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1.
Sen-itiroh Hakomori 《Glycoconjugate journal》2000,17(7-9):627-647
Our studies on glycosphingolipids (GSLs) were initiated through isolation and structural characterization of lacto-series type 1 and 2 GSLs, and globo-series GSLs. Lacto-series structures included histo-blood group ABH and I/i antigens. Our subsequent studies were focused on GSL changes associated with: (i) ontogenic development and differentiation; (ii) oncogenic transformation and tumor progression. Various novel types of GSLs such as extended globo-series, sialyl-Lex (SLex), sialyl-dimeric-Lex (SLex-Lex), dimeric-Lex (Lex-Lex), Ley-on-Lex, dimeric-Lea (Lea-Lea), Leb-on-Lea, etc. were identified as tumor-associated antigens. These studies provide an essential basis for up- or down-regulation of key glycosyltransferase genes controlling development, differentiation, and oncogenesis. GSL structures established in our laboratory are summarized in Table 1, and structural changes of GSLs associated with ontogenesis and oncogenesis are summarized in Sections 2 and 3.Based on these results, we endeavored to find out the cell biological significance of GSL changes, focused on (i) cell adhesion, e.g., the compaction process of preimplantation embryo in which Lex-to-Lex, Gb4-to-GalGb4 or -nLc4 play major roles; and (ii) modulation of signal transduction through interaction of growth factor receptor tyrosine kinase with ganglioside, e.g., EGF receptor tyrosine kinase with GM3. Recent trends of studies on i and ii lead to the concept that GSL clusters (microdomains) are organized with various signal transducer molecules to form glycosignaling domains (GSD). GSL-dependent adhesion occurs through clustered GSLs, and is coupled with activation of signal transducers (cSrc, Src family kinase, Rho A, etc.). Clustered GSLs involved in cell adhesion are recognized by GSLs on counterpart cells (carbohydrate-to-carbohydrate interaction), or by lectins (e.g., siglecs, selectins).Our major effort in utilization of GSLs in medical science has been for: (i) cancer diagnosis and treatment (vaccine development) based on tumor-associated GSLs and glycoepitopes; (ii) genetically defined phenotype for susceptibility to E. coli
infection; (iii) clear identification of physiological E-selectin epitope (myeloglycan) expressed on neutrophils and myelocytes; (iv) characterization of sialyl poly-LacNAc epitopes recognized as male-specific antigens. Utilization of these GSLs or glycoepitopes in development of anti-adhesion approach to prevent tumor metastasis, infection, inflammation, or fertilization (i.e., contraceptive) is discussed. For each approach, development of mimetics of key GSLs or glycoepitopes is an important subject of future study. 相似文献
2.
Patricia L. Stranahan Jennifer Laroe Ronda McCombs Alex Goldsmith Iffat Rahim Mark Overland David E. Pettijohn 《Glycoconjugate journal》1996,13(5):741-747
Intratumoral phenotypic diversity is well documented with regard to tumor associated carbohydrate antigens (TACA). The factors which control the expression of these cell-surface oligosaccharides on different cells of the same tumor are not understood. We investigated the expression of a panel of mucin associated oligosaccharides in cell lines growing at different surface densities (number of cells per cm2 of growth flask). Results show that the apparent expression of extended Lea-Lex, Lea and Lex, sialyl Lea, Tn and sialyl Tn varies with density of growth by an invasive human squamous cell lung carcinoma cell line (NU6-1), a benign variant (NE-18) and the human lung epithelial cell line BEAS-2B. The results indicate that one of the factors influencing the apparent expression of mucin-associated oligosaccharides is cell-cell interactions.Abbreviations Mab
monoclonal antibody
- FIT
fluorescein isothiocyanate
- TACA
tumor associated carbohydrate antigen 相似文献
3.
A variety of glycoconjugates, including glycosphingolipids (GSLs), expressed in mammalian tissues and cells were isolated and characterized in early biochemical studies. Later studies of virus-transformed fibroblasts demonstrated the association of GSL expression profiles with cell phenotypes. Changes of GSL expression profile were observed during mammalian embryogenesis. Cell surface molecules expressed on embryos in a stage-specific manner appeared to play key roles in regulation of cell-cell interaction and cell sorting during early development. Many mAbs showing stage-specific reactivity with mouse embryos were shown to recognize carbohydrate epitopes. Among various stage-specific embryonic antigens (SSEAs), SSEA-1 was found to react with neolacto-series GSL Lex, while SSEA-3 and SSEA-4 reacted with globo-series Gb5 and monosialyl-Gb5, respectively. GSL expression during mouse early development was shown to shift rapidly from globo-series to neolacto/lacto-series, and then to ganglio-series. We found that multivalent Lex caused decompaction of mouse embryos, indicating a functional role of Lex epitope in the compaction process. Autoaggregation of mouse embryonal carcinoma (EC) F9 cells provided a useful model of the compaction process. We showed that Lex-Lex interaction, a novel type of molecular interavction termed carbohydrate-carbohydrate interaction (CCI), was involved in cell aggregation. Similar shifting of GSL expression profiles from globo-series and neolacto/lacto-series to ganglio-series was observed during differentiation of human EC cells and embryonic stem (ES) cells, reflecting the essential role of cell surface glycoconjugates in early development. 相似文献
4.
Human blood group A, B, H, Ii, Lea and Leb antigens and their determinants expressed on ovarian cyst glycoproteins have been studied for over five decades. However, little is known about sialyl Lex and sialyl Lea glycotopes, which play essential roles in normal immunity, inflammation, and cancer cell metastasis. Furthermore, Lex and Ley were classified as glycotopes of unknown genes. Identification of these Lewis epitopes was hampered by the lack of specific antibodies. In this study, the occurrence of sialyl Lex, sialyl Lea, Lex and Ley reactivities in cyst glycoproteins was characterized by enzyme-linked immunosorbent assays. The results indicated that most human ovarian cyst glycoproteins carried Lex (8/25) and/or Ley (17/25) glycotopes. The expression (epitopes) of the new genes described in previous reports are Lex and Ley glycotopes; the reactivities of sialyl Lex and sialyl Lea glycotopes in secreted cyst glycoproteins may be affected by the conditions of purification; the relationship between Ley and human blood group ABH was confirmed; recognition profiles of sialyl Lex, sialyl Lea, Lex and Ley present in the carbohydrate chains of water-soluble cyst glycoproteins were illustrated; possible attachments of glycotopes to the internal carbohydrate complex of cyst glycoproteins have been reconstructed; proposed biosynthetic pathways for the formation of sialyl Lea, sialyl Lex, Lex, Ley, ALey and BLey determinant structures on Type I and Type II core structures of human ovarian cyst glycoproteins are also included in this study. 相似文献
5.
Differentiation of isomeric Lewis blood groups by positive ion electrospray tandem mass spectrometry
Lewis histo-blood group antigens are one of the major classes of biologically active oligosaccharides. In this work, underivatized Lewis blood groups were studied by electrospray tandem mass spectrometry (ESI-MSn) in the positive mode with three different mass analyzers: Q-TOF (quadrupole time-of-flight), QqQ (triple quadrupole), and LIT (linear ion trap). It was observed that, under collision-induced fragmentations, type 1 Lewis antigens (Lea and Leb) could be distinguished from type 2 (Lex and Ley) on the basis of specific fragmentations of the GlcNAc unit. Whereas O-4-linked sugars of the GlcNAc are lost as residues, the O-3-linked sugars undergo fragmentation both as sugar units and as sugar residues (unit −18 Da). Type 2 Lewis antigens also showed a characteristic cross-ring cleavage 0,2A2 of the GlcNAc. As a result, the product ions at m/z 388 and 305, characteristic of Lex, and m/z 372, characteristic of Lea, are proposed to distinguish the trisaccharide isomers Lex/Lea. Also, the product ions at m/z 534 and 305, characteristic of Ley, and m/z 372, characteristic of Leb, are proposed to distinguish the tetrasaccharide isomers Leb/Ley. These diagnostic fragment ions were further applied in the identification of Lewis type 2 antigens (Lex and Ley) in the lipopolysaccharide of the human gastric pathogen, Helicobacter pylori. 相似文献
6.
Gustafsson A Kacskovics I Breimer ME Hammarström L Holgersson J 《Glycoconjugate journal》2005,22(3):109-118
Breast-milk has a well-known anti-microbial effect, which is in part due to the many different carbohydrate structures expressed. This renders it a position as a potential therapeutic for treatment of infection by different pathogens, thus avoiding the drawbacks of many antibiotics. The plethora of carbohydrate epitopes in breast-milk is known to differ between species, with human milk expressing the most complex one. We have investigated the expression of protein-bound carbohydrate epitopes in milk from man, cow, goat, sheep, pig, horse, dromedary and rabbit. Proteins were separated by SDS-PAGE and the presence of carbohydrate epitopes on milk proteins were analysed by Western blotting using different lectins and carbohydrate-specific antibodies. We show that ABH, Lewis (Le)x, sialyl-Lex, Lea, sialyl-Lea and Leb carbohydrate epitopes are expressed mainly on man, pig and horse milk proteins. The blood group precursor structure H type 1 is expressed in all species investigated, while only pig, dromedary and rabbit milk proteins carry H type 2 epitopes. These epitopes are receptors for Helicobacter pylori (Leb and sialyl-Lex), enteropathogenic (H type 1, Lea and Lex) and enterotoxic Escherichia coli (heat-stable toxin; H type 1 and 2), and Campylobacter jejuni (H type 2). Thus, milk from these animals or their genetically modified descendants could have a therapeutic effect by inhibiting pathogen colonization and infection.
Published in 2005. 相似文献
7.
Radhia Mahfoud Adam Manis Beth Binnington Cameron Ackerley Clifford A. Lingwood 《The Journal of biological chemistry》2010,285(46):36049-36059
Glycosphingolipids (GSLs) accumulate in cholesterol-enriched cell membrane domains and provide receptors for protein ligands. Lipid-based “aglycone” interactions can influence GSL carbohydrate epitope presentation. To evaluate this relationship, Verotoxin binding its receptor GSL, globotriaosyl ceramide (Gb3), was analyzed in simple GSL/cholesterol, detergent-resistant membrane vesicles by equilibrium density gradient centrifugation. Vesicles separated into two Gb3/cholesterol-containing populations. The lighter, minor fraction (<5% total GSL), bound VT1, VT2, IgG/IgM mAb anti-Gb3, HIVgp120 or Bandeiraea simplicifolia lectin. Only IgM anti-Gb3, more tolerant of carbohydrate modification, bound both vesicle fractions. Post-embedding cryo-immuno-EM confirmed these results. This appears to be a general GSL-cholesterol property, because similar receptor-inactive vesicles were separated for other GSL-protein ligand systems; cholera toxin (CTx)-GM1, HIVgp120-galactosyl ceramide/sulfatide. Inclusion of galactosyl or glucosyl ceramide (GalCer and GlcCer) rendered VT1-unreactive Gb3/cholesterol vesicles, VT1-reactive. We found GalCer and GlcCer bind Gb3, suggesting GSL-GSL interaction can counter cholesterol masking of Gb3. The similar separation of Vero cell membrane-derived vesicles into minor “binding,” and major “non-binding” fractions when probed with VT1, CTx, or anti-SSEA4 (a human GSL stem cell marker), demonstrates potential physiological relevance. Cell membrane GSL masking was cholesterol- and actin-dependent. Cholesterol depletion of Vero and HeLa cells enabled differential VT1B subunit labeling of “available” and “cholesterol-masked” plasma membrane Gb3 pools by fluorescence microscopy. Thus, the model GSL/cholesterol vesicle studies predicted two distinct membrane GSL formats, which were demonstrated within the plasma membrane of cultured cells. Cholesterol masking of most cell membrane GSLs may impinge many GSL receptor functions. 相似文献
8.
Jo-Wen Wang Ari Asnani France-Isabelle Auzanneau 《Bioorganic & medicinal chemistry》2010,18(20):7174-7185
A Lex trisaccharide functionalized with a cysteamine arm was prepared and this synthesis provided additional information on the reactivity of N-acetylglucosamine O-4 acceptors when they are glycosylated with trichloroacetimidate donors activated with excess BF3·OEt2. In turn, this trisaccharide was conjugated to BSA lysine side chains through a squarate–mediated coupling. This BSA-Lex glycoconjugate displayed 35 Lex haptens per BSA molecule. The relative affinity of the anti-Lex monoclonal antibody SH1 for the Lex antigen and analogues of Lex in which the d-glucosamine, l-fucose or d-galactose residues were replaced with d-glucose, l-rhamnose and d-glucose, respectively, was measured by competitive ELISA experiments. While all analogues were weaker inhibitors than the Lex antigen, only the analogue of Lex in which the galactose residue was replaced by a glucose unit showed no binding to the SH1 mAb. To confirm that the reduced or loss of recognition of the Lex analogues by the anti-Lex mAb SH1 did not result from different conformations adopted by the analogues when compared to the native Lex antigen, we assessed the conformational behavior of all trisaccharides by a combination of stochastic searches and NMR experiments. Our results showed that, indeed, the analogues adopted the same stacked conformation as that identified for the Lex antigen. The identification of a trisaccharide analogue that does not cross-react with Lex but still retains the same conformation as Lex constitutes the first step to the design of a safe anti-cancer vaccine based on the dimeric Lex tumor associated carbohydrate antigen. 相似文献
9.
V. E. Piskarev T. L. Bushueva I. A. Yamskov 《Russian Journal of Bioorganic Chemistry》2007,33(1):170-174
We studied interaction of the lectin from the bark of Golden Rain shrub (Laburnum anagyroides, LABA) with a number of basic fucose-containing carbohydrate antigens by changes in its tryptophan fluorescence. The strongest LABA binding was observed for the trisaccharide H of type 6 [α-L-Fucp-(1-2)-β-D-Galp-(1-4)-D-Glc, K a = 4.2 × 103 M?1]. The following antigens were bound with a weaker affinity: H-disaccharide α-L-Fucp-(1-2)-D-Gal, a glucoanalogue of tetrasaccharide Ley α-L-Fucp-(1-2)-β-D-Galp-(1-4)-[α-L-Fucp-(1-3)]-D-Glc, and 6-fucosyl-N-acetylglucosamine, a fragment of core of the N-glycans family (K a 1.1?1.7 × 103 M?1). The lowest binding was observed for L-fucose (K a = 2.7 × 102 M?1) and trisaccharide Lea, (β-Galp-(1-3)-[α-L-Fucp-(1-4)]-GlcNAc (K a = 6.4 × 102 M?1). The Led, Lea, and Lex pentasaccharides and Leb hexasaccharide were not bound to LABA. 相似文献
10.
Sen-Itiroh Hakomori 《The Histochemical journal》1992,24(11):771-776
Summary and conclusion Lex (13 fucosylated type 2 chain) functions as an adhesion molecule capable of Ca2+-mediated homotypic binding. Cells with high surface expression of Lex therefore exhibit strong self-aggregation (based on Lex-Lex interaction) in the presence of Ca2+. In this review, I have summarized several lines of supporting data for this concept, and the role of Lex-Lex interaction in the process of embryo compaction and autoaggregation of F9 teratocarcinoma cells. In general, cell adhesion events based on Lex-Lex interaction may be followed and reinforced by integrin- or Ig receptor-based adhesion systems.SLex, the 23 sialosyl derivative of Lex, and its positional isomer SLea, have been identified as the target molecules for selectin-dependent cell adhesion. Adhesion of leukocytes or tumour cells to ECs or platelets, which express E-selectin and P-selectin respectively, is initiated by this process. The target epitopes SLex and SLea are presented mainly on transmembrane glycoproteins having many clusters of O-linked carbohydrate chains. Therefore, inhibition of O-glycosylation may be effective for blocking selectin-mediated cell adhesion. The abundant presence of Lex epitope in the central nervous system, and the physiological changes of Lex expression as described in this monograph, reflect the adhesive properties of this molecule and its sialyosylated and/or fucosylated derivatives. 相似文献
11.
Structural characterization of sulfated and sialyl Lewis (Le)-type glycosphingolipids performed by fast atom bombardment (FAB) and electrospray ionization (ESI) mass spectrometry is described. Both FAB and ESI collision-induced dissociation tandem mass spectrometry (CID-MS/MS) of acidic glycosphingolipids allowed identification of the sulfated or sialyl sugar, and provided information on the saccharide chain sequence. The negative-ion tandem FABMS of sulfated Le-type glycosphingolipids having the non-reducing end trisaccharide ion as the precursor can be used to differentiate the Lea- and LeX-type oligosaccharides. The ESI CID-MS/MS of multiple-charged ions provided even more detailed structural information, and some of the useful daughter ions appeared with higherm/z values than the precusor because of a lower charge-state. These methodologies can be applied to the structural analyses of glycoconjugates with much larger molecular masses and higher polarity, such as the poly-sulfated and sialyl analogues.Abbreviations CID
collision-induced dissociation
- ESI
electrospray ionization
- FABMS
fast atom bombardment mass spectrometry
- Fuc
fucose
- Gal
galactose
- GlcNAc
N-acetylglucosamine
- Le
Lewis
- Lea
Lewisa
- LeX
LewisX
- MS/MS
mass spectrometry/mass spectrometry
- NeuAc
N-acetylneuraminic acid
- 3-SO4-Lea
3-sulfated Lea pentaosyl ceramide
- 3-SO4-LeX
3-sulfated LeX pentaosyl ceramide
- 2,3-SO4-LeX
2,3-disulfated LeX pentaosyl ceramide
- 3-S-Lea
3-sialyl Lea pentaosyl ceramide
- 3-S-Lex
3-sialyl LeX heptaosyl ceramide
- 3-S-LeX-LeX
3-sialyl-Lex-Lex octaosyl ceramide. 相似文献
12.
Purified renal globotriaosyl ceramide (Gb3)/cholesterol mixtures, sonicated and heated in a Triton-containing buffer and placed below a discontinuous sucrose gradient,
form glycosphingolipid (GSL)-containing dense lipid structures at the 30/5% sucrose interface after centrifugation. Inclusion
of fluorescein-labeled verotoxin 1 B subunit (FITC-VT1 B) within the most dense sucrose layer results in the fluorescent labeling
of this Gb3-containing raft structure. Alternatively, inclusion of 125I-labeled VT1 and fractionation allows quantitation of binding. FITC-VT1 B effectively competes for 125I-VT1/Gb3 raft binding. This assay will allow the definition of the optimal raft composition for VT1 (or any other ligand) binding.
The effect of several potential cellular raft components are reported. Increased cholesterol content increased VT1 binding.
Addition of phosphatidylethanolamine had minimal effect while phosphatidylserine was inhibitory. Although inclusion of sphingomyelin
increased the Gb3 content of the 'raft', reduced VT1 binding was seen. Inclusion of other glycolipids can also be inhibitory. The addition
of globotetraosyl ceramide had no effect; however, addition of sulfogalactosyl ceramide, but not sulfogalactoglycerolipid,
inhibited VT1/Gb3 raft binding. These results suggest that certain GSLs can disfavour the formation of the appropriate 'raft' structure for
ligand binding and that this is dependent on both their carbohydrate and lipid structure. Such “deceptor” GSLs may provide
an as yet, unappreciated mechanism for the regulation of cellular GSL receptor activity. This model is an effective tool to
approach the dynamics and ligand-binding specificity of GSL/cholesterol-containing lipid microdomains. Published in 2004.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
13.
Minas V Mylonas I Schiessl B Mayr D Schulze S Friese K Jeschke U Makrigiannakis A 《Histochemistry and cell biology》2007,128(1):55-63
Lewis antigens belong to the blood group of antigens and mediate cellular adhesion through interaction with selectins. Invasive
trophoblasts use an array of adhesion molecules to facilitate cell–cell and cell–extracellular matrix interactions. Here,
we examined immunohistochemically the expression of Sialyl Lewis a (sLea), Sialyl Lewis x (sLex) and Lewis y (Ley) in term placentas obtained from cases of normal, intrauterine growth retardation (IUGR), preeclamptic (PE) and hemolysis,
elevated liver enzymes and low platelets syndrome (HELLP) pregnancies. We report the expression of sLex in third trimester extravillous trophoblasts (EVT). sLex was significantly decreased in IUGR and moderately decreased in PE compared to normal placentas. sLex was additionally found in syncytiotrophoblast, without however any significant differences in staining intensity between
normal and pathological cases. sLea was restricted to amnion epithelium. Finally, Ley was expressed in cytotrophoblasts and villous endothelial cells. Ley expression was significantly upregulated in IUGR and HELLP, whereas there was a trend toward increase in PE compared to normal
placentas. The present study suggests that downregulation of sLex in EVT might be associated with IUGR and PE. Furthermore, Ley, which was recently described as a potent angiogenic factor, is upregulated in placental villi in conditions associated with
placental malperfusion.
U. Jeschke and A. Makrigiannakis have contributed equally. 相似文献
14.
Noé Velázquez-Márquez Gerardo Santos-López Lucio Jiménez-Aranda Julio Reyes-Leyva Verónica Vallejo-Ruiz 《Journal of biosciences》2012,37(1):999-1004
Sialylated oligosaccharides of glycoproteins and glycolipids have been implicated in tumour progression and metastases. Altered expression of glycosidic antigens has been reported in cervical cancer. In cervix premalignant lesions, an increased expression of sialic acid has been reported. In the present study we determined the expression profiles of the glycosidic antigens Tn, sialyl Tn (sTn), Lewis a (Lea), sialyl Lewis a (sLea), Lewis x (Lex) and sialyl Lewis x (sLex) in cervical scrapes with cytological diagnoses of normal, low-grade squamous intraepithelial lesions (LGSIL) and high-grade squamous intraepithelial lesions (HGSIL). Cervical scrapings were collected to detect tumour antigens expressions by flow cytometry using monoclonal antibodies. Cytometry analysis of Tn, sTn, Lea and Lex did not reveal differences at the expression level among groups. The number of positive cells to sLea antigen increased in the HGSIL group with respect to the normal group (p?=?0.0495). The number of positive cells to sLex antigen in the samples increased with respect to the grade of squamous intraepithelial lesion (SIL) (p?<?0.001, Mann–Whitney U test). The intensity of expression of this antigen increased in the HGSIL samples with respect to normal samples (p?<?0.0068). sLex antigen could be a candidate to be used as biomarker for the early diagnosis of cervical cancer. 相似文献
15.
Xiaoxia Guo Xiaoxi Ling Fang Du Qingbing Wang Wei Huang Zhongmin Wang Xiaoyi Ding Mingfeng Bai Zhiyuan Wu 《Translational oncology》2018,11(5):1065-1073
Imaging probes targeting type 2 cannabinoid receptor (CB2R) overexpressed in pancreatic duct adenocarcinoma (PDAC) tissue have the potential to improve early detection and surgical outcome of PDAC. The aim of our study was to evaluate the molecular imaging potential of a CB2R-targeted near-infrared (NIR) fluorescent probe (NIR760-XLP6) for PDAC. CB2R overexpression was observed in both PDAC patient tissues and various pancreatic cancer cell lines. In vitro fluorescence imaging indicated specific binding of NIR760-XLP6 to CB2R in human PDAC PANC-1 cells. In a xenograft mouse tumor model, NIR760-XLP6 showed remarkable 50- (ex vivo) and 3.2-fold (in vivo) tumor to normal contrast enhancement with minimal liver and kidney uptake. In a PDAC lymph node metastasis model, significant signal contrast was observed in bilateral axillary lymph nodes with PDAC metastasis after injection of the probe. In conclusion, NIR760-XLP6 exhibits promising characteristics for imaging PDAC, and CB2R appears to be an attractive target for PDAC imaging. 相似文献
16.
Vibeke Ravn Christence Stubbe Teglbjærg Ulla Mandel Erik Dabelsteen 《Cell and tissue research》1992,270(3):425-433
Summary The blood group ABO(H) determinants are major allogenic antigens in both erythrocytes and tissue of man. These antigens and related carbohydrates are markers of cellular maturation and differentiation in many epithelial tissues and have recently attracted great interest as tumor-associated antigens. Previous studies of endometrial tissues have indicated that glycosylation in this tissue may be related to hormonal stimulation. We have investigated the immunohistochemical distribution of type-2 chain histo-blood group-related carbohydrates in specimens of normal, cycling endometria obtained from hysterectomies on women with known ABO/Lewis erythrocyte type and saliva secretor status. N-acetyllactosamine and Lex were demonstrated to be uninfluenced by the genetic background. A and Aley antigens were exclusively demonstrated in endometria from blood group A individuals, while Ley was expressed in endometria from blood group 0 individuals mainly. The precursor N-acetyllactosamine as well as the terminal H, A, and ALey antigens were shown in only a few cells. In contrast, N-acetyllactosamine substituted by sialic acid and/or fucose residues (Lex, sialosyl-Lex, Ley) were demonstrated in epithelial cells of normal, cycling endometrium, but with both quantitative and qualitative differences in staining relating to the menstrual cycle, indicating that type-2 chain antigens are expressed under both genetic and hormonal influence in human cycling endometrium. 相似文献
17.
Govind Ragupathi Payal Damani Geeta Srivastava Om Srivastava Steven J. Sucheck Yoshi Ichikawa Philip O. Livingston 《Cancer immunology, immunotherapy : CII》2009,58(9):1397-1405
Sialyl Lewisa (sLea), also termed CA19-9 antigen, is recognized by murine mAb19-9 and is expressed on the cancer cell surface as a glycolipid
and as an O-linked glycoprotein. It is highly expressed in a variety of gastrointestinal epithelial malignancies including
colon cancer and pancreatic cancer, and in breast cancer and small cell lung cancer, but has a limited expression on normal
tissues. sLea is known to be the ligand for endothelial cell selectins suggesting a role for sLea in cancer metastases and adhesion. For these reasons, sLea may be a good target for antibody mediated immunotherapy including monoclonal antibodies and tumor vaccines. However, sLea is structurally similar to sLex and other blood group related carbohydrates which are widely expressed on polymorphonucleocytes and other circulating cells,
raising concern that immunization against sLea will induce antibodies reactive with these more widely expressed autoantigens. We have shown previously both in mice and
in patients that conjugation of a variety of carbohydrate cancer antigen to keyhole limpet hemocyanin (KLH) and administration
of this conjugate mixed with saponin adjuvants QS-21 or GPI-0100 are the most effective methods for induction of antibodies
against these cancer antigens. We describe here for the first time the total synthesis of pentenyl glycoside of sLea hexasaccharide and its conjugation to KLH to construct a sLea-KLH conjugate. Groups of five mice were vaccinated subcutaneously four times over 6 weeks. Sera were tested against sLea-HSA by ELISA and against sLea positive human cell lines adenocarcinoma SW626 and small cell lung cancer (SCLC) DMS79 by FACS. As expected, mice immunized
with unconjugated sLea plus GPI-0100 or unconjugated sLea mixed with KLH plus GPI-0100 failed to produce antibodies against sLea. However, mice immunized with sLea-KLH conjugate without GPI-0100 produced low levels of antibodies and mice immunized with sLea-KLH plus GPI-0100 produced significantly higher titer IgG and IgM antibodies against sLea by ELISA. These antibodies were highly reactive by FACS and mediated potent complement mediated cytotoxicity against sLea positive SW626 and DMS79 cells. They showed no detectable cross reactivity against a series of other blood group-related
antigens, including Ley, Lex, and sLex by dot blot immune staining. This vaccine is ready for testing as an active immunotherapy for treating sLea positive cancer in clinical settings.
Govind Ragupathi and Philip O. Livingston are paid consultants and shareholders in MabVax Therapeutics, Inc., San Diego, CA
92121. The sLea vaccine is licensed to MabVax. 相似文献
18.
Streptococcus mutans is a major etiological agent in dental caries. Salivary agglutinin is one of the main salivary components binding to S.mutans. To learn more about the interaction of salivary agglutinin with S.mutans, parotid, submandibular, sublingual and palatal saliva samples were incubated with S. mutans suspension. Both depleted saliva samples and bacterial extracts were analyzed by SDS-PAGE and immunoblotting. Salivary agglutinin was present in all types of glandular saliva and in all cases bound to S.mutans, also to PC337C, a P1– mutant of S.mutans. Agglutinin was separated by SDS-PAGE under reducing and non-reducing conditions and then transferred to nitrocellulose. Non-reduced agglutinin bound S.mutans, but reduced agglutinin did not. Adhesion of S.mutans to agglutinin-coated microplates was inhibited by amine-containing components, 1 M NaCl or KCl and EDTA. Adhesion decreased with decreasing pH with no adhesion below pH 5.0. These data suggest that calcium-dependent electrostatic interactions play a role in binding. By immunoblotting was demonstrated that blood group antigens and Lewis antigens were present on agglutinin. Synthetic blood group antigens and Lewis antigens covalently coupled to polyacrylamide were tested for binding to S.mutans. Only Lea(Gal1,3(Fuc1,4)GlcNAc) bound to S.mutans, whereas the blood group antigens Leb, Lex, Ley, H1, H2, A, B and sialylated Lea did not. Lea without galactose (Fuc1,4GlcNAc) still bound to S. mutans, but Lea without fucose (Gal1,3GlcNAc) did not. Binding of agglutinin to S. mutans was not inhibited by Lea. In conclusion, S. mutans can bind to Lea carbohydrate epitopes in which the fucose is an essential residue. Lea carbohydrate epitopes are present on salivary agglutinin but play no major role in binding. 相似文献
19.
Piskarev V. E. Busheva T. L. Yamskov I. A. 《Applied Biochemistry and Microbiology》2004,40(4):415-417
The interaction of fucolectin of perch Perca fluviatilis (PFL) with a set of Lewis antigens was studied by monitoring changes in its tryptophan fluorescence. PFL bound Lec (H type 1)-pentasaccharide (K
a = 6.6 × 103 –1) and H type 6-trisaccharide (K
a = 2.5 × 103 –1); bound, although less strongly, with Leb-hexasaccharide (K
a = 4.0 × 102 –1); and failed to interact with Lea-, Lex-, and Led-containing oligosaccharides. PFL belongs to a new type of the fucolectins recognizing H-disaccharide Fuc1-2Gal within various antigens, including H type 1/2 and Leb. 相似文献
20.
Sialosyl-Lex (SLex) and its positional isomer sialosyl-Lea are the epitopes recognized by the lectin domain of E- and P-selectins. Expression of SLex in polymorphonuclear leukocytes (PMN) plays an important role in recruitment of these cells at sites of inflammation through activation of selectins. We studied expression of SLex in PMN of seven mammalian species in comparison with that in humans. Only PMN of humans (no other species) expressed SLex or other lacto-series epitopes such as Lex or Ley. The observed absence of these epitopes in rat PMN seems inconsistent with recent reports that the lung inflammation process in a rat model is inhibited by perfusion of SLex oligosaccharide (Mulligan MS,et al. (1993a)Nature
364:149; (1993b)J Exp Med
178:623). Rat selectins may be able to recognize SLex, even though this epitope is absent in rat PMN.Abbreviations FITC
fluorescein isothiocyanate
- mAb
monoclonal antibody
- PMN
polymorphonuclear leukocytes
- SLea
sialosyl-Lea antigen
- SLex
sialosyl-Lex antigen 相似文献