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1.
Immune Network ; : 392-401, 2017.
Article in English | WPRIM | ID: wpr-102687

ABSTRACT

We previously reported peritoneal innate-like integrin α4 (CD49d)highCD4+ T cells that provided help for B-1a cells. Here we analyzed the expression of various integrin chains on the peritoneal and pleural integrin α4highCD4+ T cells and investigated the functional heterogeneity of the subpopulations based on the integrin expression. Pleural cavity contained a lower ratio of integrin α4highCD4+ T cells to integrin α4lowCD4+ T cells than peritoneal cavity, but the pleural integrin α4highCD4+ T cells have the same characteristics of the peritoneal integrin α4highCD4+ T cells. Most of integrin α4highCD4+ T cells were integrin β1highβ7−, but a minor population of integrin α4highCD4+ T cells was integrin β1+β7+. Interestingly, the integrin α4highβ1highβ7− CD4+ T cells expressed high levels of integrin α4β1 and α6β1, whereas integrin α4highβ1+β7+ CD4+ T cells expressed high levels of integrin α4β1 and α4β7, suggesting an alternative expression of integrin α6β1 or α4β7 in combination with α4β1 in respective major and minor populations of integrin α4highCD4+ T cells. The minor population, integrin α4highβ1+β7+ CD4+ T cells, were different from the integrin α4highβ1highβ7− CD4+ T cells in that they secreted a smaller amount of Th1 cytokines upon stimulation and expressed lower levels of Th1-related chemokine receptors CCR5 and CXCR3 than the integrin α4highβ1 highβ7− CD4+ T cells. In summary, the innate-like integrin α4highCD4+ T cells could be divided into 2 populations, integrin α4β1+α6β1+α4β7− and α4β1+α6β1−α4β7+ cells. The functional significance of serosal integrin α4β7+ CD4+ T cells needed to be investigated especially in view of mucosal immunity.


Subject(s)
CD4-Positive T-Lymphocytes , Cytokines , Immunity, Mucosal , Integrin alpha4 , Peritoneal Cavity , Pleural Cavity , Population Characteristics , Receptors, CCR5 , Receptors, Chemokine , Receptors, CXCR3 , T-Lymphocytes , Th1 Cells
2.
IJRM-Iranian Journal of Reproductive Medicine. 2014; 12 (10): 687-694
in English | IMEMR | ID: emr-148984

ABSTRACT

Endometrial integrin expression changes might be a reason for implantation failure in polycystic ovarian syndromes [PCOS]. Assessment of integrin genes and proteins expression upon endometrium in the PCOS experimental mouse model was the main goal of this study. 30 NMRI female mice were equally divided into control, experimental [PCOS; received estradiol valerate [40 mg/kg]] and sham group [received; olive oil]. After 8 weeks, each group was hyper stimulated by 7 IU PMSG and then, after 48hrs, 7 IU HCG was injected. Vaginal plaque was checked. After 5 days, Progesterone and estradiol levels and endometrial tissues were investigated to evaluate of alpha4, alphav, beta1 and beta3 integrins gene and protein by qPCR method and immunohistochemistry, respectively. Tissue samples were assessed and showed that level of progesterone was significantly decreased in PCOS group. Results of molecular part in the amount of alphav, beta3, beta1 and alpha4 gene expressions showed a great difference in beta3 and alphav genes expressions between experimental groups, alphav, beta3, alpha4 and beta1 proteins in the endometrial stroma in the control group were expressed, but they were not detected in PCOS group. According to the results, integrins had different expression patterns in different areas of the endometrium; such as epithelial and stromal. It seems that in PCOS, this pattern has changed and the results might have a great influence on implantation failure. Therefore, this study suggests that a great attention to this problem may be essential in patients who are involved


Subject(s)
Animals, Laboratory , Integrin alpha4 , Integrin alpha5 , Integrin beta3 , Integrin beta1 , Integrins , Endometrium , Mice , Gene Expression , Embryo Implantation
3.
IJRM-Iranian Journal of Reproductive Medicine. 2014; 12 (9): 623-632
in English | IMEMR | ID: emr-149664

ABSTRACT

It has been hypothesized that blastocyst integrin expression changes can affect the spontaneous miscarriage in polycystic ovarian syndromes [PCOS]. In this study, the profile of integrin genes and proteins was investigated on blastocyst of the PCOS experimental mouse model. 30 NMRI female mice were equally divided into 3 groups: control, experimental [PCOS that was injected estradiol valerate [40 mg/kg]]. After 8 weeks, each group was hyper stimulated by PMSG and HCG. Vaginal plaque was checked, and mice were investigated 5 days after the test. Progesterone and estradiol levels were determined; alpha4, alpha v, beta 1 and beta 3 integrin genes and protein of blastocysts were examined by real time PCR method and immunohistochemistry, respectively. Estradiol level was significantly increased [p

Subject(s)
Animals, Laboratory , Integrin alpha4 , Integrin alphaV , Integrin beta3 , Integrin beta1 , Blastocyst , Mice , Integrins , Embryo Implantation
4.
Journal of Experimental Hematology ; (6): 1552-1556, 2013.
Article in Chinese | WPRIM | ID: wpr-264977

ABSTRACT

The main aim of this study was to investigate the biological activities and immune modulation changes of chorionic villi mesenchymal stem cells (CV-MSC) after long term culture. The morphology of the CV-MSC of passage 3 and passage 9 were observed by microscopy, and their phenotypes were detected by flow cytometry. CV-MSC of passage 3 and 9 were co-cultured with PHA-stimulated PBMNC, and IFN-γ concentration in culture medium was detected by ELISA. The mRNA expression of COX-2, HGF and HLA-G in CV-MSC were detected by real-time PCR. The results showed that after long term culture, the CV-MSC kept the MSC morphology and most of the phenotypes including CD31, CD34, CD44, CD45, CD62L, CD73, CD90, CD105, CD117, CD151, CD235a, CD271 and HLA-DR, while the CD49d was significantly up-regulated. Immune modulation ability of CV-MSC was reduced and the mRNA expression of COX-2 and HGF was down regulated after long term culture, but the expression of HLA-G did not found to be obvious change. It is concluded that the long term in vitro expansion changes the expression of CD49d and reduces immune modulation of CV-MSC.


Subject(s)
Female , Humans , Pregnancy , Cells, Cultured , Chorionic Villi , Allergy and Immunology , Integrin alpha4 , Metabolism , Mesenchymal Stem Cells , Cell Biology , Allergy and Immunology , Monocytes , Cell Biology , Placenta , Cell Biology
5.
Journal of Korean Medical Science ; : 815-819, 2007.
Article in English | WPRIM | ID: wpr-176606

ABSTRACT

The house dust mite (HDM) is considered to be the most common indoor allergen associated with bronchial asthma. In this study, we investigated whether crude extract of the HDM Dermatophagoides farinae could activate human eosinophilic leukemic cells (EoL-1) to induce upregulation of cell-surface adhesion molecules. When EoL-1 cells were incubated with D. farinae extract, expression of intercellular adhesion molecule-1 (ICAM-1) significantly increased on the cell surfaces compared to cells incubated with medium alone. In contrast, surface expression of CD11b and CD49d in EoL-1 cells was not affected by D. farinae extract. In addition, pretreatment of cells with NF- kappaB inhibitor (MG-132) or JNK inhibitor (SP600125) significantly inhibited ICAM-1 expression promoted by HDM extract. However, neither p38 MAP kinase inhibitor nor MEK inhibitor prevented HDM-induced ICAM-1 expression in EoL-1 cells. These results suggest that crude extract of D. farinae induces ICAM-1 expression in EoL-1 cells through signaling pathways involving both NF- kappaB and JNK.


Subject(s)
Animals , Humans , Anthracenes/pharmacology , CD11b Antigen/biosynthesis , Cell Line, Tumor , Cell Membrane/metabolism , Eosinophils/metabolism , Flow Cytometry/methods , Gene Expression Regulation , Integrin alpha4/biosynthesis , Intercellular Adhesion Molecule-1/metabolism , Leukemia/metabolism , Leupeptins/pharmacology , Mitogen-Activated Protein Kinase 8/metabolism , NF-kappa B/metabolism , Pyroglyphidae , p38 Mitogen-Activated Protein Kinases/metabolism
6.
Journal of Experimental Hematology ; (6): 318-321, 2006.
Article in Chinese | WPRIM | ID: wpr-280675

ABSTRACT

This study was purposed to explore the effect of different cytokine combinations on the expansion of the mononuclear cells drived from umbilical cord blood (CB) ex vivo and expression of CXCR4 and CD49d on CD34+ cells after expansion. Human fresh CB mononuclear cells were cultured in serum-free and stroma-free medium containing different combinations of cytokine for 7 days. At day o and 7, the total cells were counted, CD34+ cells and CD34+CXCR4+, CD34+CD49d+ cells were assayed by flow cytometry, and CFU were determined. According to the different combinations of cytokine, experiments were divided into four groups: control, SF group (SCF + FL), SFT group (SCF + FL + TPO) and SFT6 group (SCF + FL + TPO + IL-6). The results showed that the SF (SF group) combination supported only low expansion of total cells, CD34+ cells and CFU. The addition of TPO in SF group restored UCB stem/progenitors expansion to a higher level than that in SF group, while there was no difference between groups SFT and SFT6 (P > 0.05). The cytokine combinations in groups SF, SFT and SFT6 all could upregulate the expression levels of CD49d and CXCR4 on expanded cord blood CD34+ cells, but there were no significant differences between groups SF, SFT and SFT6 (P > 0.05). It is concluded that SCF + FL has no strong synergistic effects on primitive hematopoietic cells. TPO plays an important role in enhancing expansion of umbilical cord blood hematopoietic cells, while IL-6 only shows a neutral effect on it. SCF + FL + TPO combination not only promotes progenitor cells expansion but also upregulates the expression of CD49d and CXCR4 on CD34+ cells from cord blood.


Subject(s)
Humans , Antigens, CD34 , Genetics , Cytokines , Pharmacology , Drug Synergism , Fetal Blood , Cell Biology , Integrin alpha4 , Genetics , Leukocytes, Mononuclear , Cell Biology , Membrane Proteins , Pharmacology , Receptors, CXCR4 , Genetics , Stem Cell Factor , Pharmacology , Thrombopoietin , Pharmacology
7.
Chinese Journal of Hematology ; (12): 240-243, 2006.
Article in Chinese | WPRIM | ID: wpr-243966

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the biological behavior of stromal cell-derived factor-1 (SDF-1) on multiple myeloma (MM) cell migration and adhesion and it related signaling pathways.</p><p><b>METHODS</b>Expression of adhesion molecules on MM cells of RPMI8226, XG-1 and XG-7 cells was analysed by flow cytometry, the influence of SDF-1 on CD29 and CD49e distribution by immunofluorescence, the effect of SDF-1 on chemotaxis of MM cells by transwell assay. Activation of phosphoinositide-3 kinase (PI3K) in MM cells treated with SDF-1 and by immunoblotting.</p><p><b>RESULTS</b>3 strains of MM cell line expressed many adhesion molecule. RPMI8226, XG-7 cells were all high level of expression of CD29 (> 70%). XG-1, XG-7 cells were all high level of expression of CD44 (> 80%), and XG-7 cells was of CD49d (> 90%). In all of 3 strains, the levels of expression of CD49e were low (< 30%). SDF-1 could not upregulate their expression, but could trigger the establishment of polarized morphology of MM cells and the redistribution of CD29 and CD49e. SDF-1 promoted MM cells adhesion to endothelial cells, stimulated phosphorylation of P85 subunit of PI3K in MM cells and induced MM cells migration, which were inhibited by G protein inhibitor PTX and PI3K inhibitor wortmannin.</p><p><b>CONCLUSION</b>SDF-1 can promote MM cell adhesion to endothelial cells, trigger establishment of a polarized morphology of MM cells and redistribution of adhesion molecules and induce MM cells migration via PI3K signaling pathway.</p>


Subject(s)
Humans , Blotting, Western , Cell Adhesion , Physiology , Cell Adhesion Molecules , Metabolism , Cell Line, Tumor , Cell Movement , Physiology , Chemokine CXCL12 , Pharmacology , Physiology , Enzyme Activation , Flow Cytometry , Fluorescent Antibody Technique , Integrin alpha4 , Metabolism , Integrin alpha5 , Metabolism , Integrin beta1 , Metabolism , Multiple Myeloma , Metabolism , Pathology , Phosphatidylinositol 3-Kinases , Metabolism , Signal Transduction , Physiology
8.
Experimental & Molecular Medicine ; : 191-195, 2006.
Article in English | WPRIM | ID: wpr-96573

ABSTRACT

The movement of leukocytes from the blood into peripheral tissues is a central feature of immune surveillance, but also contributes to the pathogenesis of inflammatory and autoimmune diseases. Integrins are a family of adhesion and signaling molecules made up of paired alpha and beta subunits, and the integrin alpha4beta1 plays a prominent role in the trafficking of mononuclear leukocytes. We have previously described the direct interaction of the signaling adaptor molecule paxillin with the cytoplasmic domain of the alpha4 integrin subunit. This interaction is critical for alpha4beta1 integrin dependent cell adhesion under shear flow conditions as it provides a needed connection to the actin cytoskeleton. Furthermore, the alpha4-paxillin interaction is required for effective alpha4beta1 dependent leukocyte migration and does so through the temporal and spatial regulation of the small GTPase Rac. These findings make the alpha4-paxillin interaction a potentially attractive therapeutic target in controlling leukocyte trafficking.


Subject(s)
Humans , Protein Binding , Paxillin/metabolism , Models, Biological , Leukocytes/cytology , Integrin alpha4beta1/metabolism , Integrin alpha4/metabolism , Cell Movement/physiology , Cell Adhesion/physiology
9.
Journal of Experimental Hematology ; (6): 664-668, 2005.
Article in Chinese | WPRIM | ID: wpr-343909

ABSTRACT

TGF-beta, as an inhibitor of hemopoiesis, excreted by hematopoietic stem and progenitor cells, down-regulates the expression of cytokines such as Flt-3 ligand, SCF, IL-3 etc on the stem and progenitor cells. The effect of anti-TGF-beta antibody on ex vivo expansion and expression of adhesive molecules on cord blood CD34(+) cells was studied in this research. The CD34(+) cells from six units of fresh umbilical cord blood were enriched by density gradient sedimentation and purified by miniMACS cell isolation system, and plated them into the SFEM serum free culture system which containing SCF, Flt-3L, TPO and IL-3 in the condition of 37 degrees C, 5% CO2, and saturated moisture. There were three groups in this experiment: (1) blank group: same as the culture system described above; (2) control group: added with normal rabbit IgG into the mentioned culture system; (3) test group: the same culture system with anti-TGF-beta1 antibo-dy. Cultured for 6 days, the number of mononuclear cells (MNC) was counted, the expression of CD34 antigen, CD117 (c-kit) antigen, CD11a antigen, CD49d antigen and CD33 antigen was tested with FCM. Meanwhile, cells of the three groups were plated in the methylcellulose culture system for 14 days, the number of CFU-GEMM, BFU-E, CFU-GM was counted. The results indicated that the expansion multiples of MNC, CD34(+) cells, CD34(+)c-kit(+) cells, CFU-GEMM in the test group (41.82 +/- 13.49, 15.62 +/- 6.95, 13.36 +/- 6.12, 11.07 +/- 4.05) were significantly higher than in the control group (28.86 +/- 9.03, 10.40 +/- 4.98, 9.04 +/- 4.40, 6.36 +/- 2.37) (P = 0.001, 0.002, 0.003, 0.002) respectively. The expansion multiple of more primitive CD34(+)c-kit(-) subpopulation in the test group (69.10 +/- 41.06) was even higher than in the control group (27.29 +/- 10.40) (P = 0.024). Adhesion molecule expression on the CD34(+) cells after short-term expansion: the expression of CD11a on the CD34(+) cells of the original cord blood was (61.73 +/- 4.13)%, and CD49d was (55.12 +/- 5.22)%. After expansion in each group the expression of CD11a on the CD34(+) cells did not change with statistical significance (P > 0.05), the expression of CD49d increased (P < 0.05). Compared with blank group and control group, anti-TGF-beta antibody did not impact on the expression of CD11a and CD49d (P > 0.05). It is concluded that anti-TGF-beta antibody can synergize other cytokines to effectively enhance the proliferation of cord blood NC, CD34(+) cells, progenitor subpopulation of CD34(+)c-kit(-) cells, and increase the output of more primitive progenitor colony, CFU-GEMM and BFU-E. At the same time, anti-TGF-beta antibody did not depresss the expression of adhesion molecules on CD34(+) cells.


Subject(s)
Female , Humans , Pregnancy , Antibodies , Pharmacology , Antigens, CD34 , CD11a Antigen , Cell Adhesion Molecules , Cell Proliferation , Cells, Cultured , Fetal Blood , Cell Biology , Allergy and Immunology , Flow Cytometry , Integrin alpha4 , Proto-Oncogene Proteins c-kit , Transforming Growth Factor beta , Allergy and Immunology
10.
Journal of Experimental Hematology ; (6): 422-428, 2005.
Article in Chinese | WPRIM | ID: wpr-356545

ABSTRACT

This study was aimed to explore the role of human fetal bone marrow stromal cells (FBMSC) in combination with exogenous cytokines in supporting the in vitro expansion of cord blood mononuclear cells and the expression of CXCR4(+) and CD49d(+) in CD34(+) cells. Mononuclear cells (MNC) separated from cord blood (CB) were cultured in a serum-free support culture system with FBMSC or exogenous cytokines or both of them. On day 0, 6, 10 and 14, total cells were counted, CD34(+), CD34(+)CXCR4(+) and CD34(+)CD49d(+) cells were quantitated by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay. The results showed that after culturing for 14 days, CD34(+) cells, CD34(+)CXCR4(+) cells, CD34(+) CD49d(+) cells and colony forming unit (CFU) were significantly increased (P < 0.05). Compared with other groups, expansion multiple of CD34(+), CD34(+)CXCR4(+), CD34(+)CD49d(+) cells and CFU were higher than that in FBMSC and cytokine group (P < 0.05). It is concluded that the culture system used in this study can not only support the expansion of CB MNCs but also increase the number of hematopoietic stem and progenitor cells which has chemokine and adhesion capacity. This culture system may be a feasible way for in vitro culture of cord blood cells.


Subject(s)
Humans , Antigens, CD34 , Blood , Bone Marrow Cells , Cell Biology , Allergy and Immunology , Cell Proliferation , Cells, Cultured , Coculture Techniques , Cytokines , Pharmacology , Fetal Blood , Cell Biology , Fetus , Flow Cytometry , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Integrin alpha4 , Blood , Interleukin-3 , Pharmacology , Leukocytes, Mononuclear , Cell Biology , Allergy and Immunology , Receptors, CXCR4 , Blood , Stromal Cells , Cell Biology , Allergy and Immunology , Time Factors
11.
Journal of Experimental Hematology ; (6): 807-811, 2004.
Article in Chinese | WPRIM | ID: wpr-347857

ABSTRACT

The objective of this study was to explore the effect of culture system from embryonic fibroblasts and leukemia inhibitory factor (LIF) on expansion of mouse bone marrow hematopoietic progenitor cells ex vivo, and to observe its effect on the expression of homing-related cell adhesion molecules among VLA-4 (CD49e), VLA-5 (CD49e), LFA-1 (CD11a), HCAM (CD44) and L-selectin (CD62L). The culture system from the mouse embryonic fibroblasts inactivatd by mitomycin C and contained LIF was used to culture with mouse BMMNC for 7 days. The total number of BMMNC, CFC, Sca-1(+) cells, cell apoptosis rate and the expression of above cell adhesion molecules were counted. The results showed that culture system consisted of embryonic fibroblasts and LIF significantly enhanced the total number of BMMNC, CFC, Sca-1(+) cells, suppressed cell apoptosis (P < 0.05). In control without MEF and LIF, the total number of BMMNC was reduced remarkedly, CFC and Sca-1(+) cells were completely dead, the majority of cells produced apoptosis (P < 0.01). The expression of CD49d, Cd44 and CD61L on Sca-1(+) cells were similar to that befor expression (P < 0.05), but the expression of CD49e and CD11a on Sca-1(+) cells were remarkably increased (P < 0.05). It is concluded that culture system from embryonic fibroblasts and LIF can only significantly expand mouse bone marrow hematopoietic progenitor cells ex vivo, but the expanded hematopoietic progenitor may well sustain the expression of homing-related adhesion molecules. The homing functions of these expanded hematopoietic progenitors kept no change.


Subject(s)
Animals , Female , Male , Mice , Pregnancy , Antigens, Ly , Apoptosis , CD11a Antigen , Cell Differentiation , Cell Proliferation , Cells, Cultured , Coculture Techniques , Culture Media , Pharmacology , Embryonic Stem Cells , Cell Biology , Metabolism , Fibroblasts , Cell Biology , Metabolism , Hematopoietic Stem Cells , Cell Biology , Metabolism , Hyaluronan Receptors , Integrin alpha4 , Leukemia Inhibitory Factor , Pharmacology , Leukocytes, Mononuclear , Cell Biology , Metabolism , Membrane Proteins , Mice, Inbred BALB C
12.
Journal of Experimental Hematology ; (6): 816-820, 2004.
Article in Chinese | WPRIM | ID: wpr-347855

ABSTRACT

This study was aimed to investigate the expressions of adhesion molecules such as CD54, CD49d and CD62L by CD34(+) cells sampled from different stages of bone marrow (BM) and peripheral blood (PB) before/after G-CSF mobilization and after transplantation through the direct labeling with three colour-immunofluorescence and flow cytometry, and to explore the differences in expression of adhesion molecules on CD34(+) cells from different origins and their clinical significance. Mononuclear cells collected from BM and PB before mobilization, after collection of stem cells and hematopoietic recostruction of BM at the end of transplantation were marked with CD54-FITC, CD49d-FITC and CD62L-FITC separately, as well as CD34-PE and CD45PerCE. 3-color fluorescene analysis was carried out by FACS. The expression differences of CD34(+) and adhesion molecules between BM and APBSC were compared. The results showed that expression differences of CD54, CD49d and cd62Lon CD34(+) cells belore mobilization, after collection and reconstraction of transplantation were not statiscally significant, the difference of CD54, CD49d and CD62L on CD34(+) between 1st and 2nd collections of hematopoietic stem cells also were not statiscally significant. In the collected APBSC, the expression level of CD34(+) CD49d(+) was significantly lower than those in BM before mobilization (P = 0.001). It is concluded that the method of chemotherapy combined with G-CSF mobilization can down-regulate CD49d expression in BM CD34(+) cells, thus can mobilize and move theirs into peripheral blood. After the reconstitution by transplantation, the expression of CD49d on CD34(+) cells tends to normal, the clinical significance needs to be elucidated by accumulation of much more cases.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Antigens, CD , Blood , Antigens, CD34 , Blood , Antigens, Neoplasm , Blood , Antineoplastic Agents , Therapeutic Uses , Bone Marrow Cells , Allergy and Immunology , CD52 Antigen , Cell Adhesion Molecules , Blood , Combined Modality Therapy , Flow Cytometry , Glycoproteins , Blood , Hematopoietic Stem Cells , Allergy and Immunology , Integrin alpha4 , Blood , L-Selectin , Blood , Leukocytes, Mononuclear , Allergy and Immunology , Lymphoma , Blood , Allergy and Immunology , Therapeutics , Peripheral Blood Stem Cell Transplantation , Methods , Transplantation, Autologous
13.
Journal of Experimental Hematology ; (6): 59-62, 2004.
Article in Chinese | WPRIM | ID: wpr-278801

ABSTRACT

To study the mobilization of peripheral blood stem cells (PBSC) with anti-CD49d monoclonal antibody and try to find a new method for mobilization of PBSC, anti-CD49d McAb, rhG-CSF and combination of anti-CD49d McAb with rhG-CSF were administered subcutaneously to mice separately, the count of white blood cells (WBC) and percentage of CD34(+) cells in peripheral blood of donor mice were dynamically observed, CD34 positive cells obtained by above methods were transfused to recipient mice. The results showed that the count of WBC and percentage of CD34(+) cells in peripheral blood of donor mice elevated significantly after the administration of anti-CD49d McAb, rhG-CSF or combination of anti-CD49d McAb with rhG-CSF. The most effective method for mobilization is the combination of rhG-CSF with anti-CD49d McAb. Reconstitution of hematopoiesis was successful in all group recipient mice after transplantation. Most rapid hematopoietic recovery was observed in recipient mice by rhG-CSF plus anti-CD49d McAb for mabilization. In conclusion, anti-CD49d McAb is effective and synergistic with rhG-CSF in mobilization of CD34 positive cells from bone marrow into peripheral blood.


Subject(s)
Animals , Female , Male , Mice , Antibodies, Monoclonal , Pharmacology , Antigens, CD34 , Granulocyte Colony-Stimulating Factor , Pharmacology , Hematopoiesis , Hematopoietic Stem Cell Mobilization , Hematopoietic Stem Cell Transplantation , Integrin alpha4 , Allergy and Immunology , Mice, Inbred BALB C , Recombinant Proteins
14.
Journal of Korean Medical Science ; : 523-528, 2004.
Article in English | WPRIM | ID: wpr-168450

ABSTRACT

Homing-associated cell adhesion molecules (H-CAM) on the CD34+ cells play an important role for the engraftment process following hematopoietic stem cell transplantation (HSCT). However, it seems that not only CD34+ cells but also other nucleated cells (NCs) with H-CAM could be implicated in the engraftment process and the proliferation of hematopoietic stem cells. We investigated the differences of HCAM and cell cycle status on the NCs in cord blood (CB), bone marrow (BM), and mobilized peripheral blood (PB). The proportions of CXCR4+ cells within the NC populations were greater in CB than in PB or BM (p=0.0493), although the proportions of CXCR4+, CD44+, and CD49d+ cells within the CB CD34+ cell populations were same within BM or PB. A lower proportion of CD34+CD49d+ cells within the CD34+ cell populations was more noted in CB than in PB or BM (p=0.0085). There were no differences in cell cycle status between CB and BM or PB. Our results suggest that the migrating potential of CB would be enhanced with increased CXCR4 expression on the NCs, but the adhesion potential of CB CD34+ cells would be less than that of PB and BM. These findings may help explain why the lower cell dose is required and engraftment is delayed in cord blood stem cell transplantation.


Subject(s)
Humans , Antigens, CD34/metabolism , Hyaluronan Receptors/metabolism , Bone Marrow Cells/metabolism , Cell Adhesion Molecules/metabolism , Cell Cycle/physiology , Cell Proliferation , Cell Separation , Fetal Blood/cytology , Flow Cytometry , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells/metabolism , Integrin alpha4/metabolism , Receptors, CXCR4/metabolism
15.
Chinese Journal of Applied Physiology ; (6): 141-144, 2003.
Article in Chinese | WPRIM | ID: wpr-339658

ABSTRACT

<p><b>AIM</b>To study the effect of PF4 and relative peptide PF4 17-70 on the chemoattract ability, the expression of adhesion molecules and CXCR4 on the flesh cord blood CD34+ cells.</p><p><b>METHODS</b>CD34+ cells were separated from the cord blood using MACS immune magnetic beads, the chemoattract ability was assayed using the Transwell board, the expression of adhesion molecules and CXCR4 was measured by FACS.</p><p><b>RESULTS</b>(1) PF4 and PF4 17-70 increased the migration of the CD34+ cells, the chemoattract percentage of PF4 was 157.43% +/- 50.06% (P < 0.05) and PF4 17-70 was 187.02% +/- 10.69% (P < 0.05). (2) The expression of CD49d and CXCR4 on the CD34+ cells increased after PF4 incubated, but the expressions of other adherent molecules including CD31, CD44, CD11a, CD62p, CD62E did not change.</p><p><b>CONCLUSION</b>PF4 has the chemoattract ability on the umbilical blood CD34+ cells by promoting the expression of integrin CD49d and CXCR4, PF4 may help the cord stem cells homing.</p>


Subject(s)
Humans , Antigens, CD34 , Metabolism , Cell Adhesion Molecules , Metabolism , Chemotaxis , Fetal Blood , Cell Biology , Hematopoietic Stem Cells , Cell Biology , Integrin alpha4 , Metabolism , Platelet Factor 4 , Pharmacology , Receptors, CXCR4 , Metabolism
16.
Chinese Journal of Hematology ; (12): 467-469, 2003.
Article in Chinese | WPRIM | ID: wpr-354851

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of platelet factor 4 (PF4) on the adherence, and the expressions of adherent molecules CD(49d) and CXCR4 and the receptor of SDF-1 of fresh and expanded cord blood CD(34)(+) cells.</p><p><b>METHODS</b>CD(34)(+) cells were isolated from cord blood using MACS immune magnetic beads. The adherent ability was assayed by using crystal violet staining and the expression of adherent molecule CD(49d) and CXCR4 by FACS.</p><p><b>RESULTS</b>(1) PF4 could increase the adherent ability of the fresh cord blood CD(34)(+) cells, the effect being positively correlated with the dose of PF4. (2) SDF-1 at concentration of 100 ng/ml increased the adherent ability of the fresh cord blood CD(34)(+) cells. (3) The spontaneous and the SDF-1 induced adherent ability of the cord blood CD(34)(+) cells began to decrease after being cultured for 10 days without PF4, while in the presence of PF4 at 100 ng/ml, the ability of the cord blood CD(34)(+) cell adhering to the stroma layer still remained at higher level. At day 14, the adherent ability was (262.04 +/- 64.81)% and (64.35 +/- 8.29)% in PF4 group and control group, respectively, if it was defined as 100% at day 0. SDF-1 at concentration of 100 ng/ml induced adherent ability was (138.31 +/- 32.39)% and (67.66 +/- 12.44)% in PF4 group and control group, respectively. (4) The expression of CD(49d) and CXCR4 increased 13.02% and 17.33%, respectively, when incubated with PF4.</p><p><b>CONCLUSIONS</b>PF4 could increase the adherent ability and promote the expression of CD(49d) and CXCR4 of the cord blood CD(34)(+) cells, suggesting that PF4 promote the circulating stem cells homing to the marrow in the process of stem cells transplantation.</p>


Subject(s)
Humans , Antigens, CD34 , Blood , Cell Adhesion , Fetal Blood , Cell Biology , Flow Cytometry , Hematopoietic Stem Cells , Cell Biology , Integrin alpha4 , Blood , Platelet Factor 4 , Pharmacology , Receptors, CXCR4 , Blood
17.
Chinese Medical Journal ; (24): 273-277, 2003.
Article in English | WPRIM | ID: wpr-356818

ABSTRACT

<p><b>OBJECTIVE</b>To study the changes of adhesion molecules' expressions during the recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization in peripheral blood stem cell transplantation (PBSCT), and to confirm the influence of rhG-CSF on hematopoietic stem cells, which are proposed to guide mobilization in PBSCT.</p><p><b>METHODS</b>Mice were injected subcutaneously with diluted rhG-CSF or normal saline for 7 days. The blood Sca-1(+) stem cell count and bone marrow (BM) nucleated cell count were enumerated. The expressions of CD49d and CD44 and the adhesive ability of mononuclear cells to bone marrow matrix (fibronectin) were examined by flow cytometry and (51)Cr adhesive assay, respectively.</p><p><b>RESULTS</b>The mobilizing effect of rhG-CSF on mice was the same as on humans. The number of Sca-1(+) cells in peripheral blood reached the peak on the seventh day, the BM nucleated cell count was reduced, and the expressions of CD49d and the cells' adhesive ability in BM and PB declined.</p><p><b>CONCLUSIONS</b>rhG-CSF can reduce some cell adhesion molecules' expressions and the adhesive ability of hematopoietic stem cells to BM matrix, therefore mobilizing hematopoietic stem cells (HSC) from the BM to the peripheral blood.</p>


Subject(s)
Animals , Female , Mice , Bone Marrow Cells , Physiology , Fibronectins , Physiology , Granulocyte Colony-Stimulating Factor , Pharmacology , Hematopoietic Stem Cell Mobilization , Hematopoietic Stem Cell Transplantation , Hyaluronan Receptors , Physiology , Integrin alpha4 , Physiology , Leukocytes, Mononuclear , Physiology , Mice, Inbred BALB C , Recombinant Proteins
18.
Journal of Experimental Hematology ; (6): 659-661, 2003.
Article in Chinese | WPRIM | ID: wpr-278818

ABSTRACT

To explore the cell adhesion molecules (CAMs) CD11a and CD49d in patients with chronic aplastic anemia (CAA) and its clinical implications, the expression of CD11a and CD49d in mononuclear cell (MNC) of bone marrow (BM) and peripheral blood (PB) were measured using APAAP techniques in 20 patients with CAA before and after SSL/C therapy. The results showed that the expression of CD11a and CD49d in MNC of BM and CD11a in MNC of PB increased significantly (P < 0.05) after SSL/C therapy, and there was no significant change of CD49d in MNC of PB in both groups. In conclusion, the decrease of CAMs of CD11a and CD49d participated in the pathogenesis of CAA. The expression of CAMs increases with effective treatment, so the restoration or improvement of altered CAMs of CAA might be beneficial to the proliferation and differentiation of hematopoietic stem cell, and improvement of hematopoiesis in CAA.


Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Anemia, Aplastic , Blood , Blood Cell Count , CD11a Antigen , Chronic Disease , Integrin alpha4 , Leukocytes , Chemistry
19.
Journal of Experimental Hematology ; (6): 240-242, 2002.
Article in Chinese | WPRIM | ID: wpr-337601

ABSTRACT

To explore the dynamic change of CD34(+) cell expressing adhesion molecules in bone marrow and peripheral blood during mobilization with combination of chemotherapy and G-CSF and its clinical significance, mononuclear cells of bone marrow and peripheral blood from malignant hematopathy cases before and after mobilization with G-CSF were labeled by CD45-CY-Chrome, PE conjugated anti-CD34, and FITC conjugated anti-CD44, anti-CD49d, anti-CD62L and anti-CXCR4. For three-color fluorescence analysis by flow cytometry was performed on a FACScalibur. Also the relationship between the number of subpopulations in different expressions of adhesion molecules infused and the time of recovery in different blood cells after transplantation was evaluated. Results showed that a significantly lower expression of CD44(+) and CD49d(+) on CD34(+) cells in bone marrow after mobilization compared to that before mobilization, whereas great higher expression of CD44(+), CD49d(+), anti-CD62L(+) and lower of anti-CXCR4(+) in peripheral blood were observed after mobilization. No significant relations were found between expression of different adhesion molecules on CD34(+) cells infused and the time of reconstitution in blood cells after transplantation. It was concluded that this mobilizing regimen could downregulate the expressions of CD44, CD49d, CD62L, and anti-CXCR4 on CD34(+) cells in bone marrow, it may related to mobilization of CD34(+) cells from marrow to blood, and homing of blood CD34(+) cells into marrow.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Antigens, CD34 , Allergy and Immunology , Bone Marrow Cells , Allergy and Immunology , Metabolism , Cell Adhesion Molecules , Blood , Flow Cytometry , Methods , Granulocyte Colony-Stimulating Factor , Therapeutic Uses , Hematologic Neoplasms , Allergy and Immunology , Metabolism , Therapeutics , Hematopoietic Stem Cell Mobilization , Hematopoietic Stem Cell Transplantation , Hodgkin Disease , Allergy and Immunology , Metabolism , Therapeutics , Hyaluronan Receptors , Integrin alpha4 , L-Selectin , Leukocytes, Mononuclear , Allergy and Immunology , Metabolism , Lymphoma, Non-Hodgkin , Allergy and Immunology , Metabolism , Therapeutics , Multiple Myeloma , Allergy and Immunology , Metabolism , Therapeutics , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Allergy and Immunology , Metabolism , Therapeutics
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