Blockade of neutrophil recruitment to tumor sites based on sialic acid-modified nanoplatforms enhances the efficacy of checkpoint blockade immunotherapy

Checkpoint inhibitors are designed to rejuvenate depleted or suppressed T cells in the tumor microenvironment,relying on the immune system to control and kill tumors.However,accumulating evidence indicates that tumor-infiltrating neutrophils impede the proliferation and activation of T cells and determine the resistance to checkpoint blockade and chemotherapy.In this study,sialic acid ligand-modified colchicine derivative phospholipid complexes specifically targeted tumor-associated neutrophils in the peripheral blood,blocked neutrophil accumulation in tumors,and attenuated the inhibitory effect of infiltrating neutrophils on T cells.Neutrophil blocking therapy enhanced the immunotherapy effect of the PD-L1 antibody in S180 advanced tumors and 4T1 breast cancer.Our study found that PD-L1 antibody monotherapy increased the tumor infiltration of immunosuppressive neutrophils.Combination therapy with neutrophil blocking can greatly reduce tumor-infiltrating neutrophils and increase the proliferation of cytotoxic CD8^(+) T lymphocytes in the tumor.The combination therapy significantly improved the survival rate of mice with advanced S180 tumors and increased the sensitivity of immune checkpoint inhibitors to 4T1 cold tumors.

Application of sialic acid/polysialic acid in the drug delivery systems

The properties of modified biomaterial are gaining more and more importance in drug delivery systems.Sialic acid(SA)and polysialic acid(PSA)serve as endogenous substances,which are non-immunogenic and biodegradable.At the same time,SA modification of the drugs/carriers can enhance the uptake of tumor cell and retention in brain;PSA modifi-cation can reduce the immunogenicity of the proteins or polypeptides and increase circulation time of the modified drugs/carriers in the blood,thus achieving active targeting effect.These properties offer a variety of opportunities for applications in drug delivery systems.This article summarizes the biological functions of SA and PSA and presents the technologies of SA/PSA modified small molecule drugs,proteins and carriers in drug delivery systems.

Structural Aberrations of Cellular Sialic Acids and Their Functions in Cancer Metastases

Sialic acids (neuraminic acids) are a special series of 9 carbon ring negatively charged carbohydrates, which has been found to be selectively changed in malignant cells from structures (both synthesis and structure modifications) to functions (up and down regulation in cells). Sialic acids, in single forms or conjugates, have been systematically studied both in lab and in clinics by GC, GC MS, NMR, HPTLC, HPLC and other modern analytical means. Sialic acids and related conjugates are predicted to be used in cancer diagnosis, cancer prognostic forecasting, designing of cancer chemotherapy regimens, uncovering carcinogenetic processes and neoplasm metastasis. Tumor cell regulative systems and pathways are correlated with sialic acids, which can be applied to prognostic evaluation of cancer patients, and antimetastatic chemotherapy by sialic acid derivatives and analogues. Searching for new biological characteristics of sialic acids in cells have also been extensively studied these days. In this paper, main stream discoveries and advancements are provided , also discussions of possible mechanisms and hypotheses are invoked.

APOPTOSIS OF EPIDIDYMIS EPITHELIUM AND CONTENT OF EPIDIDYMIS SIALIC ACID FOLLOWING TORSED/DETORSED TESTES IN RAT

Objective To investigate the apoptosis of epididyrnis epithelium and the change of epididymissialic acid following torsed/detorsed testes. Methods Twenty four adult male Sprague-Dawley rats were subjected to unilateral 720 testicular torsion with the duration of 2h and 4h, then repaired. The ischemic epididymis were collected for detecting the content of sialic acid by using spectrophotometry and the apoptosis with TUNEL technique. Results There were no statistically significant difference in the apoptosis of epididymis epithelium [(9.51± 2.78)% vs (6.34±1. 98)%] and the content of epididymis sialic acid(23, 3851 ± 9. 2199mg/mgprot vs 19. 3661 6. 3373mg/mgprot) at 24h between following 2h-torsed/detorsed testes and those of sham group. There were statistically significant difference in the apopotosis of epididymis epithelium[ (46. 81 ±3. 55)% vs (6. 34±1. 98) % ] and the content of epididymis sialic acid (13. 7249±7. 8006mg/mgprot vs 19. 3661±6. 3373mg/mgprot) at 24h between following 4h-torsed/detorsed testes and those of sham group(P <0. 05). Conclusion The results suggest that the sialic acid-secreting-function of epididymis remain normal at 24h following 2h-torsed/detorsed testes, while the apoptosis index of epididymis epithelium do not increase. The epididymis would be injured at 24h following 4h-torsed/detorsed testes, while the apoptosis index increased.

ROLE OF SIALIC ACID RESIDUES IN IRON BINDING BY HUMAN LACTOFERRIN

Lactoferrin(Lf-α)is a major constithent of the secondary granules of neutrophils, and is thought to be involved in bacteriostasis through high affinity binding of plasma Fe required for microbial growth.Lf-αis also the major Fe binding protein in milk. Fe bound to Lf-αis available for metabolic use.but the mechanism of Fe release from Lf-αis unknown.Treatment of human Lf-α with neuraminidase to remove sialic acid residues reduced its ability to bind Fe and caused release of Fe already bound to the protein. Reduction in Fe binding capacity was dependent on time of exposure to and concentration of neuraminidase,and was due to Lf-αloss or degradation. Readdition of sialic arid to desialylated Lf -a using a-2, 6-sialyltransferase restored Fe binding.The results suggest that sialylationdesialylation reactions could play a role in physiologic binding and release of Fe from the very high affinity binding sites of Lf -α.

SERUM LIPID-BOUND SIALIC ACID IN LUNG CANCER PATIENTS

Serum lipid-bound sialic acid (LSA) assay was evaluated for differential diagnosis between benign and malignant pulmonary disease.Mean serum LSA for normal controls was 13.7±3.7 mg/dl and the upper limit of the normal value was set at 19.8 mg/dl (95% confidence limit,one side).The specificity was 97.l% (95% confidence interval was 93.8%, 100.0%).The mean for benign pulmonary disease was 1 7.l±5.l mg/dl,the specificity was 89.l% (95% confidence interval was 80.9%,97.3%), and the sensitivity was 10.9%.The mean for malignant pulmonary disease was 27.l±7.9 mg/dl, and the sensitivity was 93.3%(95% confidence interval was 88.5%, 98.1%).The serum LSA levels for different malignant pulmonary diseases were different between small cell carcinoma and adenocarcinoma. The results indicate that serum LSA assay might be used as tumor marker to differentiate lung cancer from normal subjects or benign lung disease.

Correlation of Cerebrospinal Fluid Sialic Acid with Various Enzymes in Patients with Meningitis

Meningitis is often associated with cerebral compromise which may be responsible for neurological squeal in nearly half of the survivors. Little is known about the mechanisms of Central Nervous System （CNS） involvement in bacterial meningitis. The present study was undertaken to assess the changes in biochemical parameters including glucose, protein, Sialic Acid （SA） and various enzymes like Aspartate Aminotransferase （AST）, Alanine Aminotransferase （ALT）, Creatine Kinase （CK）, Gamma Glutamyltranspeptidase 7 （GGTP） and Lactate Dehydrogenase （LDH） have been determined in the Cerebrospinal Fluid （CSF） of the meningitis patients （n = 26） and compared with control subjects （n = 26）. Total Protein, CK, SA （P 〈 0.02）, AST （P 〈 0.001） and ALT （P 〈 0.0001） of meningitis patients were significantly higher, whereas, glucose （P 〈 0.05） and Gamma Glutamyltranspeptidase （GGT） （P 〈 0.01 and P 〈 0.05, respectively） were significantly lower than those of the controls. This study suggests that loss of integrity of brain-CSF barriers, sialic acid and enzyme profiles may contribute to the severity and neurological complications of meningitis.

New Extracellular Bacillus subtilis Lectins with Sialic Acid-Specificity

The purpose of this work is to perform a detailed study of carbohydrate specificity of the new extracellular bacilli lectins which is considered to determine mechanisms of the lectins action. Sources of lectins were bacterial strains from Ukrainian collection of microorganisms. The optimized protocol of bacilli lectins isolation and purification included precipitation with ammonium sulfate with subsequent gel filtration chromatography on Sepharose CL-6B. Hemagglutinating activity of bacilli lectins and their fine carbohydrate specificity to sialic acids and their derivatives as well as sialic asid-containing and asialic glycoconjugates were studied. The ability of extracellular bacilli lectins to discriminate a- and 13-conformation of carbohydrate molecule and the type of connection between the monomers was determined. Studied lectins showed the most affinity to glycoconjugates containing both types of sialic acids （N-acetylneuraminic acid （Neu5Ac） and N-glycolylneuraminic acid （NeuGc）） and it is supposed to be a basis of their diagnostic and analytical potential.

Quantitative analysis of sialic acids in Chinese conventional foods by HPLC-FLD

Background: Sialic acids are a family of ninecarbon sugar compounds with carboxylic acyl derivatives. It exists in bacteria, fish, mammals and other living organisms, participates in and regulates many important life events, such as cell recognition, membrane flow, endocytosis and so on. Sialic acid is usually located in the outermost layer of the sugar part of the cell membrane and the key positions of secreted glycoconjugates (glycolipids, glycoprotein and lipopolysaccharide). Sialic acid (Sia) is an important material foundation for variety of the structure and founction of glycoconjugates. Sia has been known as nearly 50 members, including N-acetylneuraminic acid (Neu5Ac), N-glycoulylneuraminic acid (Neu5Gc) and deaminoneuraminic acid (KDN) as its core monomer. The rest of the sialic acids are derived from these three monomers. The contents of Sia in Chinese food products are unknown. Objective: To determine the contents of Sia in raw and cooked red meat, seafood, poultry and so on. Design: The following food products were purchased from a Chinese supermarket: pork, beef, lamb, salmon, cod, tuna, cow milk, cheese, butter, duck, chicken and chicken eggs. Human milk was collected from Xiamen Maternity and Child Health Care Hospital (Xiamen, China). All tissues were homogenized and hydrolyzed with 0.05 M, 0.1 M and 0.2 M TFA for 150 min at 80°C in dark, respectively. The concentrations of Neu5Ac, Neu5Gc and KDN were determined by using HPLC with fluorescence detector. Results: The contents of total Sia (μg/g tissue or μg/mL liquid sample) in Chinese raw meat were highest in lamb (269.60), followed by pork (254.88), duck (200.63), chicken (162.86) and beef (88.03). The percentages of Neu5Gc were 36.08%, 26.48%, 0%, 0% and 28.40%, respectively. Cod contained higher levels of Sia (171.63) than salmon (104.43) and tuna (77.98). Only Neu5Ac was 50 found in detected aquatic product. Egg yolk contained the highest level of Sia (682.04), and a higher level of Sia (390.67) was found in the egg white. Also our result showed that human milk contained extremely high level of Sia (602.55). Neu5Ac was the predominant form of Sia in all the deteced samples. KDN was found in cow milk only among the samples, the content was 1.14 μg/g. Conclusion: The highest content of Sia in examined Chinese foods was found in 56 eggs, followed by lamb, pork, duck, cod, chicken, salmon, beef and tuna. Knowledge of the Sia content in conventional foods may help us to better understand possible medical disorders involving the uptake of the “non-human” Neu5Gc from our diet.

Bitter Melon (Momordica Charantia) Reduces Serum Sialic Acid in Type2 Diabetics: Evidence to Delay the Process of Atherosclerosis

More than 1000 herbal products have been used by diverse cultures of the world to treat hyperglycemia and among them bitter melon (Momordica charantia) is one of the most popular herbal resource. The beneficial effects of bitter melon is not limited to hypoglycaemia only, but it also ameliorates diet induced obesity, insulin resistance and exhibit cardioprotective effects. The present study attempts to investigate the effect of bitter melon fruit juice on a newly investigated risk factor, sialic acid in type2 diabetics. A total of 40 type2 diabetic patients, divided into group A (n = 20) and group B (n = 20) were investigated during the present study. The patients of group A were following bitter melon fruit juice treatment along with diet control, whereas the patients of group B were on diet control only. Serum sialic acid (SSA) decreased in group A from 66.20 ± 2.30 mg/dl to 63.50 ± 2.10 mg/dl (<0.11) but, increased in group B from 66.50 ± 1.70 mg/dl to 68.20 ± 2.50 mg/dl (<0.12), compared to baseline. Post-treatment between group comparison revealed a significant difference (<0.05). The beneficial effects on fasting plasma glucose (FPG) and glycohemoglobin (HbA1-c) were also greater in group A compared to group B as was the case with blood lipids, weight and blood pressure. The study provides another mechanism for the cardioprotective effect of bitter melon and further strengthens its value in the management of type2 diabetes.

Sialic acid-mediated photochemotherapy enhances infiltration of CD8^(+)T cells from tumor-draining lymph nodes into tumors of immunosenescent mice

Immunoscenescence plays a key role in the initiation and development of tumors.Furthermore,immunoscenescence also impacts drug delivery and cancer therapeutic efficacy.To reduce the impact of immunosenescence on anti-tumor therapy,this experimental plan aimed to use neutrophils with tumor tropism properties to deliver sialic acid(SA)-modified liposomes into the tumor,kill tumor cells via SA-mediated photochemotherapy,enhance infiltration of neutrophils into the tumor,induce immunogenic death of tumor cells with chemotherapy,enhance infiltration of CD8^(+)T cells into the tumordraining lymph nodes and tumors of immunosenescent mice,and achieve SA-mediated photochemotherapy.We found that CD8^(+)T cell and neutrophil levels in 16-month-old mice were significantly lower than those in 2-and 8-month-old mice;16-month-old mice exhibited immunosenescence.The anti-tumor efficacy of SA-mediated non-photochemotherapy declined in 16-month-old mice,and tumors recurred after scabbing.SA-mediated photochemotherapy enhanced tumor infiltration by CD8^(+)T cells and neutrophils,induced crusting and regression of tumors in 8-month-old mice,inhibited metastasis and recurrence of tumors and eliminated the immunosenescence-induced decline in antitumor therapeutic efficacy in 16-month-old mice via the light-heat-chemical-immunity conversion.

Colorimetric-assisted photoelectrochemical sensing for dual-model detection of sialic acid via oxidation-power mediator integration

Development of sensitive and accurate methods for sialic acid(SA) determination is of great significance in early cancer diagnosis.Here,a colorimetric-assisted Photoelectrochemical(PEC) sensor was constructed for SA detection based on the two pairs of cis-diol groups in SA molecule.With the specific recognition of SA via the two pairs of cis-diol groups,the prepared gold-modified Bi_(2)S_(3)(AuNPs@Bi_(2)S_(3)) and metal organic framework(Au@PCN-224) were introduced to the electrode and formed a sandwich structure.Based on the properties of inert electroconductivity and nanozyme of the PCN-224,dual-readout of photocurrent and visualization was achieved with the presence of 3',3',5',5'-tetramethylbenzidine(TMB).Moreover,to intensify the visualization signal,Ce^(3+) was employed as the mediator to boost the catalysis capability by transferring energy from PCN-224 surface to the whole system.With the unique SA recognition and the mediation of Ce^(3+),both the photocurrent and the visualization signals sensitively responded with the SA concentration linearly.The present method showed greatly high sensitivity,selectivity and accuracy for SA detection with a limit of detection of 1.44μmol/L and a wide linear range of 5-1000μmol/L.This method provided a new promising platform for SA detection and a potential strategy for design of novel biosensors with dual-model readout.

Histones released by NETosis enhance the infectivity of SARS-CoV-2 by bridging the spike protein subunit 2 and sialic acid on host cells

Neutrophil extracellular traps(NETs)can capture and kill viruses,such as influenza viruses,human immunodeficiency virus(HIV),and respiratory syncytial virus(RSV),thus contributing to host defense.Contrary to our expectation,we show here that the histones released by NETosis enhance the infectivity of SARS-CoV-2,as found by using live SARS-CoV-2 and two pseudovirus systems as well as a mouse model.The histone H3 or H4 selectively binds to subunit 2 of the spike(S)protein,as shown by a biochemical binding assay,surface plasmon resonance and binding energy calculation as well as the construction of a mutant S protein by replacing four acidic amino acids.Sialic acid on the host cell surface is the key molecule to which histones bridge subunit 2 of the S protein.Moreover,histones enhance cell-cell fusion.Finally,treatment with an inhibitor of NETosis,histone H3 or H4,or sialic acid notably affected the levels of sgRNA copies and the number of apoptotic cells in a mouse model.These findings suggest that SARS-CoV-2 could hijack histones from neutrophil NETosis to promote its host cell attachment and entry process and may be important in exploring pathogenesis and possible strategies to develop new effective therapies for COVID-19.

Effects of Sialic Acid from Edible Bird Nest on Cell Viability Associated with Brain Cognitive Performance in Mice

Background:Edible bird nest(EBN)is a natural food product rich in glycoprotein such as sialic acid,which has been reported to improve brain functions.The EBN is widely consumed due to its higher nutritional contents and antioxidant status;however,an interaction of EBN on brain cell metabolic activity and viability are still unclear.Objective:The objectives of this study were to identify the effect of sialic acid from EBN on the cell viability and to determine the appropriate concentration of sialic acid on cognitive performance in mice.Materials and Methods:The viability of pheochromocytoma and neuroblastoma cell lines were tested using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay.For in vivo study,7-week-old female BALB/c mice were randomly assigned into four treatment groups and were treated with sialic acid for 21 days.At day 22,all mice were tested on cognitive performance by Y-maze test.Results:Treatment concentration of sialic acid extract and sialic acid standard at 60μg/mL(0.6 ppm)increased cell viability and showed no cytotoxicity effects in pheochromocytoma and neuroblastoma cell lines.In addition,an administration of higher dose of sialic acid at 0.6 ppm in animals improved Y-maze test performance,which they showed significantly higher number of entries and time spent in the novel arm.Conclusion:Thus,the current study shows that the sialic acid extract at 0.6 ppm improved brain cognitive performance in mice associated with an increased viability of pheochromocytoma and neuroblastoma cell lines.

Diagnosis of breast cancer by analysis of sialic acid concentrations in human saliva by surface-enhanced Raman spectroscopy of silver nanoparticles

Breast cancer is the most common type of malignant tumor among women and their second leading cause of cancer-related deaths.The most common method for screening and diagnosis is mammography.Nonetheless,two main problems have been identified.First,the dose of radiation received during the test prevents the method from the use on women who are 〈 40 years old.Second,there can be mammogram failure owing to the lack of tumor contrast with the fibrous tissue.Therefore,there is a need for screening methods that will help to identify high-risk cases.We developed a biological marker test that can help to identify them.Increased levels of sialic acid （SA） in saliva are known to correlated with breast cancer.In this study,we evaluated the feasibility of Raman spectroscopy as a method for quantification of SA in saliva,using citrate-reduced silver nanoparticles （cit-Ag-NPs） as a surface-enhanced Raman spectroscopy （SERS） substrate.Quantification of SA was accomplished by measuring its intensity in saliva and comparing it with a calibration curve of SA standards.The mean SA concentration in saliva was found to be significantly higher among 100 breast cancer patients （18.3 ＆#177; 9.4 mg＆#183;dL-1;mean ＆#177; SD） than among 106 healthy controls （3.5 ＆#177; 1.0 mg＆#183;dL-1）.The SERS test showed sensitivity of 94% and specificity 98% for detection of patients with breast cancer,assuming that SA concentration 〉 7 mg＆#183;dL-1 is a cutoff for positive test results.Our findings prove the usefulness of this SERS technique as a simple,convenient,and highly sensitive method of quantitative analysis of SA in saliva.The simplicity of this nanotechnological test may help to substantially reduce the mortality among patients with breast cancer by providing women with a simple,noninvasive screening test that can be applied regardless of age or density of breast tissue.

Thermo-responsive imprinted hydrogel with switchable sialic acid recognition for selective cancer cell isolation from blood

In this work,a sialic acid(SA)-imprinted thermo-responsive hydrogel layer was prepared for selective capture and release of cancer cells.The SA-imprinting process was performed at 37℃ using thermo-responsive functional monomer,thus generating switchable SA-recognition sites with potent SA binding at 37℃and weak binding at a lower temperature(e.g.,25℃).Since SA is often overexpressed at the glycan terminals of cell membrane proteins or lipids,the SA-imprinted hydrogel layer could be used for selective cancer cell recognition.Our results confirmed that the hydrogel layer could efficiently capture cancer cells from not only the culture medium but also the real blood samples.In addition,the captured cells could be non-invasively released by lowing the temperature.Considering the non-invasive processing mode,considerable capture efficiency,good cell selectivity,as well as the more stable and durable SA-imprinted sites compared to natural antibodies or receptors,this thermo-responsive hydrogel layer could be used as a promising and general platform for cell-based cancer diagnosis.

Metabolic fate of dietary sialic acid and its infuence on gut and oral bacteria

In recent years,the biological functions of human milk oligosaccharides and the potential toxic efects of red meat on human health have attracted considerable attention.Sialic acid is an important carbohydrate in milk and red meat,corresponding to sialylated oligosaccharides and N-glycolylneuraminic acid(NeuGc,one type of sialic acid).Herein,we reviewed the metabolic fate of dietary sialic acid in the body and their efects on gut and oral microbes.In summary,dietary NeuAc monomer is directly excreted through urine after being assimilated through the intestines and is not utilized by the human body;in contrast,dietary NeuGc from red meat is easily utilized by the human body and can be incorporated into the brain and other organs.Sialoglycans can be partially utilized by the human body,but they do not afect the cognitive development and growth of children.Dietary sialic acid may mainly regulate the growth and metabolism of gastrointestinal microbiota and human health and development through the gut–brain axis.

Glycophorin variants and contents of sialic acid and total sulfhydryl groups on erythrocyte membranes of residents in a malaria hyperendemic area

Abstract Objective To conduct a screening survey of glycophorin (GP) variants and observe the content changes of sialic acid (SA) and total sulfhydryl (SH) groups on the erythrocyte membranes among residents in a tertian malaria hyperendemic area of Guizhou Province. Methods GP variants were detected in the erythrocyte hemolysates of 173 local residents at two villages of Libo County by SDS PAGE on 10% to 15% gradients gel and Western immunoblotting. Their SA and total SH group contents were estimated in erythrocyte membranes by spectrophotometric methods. 114 healthy subjects in Changsha and 49 individuals at a neighbouring village of the above area showing low morbidity of malaria served as normal and endemic controls respectively. Results Three distinct types of GP variants were found among 19 propositi in this hyperendemic area. The incidence of GP variants was 7.9% (8/101) at Yaolu Village whose population was mainly composed of Yao ethnic group;while that of Buyi ethnic group at Maolan Village was higher (15.3%; 11/72).The erythrocyte membrane contents of SA in residents at both villages exhibited a very significant tendency of decline (P<0.01), whereas those of total SH groups increased prominently in residents of Yaolu Village only (P<0.05). Conclusions The frequency of GP variants in this hyperendemic area does not depend upon the severity of malarial prevalence. The evident reduction of SA contents in the residents may be related to the breaking down of the SA residues on membrane GPs by the invasion of Plasmodium vivax.

Effects of cadmium and copper on sialic acid levels in blood and brain tissues of Cyprinus carpio L.

Objective:To investigate the effects of cadmium(Cd)and copper(Cu)on sialic acid levels of brain and blood tissues of Cyprinus carpio.Methods:Adult carps were exposed to 0.1,0.5 mg/L Cu,0.1,0.5 and 1.0 mg/L Cd and 0.1 mg/L Cu+0.1 mg/L Cd under static experiment conditions for 1 week.At the end of exposure period,heavy metal accumulations and sialic acid levels in blood and brain tissues of the test animals were analyzed.Results:Cu and Cd accumulated in tissues in a dramatically increasing dose-dependent manner.Sialic acids level of the fish exposed to 0.1,0.5 and 1.0 mg/L Cu and Cd and control grups for 1 week were 0.834,1.427,0.672,0.934,2.968,4.714 mg/mL respectively.The results also showed that Cu has an antagonistic effect on tissue sialic acid level.Conclusions:We propose that Cd and Cu make a complex with sialic acids of membranes in the tissues researched.This complex between metal ions and sialic acid migth account for the cellular toxicity based on Cu and Cd.

Determination of cytokine regulated glycan expression by using molecularly imprinted polymers targeting sialic acid

Cancer cells often have an increased amount of glycans, such as sialic acid (SA), on the cell surface, which normally play an important role in cell growth, proliferation and differentiation. In this study, SA expression is determined by fluorescent nanoprobes, molecularly imprinted polymers, SA-MIPs. The nanoprobes are synthesized with an imprinting approach to produce tailor-made fluorescent core-shell particles with high affinity for cell surface SA. Inflammation and cytokine production are well known tumor promoters, modulating the cellular microenvironment, including an aberrant cell surface glycan pattern. The recombinant cytokines IL-4, IL-6, IL-8 and a cocktail of cytokines collected from stimulated T leukemia Jurkat cells were used to induce in vitro inflammation in two cell lines, and thereafter analyzed with the use of SA-MIPs and flow cytometry. One of the cell lines showed a different binding pattern of SA-MIPs after treatment with recombinant cytokines and the cytokine cocktail. This study shows that SA-MIPs can be an important tool in the investigation of overexpressed glycans in the tumor microenvironment.