Multiple immunoenzyme staining techniques use of fluoresceinated, biotinylated and unlabelled monoclonal antibodies

Loos, Chris M. van der; Das, Pranab K.; Houthoff, H. J.

doi:10.1016/0022-1759(89)90117-8

Cited by 74 publications

(37 citation statements)

References 20 publications

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“…19 In brief, after fixation of the sections in acetone, endogenous peroxidase was blocked. Nonspecific background was blocked by normal goat (DAKO Cytomation) or normal donkey serum (Jackson ImmunoResearch Laboratory Inc, West Grove, Penn).…”

Section: Immunohistochemistrymentioning

confidence: 99%

High Prevalence of Circulating CD4⁺CD28⁻T-Cells in Patients With Small Abdominal Aortic Aneurysms

Duftner

Seiler²,

Klein-Weigel³

et al. 2005

ATVB

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Objective-To assess the possible role of proinflammatory CD28Ϫ T cells in abdominal aortic aneurysms (AAAs). Animal studies and human tissue studies suggest a role for interferon (IFN)-␥-producing T cells in the development and progression of AAAs. Methods and Results-Fluorescence-activated cells sorter analysis of peripheral blood samples and measurement of AAA size using sonography were performed in 101 AAA patients and 38 healthy controls. Key Words: aortic disease Ⅲ aneurysms Ⅲ leukocytes Ⅲ immune system Ⅲ human A bdominal aortic aneurysm (AAA) is a common disease with a prevalence of 3% of individuals aged 60 years and older and is a potentially lethal disorder causing Ϸ15 000 deaths annually in the United States. 1 Recent human tissue studies and animal models have led now to a paradigm shift in the pathogenetic concept of AAAs. Rather than a simple degenerative process, the majority of AAAs has proven to be a complex and dynamic remodeling process. In brief, studies of human AAA tissues have identified extensive inflammatory infiltrates in both the media and the adventitia, 2 leading to an increased expression of proinflammatory cytokines and C-reactive protein (CRP) in aneurysmal tissue. [3][4][5] The presence of vascular-associated lymphoid tissue (VALT) with lymphoid follicles and lymph node-like structures in the adventitia of AAAs suggests a role for immunocompetent and antigen presenting cells not only in atherosclerosis 6 but also in AAA disease. 2 The role for T cells has been further supported by immunogenetic findings with associations between AAA and human leukocyte antigen class II molecules. 7 Infiltration of the adventitia usually occurs together with medial thinning. This inflammatory process triggers the production of metalloproteinases and apoptosis of medial smooth muscle cells thus explaining the disruption of the orderly lamellar structure in aortic aneurysms.Recently, a subgroup of proinflammatory T cells has been identified in patients with immune-mediated disorders including rheumatoid arthritis, 8 -9 ankylosing spondylitis, 10 multiple sclerosis, 11 Wegener granulomatosis, 12 and unstable angina, 13 which lack the costimulatory molecule CD28 on their surface and are considered as markers for chronic inflammation and early aging. 14 Under these circumstances the CD28 Ϫ T cells are part of the CD4 ϩ as well as the CD8 ϩ T cell compartment, persist over years, and include most of the oligoclonally expanded T cells. Phenotypically, CD4 ϩ CD28 Ϫ T cells from rheumatoid arthritis and ankylosing spondylitis patients and CD8 ϩ CD28 Ϫ T cells from aged persons, rheumatoid arthritis, and melanoma patients share the expression of various natural killer (NK) cell receptors and lack the expression of the lymphocyte marker CD7. 9,15-16 Functionally these T cells are capable to release large amounts of interferon (IFN)-␥, perforin, and granzyme B, providing them with the possibility to lyse target cells. 17 In the pathogenesis of coronary arteriosclerosis, the critical role of IFN-␥ was alread...

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Section: Immunohistochemistrymentioning

confidence: 99%

High Prevalence of Circulating CD4⁺CD28⁻T-Cells in Patients With Small Abdominal Aortic Aneurysms

Duftner

Seiler²,

Klein-Weigel³

et al. 2005

ATVB

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“…In order to confirm that the EpCAM-positive cells were also CK-positive, we performed double immunolabeling according to the technique described by Van der Loos et al 28 EpCAM antibody was revealed by an anti-mouse IgG labeled with the fluorochrome Alexa488 (green). For anti-CK labeling, A45B/B3 antibody was applied and revealed by a polyclonal anti-FITC antibody, and staining was performed by an anti-rabbit antibody labeled with fluorochrome Alexa594 (red) (Molecular Probe, Leiden, Netherlands) (excitation wavelength 594 nm).…”

Section: Immunocytochemic Stainingmentioning

confidence: 99%

Real‐time quantitative PCR determination of urokinase‐type plasminogen activator receptor (uPAR) expression of isolated micrometastatic cells from bone marrow of breast cancer patients

Pierga

Bonneton

Magdelénat

et al. 2004

Intl Journal of Cancer

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“…After washing, slides were incubated for 30 min with anti-rabbit-labelled polymers-HRP. Then, 3,3 -diaminobenzidine tetrahydrochloride (DAB chromogen) was added for revelation system [Van der Loos et al, 1989] and counterstaining with haematoxylin was performed. Finally, samples were examined under the imaging system microscope (model BX41; Olympus Italia Srl, Milan, Italy).…”

Section: Immunocytochemistry Analysismentioning

confidence: 99%

A Comparative Study of Proliferation and Hepatic Differentiation of Human Adipose-Derived Stem Cells

Coradeghini

Guida

Scanarotti

et al. 2009

Cells Tissues Organs

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Human adipose-derived stem cells possess a lot of stem cell characteristics, so they may be considered a source of stem cell population. On the basis of that, we have investigated the hepatic potential of adipose-derived stem cells, obtained from liposuction, following two differentiation protocols. In the first procedure, medium was supplemented with epidermal growth factor (EGF), basic fibroblast growth factor, hepatocyte growth factor (HGF) and nicotinamide; the second involved the addition of factors such as dexametasone, EGF, insulin-transferrin-sodium selenite, HGF, dimethyl sulfoxide and oncostatin. In parallel, we carried out our study in the Hep G2 cell line, as human hepatic differentiated in vitro model. Immunocytochemical analysis and RT-PCR were performed using hepatic markers to evaluate cell differentiation. DNA content, MTT test and carboxyl fluorescein succinimidyl ester staining were carried out to evaluate cell proliferation. We reported the evidence of basal hepatic marker in undifferentiated adipose-derived stem cells, which confirmed their multipotency. A strong expression of albumin and α-fetoprotein was observed in hepatic-induced adipose-derived stem cells following both differentiation procedures. Morphological aspects of the two types of hepatic adipose-derived stem cells were alike. Proliferation index suggested that the first differentiation procedure promoted better growth than the second. These preliminary findings suggest adipose-derived stem cells may be induced into hepatic lineage, and the most significant difference between the two standard differentiation procedures concerns proliferation rate. This aspect is to be considered when adipose-derived stem cells are employed in research and clinical studies.

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Multiple immunoenzyme staining techniques use of fluoresceinated, biotinylated and unlabelled monoclonal antibodies

Cited by 74 publications

References 20 publications

High Prevalence of Circulating CD4⁺CD28⁻T-Cells in Patients With Small Abdominal Aortic Aneurysms

High Prevalence of Circulating CD4⁺CD28⁻T-Cells in Patients With Small Abdominal Aortic Aneurysms

Real‐time quantitative PCR determination of urokinase‐type plasminogen activator receptor (uPAR) expression of isolated micrometastatic cells from bone marrow of breast cancer patients

A Comparative Study of Proliferation and Hepatic Differentiation of Human Adipose-Derived Stem Cells

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Multiple immunoenzyme staining techniques use of fluoresceinated, biotinylated and unlabelled monoclonal antibodies

Cited by 74 publications

References 20 publications

High Prevalence of Circulating CD4+CD28−T-Cells in Patients With Small Abdominal Aortic Aneurysms

High Prevalence of Circulating CD4+CD28−T-Cells in Patients With Small Abdominal Aortic Aneurysms

Real‐time quantitative PCR determination of urokinase‐type plasminogen activator receptor (uPAR) expression of isolated micrometastatic cells from bone marrow of breast cancer patients

A Comparative Study of Proliferation and Hepatic Differentiation of Human Adipose-Derived Stem Cells

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High Prevalence of Circulating CD4⁺CD28⁻T-Cells in Patients With Small Abdominal Aortic Aneurysms

High Prevalence of Circulating CD4⁺CD28⁻T-Cells in Patients With Small Abdominal Aortic Aneurysms