Tinker, tailor, soldier, cell: the role of C-type lectins in the defense and promotion of disease

C-type lectins (CTLs) represent a large family of soluble and membrane-bound proteins which bind calcium dependently via carbohydrate recognition domains (CRDs) to glycan residues presented on the surface of a variety of pathogens. The deconvolution of a cell's glycan code by CTLs underpins several important physiological processes in mammals such as pathogen neutralization and opsonization, leukocyte trafficking, and the inflammatory response. However, as our knowledge of CTLs has developed it has become apparent that the role of this innate immune family of proteins can be double-edged, where some pathogens have developed approaches to subvert and exploit CTL interactions to promote infection and sustain the pathological state. Equally, CTL interactions with host glycoproteins can contribute to inflammatory diseases such as arthritis and cancer whereby, in certain contexts, they exacerbate inflammation and drive malignant progression. This review discusses the 'dual agent' roles of some of the major mammalian CTLs in both resolving and promoting infection, inflammation and inflammatory disease and highlights opportunities and emerging approaches for their therapeutic modulation.

Application of lectins for detection of glycomic differences in the epididymal duct in water buffalo (Bubalus bubalis) and dromedary camel (Camelus dromedarius) / Aplicación de lectinas para la detección de diferencias glucómicas en el conducto epididimario en búfalos de agua (Bubalus bubalis) y dromedarios (Camelus dromedarius)

SUMMARY: The present study was conducted to detect the differences in glycohistochemical features in the epididymal duct of the dromedary camel and the water buffalo. Epididymal sections (caput, corpus and cauda) from both species were stained with fluorescein isothiocyanate (FITC) conjugated lectins. Binding sites for five lectins (DBA, GSA-1, HPA, PNA and WGA) have been found in both species. The binding sites of different lectins showed significant variations in the pattern of distribution in both a species. This included both species-specific and region-specific order. Additionally, only three (GSA-1, PNA and WGA) out the five lectins studied exhibited binding sites in all epididymal regions in both species. The other two lectins (DBA and HPA) followed the same order recorded for the other three (GSA-1, PNA and WGA) in buffalo, but failed to show any binding sites in cauda epididymis in camel. In conclusion, the variable regional and species-specific distribution features of lectins revealed that both species have diverse glycomic characteristics that may be related to their different reproductive patterns. However, the glycome-associated functional capacities remain obscured and need further profound investigations.

RESUMEN: El presente estudio se realizó para detectar las diferencias en las características glicohistoquímicas del conducto epididimal del dromedario y el búfalo de agua. Las secciones del epidídimo (cabeza, cuerpo y cola) de ambas especies se tiñeron con lectinas conjugadas con isotiocianato de fluoresceína (FITC). Se encontraron sitios de unión para cinco lectinas (DBA, GSA-1, HPA, PNA y WGA) en ambas especies. Los sitios de unión de diferentes lectinas mostraron variaciones significativas en el patrón de distribución en ambas especies. Esto incluía tanto el orden específico de la especie como el específico de la región. Además, solo tres (GSA-1, PNA y WGA) de las cinco lectinas estudiadas exhibieron sitios de unión en todas las regiones del epidídimo en ambas especies. Las otras dos lectinas (DBA y HPA) siguieron el mismo orden registrado para las tres restantes (GSA-1, PNA y WGA) en búfalos, pero no mostraron ningún sitio de union en la cola del epidídimo en camellos. En conclusión, las características de distribución regionales y específicas de especies variables de las lectinas revelaron que ambas especies tienen características glucómicas diversas que pueden estar relacionadas con sus diferentes patrones reproductivos. Sin embargo, las capacidades funcionales asociadas con el glicoma permanecen desconocidas y requieren mayor investigación.

Establishment of lectin-ELISA for sialylated fetuin-A and its diagnostic value in primary hepatocellular carcinoma / 中华检验医学杂志

Objective:To establish a lectin enzyme-linked immunosorbent assay (lectin-ELISA) for the dection of sialylated fetuin-A and to explore the clinical diagnostic value of sialylated fetuin-A in hepatocellular carcinoma (HCC).Methods:From January 2017 to December 2020, 300 HCC patients and 160 disease controls, including 36 liver cirrhosis subgroups and 124 chronic hepatitis B subgroups, were collected from Shanghai Eastern Hepatobiliary Surgery Hospital. At the same time, 100 healthy subjects were collected as healthy controls. Lectin-ELISA method for detecting sialylated fetuin A was established based on the principle that Sambucus nigra lectin (SNA) can recognize the structure of α-2, 6-linked sialic acid residues. Differences between groups were compared using t-test or analysis of variance. Logistic regression method was used to establish the multi-index joint detection model, and receiver operating characteristic curve (ROC) was used to evaluate the efficacy of single index and joint detection model in the diagnosis of HCC.Results:A lectin-ELISA method for the detection of serum Sia-fetuin A was established. The linear regression coefficient of the system was 0.978 5, and the precision evaluation and interference experiments were in line with the clinical detection requirements. Using this method to detect serum Sia-fetuin A levels in each group, the levels of HCC group, disease control group and healthy control group were 1.362±0.310, 1.199±0.370, 1.086±0.420, respectively, and the three groups decreased in turn. The areas under the curve of Sia-fetuin A, α-fetoprotein, and their combined detection models for differential diagnosis of HCC were 0.790, 0.809, and 0.860, respectively. The diagnostic model had a sensitivity of 79.3% (238/300) and a specificity of 95.0% (247/260). Among the 300 patients in the HCC group, 138 (46%) patients were negative for serum AFP (<20 μg/L), and their serum Sia-fetuin A level was 1.364±0.305. Combining the disease control group and the healthy control group into the non-Cancer group, the serum Sia-fetuin A level was 1.146±0.381. The serum level of Sia-fetuin A in AFP-negative HCC patients was higher than that in non-HCC group ( t=6.134, P<0.001). The areas under the curve of Sia-fetuin A and the combined diagnostic model for the diagnosis of AFP-negative HCC were 0.776 and 0.919, respectively. The combined diagnostic model had a sensitivity of 93.4% (129/138) and a specificity of 77.3% (201/260). Conclusion:Serum Sia-fetuin A and combined determination model can provide a new auxiliary diagnostic index for AFP-negative HCC.

Peruvian plants of traditional use as potential sources of molecules with activity against COVID-19

RESUMEN Introducción: La situación actual de la COVID-19 es un gran problema para la población humana. En la actualidad, no hay medicamentos curativos disponibles en el mercado. Los investigadores están haciendo todo lo posible para producir fármacos con que luchar contra la enfermedad. Se están considerando varios esfuerzos basados en diferentes orientaciones del conocimiento científico y en las tecnologías para el tratamiento de la enfermedad. Desafortunadamente, ninguno de estos medicamentos funciona absolutamente contra la corriente pandémica. Por lo tanto, las moléculas bioactivas de plantas, animales y microorganismos podrían ser una mejor opción para tratar la COVID-19. Objetivo: Revisar la literatura sobre especies de la flora del Perú utilizadas en el tratamiento de enfermedades respiratorias y destacar las plantas con posible producción de metabolitos secundarios y lectinas vegetales potencialmente útiles como alternativa frente a la COVID-19. Métodos: Se revisaron artículos de literatura científica relacionados con el uso de la medicina tradicional en Perú, China e India para el tratamiento de enfermedades respiratorias, así como la información sobre lectinas vegetales y metabolitos secundarios con potencial utilidad contra la COVID-19. Resultados: Se presenta una amplia relación de géneros y especies de la flora del Perú con gran potencial contra la COVID-19. La mayoría de estas especies pertenecen a las familias Asteraceae, Loranthaceae, Piperaceae, Viscaceae y Zingiberaceae. Numerosas especies son endémicas del Perú. Conclusiones: La flora del Perú tiene más de 22 000 especies de plantas. Muchas de estas especies se utilizan tradicionalmente en el tratamiento de enfermedades respiratorias y pueden ser potencialmente útiles en el tratamiento de la COVID-19.

ABSTRACT Introduction: The current situation of COVID-19 is a big issue for the human population. At present, no healing drug is available in the market. Researchers are doing their best to produce drugs to fight the disease. Various efforts are being considered based on different directions of scientific knowledge and technologies for the treatment of the disease. Unfortunately, none of these drugs works absolutely against the pandemic. Therefore, bioactive molecules from plants, animals and microorganisms could be a better option to treat COVID-19. Objective: Review the literature about species of the flora of Peru used for the treatment of respiratory diseases and highlight the plants with potential in the production of secondary metabolites and plant lectins as an alternative against COVID-19. Methods: A review was conducted of scientific articles related to the use of traditional medicine in Peru, China, and India for the treatment of respiratory diseases, as well as information about plant lectins and secondary metabolites potentially useful against COVID-19. Results: A long list is presented of genera and species of the flora of Peru with great potential against COVID-19. Most of these species belong to the Asteraceae, Loranthaceae, Piperaceae, Viscaceae and Zingiberaceae families. Numerous species are endemic to Peru. Conclusions: The flora of Peru has more than 22 000 plant species. Many of these species are traditionally used in the treatment of respiratory diseases and are potentially useful for the treatment of COVID-19.

Serum C-type lectin-like receptor-2 combined with insulin resistance predicts the outcome of patients with acute ischemic stroke after intravenous thrombolysis / 国际脑血管病杂志

Objective:To investigate the predictive value of serum C-type lectin-like receptor 2 (CLEC-2) combined with insulin resistance in the outcome of patients with acute ischemic stroke (AIS) after intravenous thrombolysis.Methods:Patients with AIS received alteplase intravenous thrombolytic therapy in the Department of Neurology, the Second Affiliated Hospital of Nantong University from October 2019 to March 2021 were enrolled retrospectively. According to the modified Rankin Scale score at 90 d after onset, they were divided into good outcome group (0-2) and poor outcome group (>2). Homeostasis model assessment of insulin resistance (HOMA-IR) was used to evaluate insulin resistance. Person correlation analysis was used to determine the correlation between CLEC-2 and HOMA-IR. Multivariate logistic regression analysis was used to determine the correlation between serum CELC-2, HOMA-IR and the outcome after intravenous thrombolysis. Receiver operating characteristic (ROC) curve was used to determine the predictive value of serum CLEC-2 combined with HOMA-IR for poor outcome after intravenous thrombolysis. Results:A total of 100 patients were enrolled (56 males, 56.0%; aged 70.6±10.86 years, range 49-83 years). The baseline National Institutes of Health Stroke Scale (NIHSS) score was 10.00±6.36. Senenty-four patients (74.0%) had a good outcome and 26 (26.0%) had a poor outcome. Person correlation analysis showed that there was a significant positive correlation between serum CLEC-2 and HOMA-IR ( r=0.523; P<0.001). Multivariate logistic regression analysis showed that after adjusting for confounding factors (C-reactive protein, baseline NIHSS score, onset-to-needle time), the highest quartile of serum CLEC-2 (compared with the lowest quartile: odds ratio [ OR] 4.836, 95% confidence interval [ CI] 1.105-21.169; P=0.036) and the highest quartile of HOMA-IR (compared with the quartile 1-3: OR 15, 95% CI 2.647-30.722; P=0.002) were the independent risk factors for the poor outcome in patients with AIS after intravenous thrombolysis. ROC curve analysis showed that the area under the curve for serum CLEC-2 combined with HOMA-IR to predict poor outcome was 0.785 (95% CI 0.688-0.883; P<0.001), the optimal cut-off value was 0.72, and the sensitivity and specificity were 76.0% and 95.0%, respectively. Conclusion:CLEC-2 combined with insulin resistance has a certain predictive value for the poor outcome of patients with AIS after intravenous thrombolysis.

Lectins in Viscum:a progress review / 中国中药杂志

Viscum plants,the evergreen perennial parasitic shrubs or subshrubs,are mainly distributed in tropical and subtropical regions. There are about 70 Viscum species around the world,including 11 species and one variety in China. Mistletoe lectins are typeⅡ ribosome-inactivating proteins( RIPs) extracted from Viscum plants with anticancer and immunoregulatory activities. Many studies have focused on the mistletoe lectins isolated from V. album in Europe and V. album var. coloratum distributed in South Korea,respectively,and several preparations,such as Iscucin Ⓡ,were developed and clinically applied for cancer treatment. Although Viscum plants are widely distributed in China,only a few studies of mistletoe lectins have been reported. The recent progress of mistletoe lectins was reviewed from extraction,purification,quantitative/qualitative detection,molecular structure,pharmacological activities,toxicities,and clinical application,aiming at providing a reference for in-depth research and utilization of mistletoe lectins produced in China.

Leguminosae Lectins as Biological Tools in Medical Research: a Review

Abstract Lectins were discovered first in plants and later in other living things, and nowadays it is known that they are present in almost all many life forms. These proteins can bind to specific carbohydrates, which make them perform a number of biological activities and can be used as tools in the study of glycoconjugate structures present on the cell surface, being effective in medical research. Plant lectins, leguminosae lectins particularly, are among the most studied plant proteins. They are very versatile molecules, which show several interesting biological properties. Thus, the present paper reviewed the advances about the leguminosae lectins biological properties studies in the last ten years, taking into account their possible applications in the fields of Clinical Microbiology, Pharmacy and Cancerology through a search in the electronic databases, providing opportunity to exchange information about the theme. Leguminosae lectins can neutralize pathogenic organisms, be they viruses, prokaryotes and eukaryotes, in addition carcinogenic cells, besides decreasing oxidative stress, conditions which increases the possibility of alternative substances for the design of new drugs to be used in current therapeutic, expanding the possibilities of diseases cure.

Evaluation of galectins 1 and 3 expression in canine melanoma / Avaliação da expressão das galectinas 1 e 3 no melanoma canino

Canine melanoma is a frequently-occuring neoplasm in dogs and presents as malignant and highly metastatic in this context, studies that contribute to the understanding of the tumor microenvironment in melanoma include the role of galectins. Galectins are proteins of the family of animal lectins that display carbohydrate recognition domains. Galectin-1 and galectin-3 are associated with neoplastic transformation, neoplastic cell survival, angiogenesis, immune system evasion, and metastasis. The goal of this study was to establish a correlation between expression patterns of galectin-1 and galectin-3 and the different degrees of aggressiveness of canine melanoma, as well as to determine serum concentration of galectin-3 in dogs with melanoma. Galectin-1 and galectin-3 expression was analyzed by immunohistochemistry in 30 canine melanomas, six melanocytomas and nine metastatic lymph nodes from patients whose primary tumors were also processed and analyzed. Serum samples from 30 dogs were collected and galectin-3 concentration was determined by ELISA and compared to the samples of 10 healthy dogs. Canine melanoma samples expressed galectin-1 in the cytoplasm and presented a variable pattern of galectin-3 staining depending on melanoma aggressiveness. We observed a decrease in the percentage of cells with cytoplasmic galectin-3 immunolabeling simultaneous to the increased nuclear staining intensity, while there was also a decrease in the percent frequency of nuclear galectin-3 immunolabeled cells according to progression of melanoma, comparing the least to the most aggressive cases. Dogs with melanoma had increased serum levels of galectin-3 when compared to healthy animals, suggesting its potential biomarker of patients with melanoma.(AU)

O melanoma canino é uma neoplasia frequente em cães que apresenta um potencial maligno e metastático. Neste contexto, investigar o microambiente tumoral é fundamental para compreender os mecanismos intercelulares e intracelulares envolvidos no desenvolvimento e progressão da doença. Neste estudo, destacamos as galectinas, proteínas da família das lectinas animais que exibem domínios de reconhecimento à carboidratos; a galectina-1 e a galectina-3 estão associadas a transformação neoplásica, sobrevivência de células neoplásicas, angiogênese, evasão do sistema immune e desenvolvimento de metástases. O objetivo deste estudo foi determinar os padrões de expressão de galectina-1 e galectina-3 em diferentes graus de agressividade do melanoma canino, bem como dosar a concentração sérica de galectina-3 em cães com melanoma e comparar com cães saudáveis. A expressão de galectina-1 e galectina-3 foi analisada em 30 melanomas caninos, seis melanocitomas e nove linfonodos metastáticos. A galectina-3 sérica foi mensurada em 30 cães com melanoma e comparada a 10 cães saudáveis. No melanoma canino a expressão de galectina-1 foi citoplasmática e a expressão de galectina-3 foi variável de acordo com o grau de agressividade. Notou-se uma redução na porcentagem de células com imunomarcação de galectina-3 citoplasmática e um aumento simultâneo da intensidade de imunomarcação nuclear, enquanto houve também uma diminuição na frequência percentual de células com imunomarcação nuclear de acordo com a progressão do melanoma comparando-se os casos menos com os mais agressivos. Cães com melanoma apresentaram níveis séricos aumentados de galectina-3 quando comparados a animais saudáveis, mostrando seu uso potencial como biomarcador em pacientes com melanoma.(AU)

Avaliação dos Efeitos Cardíacos de Lectina Solúvel em Água (WSMoL) de Sementes de Moringa Oleifera / Evaluation of the Cardiac Effects of a Water-Soluble Lectin (Wsmol) from Moringa Oleifera Seeds

Resumo Fundsamento As sementes de Moringa oleifera , que são utilizadas para clarificação de água, contêm uma lectina chamada WSMoL que tem mostrado atividade antibacteriana e imunomoduladora in vitro . Devido ao seu valor nutritivo e potencial terapêutico, as folhas e as sementes dessa árvore são consumidas em algumas comunidades. Algumas lectinas de plantas não são tóxicas para mamíferos, mas tem sido relatado que outras são prejudiciais quando ingeridas ou administradas por outros meios. Objetivo Como um dos passos necessários para determinar a segurança de WSMoL, nós avaliamos os possíveis efeitos cardiotóxicos desta proteína purificada. Métodos Durante 21 dias consecutivos, a WSMoL foi administrada a camundongos por gavagem. Foram investigadas as funções eletrofisiológicas, mecânicas e metabólicas in vivo e ex vivo por meio de registros eletrocardiográficos, ressonância magnética nuclear e respirometria de alta resolução. Resultados O tratamento com WSMoL não induziu alterações nos níveis de glicose no sangue ou peso corporal em comparação com o grupo controle. Adicionalmente, as relações peso cardíaco/peso corporal e peso cardíaco/comprimento tibial estavam semelhantes em ambos os grupos. A ingestão de lectina também não modificou a tolerância à glicose ou resistência à insulina. Não foram observadas alterações nos parâmetros eletrocardiográficos ou na duração do potencial de ação cardíaco. Os corações dos camundongos dos grupos controle e WSMoL mostraram função ventricular esquerda preservada. Além disso, a WSMoL não induziu alterações na função mitocondrial (em todos os casos, p > 0,05). Conclusões A administração de WSMoL demonstrou ter um perfil de segurança cardíaca. Estes resultados contribuem à avaliação de segurança do uso de sementes de M. oleifera para tratar água, visto que essa lectina está presente na preparação empregada por algumas populações com esse fim. (Arq Bras Cardiol. 2020; [online].ahead print, PP.0-0)

Abstract Background Moringa oleifera seeds, which are used for water clarification, contain a lectin named WSMoL which has shown in vitro antibacterial and immunomodulatory activity. Due to their nutritional value and therapeutic potential, the leaves and seeds of this tree are eaten in some communities. Some plant lectins are non-toxic to mammals, but others have been reported to be harmful when ingested or administered by other means. Objective As one of the steps needed to define the safety of WSMoL, we evaluated possible cardiotoxic effects of this purified protein. Methods: WSMoL was administered for 21 consecutive days to mice by gavage. Electrophysiological, mechanical, and metabolic cardiac functions were investigated by in vivo and ex vivo electrocardiographic recordings, nuclear magnetic resonance, and high-resolution respirometry. Results The treatment with WSMoL did not induce changes in blood glucose levels or body weight in comparison with control group. Moreover, the heart weight/body weight and heart weight/tibia length ratios were similar in both groups. Lectin ingestion also did not modify glucose tolerance or insulin resistance. No alterations were observed in electrocardiographic parameters or cardiac action potential duration. The heart of mice from the control and WSMoL groups showed preserved left ventricular function. Furthermore, WSMoL did not induce changes in mitochondrial function (in all cases, p > 0.05). Conclusions The administration of WSMoL demonstrated a cardiac safety profile. These results contribute to the safety evaluation of using M. oleifera seeds to treat water, since this lectin is present in the preparation employed by some populations to this end. (Arq Bras Cardiol. 2020; [online].ahead print, PP.0-0)

Leishmania infantum induces high phagocytic capacity and intracellular nitric oxide production by human proinflammatory monocyte

BACKGROUND The mechanism of resistance to SbIII in Leishmania is complex, multifactorial and involves not only biochemical mechanisms, but also other elements, such as the immune system of the host. OBJECTIVES In this study, putative changes in the immunological profile of human monocytes infected with wild-type (WT) and antimony (SbIII)-resistant Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum lines were evaluated. METHODS Susceptibility assays WT and SbIII-resistant L. braziliensis and L. infantum were performed using lines THP-1 human monocytic lineage. Phagocytic capacity, cytokine profile, intracellular nitric oxide (NO) production and surface carbohydrate residues profile were performed in peripheral blood monocytes by flow cytometry. FINDINGS The phagocytic capacity and intracellular NO production by classical (CD14++CD16-) and proinflammatory (CD14++CD16+) monocytes were higher in the presence of L. infantum lines compared to L. braziliensis lines. The results also highlight proinflammatory monocytes as the cellular subpopulation of major relevance in a phagocytosis event and NO expression. It is important to note that L. infantum induced a proinflammatory cytokine profile characterised by higher levels of TNF-α in culture supernatant than L. braziliensis. Conversely, both Leishmania lines induce high levels of IL-6 in culture supernatant. Analysis of the expression profile of surface carbohydrates showed that L. braziliensis presents 4.3-fold higher expression of galactose(β1,4)N-acetylglucosamine than L. infantum line. Interestingly, the expression level of α-N-acetylgalactosamine residues was 2-fold lower in the SbIII-resistant L. braziliensis line than its counterpart WT line, indicating differences in surface glycoconjugates between these lines. MAIN CONCLUSIONS Our results showed that L. braziliensis and L. infantum induce different innate immune responses and a highly inflammatory profile, which is characteristic of infection by L. infantum, the species associated with visceral disease.

Effect of ScLL and 15d-PGJ2 on viability and cytokine release in LPS-stimulated fibroblasts: an in vitro study

Abstract This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFβ-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 μg/ml and treated with 15d-PGJ2 1 and 2 μg/ml, and ScLL 2 and 5 μg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFβ-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 μg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFβ-1 release (p > 0.05). Results indicated that ScLL 5 μg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFβ-1 release.

New Rabbit Erythrocyte Specific Mycelial Lectins from Fusarium sp. with Complex Saccharide Specificity

Eight Fusarium sp. namely, F. acutatum, F. globosum, F. graminearum, F. lactis, F. nivale, F. proliferatum, F.pseudoanthophilum and F. robustum were screened for the presence of lectins by hemagglutination activityusing human ABO, porcine, ovine, goat and rabbit erythrocytes. Mycelial extracts of all the fungal culturesexcept F. graminearum displayed unique lectin activity with only rabbit erythrocytes. Enzymatic treatment ofrabbit erythrocytes with neuraminidase has significantly enhanced the titre of all the lectin-positive extractsof fungal cultures. In contrast, most of the lectins showed a decline in lectin activity with protease treatedrabbit erythrocytes. Saccharide specificity studies have shown that majority of the lectins are inhibitory towardsO-acetyl sialic acids. None of the lectins from Fusarium sp. were inhibited by dextran, meso-inositol, andN-acetyl-D-glucosamine. Most of the fungal cultures displayed highest hemagglutination activity during the10th day of growth in broth cultures. The unique saccharide specificity of Fusarium sp. lectins can be used forelucidating their clinical role in glycobiology research.

Regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer cells / 中华围产医学杂志

Objective@#To investigate the regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer (dNK) cells during early pregnancy and its association with the incidence of recurrent spontaneous abortion (RSA).@*Methods@#Decidua tissues were collected from normal pregnancies (n=16) and RSA cases (n=6) at 6-10 gestational weeks in the Department of Obstetrics and Gynecology of Peking University Third Hospital from October 2018 to May 2019. (1) Expressions of CLEC2D and CD161 in decidua from early pregnancy were detected using immunofluorescence. (2) Primary dNK cells were isolated from decidua from early pregnancy. dNK cells pre-treated with CD161 antibody (blocking CD161, B-CD161) were co-cultured with JEG-3 cells which were knocked-down by CLEC2D small interfering RNA (siCLEC2D), followed by killing capacity assessment of dNK cells by cytotoxicity assay and determination of expressions of related molecules by quantitive real-time polymerase chain reaction. (3) Western blot and flow cytometry were used to detect the expression of CLEC2D and CD161 in decidua tissues. Cytotoxicity assay was performed to analyze the killing capacity of dNK cells. T test was used for statistical analysis between normal and RSA cases.@*Results@#(1) CLEC2D was mainly expressed in extravillous trophoblast (EVT) cells and CD161 was mainly detected in dNK cells. CD161-positive dNK cells and CLEC2D-positive EVT cells were adjacently located in decidua tissues allowing their interaction. (2) Cytotoxicity assay suggested that CD161 blocking in dNK cells or CLEC2D knockdown in JEG-3 cells could enhance the cytotoxicity of dNK cells. The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5∶1 in B-CD161 group were (59.12±4.56)%, (25.96±5.44)%, (13.60±8.94)% and (12.53±8.94)%, and in IgG control group were (20.01±1.96)%, (8.51±1.32)%, (3.24±0.75)% and (3.82±1.92)%, respectively. There were significant differences between the two groups at the effector-target ratios of 40∶1 (t=13.922, P<0.01) and 20∶1 (t=5.403 P<0.05), but not at 10∶1 or 5∶1 (P>0.05). The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5∶1 in si-CLEC2D group were (43.37±2.01)%, (32.99±2.08)%, (23.47±1.36)% and (11.48±0.37)%, and in the negative control (NC) group were (15.54±1.46)%, (13.84±1.68)%, (9.94±3.01) and (5.50±0.99)%, respectively. Differences between the two groups at all effector-target ratios were statistically significant (t=19.402, 12.400, 7.093 and 9.842, all P<0.01). Moreover, the expression of dNK killing-related factor granzyme B in the siCLEC2D group was higher than that in the NC group. (3) Compared with the normal pregnancy group, the RSA group showed decreased CD161 expression and increased killing capacity of dNK cells, but no significant difference in CLEC2D expression.@*Conclusions@#At early pregnancy, CLEC2D on EVT cells can interact with CD161 on dNK cells, which inhibits the cytotoxicity of dNK cells and induces immune tolerance at the fetal-maternal interface. Decreased expression of CD161 in decidua results in increased cytotoxicity of dNK cells, which may be one of the causes of immune rejection in RSA.

Regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer cells / 中华围产医学杂志

Objective To investigate the regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer (dNK) cells during early pregnancy and its association with the incidence of recurrent spontaneous abortion (RSA). Methods Decidua tissues were collected from normal pregnancies (n=16) and RSA cases (n=6) at 6-10 gestational weeks in the Department of Obstetrics and Gynecology of Peking University Third Hospital from October 2018 to May 2019. (1) Expressions of CLEC2D and CD161 in decidua from early pregnancy were detected using immunofluorescence. (2) Primary dNK cells were isolated from decidua from early pregnancy. dNK cells pre-treated with CD161 antibody (blocking CD161, B-CD161) were co-cultured with JEG-3 cells which were knocked-down by CLEC2D small interfering RNA (siCLEC2D), followed by killing capacity assessment of dNK cells by cytotoxicity assay and determination of expressions of related molecules by quantitive real-time polymerase chain reaction. (3) Western blot and flow cytometry were used to detect the expression of CLEC2D and CD161 in decidua tissues. Cytotoxicity assay was performed to analyze the killing capacity of dNK cells. T test was used for statistical analysis between normal and RSA cases. Results (1) CLEC2D was mainly expressed in extravillous trophoblast (EVT) cells and CD161 was mainly detected in dNK cells. CD161-positive dNK cells and CLEC2D-positive EVT cells were adjacently located in decidua tissues allowing their interaction. (2) Cytotoxicity assay suggested that CD161 blocking in dNK cells or CLEC2D knockdown in JEG-3 cells could enhance the cytotoxicity of dNK cells. The target cell lysis rates at the effector-target ratios of 40 ∶ 1, 20 ∶ 1, 10 ∶ 1 and 5 ∶ 1 in B-CD161 group were (59.12±4.56)%, (25.96±5.44)%, (13.60±8.94)% and (12.53±8.94)%, and in IgG control group were (20.01±1.96)%, (8.51±1.32)%, (3.24±0.75)% and (3.82±1.92)%, respectively. There were significant differences between the two groups at the effector-target ratios of 40∶1 (t=13.922, P<0.01) and 20∶1 (t=5.403 P<0.05), but not at 10∶1 or 5∶1 (P>0.05). The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5 ∶ 1 in si-CLEC2D group were (43.37±2.01)%, (32.99±2.08)%, (23.47±1.36)% and (11.48±0.37)%, and in the negative control (NC) group were (15.54±1.46)%, (13.84±1.68)%, (9.94±3.01) and (5.50±0.99)%, respectively. Differences between the two groups at all effector-target ratios were statistically significant (t=19.402, 12.400, 7.093 and 9.842, all P<0.01). Moreover, the expression of dNK killing-related factor granzyme B in the siCLEC2D group was higher than that in the NC group. (3) Compared with the normal pregnancy group, the RSA group showed decreased CD161 expression and increased killing capacity of dNK cells, but no significant difference in CLEC2D expression. Conclusions At early pregnancy, CLEC2D on EVT cells can interact with CD161 on dNK cells, which inhibits the cytotoxicity of dNK cells and induces immune tolerance at the fetal-maternal interface. Decreased expression of CD161 in decidua results in increased cytotoxicity of dNK cells, which may be one of the causes of immune rejection in RSA.

Expressions and Roles of CLEC4D and CARD9 in Inflammatory Bowel Disease / 胃肠病学

Background: C-type lectin domain family 4 member D (CLEC4D) can induce Th17 cell differentiation and regulate IL-17A expression in systemic fungal infection, and whether CLEC4D plays a role in intestinal immunity has not been reported at home and abroad. Aims: To investigate the expressions and clinical significance of CLEC4D and CARD9 in colon tissue in patients with inflammatory bowel disease (IBD). Methods: A total of 48 IBD patients from October 2016 to June 2018 at the Second Affiliated Hospital of Zhengzhou University were enrolled, including 36 patients with ulcerative colitis (UC) and 12 patients with Crohn's disease (CD). And 30 adjacent normal colon tissues in patients with colon cancer were served as controls. Immunohistochemistry was used to detect the expressions of CLEC4D and CARD9 in colon tissue. And their correlations with clinical characteristics were analyzed. Results: The expressions of CLEC4D and CARD9 in UC group and CD group were significantly higher than those in control group (P0.05). The expressions of CLEC4D and CARD9 in UC group were positively correlated with Mayo score and Baron grade (P0.05). The expressions of CLEC4D and CARD9 in IBD patients were positively correlated with CRP (P0.05). Conclusions: The expressions of CLEC4D and CARD9 are elevated in patients with IBD and are closely related to patients' condition. CLEC4D may participate in the intestinal immunity of IBD through the CARD9-related pathway.

Changes in expression of IB4 and CGRP in neurons in dorsal root ganglia of rats with neuropathic pain / 中华麻醉学杂志

Objective To evaluate the changes in the expression of isolectin B4 (IB4) and calcium gene-related peptide (CGRP) in neurons in dorsal root ganglion (DRG) of rats with neuropathic pain (NP).Methods Forty healthy male Sprague-Dawley rats,weighing 250-280 g,were divided into 2 groups (n =20 each) using a random number table method:solvent group (group S) and group NP.NP was induced by intraperitoneally injecting resiniferatoxin 210 μg/kg,and the solvent of resiniferatoxin was intraperitoneally injected in group S.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured in 5 rats selected before establishing the model and at 1,3,7 and 42 days after establishing the model.Five rats were sacrificed at 1,3,7 and 42 days after establishing the model,and the L4-6 segments of the DRGs were removed to determine the expression of CGRP and IB4 in neurons using immunofluorescence.Results Compared with the baseline before establishing the model,the MWT was signilicantly decreased at 3,7 and 42 days after establishing the model,and the TWL was prolonged at 1,3,7 and 42 days after establishing the model in group NP (P＜0.05).Compared with group S,the MWT was significantly decreased at 3,7 and 42 days after establishing the model,the TWL was prolonged at 1,3,7 and 42 days after establishing the model,and the expression of IB4 and CGRP in neurons in DRGs was down-regulated at 1,3,7 and 42 days after establishing the model in group NP (P＜0.05).Conclusion Down-regulated expression of IB4 expression in neurons in DRGs may be involved in the development and maintenance of mechanical hypersensitivity to pain,and down-regulated expression of CGRP may be involved in the development and maintenance of thermal analgesia in rats with NP.

Expression and clinical significance of mast cells and related immune factors in interstitial cystitis/bladder pain syndrome / 中华泌尿外科杂志

Objective To investigate the significance of mast cells activation,and IL-6,IL-10,sialic acid-binding immunoglobulin-like lectins 8 (Siglec-8) expression in interstitial cystitis/bladder pain syndrome.Methods Bladder mucosal biopsy tissues of 21 patients with IC/BPS admitted to our hospital from March 2016 to October 2017 were taken as the IC/BPS group,and normal bladder mucosa biopsy tissues of 9 patients who underwent ureteroscopy were taken as the control group.In the IC/BPS group,there were 3 males and 18 females aged (56.2 ± 3.4) years,and the patients'pain symptom score (PUF) was (24.6 ± 3.6).Four males and five females in the control group were aged (63 ± 5.1) years.The infiltration of bladder mucosa mast cells and plasma cells in IC/BPS group and control group was observed by transmission electron microscopy.The expression of IL-6,IL-10 and Siglec-8 were detected by immunohistochemical staining.The relationship between the expression level of the immune index and the PUF score was analyzed.Results In the IC/BPS group,mast cells and plasma cells was observed in 18 of the 21 cases,but no mast cells or plasma cells were observed in the control group.Expression of IL-6 in IC/BPS group 1 case (-),11 cases(+) and 9 cases (+ +).IL-6 expression in the control group 9 cases were all (-).The difference of IL-6 expression between the IC/BPS group and the control group was statistically significant (P =0.001).Expression of IL-10 were 3 cases (-),7 cases (+ +),11 cases (+ +) in IC/BPS group,while 7 cases (-),2 cases (+ +),and 0 cases (+ +)in the control group,and there was significant difference between the two group (P =0.001).In the IC/BPS group,expression of Siglee-8 were 12 cases (-),5 (+),4 (+ +),while in control group 7 cases (-),2 (+),and 0 (+ +),no statistically significant difference was found between the two group (P =0.214).The PUF score of the patients with IL-10 expression (-) was 19.7 ± 2.1,lower than those with IL-10 (+ +) (27.1 + 2.5,P ＜ 0.001).PUF of patients with IL-10 (+) was 22.7 ± 1.8,lower than those with (+ +) (P =0.001).The PUF score of IL-6 expression (-) was 21.The PUF score of IL-6 expression (+) was (23.2 + 3.2),lower than (+ +) (26.7 + 3.1,P =0.025).The PUF score of (+ +) IC/BPS patients was (26.6 ± 2.4),higher than that of (+) IC/BPS patients (21.5 ± 2.1,P ＜ 0.0 l).The PUF score of Siglec-8 expression (+ +) and (+) in IC/BPS group was (21.3 ± 2.0),lower than that of Siglec-8 (-) (27.0 ± 2.3,P ＜ 0.0l).Conclusions Mast cells and plasma cells were expressed in IC/BPS tissues.The expression of IL-6 and IL-10 was positively correlated with clinical symptoms,while the expression of Siglec-8 was negatively correlated with clinical symptoms.

Resistance of maize cultivars to Sitophilus zeamais (Coleoptera: Curculionidae) / Resistência de cultivares de milho a Sitophilus zeamais (Coleoptera: Curculionidae)

Five Zea mays cultivars (BRS Caatingueiro, BRS Gorutuba, BRS Sertanejo, BRS Asa Branca and BR 106) were evaluated considering their effect on the nutrition of the maize weevil Sitophilus zeamais, by analysis of total protein in adult fed with these cultivars and for the presence of lectins and trypsin inhibitors in grains. In addition, free-choice and no-choice assays were performed to investigate the resistance of grains of the Z. mays cultivars to an attack by S. zeamais. The BR 106 cultivar showed the lowest susceptibility index, followed by BRS Caatingueiro, BRS Asa Branca, BRS Sertanejo and BRS Gorutuba. The number of emerged adults in the Z. mays cultivars ranged from 213.17 to 74.0, and the lowest number of insects was recorded for the BR 106 cultivar. The insects were able to feed on grains of all cultivars, but the BR 106 cultivar showed the least reduction in dried biomass. Lectins were detected in extracts from BR 106, BRS Asa Branca, BRS Sertanejo and BRS Gorutuba, and the highest activity was shown by BR 106. The lowest protein assimilation was detected in the insects from treatments with BRS Asa Branca. The extracts from all cultivars were able to inhibit the activity of bovine trypsin, but this effect was not related to the resistance degree of Z. mays cultivars. The results suggest the resistance of BR 160 to the attack of S. zeamais, as well as indicating that the presence of lectin in the grains is the cause of this resistance.(AU)

Foram avaliadas cinco cultivares de Zea mays (BRS Caatingueiro, BRS Gorutuba, BRS Sertanejo, BRS Asa Branca e BR 106) e seu efeito na nutrição do gorgulho-do-milho Sitophilus zeamais, por meio da análise de proteína total em adultos alimentados com esses cultivares e a presença de lectinas e inibidores da tripsina nos grãos. Além disso, foram realizados ensaios com e sem chance de escolha para investigar a resistência dos cultivares de Z. mays ao ataque de S. zeamais. O cultivar BR 106 apresentou o menor índice de susceptibilidade, seguido por BRS Caatingueiro, BRS Asa Branca, BRS Sertanejo e BRS Gorutuba. O número de adultos emergidos nos cultivares de Z. mays variou de 213,17 a 74,0, e o menor número de insetos foi registrado para o cultivar BR 106. Os insetos foram capazes de se alimentar de todos os cultivares, no entanto, o BR 106 mostrou a menor redução na biomassa seca. As lectinas foram detectadas em extratos de BR 106, BRS Asa Branca, BRS Sertanejo e BRS Gorutuba, e a maior atividade foi demonstrada pela BR 106. A menor assimilação de proteína foi detectada nos insetos que se alimentaram com BRS Asa Branca. Os extratos de todos os cultivares foram capazes de inibir a atividade da tripsina bovina, mas esse efeito não está correlacionado ao grau de resistência dos cultivares de Z. mays. Os resultados sugerem a resistência da BR 160 ao ataque de S. zeamais, além de indicar que a presença de lectina nos grãos é a causa dessa resistência.(AU)

Vía de las lectinas, una ruta del complemento en construcción / Lectin pathway, a complement route under construction

ntroducción: El sistema del complemento puede ser activado por tres vías: clásica, alternativa y de las lectinas, esta última en fase de estudio para su completamiento. Objetivo: Describir hasta donde se ha avanzado en la construcción de la vía de las lectinas, sus iniciadores, activadores, reguladores, cascada enzimática y sus funciones biológicas. Metodología: Se realizó una revisión sobre el tema en estudio empleando artículos de libre acceso en la base de datos Pubmed y los trabajos publicados por el grupo de trabajo de la Universidad de Goettigen, la Universidad de Aarhus en Dinamarca y el Laboratorio Central de Líquido Cefalorraquídeo (LABCEL) de la Universidad de Ciencias Médicas de La Habana en los últimos cinco años comprendidos en el período de enero de 2012 a marzo del 2017. Desarrollo: Los iniciadores de la vía de las lectinas son las moléculas de reconocimiento colectinas y ficolinas circulantes en sangre, que participan en muchos procesos del organismo. Los activadores de esta vía son las MASP 1, 2 presentes como proenzimas; y la MASP 3, MAp 19 y 44 actúan como reguladoras. La cascada enzimática luego del reconocimiento es similar a la ruta clásica. Conclusiones: Las colectinas y ficolinas inician la vía de las lectinas. Sus activadores son las MASP 1, 2. Los reguladores son la MASP-3, y las MAp 19 y 44. Similar a la clásica en su cascada enzimática. Es la más antigua en la filogenia por eso participa en muchos procesos en el organismo(AU)

Introduction. The complement system can be activated in three ways: classical, alternative and lectins, the latter in the study phase for its completion. Objective. To describe the progress made in the construction of the lectin pathway, its initiators, activators, regulators, enzymatic cascade and its biological functions. Methods. A review was made on the subject under study using articles of free access in the Pubmed database and the works published by the working group of the University of Goettigen, the University of Aarhus in Denmark and the Central Laboratory of Cefalorraquìdeo liquid (LABCEL) of the University of Medical Sciences of Havana in the last five years included in the period from January 2012 to March 2017. Development. The initiators of the lectin pathway are the collectin recognition molecules and circulating ficolins in blood, which participate in many processes of the organism. The activators of this pathway are MASP 1, 2 present as proenzymes; and MASP 3, MAp 19 and 44 act as regulators. The enzymatic cascade after recognition is similar to the classical route. Conclusions. Collectins and ficolines initiate the lectin pathway. Its activators are MASP 1, 2. The regulators are MASP-3, and MAp 19 and 44. Similar to the classic in its enzymatic cascade. It is the oldest in phylogeny so it participates in many processes in the body.(AU)