Lewis Antigens Research Articles

Integrated Glycome Strategy for Characterization of Aberrant LacNAc Contained N-Glycans Associated With Gastric Carcinoma.

Aberrant glycosylation is not only a feature of malignant cell transformation, but also plays an important role in metastasis. In the present study, an integrated strategy combining the lectin microarrays and lectin cytochemistry was employed to investigate and verify the altered glycopatterns in gastric cancer (GC) cell lines as well as resected tumor specimens from matched tissue sets of 46 GC patients. Subsequently, lectin-mediated affinity capture glycoproteins, and MALDI-TOF/TOF-MS were employed to further acquire precise structural information of the altered glycans. According to the results, the glycopatterns recognized by 10 (e.g., ACA, MAL-I, and ConA) and 3 lectins (PNA, MAL-I, and VVA) showed significantly variations in GC cells and tissue compared to their corresponding controls, respectively. Notably, the relative abundance of Galβ-1,4GlcNAc (LacNAc) recognized by MAL-I exhibited a significant increase in GC cells (p < 0.001) and tissue from patients at stage II and III (p < 0.05), and a significant increase in lymph node positive tumor cases, compared with lymph node negative tumor cases (p < 0.05). More LacNAc contained N-glycans were characterized in tumor sample with advanced stage compared to corresponding control. Moreover, there were 10 neo-LacNAc-contained N-glycans (e.g., m/z 1625.605, 1803.652, and 1914.671) only presented in GC tissue with advanced stage. Among these, six N-glycans were modified with sialic acid or fucose based on LacNAc to form sialylated N-glycans or lewis antigens, respectively. Our results revealed that the aberrant expression of LacNAc is a characteristic of GC, and LacNAc may serve as a scaffold to be further modified with sialic acid or fucose. Our findings provided useful information for us to understand the development of GC.

Abstract 2301: Antiglycan antibodies as predictors of therapeutic outcome in cervical cancer patients

Abstract Cervical cancer is a significant cause of cancer related death among women, with the highest mortality rates in developing countries. We aim to identify serum biomarkers for predicting outcome of radio therapy alone or combination of radiotherapy and brachytherapy. We measured natural anti-glycan IgG antibodies in the serum of stage II (n=276) and III (n=292) cervical cancer patients treated with radiation alone or in combination with brachytherapy on a suspension glycan array containing 184 glycans. Overall and progression free survival was used to determine the treatment outcome by Kaplan-Meier and Cox proportional hazards ratio. Significant protective effects were observed for Blood Group H antigens (HR=0.642, p=0.0013), Lewis Antigens (0.72, 0.015; 0.654, 0.003), Galili antigens (0.671, p=0.004), and other glycans belonging to Isoglobo (0.671,0.004) and 3fucosyllactose series (0.687,0.004). The protective effects remained significant after adjusting for stage and treatment. Our data suggest that measurement of natural anti-glycan IgG antibodies in serum can predict outcome of therapy in cervical cancer patients. Citation Format: SHARAD PUROHIT, Paul M. Tran, Lynn K. Tran, Xuezheng Song, Ferris G. Ferris, John J. Wallbillich, Boying Dun, Jin-Xiong She. Antiglycan antibodies as predictors of therapeutic outcome in cervical cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2301.

Human glycan expression patterns influence Group A streptococcal colonization of epithelial cells.

Colonization of the oropharynx is the initial step in Group A Streptococcus (GAS) pharyngeal infection. We have previously reported that the highly virulent M1T1 GAS clone attaches to oral epithelial cells via M1 protein interaction with blood group antigen carbohydrate structures. Here, we have identified that colonization of human oral epithelial cells by GAS serotypes M3 and M12 is mediated by human blood group antigens [ABO(H)] and Lewis (Le) antigen expression. Removal of linkage-specific fucose, galactose, N-acetylgalactosamine, and sialic acid modulated GAS colonization, dependent on host ABO(H) blood group and Le expression profile. Furthermore, N-linked glycans from human salivary glycoproteins, when released and purified, were potent inhibitors of M1, M3, and M12 GAS colonization ex vivo. These data highlight the important role played by human protein glycosylation patterns in GAS attachment to oral epithelial cell surfaces.-De Oliveira, D. M. P., Everest-Dass, A., Hartley-Tassell, L., Day, C. J., Indraratna, A., Brouwer, S., Cleary, A., Kautto, L., Gorman, J., Packer, N. H., Jennings, M. P., Walker, M. J., Sanderson-Smith, M. L. Human glycan expression patterns influence Group A streptococcal colonization of epithelial cells.

Biologic roles of the ABH and Lewis histo-blood group antigens part II: thrombosis, cardiovascular disease and metabolism.

The ABH and Lewis antigens were among the first of the human red blood cell polymorphisms to be identified and, in the case of the former, play a dominant role in transfusion and transplantation. But these two therapies are largely twentieth-century innovations, and the ABH and related carbohydrate antigens are not only expressed on a very wide range of human tissues, but were present in primates long before modern humans evolved. Although we have learned a great deal about the biochemistry and genetics of these structures, the biological roles that they play in human health and disease are incompletely understood. This review and its companion, which appeared in a previous issue of Vox Sanguinis, will focus on a few of the biologic and pathologic processes which appear to be affected by histo-blood group phenotype. The first of the two reviews explored the interactions of two bacteria with the ABH and Lewis glycoconjugates of their human host cells, and described the possible connections between the immune response of the human host to infection and the development of the AB-isoagglutinins. This second review will describe the relationship between ABO phenotype and thromboembolic disease, cardiovascular disease states, and general metabolism.

Biologic roles of the ABH and Lewis histo-blood group antigens Part I: infection and immunity.

The ABH and Lewis antigens were among the first of the human red blood cell polymorphisms to be identified and, in the case of the former, play a dominant role in transfusion and transplantation. But these two therapies are largely twentieth century innovations, and the ABH and related carbohydrate antigens are not only expressed on a very wide range of human tissues, but were present in primates long before modern humans evolved. Although we have learned a great deal about the biochemistry and genetics of these structures, the biological roles that they play in human health and disease are incompletely understood. This review and its companion, to appear in a later issue of Vox Sanguinis, will focus on a few of the biologic and pathologic processes which appear to be affected by histo-blood group phenotype. The first of the two reviews will explore the interactions of two bacteria with the ABH and Lewis glycoconjugates of their human host cells, and describe the possible connections between the immune response of the human host to infection and the development of the AB-isoagglutinins. The second review will describe the relationship between ABO phenotype and thromboembolic disease, cardio-vascular disease states, and general metabolism.

Reprogramming the enzymatic assembly line for site-specific fucosylation

Fucosylated carbohydrate determinants are common components of cell surface glycoconjugates and secreted unconjugated glycans, which play pivotal roles in many physiological and pathological processes. The biosynthesis of Lewis antigens involves multiple fucosyltransferases that catalyse the fucosylation of the poly-N-acetyllactosamine carbohydrate backbone in a non-site-specific manner and thus generate heterogeneous and incompletely fucosylated Lewis antigen regioisomers. In this study, an α2,6-sialylation module was used to introduce α2,6-linked sialic acid to specific sites as the protecting group against fucosylation, thus precisely controlling enzymatic fucosylation of poly-N-acetyllactosamine glycans in a site-specific manner. The sialic acid protecting group can be easily removed by sialidase after fucosylation to provide a variety of fucosylated poly-N-acetyllactosamine glycans with defined fucosylation patterns. The general applicability and robustness of this reprogrammed enzymatic assembly line was exemplified in the synthesis of 22 complex Lewis antigens and chimeric histo-blood group antigens with a total of 10 enzyme modules for the construction of 10 different glycosidic linkages. The availability of Lewis antigens allows the investigation of their important biological functions, but site-specific fucosylation for their synthesis is challenging. This work reports an enzymatic platform for the synthesis of complex Lewis antigens, offering an opportunity to explore the Lewis antigen related glycome.

Linkage and sequence analysis of neutral oligosaccharides by negative-ion MALDI tandem mass spectrometry with laser-induced dissociation

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.

Lewis and AB0 blood group-phenotypes in periodontitis, cardiovascular disease, obesity and stroke

The AB0 blood group has been linked to ischaemic heart disease, stroke, and periodontal disease, while the Lewis blood group has been linked to ischaemic heart disease and obesity, all of which have been associated with periodontitis. AB0 or Lewis blood group phenotype may therefore constitute common hereditary components predisposing to these disorders. In this study, we investigated if blood group phenotype associated with periodontitis in a subpopulation consisting of 702 participants from a Danish cross-sectional cohort and, secondarily, attempted to confirm their association with hypertension, ischaemic heart disease, stroke, and obesity. No significant association between blood group phenotype and periodontitis was detected, nor were previously reported associations between blood group phenotype and hypertension, ischaemic heart disease, stroke, and obesity confirmed. This may, at least partly, be attributed to differences in study type, outcome definitions, cohort sizes, and population attributable factors. However, our results suggested a strong association between self-reported stroke and the Lewis (a−b−) phenotype (P = 0.0002, OR: 22.28; CI 95: 4.72–131.63).

Histo-Blood Group Antigens in Children with Symptomatic Rotavirus Infection.

Group A rotaviruses are a major cause of acute gastroenteritis in children. The diversity and unequal geographical prevalence of rotavirus genotypes have been linked to histo-blood group antigens (HBGAs) in different human populations. In order to evaluate the role of HBGAs in rotavirus infections in our population, secretor status (FUT2+), ABO blood group, and Lewis antigens were determined in children attended for rotavirus gastroenteritis in Valencia, Spain. During three consecutive years (2013–2015), stool and saliva samples were collected from 133 children with rotavirus infection. Infecting viral genotypes and HBGAs were determined in patients and compared to a control group and data from blood donors. Rotavirus G9P[8] was the most prevalent strain (49.6%), followed by G1P[8] (20.3%) and G12P[8] (14.3%). Rotavirus infected predominantly secretor (99%) and Lewis b positive (91.7%) children. Children with blood group A and AB were significantly more prone to rotavirus gastroenteritis than those with blood group O. Our results confirm that a HBGA genetic background is linked to rotavirus P[8] susceptibility. Rotavirus P[8] symptomatic infection is manifestly more frequent in secretor-positive (FUT2+) than in non-secretor individuals, although no differences between rotavirus G genotypes were found.

Lectin activity of Pseudomonas aeruginosa vaccine candidates PSE17-1, PSE41-5 and PSE54

Pseudomonas aeruginosa is an opportunistic pathogen that causes nosocomial infections most commonly in immunocompromised, cystic fibrosis (CF) and burns patients. The pilin and Pseudomonas lectins 1 (PA-IL) and 2 (PA-IIL) are known glycan-binding proteins of P. aeruginosa that are involved in adherence to host cells, particularly CF host airways. Recently, new P. aeruginosa surface proteins were identified by reverse vaccinology and tested in vivo as potential vaccine antigens. Three of these, namely PSE17-1, PSE41-5 and PSE54, were screened for glycan binding using glycan arrays displaying glycan structures representative of those found on human cells. Surface plasmon resonance was used to confirm the lectin activity of these proteins, and determined affinities with several host glycans to be in the nanomolar range. PSE17-1 binds hyaluronic acid and sialyl Lewis A and X. PSE41-5 binds terminal β-linked galactose structures, Lewis and ABO blood group antigens. PSE54 binds to ABO blood group antigens and some terminal β-linked galactose. All three proteins are novel lectins of P. aeruginosa with potential roles in infection of host cells.

Blood group antigen-Binding Adhesion2 (BabA2) gene in gastric tissue biopsies as a diagnostic biomarker for Helicobacter pylori infection.

The Lewis (b) blood group antigen-Binding Adhesion2 (BabA2) has been reported to mediate the attachment of H. pylori to human. assessment the diagnostic potential of detection of (BabA2) gene compared with immunostaining of Lewis (b) by specific mouse monoclonal antibodies in gastric biopsies from Egyptian Patients as a diagnostic maker for Helicobacter pylori infection. Fifty untreated patients suffering from dyspeptic complaints were enrolled in this study and underwent for upper gastro-duodenal endoscopy. Biopsies were taken for histological examination by (H&E) and immunohistochemical analysis for Lewis b by specific mouse monoclonal antibodies, and scoring of Lewis b expression in gastric tissue biopsy as well as molecular detection of BabA2 gene of H. pylori by PCR. Biochemical analysis was performed to detect the presence of H. pylori urease activity using Rapid Urease Test (RUT). : Out of 50 gastric biopsies, 41biopsies were positive for histological, Immunostaining for Lewis b expression and urease activity test (RUT) for H pylori. RUT showed a sensitivity of 87.8%, specificity 88.9%, positive predictive value (PPV) 97.2%, and negative predictive value (NPV) 61.5%. BabA2 gene results revealed that, out of 41 positive biopsied cases, 39 (95.1%) were positive by the PCR test for BabA2 gene. And all 9 negative biopsies (100%) for H pylori negative for BabA2gene so the sensitivity and specificity of BabA2 gene detection in gastric biopsies by PCR were 95.1% and 100%; respectively. : BabA2 gene detection in gastric tissue biopsies could be suggested as a diagnostic biomarker to be included among the other biomarkers routinely performed for clinical diagnosis of H. pylori infection.

Lectin Activity of the TcdA and TcdB Toxins of Clostridium difficile.

Clostridium difficile is a major cause of hospital-acquired antibiotic-associated diarrhea. C. difficile produces two cytotoxins, TcdA and TcdB; both toxins are multidomain proteins that lead to cytotoxicity through the modification and inactivation of small GTPases of the Rho/Rac family. Previous studies have indicated that host glycans are targets for TcdA and TcdB, with interactions thought to be with both α- and β-linked galactose. In the current study, screening of glycan arrays with different domains of TcdA and TcdB revealed that the binding regions of both toxins interact with a wider range of host glycoconjugates than just terminal α- and β-linked galactose, including blood groups, Lewis antigens, N-acetylglucosamine, mannose, and glycosaminoglycans. The interactions of TcdA and TcdB with ABO blood group and Lewis antigens were assessed by surface plasmon resonance (SPR). The blood group A antigen was the highest-affinity ligand for both toxins. Free glycans alone or in combination were unable to abolish Vero cell cytotoxicity by TcdB. SPR competition assays indicate that there is more than one glycan binding site on TcdB. Host glycoconjugates are common targets of bacterial toxins, but typically this binding is to a specific structure or related structures. The binding of TcdA and TcdB is to a wide range of host glycans providing a wide range of target cells and tissues in vivo.

An update on the Lewis blood group system

This update of the Lewis blood group system (Combs MR. Lewis blood group system review. Immunohematology 2009;25:112-8) describes new information on the clinical significance of Lewis antigens regarding susceptibility of individuals to certain diseases and the possible role of bacteria in Lewis expression. This update also describes recently reported examples of Lewis antibodies causing hemolytic transfusion reactions. No new antigens have been identified in the International Society of Blood Transfusion system 7, leaving the antigen count to stand at six: Lea, Leb, LebH, ALeb, BLeb, and Leab.

The influence of Lewis, Duffy and Kidd blood groups on renal allograft rejection: Review

The blood groups antigens are not restricted to erythroid cells and are widely distributed in several tissues and fluids. The purpose of this study is to describe the influence Lewis, Duffy and Kidd blood groups on renal graft rejection.

Lewis (Le) blood group system phenotypes and genotypes

Lewis (Le) blood group system phenotypes and genotypes

Differential involvement of glycans in the binding of Staphylococcus epidermidis and Corynebacterium spp. to human sweat

Sweat is a secretory fluid that can be a source of unpleasant body odour due to interaction of resident bacteria with sweat components. Identification of glycoproteins in sweat suggests that protein-conjugated glycans may act as binding epitopes for bacteria, as found in other secretory fluids such as human milk, tears and saliva which help to protect epithelial surfaces from infection.We conducted proteomic and glycomic analysis of sweat to reveal an abundance of glycoproteins, predominantly carrying bi-antennary sialylated N-glycans with or without fucose. A fluorescent plate assay was used to determine whether glycans on sweat proteins provide binding epitopes for odour-producing skin commensals Staphylococcus epidermidis and Corynebacterium. Sialic acid and fucose were found to be important binding epitopes for S. epidermidis 3-22-BD-6, a strain recently isolated from human sweat, whereas fucose (but not sialic acid) contributed to the binding of Type strain S. epidermidis ATCC 12228. In contrast, our results indicate that sweat N-glycans do not provide binding epitopes for Corynebacterium.Synthetic sugar mimics of Lewis blood group antigens were investigated as potential inhibitors of the binding of S. epidermidis 3-22-BD-6 to sweat. Pre-incubation of the bacterium with LeB, LeX, LeY and sLeX (pentaose) resulted in a significant reduction in sweat protein adhesion indicating that terminal fucose is a key binding epitope, particularly when linked to a Type 2 chain (Galβ1-4GlcNAc) configuration (LeY).Our results form an impetus for future studies seeking to elucidate the role of glycans in sweat associated malodour, with possible implications for cosmetic and medical fields.

Contribution of Lewis blood groups molecules in biofilm formation of Pseudomonas aeruginosa isolated from atopic dermatitis patients.

Abstract &#x0D; Biofilm formation is a mechanism for bacterial community defense against insults including antibiotics .In this report we evaluated the potency of Pseudomonas aeruginosa(P. aeruginosa) isolates from atopic dermatitis patients skin as well as stool to colonize different Lewis types saliva , manifested by biofilm formation .&#x0D; The bacteria were cultured on tryptose soy broth .96-well polystyrene plate were used .Coating with heat inactivated Le (a), (b) and (c)saliva was performed. Biofilm intensity was measured using crystal violet stained films compared to non –saliva coated situation.&#x0D; The results showed a superior capability of most isolates to form biofilm on Le (a) followed by Le (b) saliva. The highest binding mean was for isolate ( 4).&#x0D; Le (a) saliva binding (mean ± SD was 0.66± 0.25 for test compared to 0.21± 0.04 for control non coated wells) , p=0.04,cl=0.041-0.864.&#x0D; Other isolates demonstrated variable degree of biofilm formation on this substrate .In contrast to Le (c) saliva, Le (b) saliva demonstrated weak biofilm formation .&#x0D; We conclude that, among atopic dermatitis patients skins, P. aeruginosa Lec (A) or Lec (B) lectins might be involved in colonization in such patients.&#x0D; Key Words:- Lewis blood groups – Pseudomonas aeruginosa -atopic dermatitis– Biofilm

Circulating pancreatic cancer exosomal RNAs for detection of pancreatic cancer.

Diagnostic biomarkers for the early diagnosis of pancreatic cancer are needed to improve prognosis for this disease. The aim of this study was to investigate differences in the expression of four messenger RNAs (mRNAs: CCDC88A,ARF6, Vav3, and WASF2) and five small nucleolar RNAs (snoRNAs: SNORA14B,SNORA18,SNORA25,SNORA74A, and SNORD22) in serum of patients with pancreatic cancer and control participants for use in the diagnosis of pancreatic cancer. Results were compared with the expression of sialylated Lewis (a) blood group antigen CA19‐9, the standard clinical tumor biomarker. Reverse transcription quantitative real‐time PCR showed that all of the mRNAs and snoRNAs, except CCDC88A, were encapsulated in exosomes and secreted from cultured pancreatic cancer cells, and present in cell culture medium. In a discovery‐stage clinical study involving 27 pancreatic cancer patients and 13 controls, the area under the receiver operating characteristic curve (AUC) of two mRNAs (WASF2 and ARF6) and two snoRNAs (SNORA74A and SNORA25) was > 0.9 for distinguishing pancreatic cancer patients from controls; the AUC of CA19‐9 was 0.897. Comparing serum levels of WASF2,ARF6,SNORA74A,SNORA25, and CA19‐9 revealed that levels of WASF2 were the most highly correlated with the risk of pancreatic cancer. The AUCs of WASF2,ARF6,SNORA74A, and SNORA25 in serum from patients in the early stages of pancreatic cancer (stages 0, I, and IIA) were > 0.9, compared with an AUC of 0.93 for the level of CA19‐9. The results of this study suggest that WASF2,ARF6,SNORA74A, and SNORA25 may be useful tools for the early detection of pancreatic cancer. Monitoring serum levels of WASF2 mRNA may be particularly useful, as it was the most highly correlated with pancreatic cancer risk.

Rotavirus Vaccine Take in Infants Is Associated With Secretor Status.

Rotaviruses bind to enterocytes in a genotype-specific manner via histo-blood group antigens (HBGAs), which are also detectable in saliva. We evaluated antirotavirus immunoglobulin A seroconversion ('vaccine take") among 166 Ghanaian infants after 2-3 doses of G1P[8] rotavirus vaccine during a vaccine trial, by HBGA status from saliva collected at age 4.1 years. Only secretor status was associated with seroconversion: 41% seroconversion for secretors vs 13% for nonsecretors; relative risk, 3.2 (95% confidence interval, 1.2-8.1; P = .016). Neither Lewis antigen nor salivary antigen blood type was associated with seroconversion. Likelihood of "take" for any particular rotavirus vaccine may differ across populations based on HBGAs.

Lipopolysaccharide Structural Differences between Western and Asian Helicobacter pylori Strains.

Recent structural analysis of the lipopolysaccharide (LPS) isolated from Helicobacter pylori G27 wild-type and O-antigen ligase mutant resulted in the redefinition of the core-oligosaccharide and O-antigen domains. The short core-oligosaccharide (Glc–Gal–Hep-III–Hep-II–Hep-I–KDO) and its attached trisaccharide (Trio, GlcNAc–Fuc–Hep) appear to be highly conserved structures among H. pylori strains. The G27 LPS contains a linear glucan–heptan linker between the core-Trio and distal Lewis antigens. This linker domain was commonly identified in Western strains. In contrast, out of 12 partial LPS structures of Asian strains, none displayed the heptan moiety, despite the presence of Lewis antigens. This raises the question of how Lewis antigens are attached to the Trio, and whether the LPS structure of Asian strains contain another linker. Of note, a riban was identified as a linker in LPS of the mouse-adapted SS1 strain, suggesting that alternative linker structures can occur. In summary, additional full structural analyses of LPS in Asian strains are required to assess the presence or absence of an alternative linker in these strains. It will also be interesting to study the glucan-heptan linker moieties in pathogenesis as H. pylori infections in Asia are usually more symptomatic than the ones presented in the Western world.