Plasma Macrophage Migration Inhibitory Factor and CCL3 as Potential Biomarkers for Distinguishing Patients with Nasopharyngeal Carcinoma from High-Risk Individuals Who Have Positive Epstein-Barr Virus Capsid Antigen-Specific IgA / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The purpose of this study was to identify novel plasma biomarkers for distinguishing nasopharyngeal carcinoma (NPC) patients from healthy individuals who have positive Epstein-Barr virus (EBV) viral capsid antigen (VCA-IgA). MATERIALS AND METHODS: One hundred seventy-four plasma cytokines were analyzed by a Cytokine Array in eight healthy individuals with positive EBV VCA-IgA and eight patients with NPC. Real-time polymerase chain reaction, Western blotting, enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry were employed to detect the expression levels of macrophage migration inhibitory factor (MIF) and CC chemokine ligand 3 (CCL3) in NPC cell lines and tumor tissues. Plasma MIF and CCL3 were measured by ELISA in 138 NPC patients, 127 EBV VCA-IgA negative (VN) and 100 EBV VCA-IgA positive healthy donors (VP). Plasma EBV VCA-IgA was determined by immunoenzymatic techniques. RESULTS: Thirty-four of the 174 cytokines varied significantly between the VP and NPC group. Plasma MIF and CCL3 were significantly elevated in NPC patients compared with VN and VP. Combination of MIF and CCL3 could be used for the differential diagnosis of NPC from VN cohort (area under the curve [AUC], 0.913; sensitivity, 90.00%; specificity, 80.30%), and combination of MIF, CCL3, and VCA-IgA could be used for the differential diagnosis of NPC from VP cohort (AUC, 0.920; sensitivity, 90.00%; specificity, 84.00%), from (VN+VP) cohort (AUC, 0.961; sensitivity, 90.00%; specificity, 92.00%). Overexpressions of MIF and CCL3 were observed in NPC plasma, NPC cell lines and NPC tissues. CONCLUSION: Plasma MIF, CCL3, and VCA-IgA combination significantly improves the diagnostic specificity of NPC in high-risk individuals.

A review on the Variable copies of C-C motif Chemokine Ligand 3 Like 1 (CCL3L1) Gene among Different Populations and Current Methods for Quantification

@#C-C motif Chemokine Ligand 3 Like 1 (CCL3L1) is characterized as a gene with copy number variable (CNV) and clustered at the segmental duplication on chromosome 17q12. CCL3L1 is responsible for the production of macrophage inflammatory protein (MIP) 1α that plays an important function in the immune system and host defense. Various copies range of CCL3L1 have been reported and associated with the diseases in different populations. Thus, this review aimed to summarise the distribution of CCL3L1 copy number from different populations according to the geographical region and highlighted the lacking of data from Malaysian population, which is one of the multi-ethnic countries due to the impacts of CCL3L1 copies on various diseases. Besides, we also outlined the methodologies available for the copy number typing. In overall, this review could provide significant information on the role of CCL3L1 copies in disease association and as well as providing evidence on the population diversity

Effect of Saposhnikovan Sulphate on the Recruitment and Re-education of Macrophages by MCF-7 Cells In Vitro through CCL3 / 中国药学杂志

OBJECTIVE: To observe the inhibition of saposhnikovan sulphate obtained by ultrasonic extraction(S-USPS) on human leukemia MCF-7 cells in vitro and the effect of S-USPS on the recruitment and re-education of macrophages by MCF-7 cells. METHODS: S-USPS was obtained from saposknikovan(SPS) by the method of chlorosulfonic acid-pyridine complex. The inhibition of S-USPS and USPS on proliferation of MCF-7 cells was measured by MTT. THP-1-derived macrophages were differentiated in vitro. The effect of S-USPS on the recruitment of macrophages by MCF-7 cells was detected by transwell migration assay. The effects of S-USPS on the expression and secretion of chemokine were detected by qRT-PCR and ELISA. The effects of S-USPS and CCL3 on the expression of inflammatory cytokines were detected by qRT-PCR. RESULTS: The inhibitory rate of S-USPS on the proliferation of MCF-7 was dose-dependent and time-dependent and was higher than that of USPS(P<0.05).S-USPS increased the transcription and secretion of CCL3 by MCF-7 cells. There were higher expressions of IL-1β, IL-6, and TNF-α mRNA and lower expressions of TGF-β, IL-10 mRNA of macrophages in the S-USPS group than in the control group and MCF-7 supernatant group(P<0.05). The promotion of S-USPS on the expressions of IL-1β and TNF-α was inhibited by CCL3 neutralizing antibody(P<0.05). CONCLUSION: There are significant evidences that the saposhnikovan sulphate effectively promotes the recruitment of macrophages by MCF-7 cells in vitro, and re-educates their secretion profile toward M1-type.

CCL3 promotes hBMSC proliferation and inhibits exosome secretion / 中国病理生理杂志

AIM:To explore the regulatory effect of chemokine CCL 3 on exosome secretion from human bone marrow mesenchymal stem cells(hBMSCs).METHODS: hBMSCs were stimulated with chemokine CCL 3 at different concentrations in vitro.The proliferation of hBMSCs was measured by CCK-8 assay and viable cell counting.Exosome se-cretion from hBMSCs was qualitatively analyzed by transmission electron microscope(TEM)and flow cytometry, and the quantitative analysis was carried out by flow cytometry and nanoparticle tracking analysis(NTA).RESULTS:Compared with control group,the viability of the hBMSCs detected by CCK-8 assay was increased when hBMSCs were treated with CCL3(P<0.05).The results of viable cell counting demonstrated that the number of hBMSCs was raised in CCL 3 group in a dose-dependent manner(P<0.05).The results of flow cytometry showed that hBMSCs expressed 3 CCL3-related spe-cific receptors,CCR1,CCR5 and CCR9.Compared with control group,the fluorescence intensity of CCR9 in CCL3 group was obviously enhanced.However,no significant difference of fluorescence intensity for CCR 5 and CCR1 was observed be-tween the 2 groups.The results of NTA demonstrated that the secretion capacity of CCL 3-induced hBMSCs was far less than that in control group(P<0.05).However, the microvesicles larger than 100 nm in CCL3 groups were increased(P<0.05).The above results indicated that the higher concentration of CCL 3 induced the lower secretion of exosomes.In addi-tion,the results of flow cytometry demonstrated that CCL 3-induced hBMSCs showed lower quantity of CD 9 +exosomes than those in control group(P<0.01).CONCLUSION:CCL3 promotes the proliferation of hBMSCs but depresses the secre-tion of exosomes in a dose-dependent manner.CCL3 affects the size distribution of exosomes and increases the number of nonfunctional microvesicles of larger than 100 nm in size.CCL3 induces the expression of CCR9 in hBMSCs.

Expression and clinical significance of MIP-1 alpha in serum of asthmatic patients / 中国医师杂志

Objective To investigate the levels of serum macrophage inflammatory protein-1α (MIP-1α),inflammatory factors and pulnonary function in patients with asthma,and the correlation between thenm.Methods Eighty patients with asthma were selected who were treated in our hospital from March 2015 to August 2016,and the data of 80 healthy adults were selected as control.The differences of serum MIP-1α,inflammatory factors,imnmunoglobulin and lung function were observed in two groups.The correlation between MIP-1α and inflammatory factors,lung function and immunoglobulin were analyzed.Results The levels of MIP-1α,interleukin (IL)-6,and tumor necrosis factor-α (TNF-α) in patients with asthma were significantly higher than those in control group (t =-207.04,-33.209,-55.132,P ＜ 0.01);The levels of forced vital capacity (FVC),the forced expiratory volume in one second (FEV1),FEV1/FVC,maximal mid expiratory flow (MMEF),and peak expiratory flow (PEF) in the asthma group were significantly lower than those in the control group (t =17.100,39.154,22.791,25.391,19.356,P ＜ 0.01).The IgG and IgM of asthma group were significantly lower than those of the control group (t =9.564,7.528,P ＜ 0.01).The IgE level was significantly higher than that of the control group (t =-82.683,P ＜0.01).The level of MIP-1α was positively correlated with the levels of IL-6,TNF-0 and IgE,and negatively correlated with FVC,FEV1,FEV1/FVC,MMEF,PEF,IgG and IgM in patients with asthma.Conclusions The level of serum MIP-1 α in asthmatic patients is high,and is closely related to the inflammatory cytokines and lung function in patients with asthma.

Association between the CCL3L1 gene copy number variation and susceptibility to ankylosing spondylitis / 中华风湿病学杂志

Objective This study aimed to investigate whether the copy numbers of the CCL3L1 (Chemokine C-C-Motif Ligand 3 Like Protein 1) gene were associated with susceptibility to ankylosing spondylitis (AS). Methods A total of 806 Chinese individuals including 405 AS patients and 401 healthy controls were enrolled. The CCL3L1 gene copy number was measured by a custom-by-design Multiplex AccuCopyTM Kit based on a multiplex fluorescence competitive polymerase chain reaction (PCR) principle, and 50 samples were randomly selected using the fluorescent quantitative PCR method to verify copy number. Main statistical method was t test, chi-square test and logistic regression model. Results There were no statistically significant differences between the case group and control group in age and gender ( t=1.77, P=0.076, χ2=1.14, P=0.289). The copy number of CCL3L1 gene ranged from 0 to 13 in both AS patients and the controls. After copy numbers were classified into 3 categories by 3, we did not find significant difference between the two groups ( χ2=0.591, P=0.669). And regression analyses also did not support the hypothesis that CCL3L1 gene copy number variation (CNV) could be an impact factor to the severity or function indexes of AS patients ( χ2=0.341, P=0.804 and χ2=0.472, P=0.774, respectively). Conclusion We suggest that the copy number of the CCL3L1 gene does not have a role in the susceptibility and the severity or function to AS.

Protective Effect of Sivelestat Sodium on Experimental Autoimmune Encephalomyelitis Model Rats / 中国药房

OBJECTIVE:To study the protective effect of sivelestat sodium on experimental autoimmune encephalomyelitis (EAE) model rats. METHODS:60 Wistar rats were randomly divided into normal control group (normal saline),model group (normal saline),positive drug group [prednisone acetate tablets 5 mg/(kg·d)] and sivelestat sodium low-dose,middle-dose and high-dose groups [5,8,10 mg/(kg·d)] with 10 rats in each group. Except for normal control group,other groups were given guin-ea pig spinal cord homogenate as antigen to produce EAE model,and then given relevant medicine ip since the same day of model-ing,for consecutive 16 d. The neurologic function of mice was scored,and pathological changes of brain and spinal cord were ob-served;the content of IFN-γ,IL-4,CCL3,chemotactic factor CCL5 regulating and activating normal T cell expression and secre-tion were determined. RESULTS:Compared with normal control group,neurological function score and the content of IFN-γ, CCL5 and CCL3 increased,while IL-4 content decreased (P0.05). Above effect depended on drug dose. CONCLUSIONS:Sivelestat sodium can relieve myelinoclasis and inflammatory cell infiltration,and the mechanism may be related to the decrease of IFN-γ content, the increase of IL-4 content,and inhibition of CCL3 and CCL5 expression in peripheral blood.

Copy Number Variation Burden on Asthma Subgenome in Normal Cohorts Identifies Susceptibility Markers

PURPOSE: Asthma is a complex disease caused by interplay of genes and environment on the genome of an individual. Copy number variations (CNVs) are more common compared to the other variations that disrupt genome organization. The effect of CNVs on asthma subgenome has been less studied compared to studies on the other variations. We report the assessments of CNV burden in asthma genes of normal cohorts carried out in different geographical areas of the world and discuss the relevance of the observation with respect to asthma pathogenesis. METHODS: CNV analysis was performed using Affymerix high-resolution arrays, and various bioinformatics tools were used to understand the influence of genes on asthma pathogenesis. RESULTS: This study identified 61 genes associated with asthma and provided various mechanisms and pathways underlying asthma pathogenesis. CCL3L1, ADAM8, and MUC5B were the most prevalent asthma genes. Among them, CCL3L1 was found across all 12 populations in varying copy number states. This study also identified the inheritance of asthma-CNVs from parents to offspring creating the latent period for manifestation of asthma. CONCLUSIONS: This study revealed CNV burden with varying copy number states and identified susceptibility towards the disease manifestation. It can be hypothesized that primary CNVs may not be the initiating event in the pathogenesis of asthma and additional preceding mutations or CNVs may be required. The initiator or primary CNVs sensitize normal cohorts leading to an increased probability of accumulating mutations or exposure to allergic stimulating agents that can augment the development of asthma.

Changes in CCL3 and CCR5 expression in spinal cord during hyperalgesia induced by remifentanil in rats with incisional pain / 中华麻醉学杂志

Objective To evaluate the changes in the expression of CC-chemokine ligand 3 (CCL3) and CC-chemokine receptor 5 (CCR5) in the spinal cord during hyperalgesia induced by remifentanil in rats with incisional pain.Methods Thirty-two male Sprague-Dawley rats,aged 2-3 months,weighing 240-260 g,were randomly divided into 4 groups (n=8 each) using a random number table:control group (group C),incisional pain group (group Ⅰ),remifentanil group (group R) and remifentanil+incisional pain group (group R+I).A 1-cm longitudinal incision was made in the plantar surface of the left hindpaw in anesthetized rats.While the model of incisional pain was established,remifentanil was infused for 60 min at 1 μg · kg-1 · min-1.At 24 h before infusion of remifentanil (baseline) and 2,6,24 and 48 h after the end of infusion,the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The rats were sacrificed after the last measurement of pain threshold,the lumbar segment (L4-6) of the spinal cord was removed for determination of CL3 and CCR5 mRNA expression (by real-time PCR) and CL3 and CCR5 expression (by Western blot).Results Compared with group C,the MWT was significantly decreased,the TWL was shortened,and the expression of CCL3 and CCR5 mRNA and protein was up-regulated in I,R and R+ I groups.Compared with I and R groups,the MWT was significantly dccreascd,the TWL was shortened,and the expression of CCL3 and CCR5 mRNA and protein was up-regulated in group R+I.Conclusion The mechanism by which remifentanil induces hyperalgesia is related to up-regulated expression of CCL3 and CCR5 in the spinal cord of rats with incisional pain.

The correlation between human chemokine type 1 chemokine ligand 3 and chemokine ligand 4 gene and the immune reconstitution of acquired immunodeficiency syndrome patients after antiretroviral therapy / 中华传染病杂志

Objective To investigate the correlation between the human chemokine type 1 chemokine ligand 3 (CCL3L1) and chemokine ligand 4 (CCL4L) gene expression and the immune reconstitution of acquired immunodeficiency syndrome (AIDS) patients after antiretroviral therapy.Methods The gene copy numbers of CCL3L1 and CCL4L were detected by real time polymerase chain reaction in 217 AIDS patients before antiretroviral therapy.And the correlation between CCL3L1 and CCL4L gene copy numbers and the level of CD4+ and CD8+ T lymphocytes were analyzed.The changes of CD4+ and CD8+ T lymphocytes were defined as mean change value per month after 48 months treatment.The change rates of CD4+ and CD8+ T lymphocytes were defined as the logarithm of the ratio of the value after 48 month to that at baseline.Comparison between groups was conducted using analysis of variance.Results The median of gene copy numbers of CCL3L1 and CCL4L were 2 (range:0-8) and 3 (range:0-7),respectively.After antiviral treatment,there were significantly different changes of CD8+ T lymphocyte level (F=3.054,P＜0.05) and change rate of CD4+/CD8+ (F=3.520,P＜0.05) among groups of high (gene copy 4-8),median (gene copy 2-3) and low (gene copy 0-1) CCL3L1 gene copy numbers.The changes of CD8+ T lymphocyte levels (P=0.023) and change rates (P=0.038) in high and low CCL3L1 gene copy groups were both significantly different.There were significant differents changes rate of ratio of CD4+/CD8+ T lymphocyte among high and median (P=0.010),high and low CCL3L1 gene copy numbers (F=4.397,P＜0.05).The significant difference of the change rates of CD4+/CD8+ were found between the gene copy 3 group and gene copy 4-7 group CCL4L (P=0.005) and between the gene copy 4-7 group and gene copy 0-2 group of CCL4L (P=0.030).The change ratio of CD4+/CD8+ T lymphocytes increased with the increase of copy numbers of CCL4L gene.Conclusions The gene expressions of CCL3L1 and CCL4L might be associated with the ability of immune reconstitution of AIDS patients after antiretroviral therapy.

The impact of methamphetamine on the expressions of macrophage inflammatory proteins in human immunodeficiency virus-infected patients / 中华传染病杂志

Objective To investigate the impact of methamphetamine (Meth) on the expressions of macrophage inflammatory protein (MIP)-1α ,MIP-1β ,interleukin (IL)-6 among human immunodeficiency virus(HIV)-infected patients .Methods The investigation was performed among 15 Meth-abuse and HIV-infected subjects (Meth + HIV ) ,15 non-Meth-abuse and HIV-infected subjects (non-Meth + HIV ) ,15 Meth-abuse and HIV-uninfected subjects (Meth) ,and 15 healthy subjects (HC) .CD4 + T lymphocyte counts in peripheral blood were detected by flow cytometry .The HIV viral loads in HIV-infected patients were detected by standard detection method .The levels of plasma MIP-1α ,MIP-1β and IL-6 from four groups were determined by enzyme-linked immunosorbent assay (ELISA ) .Intergroup difference was compared using t-test and interactive analysis was conducted using analysis of variance .Results In HIV-infected patients ,CD4 + T lymphocyte counts in Meth + HIV group was significant lower than non-Meth +HIV group (t= 5 .431 , P 0 .05) ,neither between HIV infection and the levels of cytokines (P> 0 .05) .Conclusion Meth abuse results in elevated expressions of MIP-1αand MIP-1β ,which indicates that Meth abuse may play a regulating role on promoting HIV infection .

CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS

ABSTRACT Objective: The aim of this study was to compare the production of the chemokines CCL3 and CXCL12 by cultured dental pulp fibroblasts from permanent (PDPF) and deciduous (DDPF) teeth under stimulation by Porphyromonas gingivalis LPS (PgLPS). Material and Methods: Primary culture of fibroblasts from permanent (n=3) and deciduous (n=2) teeth were established using an explant technique. After the fourth passage, fibroblasts were stimulated by increasing concentrations of PgLPS (0 – 10 µg/mL) at 1, 6 and 24 h. The cells were tested for viability through MTT assay, and production of the chemokines CCL3 and CXCL12 was determined through ELISA. Comparisons among samples were performed using One-way ANOVA for MTT assay and Two-way ANOVA for ELISA results. Results: Cell viability was not affected by the antigen after 24 h of stimulation. PgLPS induced the production of CCL3 by dental pulp fibroblasts at similar levels for both permanent and deciduous pulp fibroblasts. Production of CXCL12, however, was significantly higher for PDPF than DDPF at 1 and 6 h. PgLPS, in turn, downregulated the production of CXCL12 by PDPF but not by DDPF. Conclusion: These data suggest that dental pulp fibroblasts from permanent and deciduous teeth may present a differential behavior under PgLPS stimulation. .

Expression of macrophage inlfammatory protein 1αin mice with viral myocarditis / 临床儿科杂志

Objective To detect the expression of macrophage inlfammatory protein 1α(MIP-1α) in the myocardium of viral myocarditis (VMC) mice at different phases. Methods A total of 120 4-week-old male BALB/c mice were randomly divided into 2 groups, 80 in the VMC group and 40 in the control group. Mice in VMC group were inoculated intraperitoneally with coxsackievirus B3 to build VMC models, while mice in control group were treated with DMEM cultivate liquid. Ten mice of each group were sacriifced on days 3, 7, 15 and 30 after treatment and their heart tissues were collected for analysis. The level of MIP-1αin the myocardium was determined by immunohistochemistry. Myocardial histopathology was examined with hematoxylin and eosin stain. In addition, the relationship between the level of MIP-1αand the degree of myocardial lesion was investigated. Results The expression of myocardial MIP-1αprotein in VMC group was up-regulated in myocardium on day 3 after inoculation of virus, and slowly decreased after the peak on day 7, but still sustained a high level on day 30. Compared with the control group, the levels of MIP-1αin VMC group were increased signiifcantly at every phase (P<0.05). Furthermore, positive correlation was found between MIP-1αprotein expression levels and myocardial histopathologic scores in VMC group (r=0.94, P<0.01). Conclusion The up-regulated expression of MIP-1αmay play a critical role in the pathogenic mechanisms of viral myocarditis.

Relevancia del CCR5 co-receptor del HIV-1 y su ligando CCL3L1 en el tropismo viral en pediatría / Relevance of the CCR5 co-receptor of HIV-1 and its ligand CCL3L1 in viral tropism in pediatrics

El objetivo del trabajo fue determinar la participación del coreceptor CCR5 y su ligando CCL3L1 en relación a una de las características del fenotipo del HIV-1, el tropismo viral. Los resultados que obtuvimos así como otras investigaciones que estamos desarrollando tienen implicancias clínicas y terapéuticas en los niños HIV-1 infectados. Para lograr el objetivo hemos evaluado en forma conjunta y combinada los datos que habíamos obtenido a lo largo de casi 15 años en relación a las variantes genéticas del CCR5, en particular la mutación Δ32 y su ligando CCL3L1 en función del número de copias del gen en los niños HIV-1 infectados por transmisión vertical que son asistidos en el Hospital Garrahan, investigando la probable asociación de ellas con el tropismo viral. Hallamos que los niños en primoinfección tienen una proporción considerable de variantes HIV-1 SI que emplean como co-receptor al CXCR4 en lugar del CCR5. Otro hecho relevante fue que la presencia de las variantes SI predominaron en los niños heterocigotas para la variante genética CCR5Δ32. En este último grupo encontramos además que estaba significativamente asociado con un número de copias del CCL3L1 alto (≥2). Probablemente ambos factores participan favoreciendo la reducción en el número de moléculas del co-receptor CCR5 expresadas en la superficie celular facilitado la infección por las variantes X4. Aunque las variantes SI en la etapa crónica alcanzan a un 40% no parecieran asociadas con el genotipo CCR5Δ32 ni con el número de copias del CCL3L1. En resumen, hemos demostrado que las variantes SI X4 T-trópicas del HIV-1 pueden estar presentes en los estadios muy tempranos de la infección viral sugiriendo que puede ser transmisible verticalmente. Además, el genotipo CCR5Δ32 en el contexto de copias altas del CCL3L1 en el niño HIV-1 infectado, contribuyen a un mayor riesgo a ser infectado por variantes SI en primoinfección. Este hecho no pareciera suceder en la etapa crónica de la infección viral.

The aim of the study was to assess the role of co-receptor CCR5 and its ligand CCL3L1 in viral tropism, one of the char-acteristics of the HIV-1 phenotype. The results of this study as well as those found in our other ongoing research have clini-cal and therapeutic consequences for HIV-1 Infected children. For the aim of the study we collected and evaluated the data obtained over a period of almost 15 years on genetic variants of CCR5, specially the 32 mutation and its ligand CCL3L1 (MIP-1 P), in relation to the number of copies of the gene in HIV-1-mother-to-child infected children seen at the pediatric hospital J.P. Garrahan, to investigate a probable association with viral tropism. We found that children with a primary in-fection have a considerable number of HIV-1 SI (syncytium-inducing, i.e. cytopathic) variants that use CXCR4 instead of CCR5 as a co-receptor. Another relevant finding was that SI variants were predominant in children that were homozygous for the genetic CCR5 32 variant. In this latter group we additionally found that this was significantly associated with a high number of CCL3L1 copies ( 2). Both factors may play a favorable role in the decrease of the number of molecules of the CCR5 co-receptor expressed on the cell surface that facilitate infection through X4 variants. Although in the chronic stage SI variants reach 40%, they do not seem to be associated with either the CCR5 32 genotype or the number of CCL3L1 copies. In sum-mary, we have shown that SI X4 T-tropic variants of HIV-1 may be present in very early stages of viral infection suggesting that they may be transmitted from mother to child. In addition, the CCR5 32 genotype in the setting of a high number of CCL3L1 copies in an HIV-1 infected child contribute to a higher risk of being infected by SI variants in primary infection, however, this mechanism does not seem to occur in the chronic stage of viral infection.

Gene Expression Profiles in Genetically Different Mice Infected with Toxoplasma gondii: ALDH1A2, BEX2, EGR2, CCL3 and PLAU

Toxoplasma gondii can modulate host cell gene expression; however, determining gene expression levels in intermediate hosts after T. gondii infection is not known much. We selected 5 genes (ALDH1A2, BEX2, CCL3, EGR2 and PLAU) and compared the mRNA expression levels in the spleen, liver, lung and small intestine of genetically different mice infected with T. gondii. ALDH1A2 mRNA expressions of both mouse strains were markedly increased at day 1-4 postinfection (PI) and then decreased, and its expressions in the spleen and lung were significantly higher in C57BL/6 mice than those of BALB/c mice. BEX2 and CCR3 mRNA expressions of both mouse strains were significantly increased from day 7 PI and peaked at day 15-30 PI (P<0.05), especially high in the spleen liver or small intestine of C57BL/6 mice. EGR2 and PLAU mRNA expressions of both mouse strains were significantly increased after infection, especially high in the spleen and liver. However, their expression patterns were varied depending on the tissue and mouse strain. Taken together, T. gondii-susceptible C57BL/6 mice expressed higher levels of these 5 genes than did T. gondii-resistant BALB/c mice, particularly in the spleen and liver. And ALDH1A2 and PLAU expressions were increased acutely, whereas BEX2, CCL3 and EGR2 expressions were increased lately. Thus, these demonstrate that host genetic factors exert a strong impact on the expression of these 5 genes and their expression patterns were varied depending on the gene or tissue.

The study on the lowered tumorigenicity of hepatocellular carcinoma cells cotransfected with chemokine mMIP-1α and costimulatory molecule m4-1BB L in vivo / 中华消化杂志

Objective To investigate the effects of macrophage inflammatory protein-1α (MIP-1α) combined with molecule 4-1BB L on the tumorigenicity of hepatocellular carcinoma cells in vivo. Methods Mouse MIP-1α (mMIP-1α) expressed Hepa 1-6 cells were transfected with m4-1BBL recombinant retrovirus, the anti-histidinol cells clones were selected and amplified. The expression of m4-1BB L was confirmed by flow cytometry. The growth curve of Hepa 1-6 cells transfected with mMIP-1α and m4-1BBL alone or together was drawn and compared. C57B/L Mice were randomly divided into 7 groups, 9 mice in each group, injected with mMIP-1α+m4-1BB L Hepa 1-6 cells, m4-1BB L Hepa 1-6 cells, mMIP-1α Hepa 1-6 cells, Hepa 1-6 cells, pLXSHD Hepa 1-6 cells or PBS respectively. The tumorigenicity of hepatocellular carcinoma cells and the mice survival rate were compared between each groups. Results Hepa 1-6 mMIP-1α+m4-1BB L cells which expressed both mMIP-1α and m4-1BB L were successfully established. The expression of mMIP-1α and m4-1BB L alone or together did not affect the growth curve of Hepa 1-6 cells. Observed for 5 weeks, no tumor developed in Hepa 1-6 mMIP-1α+m4-1BB L injected mice. The tumorigenicity of Hepa 1-6 mMIP-1α+m4-1BB L was lower than that of Hepa 1-6 mMIP-1α or Hepa 1-6 m4-1BB L in vivo. The survival rate of Hepa 1-6 mMIP-1α+m4-1BBL injected mice(9/9) was higher than that of Hepa 1-6 m4-1BB L injected mice (6/9)or Hepa 1-6 mMIP-1α injected mice (1/9). Conclusion Chemokine MIP-1α combined with costimulatory 4-1BB L lowered the tumorigenicity of hepatocellular carcinoma cells in vivo, and prolonged the mice survival period.

Expression of chemokine receptor CCR5 in human follicular thyroid carcinoma / 第二军医大学学报

Objective: To study the expression of chemokine receptor CCR5 in follicular thyroid carcinoma and the serum level of CCR5 ligand, so as to assess the role of CCR5 in progression and metastasis of follicular thyroid carcinoma. Methods: Fifteen samples of follicular thyroid carcinoma, 17 samples of follicular thyroid adenoma and 12 adjacent normal samples were analyzed immunohistochemically for CCR5 expression. The sera concentrations of CCL3, CCL4 and CCL5 were measured by ELISA in all patients. Results: CCR5 was positive in follicular thyroid carcinoma samples, with the positive rate being 73.33%, and was not detected in the follicular thyroid adenoma and the normal samples (P<0.01). The concentrations of CCL3 and CCL5 in the sera of follicular thyroid carcinoma patients were significantly higher than those of the other 2 groups (P<0.05). Conclusion: CCR5 is highly expressed in follicular thyroid carcinoma tissues and the concentrations of CCL3 and CCL5 are obviously increased in the sera of patients, indicating that CCR5 may play an important role in the pathogenesis of follicular thyroid carcinoma.

The Role of the Peripheral Chemokine, CCL3, in Hyperalgesia following Peripheral Nerve Injury in the Rat / 대한통증학회지

BACKGROUND: Upregulation of one type of the pro-inflammatory chemokine (CCL2) and its receptor (CCR2) following peripheral nerve injury contributes to the induction of neuropathic pain. Here, we examined whether another type of chemokine (CCL3) is involved in neuropathic pain. METHODS: We measured changes in mechanical and thermal sensitivity in the hind paws of naive rats or rats with an L5 spinal nerve ligation (SNL) after intra-plantar injection of CCL3 or met-RANTES, an antagonist of the CCL3 receptor, CCR1. We also measured CCL3 levels in the sciatic nerve and the hind paw skin as well as CCR1 expression in dorsal root ganglion (DRG) cells from the lumbar spinal segments. RESULTS: Intra-plantar injection of CCL3 into the hind paw of naive rats mimicked L5 SNL-produced hyperalgesia. Intra-plantar injection of met-RANTES into the hind paw of rats with L5 SNL attenuated hyperalgesia. L5 SNL increased CCL3 levels in the sciatic nerve and the hind paw skin on the affected side. The number of CCR1-positive DRG cells in the lumbar segments was not changed following L5 SNL. CONCLUSIONS: Partial peripheral nerve injury increases local CCL3 levels along the degenerating axons during Wallerian degeneration. This CCL3 binds to its receptor, CCR1, located on adjacent uninjured afferents, presumably nociceptors, to induce hyperalgesia in the neuropathic pain state.

The Phospholipase-Protein Kinase C-MEK-ERK Pathway is Essential in Mycobacteria-induced CCL3 and CCL4 Expression in Human Monocytes

BACKGROUND: Little information is available on the identification and characterization of the upstream regulators of the signal transduction cascades for Mycobacterium tuberculosis (M. tbc)-induced ERK 1/2 activation and chemokine expression. We investigated the signaling mechanisms involved in expression of CCL3/MIP-1 and CCL4/MIP-1 in human primary monocytes infected with M. tbc. METHODS: MAP kinase phosphorylation was determined using western blot analysis with specific primary antibodies (ERK 1/2, and phospho-ERK1/2), and the upstream signaling pathways were further investigated using specific inhibitors. RESULTS: An avirulent strain, M. tbc H37Ra, induced greater and more sustained ERK 1/2 phosphorylation, and higher CCL3 and CCL4 production, than did M. tbc H37Rv. Specific inhibitors for mitogen-activated protein kinase (MAPK) kinase (MEK; U0126 and PD98059) significantly inhibited the expression of CCL3 and CCL4 in human monocytes. Mycobacteria-mediated expression of CCL3 and CCL4 was not inhibited by the Ras inhibitor manumycin A or the Raf-1 inhibitor GW 5074. On the other hand, phospholipase C (PLC) inhibitor (U73122) and protein kinase C (PKC)- specific inhibitors (GO6976 and Ro31-8220) significantly reduced M. tbc-induced activation of ERK 1/2 and chemokine synthesis. CONCLUSION: These results are the first to demonstrate that the PLC-PKC-MEK-ERK, not the Ras-Raf-MEK-ERK, pathway is the major signaling pathway inducing M. tbc-mediated CCL3 and CCL4 expression in human primary monocytes.

Expression of chemokine receptor CCR5 in human follicular thyroid carcinoma / 第二军医大学学报

Objective:To study the expression of chemokine receptor CCR5 in follicular thyroid carcinoma and the serum level of CCR5 ligand,so as to assess the role of CCR5 in progression and metastasis of follicular thyroid carcinoma.Methods:Fifteen samples of follicular thyroid carcinoma,17 samples of follicular thyroid adenoma and 12 adjacent normal samples were analyzed immunohistochemically for CCR5 expression.The sera concentrations of CCL3,CCL4 and CCL5 were measured by ELISA in all patients.Results:CCR5 was positive in follicular thyroid carcinoma samples,with the positive rate being 73.33%,and was not detected in the follicular thyroid adenoma and the normal samples(P