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1.
Pancreas ; 49(10): 1348-1354, 2020.
Article in English | MEDLINE | ID: mdl-33122524

ABSTRACT

OBJECTIVES: The association of Lewis antigen phenotype with survival of patients with pancreatic ductal adenocarcinoma was investigated. METHODS: A total of 1187 patients diagnosed with pancreatic ductal adenocarcinoma were evaluated in a prospective cohort. Patients were classified into 3 different groups according to Lewis antigen phenotype: Lewis antigen (1) A positive [Le(a+b-)], (2) B positive [Le(a-b+)], and (3) negative [Le(a-b-)]. Risk of mortality was analyzed with Cox regression after adjusting for other predictors. RESULTS: The risk of mortality increased in the order of Le(a+b-), Le(a-b+), and Le(a-b-) [reference; hazard ratio (HR), 1.27; 95% confidence interval (CI)], 1.03-1.57; P = 0.02; and HR, 1.65; 95% CI, 1.31-2.09; P < 0.001] after adjusting for other predictors. Among patients with serum carbohydrate antigen (CA) 19-9 lower than 37 U/mL, the association seemed more apparent (reference; HR, 1.50; 95% CI, 0.77-2.29; P = 0.22; and HR, 2.10; 95% CI, 1.10-4.02; P < 0.02). CONCLUSIONS: The risk of mortality increased in the order of Le(a+b-), Le(a-b+), and Le(a-b-). The difference in prognosis according to the Lewis antigen phenotype was more pronounced in the low CA 19-9 group, which suggests that the Lewis antigen phenotype works as a biomarker predicting the prognosis of patients with pancreatic cancer with undetectable CA 19-9 level.


Subject(s)
Carcinoma, Pancreatic Ductal/immunology , Lewis Blood Group Antigens/analysis , Pancreatic Neoplasms/immunology , Aged , Antigens, Tumor-Associated, Carbohydrate/analysis , Carcinoma, Pancreatic Ductal/mortality , Carcinoma, Pancreatic Ductal/pathology , Female , Humans , Male , Middle Aged , Neoplasm Staging , Pancreatic Neoplasms/mortality , Pancreatic Neoplasms/pathology , Phenotype , Prospective Studies , Registries , Risk Assessment , Risk Factors , Time Factors
2.
Mol Omics ; 16(3): 210-220, 2020 06 15.
Article in English | MEDLINE | ID: mdl-32149324

ABSTRACT

Glycans are known to be involved in many biological processes, while little is known about the expression of N-glycans during vertebrate development. We now report the first quantitative studies of both the expression of N-linked glycans at six early development stages and the expression of N-glycosylated peptides at two early development stages in Xenopus laevis, the African clawed frog. N-Glycans were labeled with isobaric tandem mass tags, pooled, separated by capillary electrophoresis, and characterized using tandem mass spectrometry. We quantified 110 N-glycan compositions that spanned four orders of magnitude in abundance. Capillary electrophoresis was particularly useful in identifying charged glycans; over 40% of the observed glycan compositions were sialylated. The glycan expression was relatively constant until the gastrula-neurula transition (developmental stage 13), followed by massive reprogramming. An increase in oligomannosidic and a decrease in the paucimannosidic and phosphorylated oligomannosidic glycans were observed at the late tailbud stage (developmental stage 41). Two notable and opposing regulation events were detected for sialylated glycans. LacdiNAc and Lewis antigen features distinguished down-regulated sialylation from up-regulated species. The level of Lewis antigen decreased at later stages, which was validated by Aleuria aurantia lectin (AAL) and Ulex europaeus lectin (UEA-I) blots. We also used HPLC coupled with tandem mass spectrometry to identify 611 N-glycosylation sites on 350 N-glycoproteins at the early stage developmental stage 1 (fertilized egg), and 1682 N-glycosylation sites on 1023 N-glycoproteins at stage 41 (late tailbud stage). Over two thirds of the N-glycoproteins identified in the late tailbud stage are associated with neuron projection morphogenesis, suggesting a vital role of the N-glycome in neuronal development.


Subject(s)
Glycomics/methods , Xenopus Proteins/chemistry , Xenopus/growth & development , Animals , Electrophoresis, Capillary , Embryo, Nonmammalian/metabolism , Embryonic Development , Female , Gene Expression Regulation, Developmental , Lewis Blood Group Antigens/analysis , Male , Oligosaccharides/analysis , Phosphorylation , Tandem Mass Spectrometry
3.
Anal Chem ; 91(13): 8199-8206, 2019 07 02.
Article in English | MEDLINE | ID: mdl-31070893

ABSTRACT

Human milk oligosaccharides (HMOs) are one of the most abundant ingredients in breast milk, and they play a beneficial role for newborns and are important for infant health. The peripheral fucosylated sequences of HMOs, such as the histo-blood group ABH(O) and Lewis a, b, x, and y antigens, are determined by the expression of the secretor (Se) and Lewis (Le) genes in the mammary gland, and are often the recognition motifs and serve as decoy receptors for microbes. In this work, we developed a method for determination of secretor status and Lewis blood phenotype and assignment of Lewis blood-group epitopes. The method was based on electrostatic repulsion/hydrophilic interaction chromatography coupled with tandem mass spectrometry (ERLIC-MS/MS). A specifically designed stationary phase, aspartic acid-bonded silica (ABS), was used to separate the acidic and neutral HMOs by electrostatic repulsion followed by HILIC. Negative-ion electrospray MS/MS was then used for analysis of secretor status and Lewis blood phenotypes and assignment of important epitopes of HMOs from the lactating mothers by selecting a specific set of unique fragment ions.


Subject(s)
Fucosyltransferases/genetics , Lewis Blood Group Antigens/analysis , Milk, Human/chemistry , Oligosaccharides/chemistry , Adult , Chromatography, High Pressure Liquid , Female , Humans , Hydrophobic and Hydrophilic Interactions , Lewis Blood Group Antigens/genetics , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry , Galactoside 2-alpha-L-fucosyltransferase
4.
Anal Chim Acta ; 1071: 25-35, 2019 Sep 13.
Article in English | MEDLINE | ID: mdl-31128752

ABSTRACT

Mass spectrometry (MS) has become the primary method for high-sensitivity structural determination of oligosaccharides. Fragmentation in the negative-ion MS can provide a wealth of structural information and these can be used for sequence determination. However, although negative-ion MS of neutral oligosaccharide using the deprotonated molecule [M-H]- as the precursor has been very successful for electrospray ionization (ESI), it has only limited success for matrix-assisted laser desorption/ionization (MALDI). In the present study, the features of negative-ion MALDI primary spectra were investigated in detail and the product-ion spectra using [M-H]- and [M+Cl]- as the precursors were carefully compared. The formation of [M-H]- was the main difficulty for MALDI while [M+Cl]- was proved to be useful as alternative precursor anion for MALDI-MS/MS to produce similar fragmentation for sequencing of neutral oligosaccharides. N-(1-naphthyl)ethylenediamine dihydrochloride was then used as both the matrix and the Cl- dopant to evaluate the extent of structural information that can be obtained by negative-ion fragmentation from [M+Cl]- using laser-induced dissociation (LID)-MS/MS for linkage assignment of gluco-oligosaccharides and for typing of blood-group ABO(H) and Lewis antigens on either type 1 or type 2 backbone-chains.


Subject(s)
ABO Blood-Group System/analysis , Glucans/analysis , Lewis Blood Group Antigens/analysis , Oligosaccharides/analysis , ABO Blood-Group System/chemistry , Blood Grouping and Crossmatching/methods , Carbohydrate Sequence , Glucans/chemistry , Lewis Blood Group Antigens/chemistry , Oligosaccharides/chemistry , Sequence Analysis , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Tandem Mass Spectrometry/methods
5.
Int J Mol Med ; 40(2): 293-302, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28586014

ABSTRACT

MUC1 is a type I transmembrane glycoprotein and is overexpressed in various epithelial tumor tissues. Some researchers have demonstrated that the glycosylation status of MUC1 can affect MUC1-mediated tumor growth and cell differentiation. In our previous study, we proved that the abilities of cell proliferation, adhesion, invasion and metastasis, and drug resistance were enhanced in ovarian cancer cells stably expressing Lewis(y). Therefore, we hypothesized that Lewis(y) antigen may play a central role in regulating MUC1 expression, and MUC1-mediated cell growth and differentiation may be closely associated with Lewis(y) antigen. This study aimed to examine the correlation between MUC1 expression and Lewis(y) antigen levels in ovarian cancer cell lines and tissue samples. A series of techniques, including RT-qPCR, western blot anlaysis, immunoprecipitation, immunohistochemistry and double-labeling immunofluorescence were applied to detect the expression of Lewis(y) and MUC1. In malignant epithelial ovarian tumors, the positive expression rates of Lewis(y) antigen and MUC1 were 88.33 and 86.67%, respectively, which were markedly higher than those in borderline (60.00 and 53.33%, P<0.05), benign (33.33 and 30%, P<0.01) and normal (0 and 25%, P<0.01) ovarian samples. There was no correlation between the positive expression rates of Lewis(y) or MUC1 and clinicopathological parameters in ovarian cancers (P>0.05). The expression levels of Lewis(y) and MUC1 correlated with the clinical FIGO stage (P<0.05). Both MUC1 and Lewis(y) were highly expressed in ovarian cancer tissues, and their expression levels were positively correlated (P<0.01). In α1,2-fucosyltransferase (α1,2-FT)-transfected cells, the gene and protein expression levels of MUC1 were significantly upregulated compared with the cells that did not overexpress α1,2-FT (P<0.05). The ratio of Lewis(y) immunoprecipitated with MUC1 to total MUC1 increased 1.55-fold in α1,2-FT-overexpressing cells (P<0.05). The overexpression of Lewis(y) resulted in the upregulation of MUC1. On the whole, our data indicate that both MUC1 and Lewis(y) are associated with the occurrence and development of ovarian cancers.


Subject(s)
Gene Expression Regulation, Neoplastic , Lewis Blood Group Antigens/metabolism , Mucin-1/genetics , Neoplasms, Glandular and Epithelial/pathology , Ovarian Neoplasms/pathology , Ovary/pathology , Adolescent , Adult , Carcinoma, Ovarian Epithelial , Cell Line, Tumor , Cell Proliferation , Disease Progression , Female , Humans , Lewis Blood Group Antigens/analysis , Middle Aged , Mucin-1/analysis , Mucin-1/metabolism , Neoplasms, Glandular and Epithelial/genetics , Neoplasms, Glandular and Epithelial/metabolism , Ovarian Neoplasms/genetics , Ovarian Neoplasms/metabolism , Ovary/metabolism , Up-Regulation , Young Adult
6.
Bioorg Med Chem Lett ; 27(9): 1983-1988, 2017 05 01.
Article in English | MEDLINE | ID: mdl-28351593

ABSTRACT

Cell surface carbohydrates of the Lewis blood group antigens, Lewis X (Lex), Lewis Y (Ley), Lewis A (Lea), and their sialylated derivatives, such as sialy Lewis X (sLex) and sialy Lewis A (sLea), play important roles in various recognition processes. These cell surface carbohydrates have also been associated with the development and progression of many types of cancers. Recently, we synthesized four anthracene-based fluorescent bisboronic acid sensors (compounds 2a-d) linked by 'click' chemistry with tethers of different lengths to match the epitope of various Lewis group of sugars. Among the four compounds, 2a appears to have both high sensitivity and selectivity for Ley among other carbohydrate antigens.


Subject(s)
Anthracenes/chemistry , Boronic Acids/chemistry , Fluorescent Dyes/chemistry , Lewis Blood Group Antigens/analysis , Triazoles/chemistry , Anthracenes/metabolism , Antigens, Tumor-Associated, Carbohydrate/analysis , Antigens, Tumor-Associated, Carbohydrate/metabolism , Boronic Acids/metabolism , Cell Line, Tumor , Fluorescent Dyes/metabolism , Humans , Lewis Blood Group Antigens/metabolism , Microscopy, Fluorescence , Neoplasms/metabolism , Oligosaccharides/analysis , Oligosaccharides/metabolism , Sialyl Lewis X Antigen , Triazoles/metabolism
7.
Rev Esp Enferm Dig ; 107(10): 598-607, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26437978

ABSTRACT

BACKGROUND: Although colorectal carcinogenesis has been intensively studied, the published investigations do not provide a consistent description of how different carbohydrate determinants of colorectal epithelium are modified in colorectal cancer (CRC). OBJECTIVE: This study is an attempt to characterize the terminal fucosylation steps responsible for the synthesis of mono- Le(a)/Le(x)- and difucosylated -Le(b)/Le(y)- Lewis antigens in healthy and tumour CRC tissue. METHODS: An immunohistochemical study of Lewis antigens' expression was undertaken, along with screening of the fucosyltransferase (FT) activities involved in their synthesis, on healthy and tumour samples from 18 patients undergoing CRC. RESULTS: Analysis of alpha(1,2/3/4)FT activities involved in the sequential fucosylation of cores 1 and 2 showed significant increases in tumour tissue. Expressed as microU/mg and control vs. tumour activity (pfrom Wilcoxon's test), the FT activities for Le(a)/Le(b) synthesis were: lacto-N-biose alpha(1,2)/alpha(1,4)FT, 65.4 ± 19.0 vs. 186 ± 35.1 (p< 0.005); lacto-N-fucopentaose 1 alpha(1,4)FT, 64.9 ± 11.9 vs. 125.4 ± 20.7 (p< 0.005); Le(a) alpha(1,2)FT, 56.2 ± 7.2 vs. 130.5 ± 15.6 (p< 0.001). Similarly, for Le(x)/Le(y) synthesis were: N-acetyllactosamine alpha(1,2)-/alpha(1,3)FT, 53.4 ± 12.2 vs. 108.1 ± 18.9 (p< 0.001); 2'-Fucosyl-N-acetyllactosamine alpha(1,3)FT, 61.3 ± 10.7 vs. 126.4 ± 22.9 (p< 0.001); 2'-Fucosyllactose alpha(1,3)FT, 38.9 ± 10.9 vs. 143.6 ± 28.9 (p< 0.001); 2'-Methyllactose alpha(1,3)FT, 30.9 ± 4.8 vs. 66.1 ± 8.1 (p< 0.005); and Le(x) alpha(1,2)FT, 54.3 ± 11.9 vs. 88.2 ± 14.4 (p< 0.001). Immunohistochemical Le(y) expression was increased (p< 0.01 according to Wilcoxon's test) in tumour tissue, with 84.6% of specimens being positive: 7.7% weak, 15.4% moderate and 61.5% high intensity. CONCLUSIONS: Results suggest the activation of the biosynthesis pathways of mono- and difucosylated Lewis histo-blood antigens in tumour tissue from CRC patients, leading to the overexpression of Le(y), probably at the expense of Le(x).


Subject(s)
Colorectal Neoplasms/enzymology , Colorectal Neoplasms/genetics , Lewis Blood Group Antigens/analysis , Lewis Blood Group Antigens/biosynthesis , Aged , Amino Sugars , Biomarkers/analysis , Female , Fucose/metabolism , Fucosyltransferases/metabolism , Humans , Immunohistochemistry , Male , Middle Aged , Trisaccharides
8.
Glycobiology ; 24(8): 728-39, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24789815

ABSTRACT

Human milk oligosaccharides (HMOs) are a major constituent of human breast milk and play an important role in reducing the risk of infections in infants. The structures of these HMOs show similarities with blood group antigens in protein glycosylation, in particular in relation to fucosylation in Lewis blood group-type epitopes, matching the maternal pattern. Previously, based on the Secretor and Lewis blood group system, four milk groups have been defined, i.e. Lewis-positive Secretors, Lewis-positive non-Secretors, Lewis-negative Secretors and Lewis-negative non-Secretors. Here, a rapid one-dimensional (1)H nuclear magnetic resonance (NMR) analysis method is presented that identifies the presence/absence of (α1-2)-, (α1-3)- and (α1-4)-linked fucose residues in HMO samples, affording the essential information to attribute different HMO samples to a specific milk group. The developed method is based on the NMR structural-reporter-group concept earlier established for glycoprotein glycans. Further evaluation of the data obtained from the analysis of 36 HMO samples shows that within each of the four milk groups the relative levels of the different fucosylation epitopes can greatly vary. The data also allow a separation of the Lewis-positive Secretor milk group into two sub-groups.


Subject(s)
Epitopes/analysis , Lewis Blood Group Antigens/analysis , Milk, Human/chemistry , Oligosaccharides/chemistry , Epitopes/chemistry , Epitopes/immunology , Humans , Lewis Blood Group Antigens/chemistry , Lewis Blood Group Antigens/immunology , Magnetic Resonance Spectroscopy , Milk, Human/immunology , Oligosaccharides/analysis , Oligosaccharides/immunology , Protons
10.
Anticancer Res ; 32(5): 2159-64, 2012 May.
Article in English | MEDLINE | ID: mdl-22593503

ABSTRACT

AIM: Lewis antigens and the Thomsen-Friedenreich (TF) antigen are complex glycan structures that modulate processes such as cell adhesion and proliferation and tumor metastasis. The aim of our study was to analyze the expression of sialyl Lewis A (sLeA), sialyl Lewis X (sLeX), Lewis Y (LeY), TF, galectin-1 (Gal-1) and galectin-3 (Gal-3) in human osteoblasts in vitro. MATERIALS AND METHODS: The expression of the tumor markers sLeA, sLeX, LeY, TF, Gal-1 and Gal-3 was studied by means of immunohistochemistry on cells grown on chamber slides (2D) and on paraffin sections three-dimensional scaffold-free cultures (3D). The results of the stainings were evaluated semiquantitatively with the immunoreactive scoring system (IRS). RESULTS: Analysis of sLeA expression in both types of culture, 2D and 3D showed no detectable staining. After 5 days, in the 2D culture, expression of sLeX was weak, but the 3D culture (after 56 weeks) displayed a strong expression. LeY was expressed very slightly in the 2D culture, however LeY was not detectable in the 3D culture. The TF epitope was identified in the 2D cell culture model. In the 3D model, however, TF was completely lacking. Gal-1 was expressed very strongly in 2D culture, but in the 3D culture was not detectable. In contrast, Gal-3 was expressed in 3D culture but not in 2D. CONCLUSION: Within this study, we present a systematic analysis of the expression of sLeA, sLeX, LeY, TF, Gal-1 and Gal-3 in human osteoblasts grown in 2D and in 3D scaffold-free cultures. Summarizing the results of our study, we suggest that Lewis antigens and Gal-1 and -3 might play an important role in cell-cell and cell-matrix interactions of osteoblastic cells.


Subject(s)
Biomarkers, Tumor/analysis , Osteoblasts/chemistry , Antigens, Tumor-Associated, Carbohydrate/analysis , CA-19-9 Antigen , Cells, Cultured , Galectin 1/analysis , Galectin 3/analysis , Humans , Immunohistochemistry , Lewis Blood Group Antigens/analysis , Lewis X Antigen/analysis , Oligosaccharides/analysis , Sialyl Lewis X Antigen
11.
Oncol Rep ; 27(4): 1065-71, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22138668

ABSTRACT

Lewis y is a difucosylated oligosaccharide carried by glycoconjugates on the cell surface. Elevation of Lewis y is frequently observed in epithelial-derived cancers. This study aimed to detect the expression and clinical significance of the Lewis y antigen and TGF-ß1 (transforming growth factor ß1) in ovarian epithelial tumors, and to evaluate the correlation between them. Immunohistochemical staining was used to detect the expression of Lewis y antigen and TGF-ß1 in 60 cases of ovarian epithelial malignant tumors, 20 cases of borderline ovary tumors, 20 cases of benign ovary tumors and 10 cases of normal ovarian tissues. An immunofluorescence double labeling method was also used to detect the correlation between Lewis y antigen and TGF-ß1. The positive rates of Lewis y antigen in ovarian epithelial cancer tissues was 88.33%, significantly higher compared to those of borderline ovarian tumors (60.00%) (P<0.05), benign ovarian tumors (35.00%) (P<0.01) and normal ovarian tissues (0%) (P<0.01). Its expression was not associated with clinical parameters; the positive rates of TGF-ß1 in ovarian epithelial cancers were 78.33%, significantly higher compared to those of benign ovarian tumors (65.00%) (P<0.05) and normal ovarian tissues (40.00%) (P<0.05); the positive rates of the TGF-ß1 and Lewis y were not associated with metastasis of lymph nodes and histological types, differentiation degree and clinical stage (P>0.05). Expression of Lewis y antigen and TGF-ß1 was significantly positively associated with epithelial carcinoma. Close correlation between Lewis y, TGF-ß1 and ovarian cancer was observed. Altered expression of Lewis y antigen may cause changes in TGF-ß1 expression. Lewis y can increase the growth of ovarian cancer cells and the invasion ability by promoting TGF-ß1 abnormal expression and by promoting angiogenesis and a change in its signal transduction pathway. This study provides theoretical evidence for the development of ovarian cancer biological treatments.


Subject(s)
Biomarkers, Tumor/analysis , Lewis Blood Group Antigens/analysis , Neoplasms, Glandular and Epithelial/chemistry , Ovarian Neoplasms/chemistry , Transforming Growth Factor beta1/analysis , Adolescent , Adult , Aged , Carcinoma, Ovarian Epithelial , Case-Control Studies , Cell Differentiation , Cell Proliferation , Chi-Square Distribution , China , Female , Fluorescent Antibody Technique , Humans , Immunohistochemistry , Linear Models , Lymphatic Metastasis , Middle Aged , Neoplasm Invasiveness , Neoplasm Staging , Neoplasms, Glandular and Epithelial/pathology , Ovarian Neoplasms/pathology , Prognosis , Risk Assessment , Risk Factors , Up-Regulation , Young Adult
13.
Anal Bioanal Chem ; 401(8): 2495-510, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21898157

ABSTRACT

The structural diversity of human milk oligosaccharides (HMOs) strongly depends on the Lewis (Le) blood group status of the donor which allows a classification of these glycans into three different groups. Starting from 50 µL of human milk, a new high-throughput, standardized, and widely automated mass spectrometric approach has been established which can be used for correlation of HMO structures with the respective Lewis blood groups on the basis of mass profiles of the entire mixture of glycans together with selected fragment ion spectra. For this purpose, the relative abundance of diagnostically relevant compositional species, such as Hex(2)Fuc(2) and Hex(3)HexNAc(1)Fuc(2), as well as the relative intensities of characteristic fragment ions obtained thereof are of key importance. For each Lewis blood group, i.e., Le(a-b+), Le(a+b-), and Le(a-b-), specific mass profile and fragment ion patterns could be thus verified. The described statistically proven classification of the derived glycan patterns may be a valuable tool for analysis and comparison of large sets of milk samples in metabolic studies. Furthermore, the outlined protocol may be used for rapid screening in clinical studies and quality control of milk samples donated to milk banks.


Subject(s)
Lewis Blood Group Antigens/analysis , Milk, Human/chemistry , Oligosaccharides/analysis , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Carbohydrate Sequence , Discriminant Analysis , Female , High-Throughput Screening Assays/economics , High-Throughput Screening Assays/methods , Humans , Molecular Sequence Data , Oligosaccharides/isolation & purification , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/economics
14.
Folia Histochem Cytobiol ; 48(4): 637-45, 2010 Dec.
Article in English | MEDLINE | ID: mdl-21478109

ABSTRACT

The factors that affect the progression of prostatic carcinoma are poorly understood, but it is known that carbohydrate antigens on the tumour cell surface play a role in the transforming and metastatic processes. The present report aimed to perform a comparative, lectin-histochemical study of benign and carcinomatous prostates, using a battery of lectins, in combination with monoclonal antibodies against Lewis antigens, and a semi quantitative study, to investigate the changes in glycosylation patterns that occur in prostatic carcinoma. Blocks from 27 necropsy cases of prostatic carcinoma were sectioned and stained with H+E, fifteen biotinylated lectins chosen to probe for a wide range of oligosaccharide sequences within several categories of glycoprotein glycans, using a lectin-biotin avidin-peroxidase method, and monoclonal antibodies against Lewisa, sialyl Lewisa and sialyl Lewisx antigens. The glycophenotype of prostatic carcinoma differed from that of the noncancerous prostate in revealing more intense staining with the following lectins (AAA, UEA-1, DBA, WFA, VVA, HPA, BSA-1B4, MPA, ECA, AHA, and CTA), while the binding patterns of (GNA and NPA) were almost similar in both prostatic carcinoma and the noncancerous prostate. Lewis antigens are found to be expressed in prostatic carcinomas but not in the noncancerous prostate. The observations of this study suggest that the gylcophenotype of transformed prostatic cells was modified. It showed a moderate increase in, and changing patterns of, fucosylation and galactosylation, increased branching of side chains and sharp rise in 2 deoxy, 2 acetamido galactosylation and masking process by sialylation, especially by α2-3 and α2-6 linkages. All these changes in the glycosylation pattern of the transformed prostatic cells were observed on O-glycans, no changes were observed on N-glycans.


Subject(s)
Carcinoma/metabolism , Lectins/metabolism , Lewis Blood Group Antigens/metabolism , Prostate/chemistry , Prostatic Neoplasms/metabolism , Adult , Aged , Aged, 80 and over , Carcinoma/pathology , Glycosylation , Histocytochemistry , Humans , Lectins/analysis , Lewis Blood Group Antigens/analysis , Male , Middle Aged , Oligosaccharides/chemistry , Oligosaccharides/metabolism , Phenotype , Prostate/metabolism , Prostate/pathology , Prostatic Neoplasms/pathology
15.
J Exp Med ; 206(13): 3061-72, 2009 Dec 21.
Article in English | MEDLINE | ID: mdl-20008521

ABSTRACT

Variation of surface antigen expression is a mechanism used by microbes to adapt to and persist within their host habitats. Helicobacter pylori, a persistent bacterial colonizer of the human stomach, can alter its surface Lewis (Le) antigen expression. We examined H. pylori colonization in mice to test the hypothesis that host phenotype selects for H. pylori (Le) phenotypes. When wild-type and Le(b)-expressing transgenic FVB/N mice were challenged with H. pylori strain HP1, expressing Le(x) and Le(y), we found that bacterial populations recovered after 8 mo from Le(b)-transgenic, but not wild-type, mice expressed Le(b). Changes in Le phenotype were linked to variation of a putative galactosyltransferase gene (beta-(1,3)galT); mutagenesis and complementation revealed its essential role in type I antigen expression. These studies indicate that H. pylori evolves to resemble the host's gastric Le phenotype, and reveal a bacterial genetic locus that is subject to host-driven selection pressure.


Subject(s)
Helicobacter pylori/immunology , Lewis Blood Group Antigens/analysis , Lewis X Antigen/analysis , Oligosaccharides/physiology , Adhesins, Bacterial/analysis , Animals , Antibodies, Bacterial/blood , Flow Cytometry , Fucosyltransferases/genetics , Galactosyltransferases/genetics , Lipopolysaccharides/toxicity , Mice , Mice, Transgenic , Phenotype
16.
Anticancer Res ; 29(1): 201-9, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19331151

ABSTRACT

BACKGROUND: The advantage of tissue microarray (TMA) is its ability to efficiently analyze large numbers of tissue specimens in a methodologically uniform way. The reliability of TMAs, especially with regard to clinicopathological characterizations, when compared to conventional immunohistochemistry (IHC) was evaluated. MATERIALS AND METHODS: Seventy-two embedded tissue sections from lung cancer specimens were stained with monoclonal antibodies against the tumor-associated markers TA-MUC1 and Lewis Y. Three representative cores of every tumor were embedded in a paraffin array multiblock. The IHC was evaluated by the immunoreactive score (IRS). RESULTS: The data for the TMA IHC and the conventional IHC were concordant (kappa > or = 80%) for both markers. Likewise, discordance (McNemar's test) was low, and sensitivity and specificity were above 80% for both markers. In the samples with high positive expression, the concordance increased (kappa > or = 90%), discordance disappeared (McNemar p = 1.0), and sensitivity and specificity increased above 90% for both markers. Using Cox regression models, all the clinicopathological dependencies were equivalent for both techniques and both markers. CONCLUSION: Immunohistochemistry with tissue microarrays is valid and provides results equivalent to conventional immunohistochemistry with respect to expression patterns and clinicopathological characterizations.


Subject(s)
Lung Neoplasms/pathology , Tissue Array Analysis/methods , Aged , Antibodies, Monoclonal/chemistry , Carcinoma, Non-Small-Cell Lung/pathology , Carcinoma, Small Cell/pathology , Female , Humans , Immunohistochemistry , Lewis Blood Group Antigens/analysis , Male , Middle Aged , Mucin-1/analysis , Prognosis , Staining and Labeling/methods
17.
Sud Med Ekspert ; 52(1): 40-3, 2009.
Article in Russian | MEDLINE | ID: mdl-19371010

ABSTRACT

A one-step modification ofimmunoenzyme assay (dot variant) is proposed for the detection of ABH-Lewis antigens in human discharges (saliva, sperm, vaginal discharge). Sensitivity and specificity of the new method is higher than those of other methods currently used in forensic medicine for the detection of antigens in human body discharges; its other advantages over routine techniques include procedural simplicity, high performance, and the possibility of conducting series measurements with minimal labour inputs. Moreover, it provides information not only about group characteristic but also about category of discharge. The analysis is carried out in supernatant fractions which permits to reserve cellular material for molecular-genetic studies.


Subject(s)
ABO Blood-Group System/analysis , Bodily Secretions/immunology , Forensic Medicine/methods , Immunoenzyme Techniques/methods , Lewis Blood Group Antigens/analysis , ABO Blood-Group System/immunology , Female , Humans , Lewis Blood Group Antigens/immunology , Male , Saliva/chemistry , Saliva/immunology
18.
Proteomics ; 7(20): 3699-717, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17880005

ABSTRACT

Expression of sialyl Lewis x (sLe(x)) and sialyl Lewis a (sLe(a)) on cell-surface glycoproteins endows cells with the ability to adhere to E-, P-, and L-selectins present on endothelia, platelets, or leukocytes. Special arrangements of these glycotopes in cancers are thought to play a key role in metastasis. Previous studies have mostly described membrane-bound sLe(x) and sLe(a) activities. In this report, the major O-glycans of the secreted human ovarian cyst sialoglycoproteins from a Le(a+) nonsecretor individual (human ovarian cyst sample 350) were characterized by MS/MS analyses and immuno-/lectin-chemical assays. The results showed that HOC 350 carries a large number of epitopes for sLe(x), sLe(a), and Le(a) reactive antibodies. Advanced MS/MS sequencing coupled with mild periodate oxidation and exoglycosidase digestions further revealed that the O-glycans from HOC 350 are mostly of core 1 and 2 structures, extended and branched on the 3-arm with both type I and type II chains, complete with variable degrees of terminal sialylation and/or fucosylation to yield the sLe(x) or sLe(a) epitopes. Thus, the underlying core and peripheral backbone structures are similar to that of a previously proposed composite structural model for nonsialylated human ovarian cysts O-glycans, but with some notable distinguishing structural features in addition to sialylation.


Subject(s)
Epitopes/analysis , Lewis Blood Group Antigens/analysis , Ovarian Cysts/metabolism , Sialoglycoproteins/analysis , Antigens, Tumor-Associated, Carbohydrate/analysis , Biomarkers, Tumor/analysis , CA-19-9 Antigen , Carbohydrate Sequence , Epitopes/chemistry , Female , Gangliosides/analysis , Humans , Lewis Blood Group Antigens/chemistry , Molecular Sequence Data , Oligosaccharides/analysis , Ovarian Cysts/pathology , Polysaccharides/analysis , Sialoglycoproteins/chemistry , Sialyl Lewis X Antigen , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
19.
Ann Surg Oncol ; 14(6): 1880-9, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17375356

ABSTRACT

BACKGROUND: We have previously demonstrated tumor-specific alpha1,2fucosylation, which is associated with resistance of tumor cells to anticancer treatment in human colorectal tumor tissues. By using the YB-2 monoclonal antibody, the resulting products have been identified as Y, Le(b), and H type 2 antigens in colorectal tumor tissues. METHODS: Immunohistochemical analyses of colorectal cancer tissues (74 specimens) were performed with a newly established mouse monoclonal antibody, YB-3 specifically recognizing H disaccharide (Fucalpha1,2Galbeta) structures, and anti-A, anti-B, YB-2, and anti-sialyl Lewis X (SLX) antibodies, together with the analyses of glycosyltransferases involved in the synthesis of ABH antigens in the same tissues. RESULTS: The YB-3 antibody enabled us to detect colorectal tumors, particularly tumors in the distal large intestine and the rectum, with high sensitivity (74.3%) and specificity (100%). From immunohistochemical and enzymatic analyses of colorectal tissues, we found that once alpha1,2fucosylation had proceeded in tumor tissues, blood group A or B antigen was also synthesized in approximately half of the tissues of A or B blood type, but not in their normal tissues. A correlation of survival rate with immunostaining of tissues was found only by YB-3 antibody and not by anti-A, anti-B, or anti-SLX antibody. CONCLUSIONS: As a predictor of postoperative prognosis of patients with colorectal cancer, immunodetection of alpha1,2fucosylated antigens with the YB-3 antibody seemed to be superior to blood groups A, B, or SLX antigen in colorectal tumor tissues.


Subject(s)
ABO Blood-Group System/analysis , Antigens, Neoplasm/analysis , Biomarkers, Tumor/analysis , Colonic Neoplasms/pathology , Lewis Blood Group Antigens/analysis , Lewis X Antigen/analysis , Oligosaccharides/analysis , Rectal Neoplasms/pathology , Antibodies, Monoclonal , Colonic Neoplasms/surgery , Disaccharides/analysis , Female , Forecasting , Fucosyl Galactose alpha-N-Acetylgalactosaminyltransferase/analysis , Glycosyltransferases/analysis , H-2 Antigens , Humans , Male , Prognosis , Rectal Neoplasms/surgery , Sialyl Lewis X Antigen
20.
Transplantation ; 82(4): 479-85, 2006 Aug 27.
Article in English | MEDLINE | ID: mdl-16926591

ABSTRACT

BACKGROUND: In the revived interest in crossing ABO barriers in organ transplantation renal A/B antigen expression has been correlated with donor ABO, Lewis, and secretor subtype to predict antigen expression. METHODS: A/B antigen expression was explored by immunohistochemistry in LD renal biopsies. Donor A1/A2/B, Lewis, and secretor status were determined by serology and polymerase chain reaction. RESULTS: In the renal vascular bed, three distinct A antigen expression patterns with a major, minor, and minimal staining distribution, and intensity (designated as types 3+, 1+ and (+) respectively) were identified. Type 3+ had a strong A antigen expression in the endothelium of arteries, glomerular/peritubular capillaries and veins. The type 1+ showed an overall weaker antigen expression, whereas type (+) had faint staining of peritubular capillaries only. In all cases, distal tubular epithelium was focally stained, whereas proximal tubules were negative. Type 3+ were all from blood group A1 subtype individuals while A2 cases expressed either a 1+ or (+) pattern. The secretor gene did not appear to influence renal A antigen expression. All B kidneys examined showed a B antigen pattern slightly weaker but otherwise similar to A type 3+. CONCLUSION: Renal vascular A antigen expression correlates to donor A1/A2 subtypes, whereas B individuals show one singular antigen pattern. From antigen perspective, A1 and B donors are a "major" and A2 individuals a "minor" antigen challenge in ABO-incompatible renal transplantation.


Subject(s)
ABO Blood-Group System/analysis , Blood Group Incompatibility , Kidney Transplantation/immunology , Kidney/immunology , Lewis Blood Group Antigens/analysis , ABO Blood-Group System/immunology , Humans , Immunohistochemistry , Tissue Donors
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