scholarly journals Immunohistochemical and Flow Cytometric Analysis of Intracellular and Membrane-Bound Hsp70, as a Putative Biomarker of Glioblastoma Multiforme, Using the cmHsp70.1 Monoclonal Antibody

2017 ◽  
pp. 307-320
Author(s):  
Stefan Stangl ◽  
Gemma A. Foulds ◽  
Helena Fellinger ◽  
Geoffrey J. Pilkington ◽  
A. Graham Pockley ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Glioblastoma Multiforme ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Membrane Bound

scholarly journals Fas Ligand Is Present in Human Erythroid Colony-Forming Cells and Interacts With Fas Induced by Interferon γ to Produce Erythroid Cell Apoptosis

Blood ◽  
1998 ◽  
Vol 91 (4) ◽  
pp. 1235-1242 ◽  
Author(s):  
Chun-Hua Dai ◽  
James O. Price ◽  
Thomas Brunner ◽  
Sanford B. Krantz
Keyword(s):  
Monoclonal Antibody ◽  
Cell Surface ◽  
Fas Ligand ◽  
Flow Cytometric Analysis ◽  
Interferon Γ ◽  
Cell Surface Receptor ◽  
Erythroid Cell ◽  
Erythroid Progenitors ◽  
Flow Cytometric ◽  
Fas Mrna

Abstract Interferon γ (IFNγ) inhibits the growth and differentiation of highly purified human erythroid colony-forming cells (ECFCs) and induces erythroblast apoptosis. These effects are dose- and time-dependent. Because the cell surface receptor known as Fas (APO-1; CD95) triggers programmed cell death after activation by its ligand and because incubation of human ECFCs with IFNγ produces apoptosis, we have investigated the expression and function of Fas and Fas ligand (FasL) in highly purified human ECFCs before and after incubation with IFNγ in vitro. Only a small percentage of normal human ECFCs express Fas and this is present at a low level as detected by Northern blotting for the Fas mRNA and flow cytometric analysis of Fas protein using a specific mouse monoclonal antibody. The addition of IFNγ markedly increased the percentage of cells expressing Fas on the surface of the ECFCs as well as the intensity of Fas expression. Fas mRNA was increased by 6 hours, whereas Fas antigen on the cell surface increased by 24 hours, with a plateau at 72 hours. This increase correlated with the inhibitory effect of IFNγ on ECFC proliferation. CH-11 anti-Fas antibody, which mimics the action of the natural FasL, greatly enhanced IFNγ-mediated suppression of cell growth and production of apoptosis, indicating that Fas is functional. Expression of FasL was also demonstrated in normal ECFCs by reverse transcriptase-polymerase chain reaction and flow cytometric analysis with specific monoclonal antibody. FasL was constitutively expressed among erythroid progenitors as they matured from day 5 to day 8 and IFNγ treatment did not change this expression. Apoptosis induced by IFNγ was greatly reduced by the NOK-2 antihuman FasL antibody and an engineered soluble FasL receptor, Fas-Fc, suggesting that Fas-FasL interactions among the ECFCs produce the erythroid inhibitory effects and apoptosis initiated by IFNγ.

Monitoring the Chimerism Status of Subpopulation in HLA Haplo-Identical Stem Cell Transplantation Using the Anti HLA Antibodies: Based on the Study of HLA Distribution in Korean Population

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4706-4706
Author(s):  
Won Ho Choe ◽  
Seongsoo Jang ◽  
Hyun-Sook Chi ◽  
Chan-Jeoung Park ◽  
Ho Joon Im
Keyword(s):  
Monoclonal Antibody ◽  
T Cells ◽  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Flow Cytometric Analysis ◽  
Korean Population ◽  
Hla Antibodies ◽  
Flow Cytometric ◽  
Donor Type ◽  
Cell Subpopulations

Abstract Abstract 4706 Background: Determining the chimerism in stem cell transplantation (SCT) is important in the monitoring of engraftment. Conventional monitoring methods such as short tandem repeat (STR) PCR are labor intensive and difficult in showing the changes of cell subpopulations. Nowadays, HLA-haploidentical SCT is performed actively and flow cytometric analysis using anti HLA antibody targeted for the different HLA between donor and recipient in HLA-haploidentical SCT can be useful by observing changes of cell subpopulations and determining the chimerism. Methods: Based on the study of HLA distribution in Korean population, we designed the panel of HLA monoclonal antibody that can detect and differentiate major HLA of Korean population. This panel was verified against 23 cases of HLA-haploidentical SCT. Flow cytometric analysis was performed using anti-HLA-antibodies on one pediatric patient with aplastic anemia who underwent HLA-haploidentical SCT. Tests were performed every day since SCT. The appearance or disappearance of donor and/or recipient HLA was observed using antibodies against mismatched HLA. Donor and/or recipient T cells, B cells, NK cells, and monocytes were observed using CD3, CD19, CD56, and CD13 respectively. The flow cytometric results were interpreted by observing the changes in subpopulations of detected cells and determining the engraftment status of patients. Results: A total of 11 anti-HLA-A-antibodies and anti-HLA-B-antibodies were selected. They cover 97.9% of HLA-A and 8.6% of HLA-B in Korean population. In the verification of our panel of HLA monoclonal antibody, we distinguished donor and recipient cells in 21 of 23 HLA-haploidentical SCT cases. In our case, the patient had HLA-A2/A24 while the donor had HLA-A2/A33. Recipient type of CD3(+) and HLA-A33(-) T cells appeared in first few days but donor type CD3(+) and HLA-A33(+) T cells appeared at day 20 and were increased. Donor type CD56(+) and HLA-A33(+) cells were seen at analysis done in day 8 and continued to appear. HLA-A33(+) cells were consistently observed and increased throughout the follow up period, showing the process of engraftment of donor stem cells. Conclusion: We were able to apply our flow cytometric analysis design using HLA monoclonal antibody for the detection of chimerism in HLA-haploidentical SCT. This method was more simple and sensitive than conventional monitoring techniques. Moreover, this method allowed us to observe the dynamics of changes in cell subpopulations after HLA-haploidentical SCT. Flow cytometric analysis can be considered as a strong tool for observing the changes in cell subpopulations and the monitoring the engraftment of HLA-haploidentical SCT. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Flow cytometric analysis of in vivo polyamine deprivation in Lewis lung carcinoma (3LL) cells using the monoclonal antibody SPM8-2

Cytometry ◽  
1997 ◽  
Vol 27 (3) ◽  
pp. 255-261 ◽  
Author(s):  
Jean-Guy Delcros ◽  
Laurence L?uillet ◽  
Laura Chamaillard ◽  
Anne Royou ◽  
Nathalie Bouill� ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Lung Carcinoma ◽  
Lewis Lung Carcinoma ◽  
Flow Cytometric Analysis ◽  
Lewis Lung ◽  
Flow Cytometric

scholarly journals Flow cytometric analysis and sorting of human endometrial cells after immunocytochemical labeling for cytokeratin using a monoclonal antibody

Cytometry ◽  
1985 ◽  
Vol 6 (2) ◽  
pp. 159-164 ◽  
Author(s):  
Peter S. Oud ◽  
Jos B. J. Henderik ◽  
Hans L. M. Beck ◽  
José A. M. Veldhuizen ◽  
G. Peter Vooijs ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometric Analysis ◽  
Endometrial Cells ◽  
Flow Cytometric

scholarly journals An Immunoglobulin G1 Monoclonal Antibody Highly Specific to the Wall of Cryptosporidium Oocysts

2000 ◽  
Vol 7 (5) ◽  
pp. 745-750 ◽  
Author(s):  
C. Weir ◽  
G. Vesey ◽  
M. Slade ◽  
B. Ferrari ◽  
D. A. Veal ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Soluble Protein ◽  
Flow Cytometric Analysis ◽  
Oocyst Wall ◽  
Protein Preparation ◽  
Water Testing ◽  
Flow Cytometric ◽  
Cryptosporidium Oocysts ◽  
Specific Igg

ABSTRACT The detection of Cryptosporidium oocysts in drinking water is critically dependent on the quality of immunofluorescent reagents. Experiments were performed to develop a method for producing highly specific antibodies to Cryptosporidium oocysts that can be used for water testing. BALB/c mice were immunized with six different antigen preparations and monitored for immunoglobulin G (IgG) and IgM responses to the surface of Cryptosporidiumoocysts. One group of mice received purified oocyst walls, a second group received a soluble protein preparation extracted from the outside of the oocyst wall, and the third group received whole inactivated oocysts. Three additional groups were immunized with sequentially prepared oocyst extracts to provide for a comparison of the immune response. Mice injected with the soluble protein extract demonstrated an IgG response to oocysts surface that was not seen in the whole-oocyst group. Mice injected with whole oocysts showed an IgM response only, while mice injected with purified oocyst walls showed little increase in IgM or IgG levels. Of the additional reported preparations only one, BME (2-mercaptoethanol treated), produced a weak IgM response to the oocyst wall. A mouse from the soluble oocyst extract group yielding a high IgG response was utilized to produce a highly specific IgG1 monoclonal antibody (Cry104) specific to the oocyst surface. Comparative flow cytometric analysis indicated that Cry104 has a higher avidity and specificity to oocysts in water concentrates than other commercially available antibodies.

Contribution of CD54 to human eosinophil and neutrophil superoxide production

2001 ◽  
Vol 91 (2) ◽  
pp. 613-622 ◽  
Author(s):  
Shuji Takashi ◽  
Yoshio Okubo ◽  
Shiro Horie
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometric Analysis ◽  
Superoxide Production ◽  
Human Eosinophil ◽  
Flow Cytometric ◽  
Leukocyte Aggregation ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Neutrophil Superoxide

We have reported that CD54 on eosinophils is involved in eosinophil degranulation. However, the role of CD54 in eosinophil and neutrophil superoxide production is still uncertain. We assessed the effect of CD54 on eosinophils and neutrophils in recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF)- or phorbol myristate acetate (PMA)-induced superoxide production through CD18. Anti-CD54 monoclonal antibody attenuated leukocyte aggregation and superoxide production of rGM-CSF- or PMA-stimulated neutrophils and PMA-stimulated eosinophils. Anti-CD18 monoclonal antibody or theophylline attenuated superoxide production of eosinophils and neutrophils stimulated by either stimuli. Flow cytometric analysis demonstrated CD54 expression on freshly isolated neutrophils but not on freshly isolated eosinophils. CD54 newly expressed on eosinophils reached its peak expression 30 min after PMA stimulation. The increase in CD18 and CD54 expression on neutrophils caused by rGM-CSF stimulation was partially inhibited by theophylline. These data demonstrated that CD54 and CD18 interaction of eosinophils or neutrophils is involved in superoxide production and that the inhibition of superoxide production by theophylline may be at least partly due to the inhibition of CD54 and CD18.

Therapeutic targeting of CD70 and CD27-CD70 interactions with the monoclonal antibody SGN-70 in Waldenstrom’s Macroglobulinemia (WM)

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 2509-2509 ◽  
Author(s):  
A. W. Ho ◽  
E. Hatjiharissi ◽  
A. Branagan ◽  
Z. Hunter ◽  
J. McEarchern ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Mast Cells ◽  
Functional Role ◽  
Cell Activation ◽  
Critical Role ◽  
Flow Cytometric Analysis ◽  
Soluble Form ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Flow Cytometric

2509 Background: WM represents a lymphoplasmacytic lymphoma characterized by a monoclonal IgM gammopathy and possesses a mast cell component that may contribute to its pathogenesis. The tumor necrosis factor (TNF) receptor family member, CD27, is a transmembrane co-stimulatory molecule that is also secreted in a soluble form (sCD27). Recent evidence has suggested that interactions between CD27 and its TNF-like ligand, CD70, play a critical role in regulating B-cell activation and survival, and therefore, may provide a viable therapeutic target for the treatment of WM. Methods: Patients with the consensus panel diagnosis of WM who provided written consent were evaluated. ELISA assay kits were obtained from Bender MedSystems. Flow cytometric analysis, RT-PCR, and calcein ADCC assays were performed as previously described (Santos et al, in press). Results: By ELISA, WM patients displayed dramatically higher levels of sCD27 in their sera (median 7.45, range 0–19.42 U/ml) versus healthy donors (median 0, range 0–2.78 U/ml; p = 2.5 × 10−7). CD27 was expressed in 7/7 patients using RT-PCR analysis, and on the tumor cell surface of 5/12 patients. CD70, the target for CD27, was widely expressed on tumor cells (6/6) and mast cells (10/11) using flow cytometric analysis. To define the functional role of sCD27 in WM, we cultured BCWM.1 (CD27−CD70+) WM cells, LAD1 (CD27−CD70+) mast cells, and primary tumor and mast cells (CD70+) isolated from WM patients with sCD27 (0.1–50 ug/mL), and observed no effect on their proliferation or induction of apoptosis. However, culture of LAD1 and primary WM mast cells (10/10) with sCD27 resulted in marked upregulation of two TNF family ligands which support the growth and survival of WM cells: CD40L (CD154) and a proliferation induction ligand (APRIL). Importantly, the anti-CD70 mAb SGN-70 (1 ug/ml) inhibited this upregulation by sCD27, establishing the specificity of the CD27-CD70 interaction. In addition, SGN-70 demonstrated significant ADCC against BCWM.1 WM cells at dose levels up to 20 ug/ml. Conclusions: Taken together, these studies suggest a novel functional role for sCD27 in the pathogenesis of WM, and demonstrate the feasibility of targeting CD70 and sCD27-CD70 interactions with the SGN-70 monoclonal antibody. [Table: see text]

Flow Cytometric Analysis of Circadian Changes in Platelet Activation Using Anti-Gmp-140 Monoclonal Antibody

1999 ◽  
Vol 16 (3) ◽  
pp. 335-342 ◽  
Author(s):  
Bülent Ündar ◽  
Nurullah Akkoç ◽  
Mehmet N. Alakavuklar ◽  
Cavit ĉehreli ◽  
Levent Ündar
Keyword(s):  
Monoclonal Antibody ◽  
Platelet Activation ◽  
Flow Cytometric Analysis ◽  
Circadian Changes ◽  
Flow Cytometric
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