Cytotoxic T lymphocyte-associated antigen 4 in acute rejection after renal transplantation / 中国组织工程研究

BACKGROUND:Cytotoxic T lymphocyte-associated antigen 4 is a newly discovered costimulatory molecule. It has been studied more in tumor and autoimmune diseases, less in the field of kidney transplantation. OBJECTIVE:To explore the role of cytotoxic T lymphocyte-associated antigen 4 in acute rejection after renal transplantation. METHODS:Fifty patients undergoing renal transplantation were divided into acute rejection group (20 cases) and stable graft function group (30 cases). Another 30 healthy persons served as control group. Blood samples were extracted from the peripheral blood. Cytotoxic T lymphocyte-associated antigen 4 was detected by enzyme linked immunosorbent assay and flow cytometry. RESULTS AND CONCLUSION:The expression of cytotoxic T lymphocyte-associated antigen 4 in the serum showed significant differences in the acute rejection group, stable graft function group and healthy control group (F=70.008 1, P=0.000 0), but showed no difference in peripheral blood lymphocytes of three groups (F=1.865 6, P=0.161 7). Compared with the healthy control group, the expression levels of cytotoxic T lymphocyte-associated antigen 4 in peripheral blood lymphocytes of acute rejection group and stable graft function group were significantly decreased (P=0.000 0). In addition, the acute rejection group had a lower cytotoxic T lymphocyte-associated antigen 4 expression than the stable graft function group (P=0.000 0). In renal transplant rejection, the expression of cytotoxic T lymphocyte-associated antigen 4 in serum was reduced, showing some correlation with acute rejection after renal transplnatation. Cytotoxic T lymphocyte-associated antigen 4 might be involved in the rejection.

Expression of leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) on osteoclasts and its potential role in rheumatoid arthritis

OBJECTIVE: Leukocyte-associated immunoglobulin-like receptor-1 is an inhibitory receptor primarily expressed by immune cells. This study was undertaken to define the role of this molecule in osteoclast differentiation and rheumatoid arthritis. METHODS: In vitro osteoclast assays were performed to characterize the role of Leukocyte-associated immunoglobulin-like receptor-1 in murine and human osteoclastogenesis. Human Leukocyte-associated immunoglobulin-like receptor-1 expression was assessed by immunohistochemistry staining in the synovium of patients with rheumatoid arthritis. The levels of soluble Human Leukocyte-associated immunoglobulin-like receptor-1 were determined by enzyme-linked immunosorbent assay. RESULTS: We found that multinucleated osteoclast formation from mouse bone marrow cells was inhibited by treatment with a monoclonal antibody against mouse Leukocyte-associated immunoglobulin-like receptor-1 in vitro. By immunohistochemistry, we found that Leukocyte-associated immunoglobulin-like receptor-1 was mainly expressed by macrophages in the inflamed synovial tissue of rheumatoid arthritis patients. In addition, serum and synovial fluid levels of soluble Leukocyte-associated immunoglobulin-like receptor-1 were higher in rheumatoid arthritis patients compared to healthy controls or osteoarthritis patients. Moreover, overexpression of Leukocyte-associated immunoglobulin-like receptor-1 in CD14+ monocytes from healthy volunteers also inhibited human osteoclastogenesis. CONCLUSION: Collectively, these data demonstrate for the first time that Leukocyte-associated immunoglobulin-like receptor-1 inhibits osteoclastogenesis. Therefore, these results may have therapeutic implications for the treatment of rheumatoid arthritis. .

Expression of ligands of DNAM-1 and NKG2D in colonic cancer / 中华消化外科杂志

Objective To investigate the expression of ligands of DNAM-1 and NKG2D in the colonic cancer.Methods The colonic cancer tissue and adjacent normal colonic tissues were collected from 42 colonic cancer patients who were admitted to the Tangdu Hospital of Fourth Military Medical University from June 2010 to January 2011 were retrospectively analyzed.The expressions of CD155,CD112 and MICA/B in the colonic cancer tissues and the normal colonic tissues were detected by immunohistochemistry.The expressions of CD155,CD112 and MICA/B in the colonic cell line SWll6,SW480,SW620 and Colo205 in the Duke's A,B,C and D phases were detected by cell cytometry.The relationship of the expressions of the 3 ligands and the clinicopathological parameters was analyzed using the Mann-Whitney U test,chi-square test and Fisher exact probobility.Results Week expression of CD155 was found in the normal colonic tissues,while the expressions of CD112 and MICA/B were not found.In the colonic cancer tissues,the expressions of CD155,CD112 and MICA/B were 81.0％,52.4％ and 47.6％,which were significantly increased.The expressions of CD155,CD112 and MICA/B were not correlated with the gender,tumor differentiation,lymph node metastasis and Duke's staging (P ＞ 0.05).The overall expression rates of CD155,CD112 and MICA/B in the colonic cancer cell line SWll6,SW480,SW620 and Colo205 were 88.9％,67.4％ and 42.3％,respectively.The overall expression of CD155 was significantly higher than CD112 and MICA/B (F =23.17,P ＜ 0.05).Conclusion CD155,CD112 and MICA/B express in the colonic cancer tissues and colonic cancer cell line SW116,SW480,SW620 and Colo205,and the expression of CD155 is the highest.

Role of monitoring of bile soluble inhibitory receptor LAIR-1 and interleukin-2 receptor expression in liver acute rejection / 中华肝胆外科杂志

Objective Based on detection of the soluble LAIR-1 (sLAIR-1) and sIL-2R in the bile from recipient after liver transplant, the role of sLAIR-1 and sIL-2R in graft acute rejection were analyzed. Methods Bile sLAIR-1 level and sIL-2R were determined by double mAb sandwich enzyme linked immunosorbent assay in 55 cases of liver transplantation. Results In 22 recipients with normal graft function, sLAIR-1 and sIL-2R were detected at low level in the bile. In the 29 cases of liver acute rejection (AR), significant increase of bile sIL-2R level was detected on the lst and 2nd d before final diagnosis. With the effective methylprednisolone pulse therapy, sIL-2R level was decreased significantly on the 3rd d. On the other hand, remarkable increase of bile sLAIR-1 was found on the lst,2nd and 3rd d before final diagnosis. After of methylprednisolone pulse therapy for 3 d, bile sLAIR-1resturned to the control level. Conclusion Both bile sIL-2R and sLAIR-1 are detected at high level in the recipients suffering from liver acute rejection. The level of bile sLAIR-1 changes dramatically and responsively according to liver acute rejection. Therefore, detecting these two markers synergistically may be a promising monitor for rejection after liver transplantation.

Immunologic function of palatine tonsil B lymphocyte / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study cellular immune function of palatine tonsil B lymph cell.@*METHOD@#The phenotype of palatine tonsil cells (PTC) and that of peripheral blood mononuclear cell (PBMC) were compared using fluorescence staining and flow cytometry (FCM) analysis, then immunomagnetic beads were used to separate CD3- cell in PTC and PBMC. The proliferation function of CD3- lymph cell of PTC and PBMC was tested after stimulated by CD20mAb.@*RESULT@#FCM analysis founding that 71.2% PTC express CD20 with higher mean fluorescence intensity, MFI, compared to the 15.5% in PBMC. There's no significant difference between the proliferation of PTC and PBMC B lymph cell.@*CONCLUSION@#CD20 expression is different in PTC and PBMC, but corresponding function is still unknown.

Establishment of leukocyte-associated immunoglobulin like-receptor 2 (CD306) eukaryotic expression vectors and purification and identification of fusion protein / 中国组织工程研究

BACKGROUND: Biological function of leukocyte-associated immunoglobulin like-receptor 1 (LAIR 1) has clearly researched in China and abroad, but the in vivo biological function of LAIR is poorly understood. OBJECTIVE: To establish LAIR-2 (CD306) eukaryotic expression vectors and to purify and identify the fusion protein. DESIGN, TIME AND SETTING: A single sample observation experiment was performed at the Fourth Military Medical University of Chinese PLA between June 2007 and June 2008.MATERIALS: plg/3c vector was offered by Oxford University, pcDNA3.1 vector was provided by Meyaard doctor. Chinese hamster ovary (CHO) cell lines were preserved by the Department of Immunology, Fourth Military Medical University of Chinese PLA.METHODS: Two eukaryotic expression vectors plg/3c-LAIR-2 and pcDNA3.1-LAIR-2 were constructed and were transfected into CHO cells. The binding activities of LAIR-2 fusion protein to LAIR-2 mAbs were identified by Western blot, immunocytochemistry and flow cytometry assay.MAIN OUTCOME MEASURES: The construction of stably transfected cell lines, and the purification and identification of fusion protein. The activity of LAIR 2 protein combined to corresponding monoclonal antibody.RESULTS: Eukaryotic expression vectors were constructed and trasnsfected into CHO cells successfully. Two cells lines CHO/LAIR-2-Fc and CHO/LAIR-2 that steadily expressed LAIR-2-Fc fusion protein and LAIR-2 protein were established. Western blot assay showed that LAIR-2 protein could bind specially to LAIR-2 mAb 1A7, 3H12 and 4A9. Immunocytochemistry and flow cytometry assay demonstrated that 3H12 and 4A9 could bind to LAIR-2 expressed in the transfected CHO cells. CONCLUSION: Two ceils lines CHO/LAIR-2-Fc and CHO/LAIR-2 were successfully constructed, which can transfected to CHO cells. The eukaryotic expressed LAIR-2 protein has good binding activity to LAIR-2 mAbs.

Role of an inhibitory receptor CD305 in renal allograft rejection / 中华泌尿外科杂志

Objective To investigate the role of soluble CD305(sCD305)in renal allograft rejection.Methods Concentration of serum sCD305 was detected on 20 healthy volunteers and 153 cases of recipients after kidney transplantation by using double monoclonal antibody sandwich enzyme linked immunosorbent assay.Results In the healthy volunteers and 98 recipients with normal renal funetion,sCD305 was detected at low levels of(4.3±2.3)μg/L and(6.3±3.7)μg/L.In 20 cases of acute rejection and 5 cases of graft loss,serum sCD305 levels were(36.3±14.7)μg/L and(28.8±9.4)μg/L,and significantly higher than those in the healthy volunteers and recipients with normal renal function.Meanwhile,in the 30 cases of chronic rejection and 6 cases under dialysis treatment,the levels of sCD205were(13.1±5.5)ttg/L and(11.2±4.6)μg/L and significantly higher than those in the healthy volunteers and recipients with normal renal function.Conclusions CD305 was presented at high level in the recipients with renal acute or chronic rejection,and it might be a potential marker for monitoring graft rejection after transplantation.

Role of an inhibitory receptor LAIR-1 expression in graft rejection / 中华器官移植杂志

Objective To investigate the relationship between the soluble LAIR-1(sLAIR-1)in the serum from recipients after transplant and graft rejection.Methods Serum sLAIR-1 level was determined by double mAb sandwich enzyme linked immunosorbent assay on 23 cases of liver transplantation and 139 cases of kidney transplantation.Results In healthy volunteers and 98 recipients with normal graft function,sLAIR-1 was detected at low level [(4.3±2.3)μg/L and(6.3±3.7)μg/L],with the difference being not significant.In 6 cases of liver acute rejection,20 cases of kidney acute rejection and 5 cases of graft loss,serum sLAIR-1 levels were increased remarkably at high 1evels [(47.2±25.9)μg/L,(36.3±14.7)μg/L,and(28.8±9.4)μg/L respectively]as compared with the two groups of healthy volunteers and the recipients with normal graft function,even peaked at 117.3 μg/L in one case of severe liver rejection.Meanwhile,in 5 cases of liver chronic rejection,27 cases of kidney chronic rejection and 6 cases under dialysis treatment.the levels of sLAIR-1 were(16.1±6.4)μg/L,(13.1±5.5)μg/L and(11.2±4.6)μg/L respectively,significantly higher than those of the healthy volunteers and the recipients with normal graft function.Conclusion sLAIR_1 was detected at high level in the recipients suffered graft acute or chronic rejection and might be a promising monitor of rejection after transplantation.

Expression and significance of soluble LAIR in sera of renal transplantation recipients with cytomegalovirus pneumonitis / 中华泌尿外科杂志

Objective To study the relationship of soluble LAIR (sCD305 and CD3060) expression in recipient serum with cytomegalovirus (CMV) pneumonitis after renal transplantation. Methods Nineteen serum specimens from recipients were divided into CMV pneumonitis group (n=10) and control group (n=9). Then the concentrations of sCD305 and CD3060 were quantitated with sandwich ELISA. The data were analyzed by using student t test. Results sCD305 was skewness distributed in both 2 groups, was 0.000-3.039 μg/L in CMV pneumonitis group and 0.000-8.375 μg/L in con-trol group. CD3060 was skewness distributed in CMV pneumonitis group and the concentration was 0.000-0.017μg/L. CD3060 was mormally distributed in control group and the concentration was 0.046±0.035 μg/L. There was significant difference of CD3060 (P=0.000) concentrations and no sig-nificant difference of sCD305(P=0.316) concentrations in 2 groups, respectively. Conclusions The concentration of CD3060 is low in CMV pneumonitis patients. The combination of CMV PP65 antigen detection and CD3060 detection is helpful for the early and precise diagnosis of CMV pneumonitis in renal transplantation patients.

Eukaryotic expression and identification of human pSecTag2B-CD226 fusion protein / 医学研究生学报

Objective: To construct and express the eukaryotic expression vector of human pSecTag2B-CD226(PTA1).Methods: The gene fragment encoding extracellular region of human CD226 was cloned into the eukaryotic expression vector pSecTag2B.After sequencing,the vector was transfected into COS-7 cells,and the expressed molecule was purified by affinity chromatography.Finally,the product was characterized by ELISA.Results: hCD226-6His was successfully expressed.After purification,the concentration of hCD226-6His was 50?g/ml.Conclusion: Human CD226-6His fusion protein involving the extracellular region of CD226cDNA has been successfully expressed and purified,which helps prepare the ground for further functional studies of this molecule.

The expressions of decoy receptors of TRAIL in carcinoma of endometrium / 医学研究生学报

Objectives:To determine the expressions of decoy receptors (DcR1 and DcR2) of TRAIL in carcinoma of endometrium. Methods:The expressions of DcR1 and DcR2 in endometrium tissues from 13 carcinoma of endometrium and 7 normal endometrium were detected by immunohistochemical staining.Results: The expressions of DcR1 and DcR2 in carcinoma of endometrium were much lower than in normal endometrium. Conclusions:The decreasing of DcR1 and DcR2 in carcinoma of endometrium may be concerned with its pathogenesis, which may be related to the prevention of endometrium from carcinomatous change.

Correlation between IL-6 levels and polymerized human serum albumin ceptor (PHSAR) in serum of chronic hepatitis patients / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To investigate the relationship between interleukin-6 (IL-6) levels in serum and HBV replication and the role of IL-6 in liver damage in chronic hepatitis B patients.</p><p><b>METHODS</b>Detected IL-6 levels and polymerized human serum albumin receptor (PHSA-R) in serum of different type of HBV markers positive patients and of different degree of chronic hepatitis B patients.</p><p><b>RESULTS</b>The results showed that the serum IL-6 levels were increased significantly in PHSA-R positive patients than in PHSA-R negative patients in chronic hepatitis B (P<0.01). The serum IL-6 levels were correlated with the levels of PHSA-R (r=0.694, P<0.01), and the degree of symptoms.</p><p><b>CONCLUSIONS</b>This study suggested that the IL-6 levels in serum of hepatitis B patients correlated HBV replication and the degree of liver damage, serum IL-6 levels may be used as a value indicating of HBV replication, the degree of symptoms and the effect of treatment.</p>

Identification of the Peptides that Bind to PTA1 mAb(LeoA1) by Phage Displayed Library / 细胞与分子免疫学杂志

Aim To screen peptides binding specifically to anti-human PTA1mAbs from a random twelve-peptide phage-disp-layed library. Methods Series of PTA1mAbs(LeoA1、 1B11、 C9、 2D1、 2E9、 2G8、 2H2 and E8)were purified using protein-A affinity column. PTA1mAbs which could bind PTA1-Fc fusion protein binding to its ligand were confirmed by flow cytometry, and then used as target to screen phage library. After three rounds of affinity screening, the peptide sequences of positive phage clones were determined and analyzed. Results LeoA1 could block PTA1-Fc fusion protein binding to its ligand. 13 phages which could bind specifically to LeoA1 were isolated from phage library and further confirmed by ELISA. Conserved motifs were found among the sequences of the peptides. Conclusion It was shown that the conserved motifs were candidafe regions binding to PTA1 ligand,which is important to identify functional epitopes for seeking ligand of PTA1 and further investigation of biological function of PTA1.

Chemokine receptors in regulating immune response and therapy / 中国病理生理杂志

Although adhesion molecules have long been recognized to be differentially expressed on naive and effector/memory T cells,it has recently been found that a number of chemokine receptors are also differentially expressed on T cells,depending on their Ag experience and type of polarization. Recent data suggests that chemokines and their receptors are essential elements that regulate the positioning of T cells and their partners for priming and T helper 1 (Th1)- or Th2-mediated responses,therefore,are probably the most promising targets for treating immune diseases.

Identification of 5-flank upstream regulation region of CD226 / 中国免疫学杂志

Objective:To identify the function of the 5′-flank upstream regulation region of human CD226 gene.Methods:The upstream regulation region of CD226 was cloned by PCR and ligated into pGL3 vector. Then the vector was transfected into Jurkat cell and luciferase activity was detected after 48 h culture.Results:CD226 gene may have two promoters, P1 and P2,which were located at the region of -843--319 bp and +1-+181 bp respectively, and PMA can up-regulate P1 while down-regulate P2. Both P1 and P2 can be up-regulated by A23187, especially P2.Conclusion:CD226 gene may have two promoters, and PMA and A23187 can regulate CD226 promoter activity in the similar pattern of protein level.

Expression and function of CD226 with CD16 on NK subsets / 中国免疫学杂志

Objective:To investigate the expression and function of CD226 on NK s ubsets and its coexpression with other activation receptor and inhibition recept or on NK cells. Methods:The expression of CD226 on CD56 bright and CD56 dim NK subsets and coe xpress ion with CD16 and NKG2A in PBMC and MLC in the presence or absence of IL-2 or IL-15 were detected by double fluorescent staining and flow cytometry analysis. The level of IFN-? in the supernatants of PBMC culture and MLC treated with o r without IL-2 or IL-15 were evaluated by ELISA. 51Cr release assay w as employed to measure the specific lysis of NK cells killing target K562 cells. Results:CD226 was mainly expressed on CD56 dim NK subsets in PBMC. When s imulated by IL-2, CD226 expression was shifted to CD56 bright NK subsets, while IL-15 increased the subpopulation of NKG2A+CD226+double positive cell s. In MLC-generated NK cells, CD226 was mainly expressed on CD56 dim NK su bsets, and also shifted to CD56 bright NK subsets in the addition of IL-15 . Furt hermore, the percentage of CD16+CD226+和NKG2A+CD226+ subsets were increa sed when stimulated by IL-2 or IL-15. There was great increase in IFN-? lev el in the supernatants of PBMC culture in the presence of IL-2 or IL-15, but no difference in the supernatants of MLC treated with or without two cytokine s. Moreover, the cytotoxicity of NK cells in PBMC and MLC were greatly enhanced by IL-2 or IL-15. Conclusion:CD226 is mainly expressed on CD56 bright NK subsets in IL-2 or IL-1 5 activated NK cells, and is coexpressed with CD16 and NKG2A preferentiatly, whi ch maybe involve in the modulation of cytotoxicity of NK cells based on the balance of coexpressed activation and inhibition receptors. [

The effects of alanyl-glutamine dipeptide-enriched parenteral nutrition on outcome in radiation enteritis rats / 肠外与肠内营养

Objective:To investigate the impact of alanyl-glutamine(Ala-Gln) on outcome in radiation enteritis rats.Methods: Male SD rats(n=70)were separated randomly into four groups: control group(n=10),AR+pseudosurgery group(n=20),AR+TPN group(n=20) and AR+TPN+Ala-Gln group(n=20).Rats were observed for mortality,changs of body weight,villous hight and area,the bcteriral translocation(BT)in mesenteric lymph nodes(MLNs),liver,spleen and peritoneal cavity.Serum TNF-? and sIL-2R level were determined by sandwich ELISA.Results: When Ala-Gln was administered in radiation enteritis rat,the mortality,body weight loss and bacterial translocation were decreaded,the villous hight and area was increased and the TNF-? and sIL-2R levels were reduced.Conclusion: Parenteral Ala-Gln nutrition can improves the results of radiation enteritis rats.

Detection of Serum Soluble CD226/PTA1Level in Patients with Psoriasis Vulgaris / 中华皮肤科杂志

Objective To determine the serum soluble CD226/PTA1(sCD226/PTA1)level in pa-tients with psoriasis vulgaris.Methods Serum sCD226/PTA1level was measured by sandwich ELISA in30patients with psoriasis vulgaris before treatment(17patients in active stage and13in inactive stage),10patients after treatment,and15healthy individuals as a control.Results Serum sCD226/PTA1level was significantly increased in patients with psoriasis compared with that in healthy controls(P

The study on TLSF_(JM)mRNA translation in Xenopus Lavevis Oocytes / 中国免疫学杂志

Total RNA from JM cell lines secreting T leukmia derived suppressor factor was isolated using single—step method of acid guanidium followed by oligo—d(T)—cellulose affinity chromotogra-phy for purification of mRNA.TLSF_(JM)mRNA was mieroinjected into Xenopus Laevis Oocytes.The inhibitory activity in Xenopus Laevis Oocytes culture supernatant or lysate was detected by PHA plus IL—2 induced—mouse thymocyte proliferation assay The results showed that Xeno-pus Laevis Oocytes could efficiently translate exogenous mRNA,the trans ated products mainly existed inside oocytes and approximately2% of the products was secreted into the culture super-natant.Success in TLSF_(JM)mRNA translation in vitro might provide the basis for establishment of cDNA library and TLSF_(JM)gene cloning and expression.

A bioassay method for human Granulocyte—Macrophage Colony stimulating factor with DMSO—induced HL60 cell line / 中国免疫学杂志

Based on the high level of human granulocyte—macrophage colony—stimulating factor(HGM—CSF)re-ceptor of HL60 cell line upon induction by DMSO,we developed a hioassay method for HGM—CSF with ahigher specificity and sensitivity.The sensitivity of this assay reached to 0.3ng/ml by selecting the optimalHL60 cell—induced time,cell density and GM—CSF—stimulated time.With this assay,we are able to quantifythe level of rHuGM—CSF as well as conditioned media from human bladder carcinoma cell line U5637 screatinga high level of GM—CSF.