro, but it couldn’t influence the rest mononuclear cells. This suggests that the rest mononuclear cells might not express the receptor of ALR.

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Association between CD4~+CD25~+ regulative T cells and systemic lupus erythematosus disease active stage

Chaojun HU ; Yongzhe LI ; Dinghua LIU ; Dawei TONG ; Shulan ZHANG

Chinese Journal of Immunology.1985;0(05):-.

Objective: To investigate the ratios of peripheral blood CD4+CD25+,CD4+CD8+ regulative T cells of systemic lupus erythematosus(SLE) patients, and explore the association with disease active stage,nephropathy,serum anti-ds-DNA antibody,and both IgG and C3 levels. Methods: The percentage of CD4+CD25+T cells and CD4+CD8+T cells of peripheral blood from patients with systemic lupus erythematosus(SLE)(30 females and 7 males),30 rheumatism controls and 30 normal individuals were measured by flowcytometry. Results: Patients with active disease had statisitically lower levels of CD4+CD25+T cells than did normal controls(P

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The study on expression and activity of human mutant CD59 gene

Ning WU ; Meihua GAO

Chinese Journal of Immunology.1985;0(05):-.

Objective: To amplify two human mutant CD59 eukaryotic expressing systems and investigate mutant CD59 functional activity. Methods: Mammalian expression vector PLATER of mutant CD59 cDNAs was transfected into CHO together with the pcDNA by lipofectamine,which confered resistance to G418(400 ?g/ml). The positive clones were tested by FIH. Activity of both mutants CD59 was determined by BCECF release assay. Results: Mutant CD59 cDNAs subcloned into the mammalian expression vector PLATER and transfected CHO together with the pcDNA,which confered resistance to G418. The positive clones were tested by FIH.Activity of both mutants CD59 before and after glycation was determined by BCECF release assay,both of them could restrict MAC formation ,and glycation could inhibit CD59. Conclusion: A eukaryotic system that expressing mutant CD59 cDNA was successfully set up.It was found that mutant CD59 could restrict MAC formation,and glycation could inhibit mutant CD59. These would be helpful for the furthur study of link mutant CD59 and the vascular proliferative of diabetes.

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CD4~+CD25~+/CD4~+CD25~(high) regulatory cells in peripheral blood of cancer patients

Junxia YAO ; Li LIU ; Shiang HUANG

Chinese Journal of Immunology.1985;0(05):-.

Objective: To investigate the frequencies of CD4+ regulatory T(Treg) cells(CD4+CD25+/CD4+CD25high) in peripheral blood of cancer patients,providing the opportunity to determine whether cancer patients exhibit an expanded CD4+CD25high pool. Methods: The frequency of CD4+CD25+/CD4+CD25high Treg in the peripheral blood of 62 cancer patients and 15 controls was determined by flow cytometry. Results: Compared with those of healthy control, the frequency of CD4+CD25+/CD4+CD25high in the peripheral blood of cancer patients showed a significant increase[(19.61?8.17)%/(4.20?1.90)%,P

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Studies on the effects of Combined IL-12 and CD40L gene therapy for murine melanoma

Jianhua ZHANG ; Dekuang ZHAO ; Qingqing WANG ; Fenping SHEN ; Yuesong WENG

Chinese Journal of Immunology.1985;0(05):-.

Objective: To observe the therapeutic efficiency of adenovirus-mediated interleukin-12 gene(AdmIL-12) and CD40 ligand gene(AdmCD40L) intratumoral transfer in established murine melanoma in vivo. Methods: C57BL/6 mice were inoculated subcutaneously with B16 cells to establish the murine melanoma model. The tumor-bearing mice were injected intratumorally with murine IL-12 gene and CD40L gene recombinant adenovirus. Tumor growth and the survival of tumor-bearing mice were observed. The CTL activity was measured in vitro by lactate dehydrogenase(LDH) release assay. Results: Both AdmIL-12 and AdmCD40L can be efficiently expressed in vitro and in vivo. The treatment with AdmIL-12 could significantly inhibit the tumor growth and prolong the survival period of the tumor-beraing mice. Splenic CTL activity of the mice was also enhanced after IL-12 gene transfer. But the anti-tumor effects of AdmCD40L gene were not significant. In contrast, Co-delivery of IL-12 gene and CD40L gene lead to stronger antitumor effects than IL-12 gene alone. Conclusion: Adenovirus-mediated interleukin-12 gene and CD40 ligand gene transfer together intratumorally has significant therapeutic effects on mice melanoma in vivo.

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The study on inhabitory effects of chitosan oligosaccharide on tumor growth in vivo and in vitro

Jie GUAN ; Xiaoqing LUO ; Qi WANG ; Dianjun LI

Chinese Journal of Immunology.1985;0(05):-.

Objective: To investigate the inhibitory effects of chitosan oligosaccharide on tumor growth in vivo. Methods: Murine tumor cell line H22 was inoculated into. Then different doses of chitosan oligosaccharide were injected into the mice-bearing H22 liver carcinoma, IL-2 and IFN-? in the sera were measured by ELISA assay. The in vitro anti-proliferation activity of chitosan oligosaccharide on the lung carcinoma LH-7 cells was evaluated by MTT assay. Results: Chitosan oligosaccharide inhibited in vivo the growth of H22 tumor cells,with increasing the content of IL-2 and IFN-? in sera of the tumor-bearing mice. The weight of spleen and thymus of the mice were increased when compared with those of control group; tissue necrosis was observed in the tumor in situ; chitosan oligosaccharide also inhibited the growth of LH-7 cells in vitro. The inhibitory effect was shown in a concentration dependent pattern, but not correrlated with incubation time. The early characteristics of apoptosis of LH-7 cell could be observed under the transmission electronic microscopy. Conclusion: Chitosan oligosaccharide inhibits proliferation of tumor and improves immunity function of the hosts, and might induce apoptosis of LH-7 cells.

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Expression of IL-29 cDNA in cos-7 cells and its anti-HBV activity in vitro

Bing CHEN ; Xin LUO ; Yunxia LU ; Congzhen XU ; Shengquan ZHANG

Chinese Journal of Immunology.1985;0(05):-.

Objective: To clone cDNA of human interleukin-29(IL-29) and express it in cos-7 cells, and to study its anti-HBV activity in vitro. Methods: Total RNA was extracted from PBMCs which had been infected with vesicular stomatitis virus(VSV) in vitro.IL-29 cDNA was amplified using one-step RT-PCR technique. The recombinant expressing plasimd of psectagB/His-myc-IL29 was constructed by inserting IL-29 cDNA into the vector and was then transfected into cos-7 cells. Stable expression stains were screened by Hygromycin B and limited dilution method. The target protein was purified through Ni2+-chelating Sepharose Fast Flow. Anti-viral bioactivity of the recombinant IL-29 fusion protein was analyzed through an in vitro model of production of HBV by the HepG2.2.2.15 cell line.ELISA was used for detection of the viral titers in the cell cultural supernants. Results: IL-29 was cloned and stably expressed in cos-7 cells successfully. SDS-PAGE and Western blot analysis showed multiple bands of the target protein with the molecular weights between 20 000 and 33 000, and the major band was located at about 33 000, indicating the fused IL-29 modified by additional glycosylation. The rhIL-29 was shown to dose-dependently inhibit secretion of HBsAg and HBeAg accompanied by the reduction of HBV genomic DNA in the cells tested. The inhibition ratio of HBsAg and HBeAg production was attained 85% at a concentration of 160 ?g/L of rhIL-29. Conclusion: The rhIL-29 with anti-HBV activity has been obtained.

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