Discovery of highly immunogenic spleen-resident FCGR3^(+)CD103^(+)cDC1s differentiated by IL-33-primed ST2^(+)basophils

Recombinant interleukin-33(IL-33)inhibits tumor growth,but the detailed immunological mechanism is still unknown.IL-33-mediated tumor suppression did not occur in Batf3^(−/−)mice,indicating that conventional type 1 dendritic cells(cDC1s)play a key role in IL-33-mediated antitumor immunity.A population of CD103^(+)cDC1s,which were barely detectable in the spleens of normal mice,increased significantly in the spleens of IL-33-treated mice.The newly emerged splenic CD103^(+)cDC1s were distinct from conventional splenic cDC1s based on their spleen residency,robust effector T-cell priming ability,and surface expression of FCGR3.DCs and DC precursors did not express Suppressor of Tumorigenicity 2(ST2).However,recombinant IL-33 induced spleen-resident FCGR3^(+)CD103^(+)cDC1s,which were found to be differentiated from DC precursors by bystander ST2+immune cells.Through immune cell fractionation and depletion assays,we found that IL-33-primed ST2^(+)basophils play a crucial role in the development of FCGR3^(+)CD103^(+)cDC1s by secreting IL-33-driven extrinsic factors.Recombinant GM-CSF also induced the population of CD103^(+)cDC1s,but the population neither expressed FCGR3 nor induced any discernable antitumor immunity.The population of FCGR3^(+)CD103^(+)cDC1s was also generated in vitro culture of Flt3L-mediated bone marrow-derived DCs(FL-BMDCs)when IL-33 was added in a pre-DC stage of culture.FL-BMDCs generated in the presence of IL-33(FL-33-DCs)offered more potent tumor immunotherapy than control Flt3L-BMDCs(FL-DCs).Human monocyte-derived DCs were also more immunogenic when exposed to IL-33-induced factors.Our findings suggest that recombinant IL-33 or an IL-33-mediated DC vaccine could be an attractive protocol for better tumor immunotherapy.

NOD2 and TLR2 ligands trigger the activation of basophils and eosinophils by interacting with dermal fibroblasts in atopic dermatitis-like skin inflammation

The skin of patients with atopic dermatitis （AD） has a unique predisposition for colonization by Staphylococcus aureus （S. aureus）, which contributes to the inflammation and grim prognosis of AD. Although the mechanism underlying the S. aureus-induced exacerbation of AD remains unclear, recent studies have found a pivotal role for pattern recognition receptors in regulating the inflammatory responses in S. aureus infection. In the present study, we used a typical mouse model of AD-like skin inflammation and found that S. aureus-associated nucleotide-binding oligomerization domain-containing protein 2 （NOD2） and toll-like receptor 2 （TLR2） ligands exacerbated AD-like symptoms, which were further deteriorated by the in vivo expansion of basophils and eosinophils. Subsequent histological analyses revealed that dermal fibroblasts were pervasive in the AD-like skin lesions, Co-culture of human dermal fibroblasts with basophils and eosinophils resulted in a vigorous cytokine/chemokine response to the NOD2/TLR2 ligands and the enhanced expression of intercellular adhesion molecule-1 on the dermal fibroblasts. Basophils and eosinophils were primarily responsible for the AD-related cytokine/chemokine expression in the co-cultures. Direct intercellular contact was necessary for the crosstalk between basophils and dermal fibroblasts, while soluble mediators were sufficient to mediate the eosinophil-fibroblast interactions. Moreover, the intracellular p38 mitogen-activated protein kinase, extracellular signal-regulated kinase, and nuclear factor-kappa B signaling pathways were essential for NOD2/TLR2 ligand-mediated activation of basophils, eosinophils, and dermal fibroblasts in AD-related inflammation. This study provides the evidence of NOD2/TLR2-mediated exacerbation of AD through activation of innate immune cells and therefore sheds light on a novel mechanistic pathway bv which S. aureus contributes to the DathoDhvsiology of AD.

New insights into roles of basophils in initiating T helper type 2 immunity

Allergic diseases,mainly mediated by T helper type 2(Th2)immunity,have become a worldwide public health problem.Traditional Chinese medicine(TCM)has long been used in treating and preventing allergic symptoms.As the new target of anti-allergy TCM,basophils,after approximately 140 years since their discovery,are just now gaining respect as important contributors in the pathogenesis underlying allergic inflammation and disease.In addition to their role as effector cells,basophils can release early IL-4,migrate from circulatory system into draining lymph nodes,present antigen to naive CD4^+T cells,and promote the differentiation of Th2 cells.Herein,we briefly summarized the recent research advances of the essential contributions of basophils in the initiation of Th2 immune responses.

Basophils as a potential therapeutic target in cancer

Basophils,which are considered as redundant relatives of mast cells and the rarest granulocytes in peripheral circulation,have been neglected by researchers in the past decades.Previous studies have revealed their vital roles in allergic diseases and parasitic infections.Intriguingly,recent studies even reported that basophils might be associated with cancer development,as activated basophils synthesize and release a variety of cytokines and chemokines in response to cancers.However,it is still subject to debate whether basophils function as tumor-protecting or tumor-promoting components;the answer may depend on the tumor biology and the microenvironment.Herein,we reviewed the role of basophils in cancers,and highlighted some potential and promising therapeutic strategies.

Adenosine cyclic phosphate with ultrasonic-assisted pectinase extraction alleviated allergic reactions in RBL-2H3 through inhibiting the influx of intracellular Ca^(2+)

Jujube contains abundant cyclic adenosine monophosphate(cAMP)and the ultrasonic-assisted pectinase extraction(UAPE)conditions for obtaining the maximum cAMP yield from jujube were optimized.Orthogonal array design was applied to evaluate the effects of 4 variables by UAPE on cAMP yield.The results showed that the optimal cAMP yield(783.0μg/g)was derived at ratio of liquid to solid 5 mL/g,ratio of pectinase to raw material 1.5%,time 60 min and temperature 40℃.Moreover,the effect of cAMP on the anti-allergic function of action induced by immunoglobulin E(IgE)and its meschanism was investigated through establishing the sensitized cell model in rat basophilic leukemia(RBL-2 H3)cells using dinitrophenylated(DNP)-bovine serum albumin(BSA)-IgE.The results showed that cAMP interfered with sensitized cells,effectively inhibited the occurrence of basophil degranulation in dose dependence,and significantly reduced the activity ofβ-hexosamindase(β-hex),at the optimal concentration of 50μg/mL.The level of anti-inflammatory factor interleukin-10(IL-10)was promoted and the content of pro-inflammatory factor tumor necrosis factor-α(TNF-α)was suppressed by cAMP.In addition,influx of intracellular Ca^(2+) was repressed effectively.Our results demonstrate that jujube cAMP regulated the cytokine balance in the allergy pathway through blocking the influx of extracellular Ca^(2+),with the prevention of allergy symptoms.

Autoimmune pancreatitis and pancreatic cancer:Epidemiological aspects and immunological considerations

Ordinary chronic pancreatitis is a well-known risk factor for pancreatic cancer,whereas such an association with autoimmune pancreatitis(AIP)is widely debated.Due to the rarity of the latter disorder,there are few specific clinical and epidemiological studies investigating the relation between AIP and pancreatic cancer,which do not seem to support it.However,these studies are affected by several limitations and,therefore,a link between AIP(and,specifically,type 1 AIP)and pancreatic cancer cannot be ruled out definitively on this basis.Moreover,several immunopathological aspects of type 1 AIP and,in general,immunoglobulin G4-related disease can create an immunological context that may impair the tumoral immunosurveillance and promote the pancreatic carcinogenesis and its progression.In detail,Th2 immunological dominance,type 2 macrophage polarization and basophil infiltration observed in type 1 AIP,may play a permissive role in creating a favorable immunological environment for pancreatic carcinogenesis,in addition to the immunosuppressive therapies that can be used in these patients.

Experimental Researches on Inhibitory Effect of Huoxiang Zhengqi Liquid (藿香正气水) on Histamine Release

Objective: To observe the blocking effect of Huoxiang Zhengqi liqu id (HXZQL) on histamine release of basophils. Methods: The study was conducted using HXZQL to neutralize and b lock IgE and antagonize the effect of IL 3 induced enhancing histamine relea se of bosaphils. Concentration and percentage of histamine release of bosaphils were tested by the basophil histamine release test. Results: The HXZQL drug serum had obvious inhibitive effect on degranulation of bosaphils induced by IgE antigen antibody complex ( P <0.0 5) and can successfully weaken the histamine release of basophils stimulated by IL 3. When compared with those in the untreated group, it showed P <0.01. Conclusion: HXZQL drug serum may neutralize and block IgE and has inhibitive effects on degranulation of cells when resensitized by antigens. It can also block allergic response type Ⅰ by antagonizing the effect of IL 3 induced enhancing histamine release.

Effect of brimonidine tartrate on basophil activation in glaucoma patients

AIM:To evaluate the mechanism of which brimonidine tartrate 0.15%causes clinical hypersensitivity.METHODS:A prospective case-control study comparing 8 glaucoma patients with clinical hypersensitivity to brimonidine to a control group consisting 13 healthy volunteers.Blood samples were stimulated with brimonidine 0.15%,timolol 0.5%or brimonidine tartrate/timolol maleate 0.2%/0.5%.Premixed antibodies(CD63/FITC and aIgE/PE)were added for direct staining and whole-blood samples were lysed,fixed and analyzed by a flow cytometer.The basophil population was defined by high IgE cell expression.Degranulation was identified by the expression of the activation molecule CD63.RESULTS:Basophil activation was not significant when comparing percent of activated basophils of patients and healthy controls after exposure to brimonidine(2.58%,2.45%,respectively,P=0.72).There was a significant suppression of basophil activation when a combination of brimonidine-timolol(0.87%)was compared to timolol(2.27%;P=0.012)and to brimonidine alone(2.58%;P=0.017).CONCLUSION:The results of our study do not support the hypothesis that brimonidine induces an immediate allergic reaction.Basophil activation was suppressed by the presence ofβ-blockers in patients hypersensitive to brimonidine and in healthy individuals.This finding indicates that timolol suppress brimonidine drug reaction by a different mechanism.

Inflammatory Cytokine Secretion Status of Bone Marrow Cells and Clinical Significance in Immune-related Hematocytopenia

Objective To observe the expression of inlfammatory molecules in bone marrow immune cells of patients with immune-related hematocytopenia (IRH), and to investigate the immune mechanism and clinical signiifcance of the disease. Methods Total of 36 IRH patients were selected as observation group and 30 healthy people were taken as control group. Serum cytokines levels, activity of immunocytes and expression of HLA-DR were detected. Immune lfuorescence was applied to observe the expression state of immunologic molecules and cytokines in IRH patients. Results Serum cytokines were elevated in various degrees in observation group. Compared with the control group, the cytokines levels were significantly higher (P < 0.05). After treatement with immunosuppressive drugs, the serum levels of cytokines in observation group reduced to a level close to the control group. HLA-DR were upregulated in activated tissue basophils, eosinophils, dendritic cells (DC) and macrophages of bone marrow in IRH patients, and POX activity in these immunocytes of IRH was higher than that of the control group. Immune molecules were highly expressed in eosinophils, DC and macrophages. Conclusions It is demonstrated that antibodies or self-reactive lymphocytes were produced in IRH marrow, which would cause lesions of hemocytes, and lead to pathological process ifnally. Structure of hematopoietic cells mutated and these cells might be acted as target cells of immunocytes in the pathological process. Immunocytes could secrete inlfammatory factors and lead to immunologic injury of hemocyte.

The Cbl family of ubiquitin ligases regulates FcεRI expression and mast cell activation

Antigen interaction with specific IgE bound to the high-affinity Fc receptor for IgE, constitutively expressed on the cell-surface of mast cells, generates signals that cause a shift in the resting state equilibrium of phosphorylation and dephosphorylation events that serves to maintain homeostasis. The outcome of this activated state is the release of a wide array of preformed and newly synthesized pro-inflammatory mediators. During the past few years, the existence of a negative feedback loop initiated upon FcεRI engagement has also been envisaged. This negative signal involves the coordinated action of adaptors, phosphatases and ubiquitin ligases that limits the intensity and duration of positive signals, thus modulating mast cell functions. Relevant to this, others and we have demonstrated that Cbl family proteins control the amplitude of FcεRI-generated signals by specific ubiquitin modification of activated receptor subunits and associated protein tyrosine kinases. In this article, we review advances in our understanding of the molecular mechanisms through which Cbl proteins regulate FcεRI expression and signaling.

Identification of CD13,CD107a,and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-de-pendent upregulation

Using two-colour flow cytometry>200 antibodies submitted to the 8^(th) International Workshop of Human Leukocyte Differentiation Antigens(HLDA8)have been analyzed for their reactivity with resting and activated CD203c^(+)basophils.Four antibodies either non-reactive or weakly reactive with resting basophils exhibited an increased reactivity with basophils activated by anti-IgE-mediated cross-linking of the high affinity IgE receptor(FcεRI).These include antibod-ies against CD164(WS-80160,clone N6B6 and WS-80162,clone 67D2),as well as two reagents with previously unknown specificities that were identified as CD13(WS-80274,clone A8)and CD107a(WS-80280,clone E63-880).The activation patterns followed either the“CD203c-like”or“CD63-like”activation profile.The CD203c profile is characterized by a rapid and significant upregulation(of CD13,CD164,and CD203c),reaching maximum levels after 5-15 min of stimulation.The phosphoinositide-3-kinase(PI3K)-specific inhibitor wortmannin inhibited the upregulation of these markers whereas 12-O-tetradecanoyl-phorbol-13-acetate(TPA)induced a rapid and FcεRI-independent upregulation within 1-2 min.In the CD63 profile,maximum upregulation(of CD63 and CD107a)was detected only after 20-40 min,and upregulation by TPA reached maximum levels after 60 min.In summary,our data identify CD13,CD107a,and CD164 as novel basophil-activation antigens.Based on time kinetics of upregulation,we hypothesize that molecules of the“CD203c group”and the“CD63 group”are linked to two different mechanisms of basophil activation.

RELATIONSHIP BETWEEN HYPERSENSITIVITY TYPE I AND WHEEZING BY RESPIRATORY SYNCYTIAL VIRUS (RSV) INFECTION

We studied the RSV specific IgE antibody, histamine and basophil from infants with RSV bronchiolitis and found during the acute phase either the titers of RSV-IgE or the concentration of histamine increased significantly, the number of basophil and basophil degranulation in the presence of RSV antigen also increased. In vitro studies revealed hypersensitivity participates in the pathogenesis of RSV bronchiolitis. We also found that infants with RSV bronchiolitis, the RSV-IgE persisted for a long time presumably this plays an important role in recurrent wheezing after RSV infection for years.

Staining Effect of Pomegranate Flower Extract on Human Blood Cells： First Results

Pomegranate flower plant, popularly known using for the treatment of various diseases, was not investigated as a source of dye for cytological studies using human blood cells. The importance of this study is to appear dyeing result of pomegranate flower extract on human blood cells. The natural dye source was pomegranate flower known as roselle and potassium aluminum sulfate （alum = KAISO4＇ 12H20） was used as mordant or metal salt. Distilled water was used as solvent. Fresh, clean and air-dried flowers were extracted with distilled water at 100℃ for 30 minutes and then filtered. One drop blood from a healthy 20-year woman was spread as a peripheral on to ten plates and dried at nearly 25℃. These slides were stained by soaking in pomegranate flower extract with/without alum （KA1SO4·12H2O） at 100℃ for 60 minutes. Slides were washed with distilled water, dried and done microscobic examination. The different blood cells dyed dark orange in alum mordant media at 100 ℃. As a result, pomegranate flower has the capacity to use dyeing human blood cells such as eosinophil, basophil and neutrophil.

Food allergy in children-the current status and the way forward

Food allergy in children is a major health concern,and its prevalence is rising.It is often over-diagnosed by parents,resulting occasionally in unnecessary exclusion of some important food.It also causes stress,anxiety,and even depression in parents and affects the family’s quality of life.Current diagnostic tests are useful when interpreted in the context of the clinical history,although cross-sensitivity and inability to predict the severity of the allergic reactions remain major limitations.Although the oral food challenge is the current gold standard for making the diagnosis,it is only available to a small number of patients because of its requirement in time and medical personnel.New diagnostic methods have recently emerged,such as the Component Resolved Diagnostics and the Basophil Activation Test,but their use is still limited,and the latter lacks standardisation.Currently,there is no definite treatment available to induce life-long natural tolerance and cure for food allergy.Presently available treatments only aim to decrease the occurrence of anaphylaxis by enabling the child to tolerate small amounts of the offending food,usually taken by accident.New evidence supports the early introduction of the allergenic food to infants to decrease the incidence of food allergy.If standardised and widely implemented,this may result in decreasing the prevalence of food allergy.

Study on the Effect of Dental Metal Alloys on the Histamine Liberation of Blood Basophiles in Vitro

Biomaterials for restoration or replacement of diseased tissues may have any origin.The major characteristic for biomaterials is biocompatibility.All biomaterials,used in medicine (dentistry,in particular),interreact with the organism tissues.And the changes occur both in the materials and the organism tissues.It is considered that there are no "inert biomaterials." The number of allergic diseases and complications is constantly growing all over the world,taking an important place in the structure of infectious and noninfectious pathology[1].Pollen,household,epidermal,and food-borne allergens,and haptens are the most frequent sources of sensibilization.

变应性鼻炎与支气管哮喘的相关性研究进展

近年来,变应性鼻炎和支气管哮喘的流行率有逐年增加的趋势,而且这种趋势是全球性的。由于上、下呼吸道炎症反应的一致性,Th2细胞、嗜酸性粒细胞在呼吸道变态反应中的共同作用,变应性鼻炎与支气管哮喘密切相关,对变应性鼻炎患者应重视并评估支气管哮喘,对支气管哮喘亦应注意评估变应性鼻炎。

嗜碱粒细胞与变应性鼻炎

本文就嗜碱粒细胞在IgE介导的变应性鼻炎中的作用，嗜碱粒细胞中IgE信赖性信号传导和调控因子的作用，嗜碱粒细胞的生长和发育及在变态反应受累组织中的汇集，嗜碱粒细胞对天然免疫的识别等方面，阐述关于人嗜碱粒细胞在变应性鼻炎中地位的新认识。

Signaling transduction pathways involved in basophil adhesion and histamine release

Background Little is known about basophil with respect to the different signaling transduction pathways involved in spontaneous, cytokine or anti-IgE induced adhesion and how this compares to IgE-dependent and IgE-independent mediator secretion. The purpose of the present study was to investigate the roles of β1 and β2 integrins in basophil adhesion as well as hosphatidylinositol 3-kinase （PI3K）, src-kinases and extracellular signal regulated kinase （ERK）1/2 in basophil adhesion and histamine release （HR）. Methods Basophils （purity of 10%-50%） were preincubated with anti-CD29 or anti-CD 18 blocking antibodies before used for adhesion study. Basophils were preincubated with the pharmacological inhibitors wortmannin, PP1, PD98059 before used for adhesion and HR study. Cell adherence to bovine serum albumin （BSA） or fibronectin （Fn） was monitored using cell associated histamine as a basophil marker and the histamine was measured by the glass fiber assay. Results Basophil spontaneous adhesion to Fn was inhibited by anti-CD29. Interleukin （IL）-3, granulocyte/macrophage colony stimulating factor （GM-CSF） induced adhesion to BSA was inhibited by anti-CD18. Wortmannin at 1 μmol/L and PP1 at 20 μmol/L strongly interfered with, whereas PD98059 at 50μmol/L weakly inhibited basophil spontaneous adhesion to Fn. One μmol/L wortmannin strongly inhibited IL-3, IL-5, GM-CSF and anti-IgE induced adhesion to BSA. PP1 at 20 μmol/L partly inhibited anti-IgE induced adhesion. Fifty μmol/L PD98059 marginally inhibited IL-5, weakly inhibited anti-IgE, partly inhibited GM-CSF induced adhesion. Wortmannin, PP1 and PD98059 inhibited anti-IgE （1：100 or 1：1000） induced basophil HR in a dose dependent manner. They inhibited calcium ionophore A23187 （10 μmol/L, 5 μmol/L） induced basophil HR in a dose dependent manner, but to different extend with PP1 being the most efficient. Conclusions Basophil spontaneous adhesion to Fn is mediated by β 1-integrins whereas cytokine induced adhesion to BSA is mediated by β 2-integrins. PI3K, src-kinases and ERK1/2 play distinct signaling roles in basophil adhesion and HR. PI3K is the key player while ERK1/2 is the weakest participant.

Arsenic trioxide in treatment of de novo acute basophilic leukemia

Acute basophilic leukemia （ABL） is a rare subtype of .acute myeloid leukemia （AML）, accounting for 4%-5% of AML and less than 2% of all hematopoietic malignancies. It is usually characterized by a very rapid clinical course, symptoms of hyperhistaminemia, peptic ulceration, gastrointestinal cerebrovascular bleeding and resistance to therapy.^1 However, the clinical outcome of ABL remains disapp2ointing. Most patients died within 1 year after diagnosis.

嗜碱性粒细胞升高与贫血形态学分类的观察

目的：是为了贫血患者找到治疗依据,找到治疗贫血用药的效果,同时在群众中做好贫血的预防工作.方法：用五分类血球仪进行细胞分析,测出血红蛋白﹑嗜碱性粒细胞﹑红细胞三个平均值和红细胞体积分布宽度.结果：从五分类检验报告中不难看出,75%贫血患者嗜碱性粒细胞升高,不贫血患者嗜碱性粒细胞升高只有30%.结论：贫血患者是否还在发展,贫血通过治疗,疗效怎样,这些都必须以嗜碱性粒细胞升高为依据,并结合形态学分类,把二者有机的结合起来,找到贫血的真正原因.