Evaluation of IL-4, MIP-1α, and MMP-9 gene expression levels in peri-implant tissues in peri-implantitis

Abstract This case-control study evaluated the gene expression levels of interleukin (IL)-4, macrophage inflammatory protein type 1 alpha (MIP-1α), and metalloproteinase (MMP)-9, factors involved in the formation of giant cells in healthy peri-implant tissue and peri-implantitis. Thirty-five subjects (15 healthy and 20 with peri-implantitis), who met the inclusion and exclusion criteria, were included in this study. The peri-implant tissue biopsies were subjected to total RNA extraction, DNAse treatment, and cDNA synthesis. Subsequently, the reaction of real-time PCR was performed to evaluate the gene expression levels of IL-4, MIP-1α, and MMP-9 concerning the reference gene. IL-4 gene expression showed higher (18-fold) values in the Peri-Implantitis Group of Patients when compared with the Healthy (Control) Group (p<0.0001). Although MIP- 1α and MMP-9 gene expression levels were higher in diseased implants, they showed no significant differences (p=0.06 and p=0.2337), respectively. Within the limitations of this study, the results showed that in tissues affected by peri-implantitis, only levels of Il-4 were increased when compared with tissues in the control group.

Resumo Este estudo caso-controle teve como objetivo avaliar a expressão gênica dos níveis de interleucina (IL)-4, proteína inflamatória de macrófagos tipo alfa 1 (MIP-1α) e metalopreoteinase (MMP)-9, todos fatores envolvidos na formação de células gigantes em tecidos peri-implantares saudáveis e com peri-implantite. Trinta e cinco indivíduos (15 saudáveis e 20 com peri-implantite) foram incluídos nesse estudo seguindo os critérios de inclusão e exclusão. Os tecidos peri-implantares foram submetidos a extração do RNA total, tratamento de DNAse e síntese de cDNA. Subsequentemente, a reação de PCR em tempo real foi realizada para avaliar os níveis da expressão de IL-4, MIP-1α, e MMP-9 em relação ao gene de referência. O nível de expressão de IL-4 foi estatisticamwente maior (18 vezes) nos tecidos de pacientes com peri-implantite quando comparados aos pacientes saudáveis (grupo controle) (p<0,0001). Embora os níveis de expressão de MIP- 1α e MMP-9 apresentassem maiores valores nos implantes doentes, esses níveis não foram estatisticamente significantes (p=0.06 and p=0.2337) respectivamente. Dentro das limitações desse estudo, os resultados mostraram que nos tecidos afetados pela peri-implantite, apenas os nívies de IL-4 estavam aumentados quando comparados ao grupo controle.

Effect of Jiedu Hugan Decoction on Protein Expression of MIP-2 and MIP-1β in Rats with Drug-induced Liver Injury / 中国实验方剂学杂志

Objective: To observe the effect and mechanism of Jiedu Hugan decoction on drug-induced liver injury in rats by detecting serum liver function, serum biomarkers, inflammatory factors, macrophage inflammatory protein-2 (MIP-2) and macrophage inflammatory protein-1β (MIP-1β). Method: The rat model of drug-induced liver injury was induced by acetaminophen (1 g·kg-1) orally once daily for 30 days. The sixty male adult Wistar rats were divided into five groups, control group,model group,administered silybin group(44.1 mg·kg-1), Jiedu Hugan decoction high, medium and low dose groups (63,31.5,15.75 g·kg-1), normal group and model group were given normal saline gavage, and the other groups were given corresponding liquid gavage for 30 days. After the experiment, the abdominal aorta separation take blood serum aspartate amino transferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), direct bilirubin (DBIL), malate dehydrogenase (MDH), enzyme for oxygen p1 (PON1) and glutamate dehydrogenase (GLDH), arginine (ARG), purine nucleotide phosphorylase (PNP), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) content. Pathological morphological changes of liver tissues were observed by hematoxylin-eosin (HE) staining. The protein expression of MIP-1β was observed by immunohistochemistry. The protein expression of MIP-2 was observed by single fluorescence immunohistochemistry, and the contents of TNF-α and IL-6 in liver homogenate were detected by enzyme-linked immunosorbent assay (ELISA). Result: Compared with normal group, levels of AST, ALT, DBIL, PON1, ARG, GLDH, MDH, PNP and TNF-α in model group were significantly increased (PPPPα in liver injury rats(PPβ protein expression, detoxification protect liver soup effect of the optimal dose group, the pathological morphology of liver cell dosage group were with different degree of protection. Conclusion: The effect of Jiedu Hugan decoction in medium dose group is better, and its mechanism may affect the chemotaxis of neutrophils induced by MIP-2 and MIP-1β by reducing the content of TNF-α, thus inhibiting the release of inflammatory factors and preventing inflammation.

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

Clinical Significance of MIP-1α and VEGF Levels in Serum and CSF of ALS Patients / 昆明医科大学学报

Objective Through the detection of MIP-1α and VEGF levels in serum and CSF of ALS patients, we evaluated the clinical value of MIP-1 and VEGF levels in ALS patients. Methods A total of 80 patients with ALS were collected from Kuangwu Hospital of Tongchuanin from Jan, 2012 to Jun, 2015, and 67 patients with non-inflammation neurological diseases were chosen as control group. We obtained CSF and serum samples,and the MIP-1α and VEGF levels were measured by ELISA method. Results The MIP-1α levels in serum and CSF of ALS patients were statistically and significantly higher than those in the control group (P＜0.05) . The VEGF levels in serum of ALS patients were statistically and significantly higher than those in the control group (P＜0.05).The VEGF levels in CSF of ALS patients were not higher than those in the control group,statistically insignificant (P＞0.05).MIP-1 alpha and VEGF levels was positively correlated with ALS course. The MIP-1αand VEGF levels of the long duration group were greater than the short duration group (P＜0.05) . Conclusion The rising of MIP-1α and VEGF may indicate an activation of compensatory responses in ALS which suggested that MIP-1 alpha and VEGF are involved in the pathophysiology of amyotrophic lateral sclerosis. We propose that MIP-1α and VEGF may be a useful biomarker witha prognostic and evaluating potential for ALS.

Expression and variation of MIP-1β, MIP-2, and IL-12p70 in mouse models with bloodstream infection caused by different bacteria / 中国感染控制杂志

Objective To investigate the expression and variation of MIP 1β,MIP-2,and IL-12p70 in mice with bloodstream infection caused by 4 kinds of bacteria.Methods CD-1 (ICR) mouse models of bloodstream infection with Staphylococcus aureus (S.aureus),Enterococcus f aecalis (E.f aecalis),Escherichia coli (E.coli),and K lebsiella pneumoniae (K.pneumoniae) were established.After mice in each trial group and PBS control group were infected by bacteria for 0.5h,1h,3h,6h,12h,24h,and 48h,concentrations of MIP-1β,MIP-2,and IL-12p70 were detected by Luminex liquid suspension chip system.Results Concentrations of MIP-1β increased significantly 1h after bacteria was in blood,S.aureus,E.faecalis,E.coli,K.pneumoniae,and control groups were (134.5 ± 18.3),(61.5 ± 15.4),(3 354.0 ±809.0),(6 888.4 ± 1 100.2),and (28.9 ± 4.6) pg/mL respectively;the peak values of IL-12p70 were (389.3 ± 118.1),(127.6 ± 10.0),(42.2 ± 3.5),(62.8 ± 8.4),and (4.8 ± 0.3) pg/mL respectively.Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were significantly higher than other trial groups and control group (all P＜0.01),while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than E.coli,K.pneumoniae,and control groups (all P＜0.01).Conclusion Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were both significantly higher than those in S.aureus and E.faecalis groups,while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than those in E.coli and K.pneumoniae groups.The combination detection of multiple cytokines or chemokines are valuable in predicting gram-positive or gram-negative bacterial infection,and can provide basis for treatment of early infection.

The brain edema and Mip1 expression of perihematoma after intracerebral hemorrhage in rat model / 天津医药

Objective To explore the protective effect of Mip1 on cerebral edema by observing the Mip1 expression of perihematoma in intracerebral hemorrhage (ICH) rat model. Methods Ninety male SD rats were randomly divided into sham-operated group (n=15) and ICH group (n=75). ICH group was equally subdivided into 6 h, 12 h, 1 d, 3 d, and 7 d groups according to cerebral hemorrhage time, 15 in each group. Hematoma was formed by infusing autologous femoral artery blood into the right caudate nucleus. Rats were killed at the above time points, brain tissues around hematoma were taked. The pathological morphological changes in brain tissue around hematoma were observed using HE staining. The water content of brain (BWC) was measured with dry/wet weight method, and immunohistochemistry was used to analyze the average optical density value of Mip1 protein in perihematoma. Results Neuronal cell body of perihematoma was significantly decreased after ICH, the cytoplasm light staining, Nissl body reduced obviously in cytoplasm, and edema fissure appeared around cells, however, no obvious changes of nerve cells were detected in sham-operated group. Brain edema gradually increased at 6 h, and reached peak at 3 d (all P0.05). A large amount of brown Mip1 positive cells were found to distribute in cytoplasm and nucleus of perihematoma after intracerebral hemorrhage. The Mip1 protein expression level was higher in 6 h after intracerebral hemorrhage than that in sham-operated group (all P0.05). Conclusion The high expression of Mip1 after ICH is in line with the trend of the brain edema development, and shows that the activation of Mip1 may participate in the pathological process of hemorrhagic cerebral edema.

Effect of naftopidil on levels of MIP-2 and MIP-1αin serum and succus prostaticus in the treatment of prostatitis in elderly / 中国生化药物杂志

Objective To investigate the effect of naftopidil in the treatment of prostatitis in the elderly on MIP-2 and MIP-1αin serum and succus prostaticus.Methods 78 elderly patients with chronic prostatitis were randomly divided into the control group and the observation group, 39 cases in each group.The control group were treated with Diosmin routine treatment, the observation group were treated on the basis of the control group combined with naftopidil tablets in the treatment,2 groups were treated continuous for 8 weeks.MIP-2 and MIP-1αin serum and prostatic fluid before and after treatment were compared, and the maximum urine flow rate and clinical efficacy were compared after the treatment.Results compared with pre-treatment, MIP-2, MIP-1αin serum and prostate fluid in 2 groups after treatment decreased, NIH-CPSI and QOL scores in 2 groups decreased, the maximum urinary flow rate increased (P<0.05);compared with the control group, the levels of MIP-2, MIP-1αin the serum and prostatic fluid, NIH-CPSI and QOL of observation group were lower, and the maximum urine flow rate was higher (P<0.05);Compared with the control group, the total efficiency of the observation group was higher (P<0.05).Conclusion Naftopidil can significantly reduce the elderly patients with chronic non bacterial prostatitis and serum of patients with prostatic fluid MIP-2, MIP-1αlevel and improve pain in urination, dysuria symptoms.

Evaluation of immunostimulatory activity of Chyawanprash using in vitro assays.

Chyawanprash is an ayurvedic formulation used in Indian traditional medicinal system for its beneficial effect on human health. We investigated the immunostimulatory effects of Chyawanprash (CHY) using in vitro assays evaluating the secretion of cytokines such as Tumor Necrosis Factor-alpha (TNF-α), Interleukin-1beta (IL-1β) and Macrophage Inflammatory Protein-1-alpha (MIP-1-α) from murine bone marrow derived Dendritic Cells (DC) which play pivotal role in immunostimulation. The effects of CHY on phagocytosis in murine macrophages (RAW264.7) and Natural Killer (NK) cell activity were also investigated. At non-cytotoxic concentrations (20–500 µg/ml), CHY enhanced the secretion of all the three cytokines from DC. CHY also stimulated both, macrophage (RAW264.7) as well as NK cell activity, in vitro. In conclusion, the data substantiates the immunoprotective role of CHY at cellular level mediated by immunostimulation in key immune cells viz. dendritic Cells, macrophages and NK cells.

Effect of diosmin on immune mediated factors in serum and prostatic fluid and clinical effect in patients with chronic prostatitis / 中国生化药物杂志

Objective To explore effect of diosmin on serum and prostatic fluid immune mediated factors and clinical effect in patients with chronic prostatitis.Methods 89 cases of old male patients with chronic non bacterial prostatitis were selected, and divided into two groups.The control group were treated with universal tablets, the experiment group were treated on the base of the control group with diosmin.12 weeks as a course.Clinic effect, adverse reaction rate, prostatic fluid MIP-2 and MIP-1αwere compared after treatment.Results Compared with control group, MIP-1αand MIP-2 in serum and prostatic fluid were lower(P<0.05),NIH-CPSI score and QOL scores were lower(P<0.05), total effective rate was higher than control group(P<0.05),the incidence of adverse reactions was lower(P<0.05).Conclusion Diosmin can reduce MIP-1α, MIP-2 in serum and prostate fluid of patients with chronic none bacterial prostatitis, and improve the pelvic pain, dysuria, sexual dysfunction and other symptoms, and has less adverse reaction.

Tuftsin-derivated TP affects MIP-1αexpression of tumor associated macrophages / 中国药理学通报

Aim To investigate the effect of TP on the expression of macrophages inflammatory protein ( MIP-1α) . Methods Total RNA of mouse Ana-1 cells and tumor associated macrophages were extracted, and MIP-1α mRNA was detected by RT-PCR. Mouse S180-xenografts were established by injecting S180 cells subcutaneously into the double abdominal flanks of the mice. The postoperative residual tumor models were generated in the right abdominal tumors when tumors grew into 250 mm3 . Animals were treated with TP or CTX, and tumor tissues were separated and MIP-1α was detected by immunohistochemistry. Results There was no significant difference of the expression of MIP-1α between Ana-1 cells and TAMs. TP couldn’ t affect MIP-1αexpression in Ana-1 cells while it signifi-cantly decrease MIP-1α expression in TAMs in a dose-dependent manner. TP significantly decreased MIP-1αexpression of tumor tissue compared with control group. Conclusions MIP-1α will be a new target of TP anti-cancer. Simple cell line tests in vitro couldn’ t reveal the real state in vivo.

Expression of resistin, CXCR3, IP-10, CCR5 and MIP-1α in obese patients with different severity of asthma

Asthma studies suggest that alteration in the inflammation pattern may be associated with the severity of asthma. The aim of this study was to compare in vitro the expression of chemokines, chemokine receptors and cytokine production from CD4+ T human lymphocytes of asthmatic, both obese and non-obese patients with different severity levels of asthma. Lymphocytes were labeled with monoclonal anti-human CXCR3/IP-10, MIP-1a/CCR5 antibodies and were analyzed by flow cytometry. Cell culture supernatants were used to measure production of interleukin IL-6 and resistin by ELISA. CXCR3/IP-10 expression increased in non-obese patients with mild persistent asthma (2.2%, p<0.05), moderate persistent asthma (3%, p<0.003) and severe persistent asthma (4%, p<0.004); this effect was stronger in obese patients with severe persistent asthma (35%, p<0.004). MIP-1 α / CCR5 increased in non-obese patients with intermittent asthma (0.65%, p<0.05) and severe asthma (1.4%, p<0.03); in obese patients, this expression was greater in intermittent asthma (8%, p<0.05) and severe persistent asthma (12%, p<0.04). Resistin production strongly increased in obese patients with intermittent (976 ng/ml) and severe persistent asthma (795 ng/ml). IL-6 increased in both lean and obese persons; however, the highest value was registered in the group of severe persistent obese asthmatics (992 pg/ml). Obesity per se increased the inflammatory profile of chemokines / cytokines secreted by cells of the blood, increasing the inflammatory status in asthmatic patients. Resistin showed characteristics of a pro-inflammatory cytokine mainly in severely obese asthmatics.

Study on inducing immune response by cotransfection of rat breast cancer cells with MIP-1α and B7-1 / 重庆医学

Objective To investigate the ability of immune response of cotransfection of rat breast cancer cells with MIP-1αand B7-1 .Methods Tumor growth was observed every day after the SHZ-88 breast cancer cells were inoculated in the rats .The tumor-bearing rats were randomly divided into control group ,MIP-1αgroup ,B7-1 group and combination group ,which were injected with corresponding transgenic cells .The change of tumor volume in 1~4 weeks and the survival time were observed and compared .The peripheral blood was collected .CD4+ ,CD8+ percentage and CD4+ /CD8+ ratio and the levels of IL-2 and IFN-γof the peripheral blood were detected and compared .Results The tumorigenic rate were 100% ,20% ,30% and 0% after the SD rats were inoculated with SHZ-88 ,SHZ-88/MIP-1α,SHZ-88/B7-1 and SHZ-88/MIP-1α+B7-1 cells for 10 days .After the transgenic therapy ,the tumor volume of the combination group in 1~4 weeks were significantly smaller than the other groups .The survival time of the combina-tion group was significantly longer than the other groups .The CD4+ ,CD8+ percentage and CD4+ /CD8+ ratio and the levels of IL-2 and IFN-γof peripheral blood in the combination group were significantly higher than the other groups .The differences above were all statistically significant (P<0 .05) .Conclusion Cotransfection of MIP-1αand B7-1 could induce immune response in breast cancer rats and play anti-cancer effects .

The Cholesterol-Binding Antibiotic Nystatin Induces Expression of Macrophage Inflammatory Protein-1 in Macrophages

Nystatin, a polyene antifungal antibiotic, is a cholesterol sequestering agent. The antifungal agent alters composition of the plasma membrane of eukaryotic cells, whereas its effects on cells are poorly investigated. In the current study, we investigated the question of whether nystatin was able to induce expression of macrophage inflammatory protein-1 (MIP-1). THP-1 cells rarely express MIP-1alpha and MIP-1beta, however, upon exposure to nystatin, significantly elevated expression of MIP-1alpha and MIP-1beta was observed in a dose-dependent fashion at the messenger and protein levels. Cellular factors activated by nystatin as well as involved in nystatin-induced expression of MIP-1 proteins were identified in order to understand the molecular mechanisms of action of the anti-fungal agent. Treatment with nystatin resulted in enhanced phosphorylation of Akt, ERK, p38 MAPK, and JNK. Abrogation or significant attenuation of nystatin-induced expression of MIP-1alpha and MIP-1beta was observed by treatment with Akt inhibitor IV, LY294002, and SP6001250. Inhibition of ERK or p38MAPK using U0126 and SB202190 did not lead to attenuation of MIP-1 expression. In addition, inhibitors of protein kinase C, such as GF109203X and Ro-318220, also attenuated expression of MIP-1. These results indicate that nystatin is able to activate multiple cellular kinases and, among them, Akt and JNK play primary roles in nystatin-induced expression of MIP-1 proteins.

An immunohistochemical study on the time-dependent expression of MIP-1?during healing of skin incised wound in mice / 重庆医科大学学报

Objective:To explore the expression of MIP-1? during healing of skin incised wound and the applicability of time-dependent expressions of MIP-1? to determination of wound age. Methods:An incised wound was made in the dorsal skin of mouse. Immunohistochemical and image-analysis techniques were employed in vital skin wounds 1h~14d after incision and postmortem incision 0.5~6h after death, and the non-incised mouse skin was used as contro1.Results:In the vital skin incisions,expression of MIP-1? began at 6h after incision,which increased subsequently,and peaked at 3d.The quantity of MIP-1?expression decreased and minimized in the specimens aged 14 days. In the non-wounded groups and postmortem incision groups MIP-1? was only detected as weak expression in the epidermic cells,sweat gland cells and endothelial cells. Conclusion:The time-dependent expression of MIP-1?during healing of skin incised wound may be used as a useful reference marker.

Influence of MIP-1 Alpha on the CD4+ Th Lymphocytes

PURPOSE: MIP-1 alpha, initially identified as a substance to promote inflammation, was recently discovered to also suppress proliferation of many kinds of cells. In this study, the influence of MIP-1 alpha on CD4+ Th cells to secrete cytokines (IL-10, IL-2, IFN-gamma) and to express CD25 molecules was investigated. METHODS: Peripheral blood mononuclear cells (PBMCs) were prepared from five normal volunteers and initially divided into 4 groups: IL-10, IL-2, IFN-gamma, and CD25. Each group was further divided into 4 subgroups according to the incubation with or without MIP-1 alpha and to the incubation for 30 minutes or 3 hours. Analysis was performed by flow cytometry. RESULTS: Incubation of CD4+ Th lymphocytes with MIP-1 alpha showed a tendency to increase Th1 cytokine (IL-2, IFN-gamma) secretion and to decrease Th2 cytokine (IL-10) release, but there was no significant difference between any of the experimental groups. Among the CD4+ Th lymphocyte groups cultured with MIP-1 alpha, the expression of CD25 was significantly lower in the 3-hour incubation group than in the 30-minute group (P=0.008). CONCLUSION: MIP-1 alpha may play a role in facilitating immune response by increasing Th1 and decreasing Th2 cytokine secretion from CD4+ Th cells, and also by decreasing the proportion of CD4+CD25+ Th cells in the peripheral blood. However, in vivo study is necessary to verify the function of MIP-1 alpha in the living body.

Expression of MIP-1alpha mRNA in Kupffer Cells and Serum MIP-1alpha Post Portal Vein Transfusion / 대한이식학회지

PURPOSE: Portal vein transfusion (PVT) has been known to induce immunosuppression or tolerance and Kupffer cell was identified to play an important role in the phenomenon. The purposes of this study were investigating PVT effect on gene regulation in Kupffer cells and subsequent change in serum cytokine. METHODS: For investigating the effect of PVT, Kupffer cells were isolated from the mice (BalbC) of six groups; 1 hour sham operation (S), 1 hour portal vein saline injection (PVS), 1 hour PVT, 24 hour S, 24 hour PVS, and 24 hour PVT groups. Each group was composed of 3 mice. Total RNAs isolated from Kupffer cells were subjected to RT-PCR differential display. The bands of 24 hour group showing increased expression was cloned for the sequencing analysis. RESULTS: Macrophage inflammatory protein 1 alpha (MIP-1alpha) was identified from the bands of increased expression. In PVT groups, increased expression of MIP-1alpha mRNA in Kupffer cells coincided with elevated serum level of MIP-1alpha. CONCLUSION: MIP-1alpha may be one of the important cytokines involved in PVT induced immunosuppression or tolerance.

Expression of Monocyte Chemotactic Protein-1 and Macrophage Inflammatory Protein-1alpha in Intraabdominal Abscess Induced by Bacteroides fragilis Infection

OBJECTIVE: Bacteroides fragilis is the most frequently isolated anaerobes in tissue of intraabdominal infection, particularly in intraabdominal sepsis or abscess. In acute experimental model with an intraabdominal infection, the response to B. fragilis is characterized by infiltration of neutrophils and monocytes. To understand the pathogenesis of B. fragilis infection, it is important to explore the mechanism for inflammatory signals such as chemokines induced by this bacteria. The goal of this study was to determine whether peritoneal monocytes or fibroblasts surrounding with intraabdominal abscess induced by B. fragilis could express chemokines such as monocyte chemotactic protein-1 (MCP-1) or macrophage inflammatory protein-1a (MIP-1a). METHODS: 1) After C57BL/6 mice were intraperitoneally inoculated with abscess-inducing agents containing B. fragilis, RNA was extracted from the intraperitoneal tissues of the mice using Ottawa sand and the guanidinium thiocyanate-phenol-chloroform method in 3 days later. 2) After C57BL/6 mouse peritoneal monocytes were infected with B. fragilis for 1, 4 and 9 hours, cellular RNA was extracted from the cells. 3) Fibroblasts isolated from intraabdominal abscess nodules induced by B. fragilis infection were growth in tissue culture for 3 to 4 weeks. After the fibroblasts were stimulated with IL-1alpha (0.1-10 ng/ml) or TNFalpha (0.1-10 ng/ml) for 24 hours, total cellular RNA was extracted. MCP-1 or MIP-1alpha mRNA expression was assessed using RTPCR. MCP-1 or MIP-1alpha proteins in cluture supernatants or tissue extracts were also measured by ELISA. RESULTS: 1) MCP-1 or MIP-1alpha mRNA was highly expressed in peritoneal tissue of C57BL/6 mice bearing with intraabdominal abscess induced by B. fragilis. 2) Expression of MCP-1 mRNA increased at 9 hours in mouse peritoneal monocytes infected with B. fragilis. MIP-1alpha mRNA was initially expressed and perisisted in the monocytes infected with B. fragilis for 9 hours. MCP-1 or MIP-1alpha proteins was also parallel to the expression of those chemokines. 3) The fibroblasts isolated from intraabdominal abscess nodules by B. fragilis infection constitutively expressed MCP-1, MIP-1alpha, production in the fibroblasts was significantly upregulated in response to proinflammatory cytokines produced in the monocytes, including IL-1alpha and TNFalpha, but MCP-1 production were not. The normal fibroblasts from uninfected mice didnot show significant production of MCP-1 or MIP-1a in response to IL-1a or TNFalpha. CONCLUSION: These results suggest that peritoneal monocytes and fibroblasts surrounding with abscesses induced by B. fragilis produce MCP-1 or MIP-1a. Furthermore, it could be extrapolated that those effects may play a role in the formation of intraabdominal abscess nodules.

Chemokine Secretion From Alveolar Macrophages in Patientswith Diffuse Interstitial Lung Diseases(DILD) / 결핵

BACKGROUND: The type of the infiltrating cells in alveolitis may be determined by the chemokines in the lesion. MIP-1alpha, a C-C type chemokine, stimulates proliferation and cytokine secretion from macrophages and induces early neutrophilic and later monocytic inflammation in vivo. IL-8, a C-X-C type chemokine is known to attract neutrophils and T-lymphocytes. This study is performed to find out the relative role of two different chemokines in diffuse interstitial lung disease. SUBJECT AND METHOD: We measured the secretion of MIP-1alpha and IL-8 from alveolar macrophages(AM), and their level in BAL fluid of 26 patients with DILD (10 IPF, 4 collagen disease, 10 sarcoidosis, and 2 hypersensitivity pneumonitis) and 7 normal control. RESULT: IL-8 secretion was significantly increased in patients with DILD (8.15+/-4.58 ng/ml) than in normal (1.10+/-0.93 ng/ml, p=0.0003). Significant correlation was found between IL-8 secretion and total cell number in BAL fluid (r=0.484, p=0.0068), %(r=0.592, p=0.0004) and No. (r=0.516, p=0.0042) of lymphocyte, and % of AM (r=-0.505, 0.0032). MIP-1alpha secretion was also increased in DILD (2.41+/-1.45 ng/ml) compared to control (0.63+/-0.30 ng/ml, p=0.0031), and showed a tendency of correlation with total cell number (r=0.368, p=0.0456) and No. of alveolar macrophages (r=0.356, p=0.0579) in BAL fluid. The concentration of IL-8 in BAL fluid was significantly increased in the patients with DILD (40.4+/-34.5 pg/ml) compared to control (3.90 +/- 2.47 pg/ml, p=0.0094) and it showed a significant correlation with the total cell number(r=0.484, p=0.0068), %(r=-0.505, p=0.0032) of AM, and % (r=0.592, p=0.0004) and No. (r=0.516, p=0.0042) of lymphocyte in BAL fluid. But there was a no significant difference in MIP-1alpha concentration in BAL fluid between normal control group and the patients with DILD. Conclusion: From the above results, we concluded that AM of DILD releases increased amount of both IL-8 and MIP-1alpha but IL-8 has better correlation with the type of alveolitis.

Establishment of a mouse hepatocellular carcinoma cell line producing mMIP-1? chemokines and the tomorigenicity of mMIP-1? transfected Hepa1-6 / 第二军医大学学报

Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP 1? and to evaluate the possibility of cancer gene therapy by mMIP 1?. Methods: mMIP 1? cDNA was cloned into retrovirus vector pBabe puro and pBabe puro mMIP 1? was constructed, then pBabe puro mMIP 1? was used to transfect packaging cells, anti puromycin cells was proliferated, the supernatant was used to infect hepa1 6, the anti puromycin clone (hepa1 6 mMIP 1?) and hepa1 6 were analysed for the expression of mMIP 1? mRNA and protein by RT PCR and immunohistochemistry respectively. The growth curve of hepa1 6 and hepa1 6 mMIP 1? was drawn. The chemotaxis of mMIP 1? produced by hepa1 6 mMIP 1? to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro mMIP 1? with mMIP 1? cDNA was constructed. Hepa1 6 did not produce mMIP 1? mRNA and protein, while hepa1 6 mMIP 1? could produce mMIP 1? mRNA and protein. The growth curve of hepa1 6 and hepa1 6 mMIP 1? showed no difference. The chemotaxis of mMIP 1? produced by hepa1 6 mMIP 1? to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1 6 mMIP 1? is established and mMIP 1? can affect the tumorigenecity of hepa1 6.