A Rat Model of Rheumatoid Arthritis: TDZD-8 is Associated with the Protection Against the Induction of the Synovium Knee Joint IL-17A/GSK3β/ROS/α-SMA Axis of Fibrosis

SUMMARY: Rheumatoid arthritis (RA) that affects the synovial knee joint causes swelling of the synovial membrane and tissue damage. Interleukin-17A (IL-17A) and the enzyme glycogen synthase kinase-3β (GSK3β) are involved in the pathogenesis of RA. The link between IL-17A, GSK3β, the oxidative stress, and the profibrogenic marker alpha-smooth muscle actin (α-SMA) with and without TDZD-8, GSK3β inhibitor has not been studied before. Consequently, active immunization of rats was performed to induce RA after three weeks using collagen type II (COII) injections. The treated group received daily injection of 1 mg/kg TDZD-8 for 21 days following the immunization protocol (COII+TDZD-8). Blood and synovium tissue samples were harvested at the end of the experiment. RA development was confirmed as corroborated by a substantial increase in blood levels of the highly specific autoantibody for RA, anti-citrullinated protein antibody as well as augmentation of reactive oxidative species (ROS) levels measured as lipid peroxidation. RA induction also increased synovium tissue levels of IL-17A and the profibrogenic marker, α-SMA. All these parameters seemed to be significantly (p<0.0001) ameliorated by TDZD-8. Additionally, a significant correlation between IL-17A, ROS, and α-SMA and biomarkers of RA was observed. Thus, knee joint synovium RA induction augmented IL-17A/GSK3β/ROS/α-SMA axis mediated arthritis in a rat model of RA, which was inhibited by TDZD-8.

La artritis reumatoide (AR) que afecta la articulación sinovial de la rodilla provoca inflamación de la membrana sinovial y daño tisular. La interleucina-17A (IL-17A) y la enzima glucógeno sintasa quinasa-3β (GSK3β) están involucradas en la patogenia de la AR. No se ha estudiadol vínculo entre IL-17A, GSK3β, el estrés oxidativo y el marcador profibrogénico actina de músculo liso alfa (α-SMA) con y sin inhibidor de TDZD-8, GSK3β. En consecuencia, se realizó una inmunización activa de ratas para inducir la AR después de tres semanas usando inyecciones de colágeno tipo II (COII). El grupo tratado recibió una inyección diaria de 1 µg/ kg de TDZD-8 durante 21 días siguiendo el protocolo de inmunización (COII+TDZD-8). Se recogieron muestras de sangre y tejido sinovial al final del experimento. El desarrollo de AR se confirmó como lo corroboró el aumento sustancial en los niveles sanguíneos del autoanticuerpo altamente específico para AR, el anticuerpo antiproteína citrulinada, así como el aumento de los niveles de especies oxidativas reactivas (ROS) medidos como peroxidación lipídica. La inducción de AR también aumentó los niveles de tejido sinovial de IL-17A y el marcador profibrogénico, α-SMA. Todos estos parámetros parecían mejorar significativamente (p<0,0001) con TDZD-8. Además, se observó una correlación significativa entre IL- 17A, ROS y α-SMA y biomarcadores de AR. Por lo tanto, la inducción de AR en la sinovial de la articulación de la rodilla aumentó la artritis mediada por el eje IL-17A/GSK3β/ROS/α-SMA en un modelo de rata de AR, que fue inhibida por TDZD-8.

A rat model of rheumatoid arthritis: TDZD-8 is associated with the protection against the induction of the synovium knee joint IL-17A/GSK3beta/ROS/alpha-SMA axis of fibrosis / Un modelo de rata de artritis reumatoide: TDZD-8 está asociado con la protección contra la inducción del eje de fibrosis de la articulación sinovial de la rodilla IL-17A/GSK3ß/ROS/α-SMA

SUMMARY: Rheumatoid arthritis (RA) that affects the synovial knee joint causes swelling of the synovial membrane and tissue damage. Interleukin-17A (IL-17A) and the enzyme glycogen synthase kinase-3β (GSK3β) are involved in the pathogenesis of RA. The link between IL-17A, GSK3β, the oxidative stress, and the profibrogenic marker alpha-smooth muscle actin (α-SMA) with and without TDZD-8, GSK3β inhibitor has not been studied before. Consequently, active immunization of rats was performed to induce RA after three weeks using collagen type II (COII) injections. The treated group received daily injection of 1 mg/kg TDZD-8 for 21 days following the immunization protocol (COII+TDZD-8). Blood and synovium tissue samples were harvested at the end of the experiment. RA development was confirmed as corroborated by a substantial increase in blood levels of the highly specific autoantibody for RA, anti-citrullinated protein antibody as well as augmentation of reactive oxidative species (ROS) levels measured as lipid peroxidation. RA induction also increased synovium tissue levels of IL-17A and the profibrogenic marker, α-SMA. All these parameters seemed to be significantly (p<0.0001) ameliorated by TDZD-8. Additionally, a significant correlation between IL-17A, ROS, and α-SMA and biomarkers of RA was observed. Thus, knee joint synovium RA induction augmented IL-17A/GSK3β/ROS/α-SMA axis mediated arthritis in a rat model of RA, which was inhibited by TDZD-8.

La artritis reumatoide (AR) que afecta la articulación sinovial de la rodilla provoca inflamación de la membrana sinovial y daño tisular. La interleucina-17A (IL-17A) y la enzima glucógeno sintasa quinasa-3β (GSK3β) están involucradas en la patogenia de la AR. No se ha estudiadol vínculo entre IL-17A, GSK3β, el estrés oxidativo y el marcador profibrogénico actina de músculo liso alfa (α-SMA) con y sin inhibidor de TDZD-8, GSK3β. En consecuencia, se realizó una inmunización activa de ratas para inducir la AR después de tres semanas usando inyecciones de colágeno tipo II (COII). El grupo tratado recibió una inyección diaria de 1 µg/ kg de TDZD-8 durante 21 días siguiendo el protocolo de inmunización (COII+TDZD-8). Se recogieron muestras de sangre y tejido sinovial al final del experimento. El desarrollo de AR se confirmó como lo corroboró el aumento sustancial en los niveles sanguíneos del autoanticuerpo altamente específico para AR, el anticuerpo antiproteína citrulinada, así como el aumento de los niveles de especies oxidativas reactivas (ROS) medidos como peroxidación lipídica. La inducción de AR también aumentó los niveles de tejido sinovial de IL-17A y el marcador profibrogénico, α-SMA. Todos estos parámetros parecían mejorar significativamente (p<0,0001) con TDZD-8. Además, se observó una correlación significativa entre IL- 17A, ROS y α-SMA y biomarcadores de AR. Por lo tanto, la inducción de AR en la sinovial de la articulación de la rodilla aumentó la artritis mediada por el eje IL-17A/GSK3β/ROS/α-SMA en un modelo de rata de AR, que fue inhibida por TDZD-8.

Secukinumab: A Silver Lining in Dermatology

Secukinumab, a fully human monoclonal antibody, is a biological agent that targets interleukin-17A. Secukinumab is used in the management of the common dermatological entity - plaque-type psoriasis and its various types, namely psoriatic arthritis, hypertrophic palmoplantar psoriasis and generalized pustular psoriasis. Other less common indications of this popular interleukin -17A inhibitor, secukinumab include ankylosing spondylitis, rheumatoid arthritis, systemic lupus erythematosus, Familial Mediterranean fever, and tumor necrosis factor receptor-associated periodic syndrome (TRAPS). This review article was written by referring to various review articles, original articles, and some books related to highly regarded databases, such as the Web of Science, PubMed, and Scopus. The keywords explored during review of literature consisted of combinations of the following words: human monoclonal antibody, IL-17A, and biologicals. The authors with this in-depth review hope to explore the working of this versatile biological, Secukinumab, especially as a silver lining in dermatology

A novel PI3K inhibitor XH30 suppresses orthotopic glioblastoma and brain metastasis in mice models

Glioblastoma is carcinogenesis of glial cells in central nervous system and has the highest incidence among primary brain tumors. Brain metastasis, such as breast cancer and lung cancer, also leads to high mortality. The available medicines are limited due to blood-brain barrier. Abnormal activation of phosphatidylinositol 3-kinases (PI3K) signaling pathway is prevalent in glioblastoma and metastatic tumors. Here, we characterized a 2-amino-4-methylquinazoline derivative XH30 as a potent PI3K inhibitor with excellent anti-tumor activity against human glioblastoma. XH30 significantly repressed the proliferation of various brain cancer cells and decreased the phosphorylation of key proteins of PI3K signaling pathway, induced cell cycle arrest in G1 phase as well. Additionally, XH30 inhibited the migration of glioma cells and blocked the activation of PI3K pathway by interleukin-17A (IL-17A), which increased the migration of U87MG. Oral administration of XH30 significantly suppressed the tumor growth in both subcutaneous and orthotopic tumor models. XH30 also repressed tumor growth in brain metastasis models of lung cancers. Moreover, XH30 reduced IL-17A and its receptor IL-17RA in vivo. These results indicate that XH30 might be a potential therapeutic drug candidate for glioblastoma migration and brain metastasis.

Role of C5a/C5aR1 pathway in imiquimod-induced chronic psoriasis skin inflammation and the underlying mechanism / 中华微生物学和免疫学杂志

Objective:To establish a mouse model of imiquimod (IMQ)-induced chronic psoriasis skin inflammation and to investigate the role of C5a/C5aR1 pathway in this process and the underlying mechanism.Methods:BALB/c mice were treated with IMQ cream or Vasline cream (control group) every other day for eight times to mimic the chronic skin inflammation. Psoriatic skin inflammation and pathological changes in the wild-type (C5aR1 + /+ ) mice and the mice with C5aR1 gene deletion (C5aR1 -/-) were monitored and analyzed. Epidermal proliferation (Ki67), keratin 6/14 expression and neutrophil infiltration in the skin lesions were observed with immunohistochemical (IHC) staining. qRCR was used to detect the expression of keratin 6/14 and related inflammatory cytokines. Flow cytometry was performed to measure the percentages of leukocytes, CD3 + T cells and IL-17A + γδTCR + T cells in local skin samples as well as IL-17A responses in draining lymph nodes. Results:IMQ treatment induced typical psoriatic skin inflammation, including scaling, erythema and thickness. Dysregulated epidermal proliferation, acanthosis, micro-abscesses, inflammatory cell infiltration and abnormal hyperplasia of dermal capillaries were observed after HE staining. Compared with the C5aR1 + /+ mice, the C5aR1 -/- mice showed significantly attenuated chronic skin inflammation, evidenced by decreased epidermal proliferation, down-regulated keratin 6/14 expression and alleviated neutrophil infiltration. Results of qPCR also indicated decreased expression of keratin 6/14, inflammatory cytokines (IFN-γ, MIP-1α, IL-1β and TNF-α) and IL-17-related cytokines (IL-6, IL-17A and IL-23) in skin samples of C5aR1 -/- mice. Moreover, the infiltration of leukocytes (CD45), CD3 + T cells and IL-17A + γδTCR + T cells in skin lesions as well as the percentages of IL-17A + , IL-17A + CD3 + T and IL-17A + γδTCR + T cells in draining lymph nodes were also decreased in C5aR1 -/- mice. Conclusions:This study suggested that IMQ treatment could induce chronic psoriasis skin inflammation in mice. C5a/C5aR1 signaling pathway mediates IMQ induced chronic skin inflammation via activating IL-17 producing cells. Targeting C5a/C5aR pathway would be a new strategy for the management of psoriasis.

Potentiality of curcumin on ISHAK scoring system and the expressions of BAX, IL-17A, and EGF in the treatment of Schistosoma mansoni infection using Swiss albino mice

@#The present study evaluated the antiparasitic effect of curcumin extract on Schistosoma mansoni in Swiss albino mice. The experimental design included four groups of S. mansoniinfected mice; without treatment (controls), curcumin-treated, Praziquantel (PZQ)-treated, and PZQ +curcumin treated mice. The results showed that curcumin improved ISHAK confluent necrosis score up to zero. PZQ +curcumin showed a significant reduction in portal inflammation. Both activity and fibrosis demonstrated lower scores in all treated groups, however, PZQ revealed a marked increase in confluent necrosis and interface hepatitis. Besides, the lobular inflammation revealed worsening in the overall ISHAK score in all treated groups compared with the control. Few periocular granulomas were recovered by PZQ +curcumin treatment at day 35 post-treatment (6±1.2), P-value <0.05. Curcumin revealed a mild reduction (60±7.376). Curcumin-treated groups, with and without PZQ, resulted in higher significant Immunoreactivity score (IRS) for Bcl-2-associated X (BAX) and lower Interleukine17A (IL-17A), and Human epidermal growth factor (EGF), compared to the control. However, PZQ revealed a lower mean IRS value in BAX, higher IL-17A and EGF in the periovulatory granuloma. It was concluded that PZQ +curcumin treatment had a potent synergistic outcome through lessening the number of granulomas, the inflammatory events, and the expression of EGF, and amelioration of apoptosis in the periovulatory granulomas if compared with either PZQ or curcumin alone.

Polydatin Ameliorated Colonic Inflammation via Precluding Phosphorylation of PKCθ/STAT3 Pathway to Inhibit Th17 Cells in Mice with Ulcerative Colitis / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect of polydatin on ulcerative colitis （UC） in mice and its regulation of protein kinase C<italic>θ</italic>（PKC<italic>θ</italic>）/signal transducer and activator of transcription 3（STAT3） signaling on T helper cell 17（Th17） and its mechanism in the treatment of UC. Method:The 32 male C57BL/6 mice were randomly divided into normal group， model group， polydatin group （0.045 g·kg^-1） and sulfasalazine group （0.5 g·kg^-1）. The UC model was established by giving 3% dextran sodium sulfate （DSS） solution to free drinking water in mice. Polydatin and sulfasalazine groups were given by gavage 0.5 h before modeling for 7 days. The normal group and model group were given the same amount of normal saline. After the last administration， the colonic tissue was taken and hematoxylin-eosin （HE） was used to observe the pathological changes of colonic tissue. Flow cytometry was used to detect the proportion of Th17 in the lamina propria of colonic mucosa. The expression of interleukin-17A （IL-17A） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. Polydatin was added to CD4⁺T cells purified from spleen of C57BL/6 mice by magnetic-activated cell sorting （MACS） under the stimulation of cell stimulation cocktail <italic>in vitro </italic>in order to detect its impact on PKC<italic>θ</italic> and STAT3 phosphorylation. Result:Compared with normal group， the body weight was significantly decreased， and disease activity index （DAI） scores of the model group was significantly increased （<italic>P</italic><0.01）， the colonic mucosal epithelium was damaged and inflammatory cells infiltration in the mucosa and submucosa was obvious， the proportion of Th17 in the lamina propria of colonic mucosa was significantly increased （<italic>P</italic><0.01）， and the content of serum IL-17A was significantly increased （<italic>P</italic><0.01）. Compared with the model group， the weight and DAI score of polydatin and sulfasalazine groups were significantly improved （<italic>P</italic><0.01）， the degree of colon tissue damage was significantly improved， the proportion of Th17 in colon mucosa lamina propria was significantly decreased （<italic>P</italic><0.01）， and the content of IL-17A in serum was significantly decreased （<italic>P</italic><0.01）. <italic>In vitro</italic> experiments showed that polydatin could significantly inhibit the phosphorylation of PKC<italic>θ</italic> and STAT3 in Th17 （<italic>P</italic><0.01） as well as IL-17A secretion. Conclusion:Polydatin can improve the ulcerative colitis in mice via inhibiting the phosphorylation of PKC<italic>θ</italic> and STAT3 to preclude IL-17A secreting in Th17.

Role of IL-17A in Chronic Rhinosinusitis With Nasal Polyp

PURPOSE: Th17-associated inflammation is increased in chronic rhinosinusitis with nasal polyp (CRSwNP), and is associated with disease severity and steroid resistance. Overexpressed interleukin (IL)-17A affects CRSwNP by tissue remodeling, eosinophilic accumulation, and neutrophilic infiltration. We aimed to identify the role of IL-17A in CRSwNP and to evaluate the effects of anti-IL-17A blocking antibody on nasal polyp (NP) formation using a murine NP model. Moreover, we sought to investigate whether the inhibition of mechanistic target of the rapamycin (mTOR) signal pathway could suppress IL-17A expression and NP formation.METHODS: Human sinonasal tissues from control subjects and patients with chronic rhinosinusitis (CRS) were analyzed using immunohistochemistry (IHC) and immunofluorescence staining. The effects of IL-17A neutralizing antibody and rapamycin were evaluated in a murine NP model. Mouse samples were analyzed using IHC, quantitative real-time polymerase chain reaction, and enzyme-linked immunosorbent assay.RESULTS: IL-17A+ inflammatory cells were significantly increased in number in NP from patients with CRSwNP compared to that in uncinate process tissues from control subjects and patients with CRS without NP or CRSwNP. CD68+ M1 macrophages dominantly expressed IL-17A, followed by neutrophils and T helper cells, in NP tissues. Neutralization of IL-17A effectively reduced the number of NPs, inflammatory cytokines, and IL-17A-producing cells, including M1 macrophages. Inhibition of IL-17A via the mTOR pathway using rapamycin also attenuated NP formation and inflammation in the murine NP model.CONCLUSIONS: IL-17A possibly plays a role in the pathogenesis of CRSwNP, the major cellular source being M1 macrophage in NP tissues. Targeting IL-17A directly or indirectly may be an effective therapeutic strategy for CRSwNP.

In silico molecular docking and ADME/T analysis of plant compounds against IL17A and IL18 targets in gouty arthritis

Accumulation of urate crystals and subsequent inflammation are the major cause of pathogenesis of gout. Twopro inflammatory cytokines IL17A and IL18 are upregulated in the serum of gout patients and plays a major rolein promoting inflammation. Inhibition of these cytokines by plant phytochemicals would reduce the severity ofinflammation in gout. In the present study, in silico analysis of inhibition of IL17A and IL18 by 10 plant phytochemicalswere studied using the AutoDock 4.2 based on the principles of Lamarckian genetic algorithm. The results revealed abinding energy in the range of −6.32 kcal/mol to −3.5 kcal/mol and interacted with the amino acids in active pocketof IL17A and IL18. Among all the compounds, syringaresinol showing the least binding energy of −6.05 kcal/molwith IL17A and −6.32 kcal/mol with IL18. The control drug, allopurinol showed a binding energy of −3.32 and −3.18kcal/mol with IL17A and IL18, respectively. In addition, ADME/T properties of the compounds were also analyzed topredict their drug likeliness. This docking study can be used for developing potent inhibitors of IL17A and IL18 forthe treatment of gout.

Establishment of Modified Model of Psoriasis Mice / 中国药房

OBJECTIVE： To establish psoriasis animal model with a longer duration and more typical psoriatic characteristics by modifying psoriasis model induced by imiquimod. METHODS： Mice were randomly divided into normal group （treated with vaseline）， model group [treated with Imiquimod cream 60 mg/（2×3 cm2·d）] and modified group [treated with Imiquimod cream 60 mg/（2×3 cm2·d）+subcutaneous injection of rmIL-12 and LPS once a week]， for consecutive 21 d， with 10 mice in each group. The skin of the model site was observed daily from the first day of modeling. Psoriasis area and severity index （PASI） score was conducted for skin lesions such as erythema， scales and thickening. 21 d after modeling， mice were sacrificed， and histopathological examination of the skin lesions was performed. The spleen index was calculated， and the contents of IL-17A and IL-12 in the skin were detected by ELISA. RESULTS： From the third and second day of medication， erythema， scales and thickening were both observed in model group and modified group respectively. The PASI score reached peak on the 12th day. From the 11th day， erythema， scales and thickening of the modified group were more serious than that in model group. PASI scores of modified group was significantly higher than that of model group for 9 consecutive days （P＜0.05）. Histopathological observation showed that Munro microabscess， acanthosis and dermal inflammatory cell infiltration occurred in both model group and modified group. Compared with model group， spleen indexes of modified group were higher （P＜0.05）. Compared with normal group， the contents of IL-17A and IL-12 in mice skin of model group and modified group were both increased， and there was statistical significance in modified group （P＜0.05）. CONCLUSIONS： The estbalished modified model has the longer duration of typical characteristics of psoriasis.

IL-17A involved in respiratory syncytial virus-associated pathogenesis by promoting IFN-γand inhib-iting viral clearance in mice / 中华微生物学和免疫学杂志

Objective To identify the role of IL-17A during respiratory syncytial virus (RSV) in-fection in a mouse model. Methods Female wild-type C57BL/6 mice and IL-17A knockout ( IL-17A-/-) mice at the age of 6 to 8 weeks were both randomly divided into two groups:control and RSV groups. Mice in the control groups were given the supernatant of Hep-2 cell culture, while those in the RSV groups were treated with RSV A2 through intranasal administration. Leukocytes in bronchoalveolar lavage fluid ( BALF) samples were counted. Left lung tissues were stained with hematoxylin and eosin ( HE ) to evaluate his-topathological scores. Airway hyperresponsiveness ( AHR) was measured by whole-body plethysmography. The concentrations of IFN-γ were determined with ELISA. RSV titers were measured by plaque assay. To assess the effects of IL-17A on IFN-γproduction and its role in RSV infection, IL-17A-/- mice were treated with exogenous recombinant murine IFN-γ or IL-17A, while wild-type mice were given IFN-γ neutralizing antibody intervention. Results The counts of inflammatory cells and neutrophils in BALF, lung tissue his-topathological scores, AHR, IFN-γlevels and virus titers of the wild-type group were higher than those of the IL-17A-/-group after RSV infection. IFN-γlevels, inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly increased in RSV-infected IL-17A-/- mice after the intervention of recombinant murine IL-17A or IFN-γ. RSV titers were much higher in the recombinant murine IL-17A-trea-ted group, but not affected by the recombinant murine IFN-γ intervention. Inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly decreased in RSV-infected wild-type mice following IFN-γ neutralizing antibody treatment, but no significant changes were found in RSV titers. Conclusions IL-17A might be involved in the pathogenesis of pulmonary diseases during RSV infection through promoting IFN-γ production and inhibiting viral clearance in mice.

Mechanism of recombinant Mycobacterium smegmatis in alleviating airway inflammation in mice with non-eosinophilic asthma / 国际儿科学杂志

Objective To observe the effects of intranasal inoculation with recombinant Mycobacteri-um smegmatis ( rMS) on the airway inflammation of non-eosinophilic asthmatic mouse and further investigate the relative mechanism. Methods DO11. 10 T-cell receptor transgenic mice were divided randomly into three groups. Non-eosinophilic asthmatic model was established via OVA challenge, rMS were administrated into mice before challenge. Anti-IL-17A autoantibody in sera, IL-6 and IL-17A in BALF were measured by ELISA, the proportion of neutrophil in BALF was measured by FCM, MPO activity in lung tissue was detected by colorimetry, and the mRNA expression of CXCL2 and CXCL5 was measured by real-time PCR. Results Compared to control group, the number of neutrophils, IL-6 and IL-17A levels in BALF from asthmatic mice was significantly increased, meanwhile MPO activity and the expression of CXCL2, CXCL5 in lung tissue were both significantly upregulated. The results showed that high titer of autoantibody of IL-17A in sera of mice vaccined with recombinant Mycobacterium smegmatis was detected. Compared to asthma group, neutrophils and IL-6, IL-17A in BALF were significantly decreased, meanwhile MPO activity and CXCL2, CXL5 mRNA expression were significantly down-regulated. Conclusion Recombinant Mycobacterium smegmatis exhibits anti-inflammatory activity in murine neutrophilic asthma model and it may have protective effects on asthma.

Correlations between Ape1/Ref-1, ICAM-1 and IL-17A Levels in Serum and Radiation Pneumonitis for Local Advanced Non-small Cell Lung Cancer Patients / 中国肺癌杂志

BACKGROUND@#The main manifestations of radiation pneumonitis are injury of alveolar epithelial and endothelial cells, abnormal expression of cytokines, abnormal proliferation of fibroblasts and synthesis of fibrous matrix. The occurrence of radiation pneumonitis is associated with multiplecytokine level abnormality. These cytokines can also be used as bio-markers to predict the occurrence of radiation pneumonitis. This study was to evaluate the correlation between the change of apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ape1/Ref-1), intercellular adhesion molecules 1 (ICAM-1) and interleukin-17A (IL-17A) before and after radiotherapy and radiation pneumonitis for local advanced non-small cell lung cancer (NSCLC) patients with concurrent chemoradiotherapy.@*METHODS@#NSCLC patients (68 cases) were treated with concurrent radiotherapy and chemotherapy, every patient's normal tissue were controlled with a same radation dose. 68 local advanced NSCLC patients with concurrent chemoradiotherapy were detected the levels of Ape1/Ref-1, ICAM-1 and IL-17A in serum by ELISA before radiotherapy and in the 14th week after radiotherapy. Acute and advanced radiation pulmonary injury was graded according to Radiation Therapy Oncology Group/European Organization For Research and Treatment (RTOG/EORTC) diagnostic and grading criteria. Grade 2 or more radiation pneumonitis was taken as the main end point.@*RESULTS@#Eighteen cases out of 68 developed radiation pneumonitis, 50 of 68 cases have no radiation pneumonia development. There was no significant change of Ape1/Ref-1 levels before and after radiotherapy in radiation pneumonitis group (P>0.05). There was no significant change of Ape1/Ref-1 concentration in serum after radiotherapy between radiation pneumonitis group and non-radiation pneumonitis group (P>0.05). Compared with before radiotherapy, upregulation degree of ICAM-1 levels in radiation pneumonitis group was significantly higher than that in non- radiation pneumonitis group (P<0.05). There was no significant change of IL-17A concentration before and after radiotherapy in radiation pneumonitis group, but after radiotherapy IL-17A concentration in serum were remarkably higher than that in non-radiation pneumonitis group (P<0.05). Correlation analysis found that the change of ICAM-1 before and after radiotherapy has no obvious correlation with the incidence of radiation pneumonitis, and IL-17A change has obvious correlation with the incidence of radiation pneumonitis.@*CONCLUSIONS@#On the basis of strictly controlling radiation dose on normal tissue, IL-17A in serum could be the predictive factors of radiation pneumonitis for local advanced NSCLC patients with concurrent chemoradiotherapy.

The recent progress of Th17 and their effects in tumor immunity / 中国肿瘤生物治疗杂志

@#[摘 要] 人体免疫系统是由固有免疫和适应性免疫应答组成。适应性免疫应答在抗原入侵时扮演着至关重要的角色， 而CD4+ 辅助性T（CD4+ T helper, CD4+Th）细胞是适应性免疫应答的主要组成部分。最近新发现的一类不同于Th1和Th2细胞亚群且能 特征性分泌白细胞介素17 (interleukin-17, IL-17)的辅助性T细胞亚群，被命名为Th17细胞。Th17 细胞参与很多炎症性疾病、 自 身免疫性疾病和肿瘤等的发展过程，可通过分泌IL-17A、IL-17F、IL-21、IL-22、IL-23、粒细胞-巨噬细胞克隆刺激因子(granulocytemacrophage colony stimulating factor，GM-CSF)和干扰素γ（interferon-gamma，IFN-γ）等炎症细胞因子来发挥免疫效应和炎症效 应。但是Th17细胞是否参与肿瘤的发生发展、具体作用机制以及发挥促肿瘤还是抑肿瘤效应等问题存在很多争议。本文综述 了近年来Th17细胞分化调节过程的相关机制，以及其在肿瘤发生发展的作用，旨在为肿瘤的诊断和治疗提供新的思路。

Bcl11b Regulates IL-17 Through the TGF-β/Smad Pathway in HDM-Induced Asthma

PURPOSE: T helper (Th) 17 cells play a critical role in the development of asthma, but the underlying mechanism of how interleukin (IL)-17 is regulated in allergic airway inflammation is poorly understood. In this study, we investigated the impact of Bcl11b on Th17 response in asthma. METHODS: Blood samples from patients with mild asthma (MA) and severe asthma (SA) were collected. Expression of Bcl11b, IL-4, IL-5, IL-13, IL-17A and transforming growth factor (TGF)-β1 were determined in CD4+ T cells and plasma by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Relative mRNA and protein levels of Bcl11b, IL-17A and genes involved in the TGF/Smad signaling pathway were examined by PCR, ELISA and western blot analysis in house dust mite (HDM)-challenged mice. Ectopic expression of Bcl11b in HDM-stimulated primary mouse splenocytes was achieved by nucleofection of Bcl11b expression plasmid. RESULTS: We found significantly decreased Bcl11b but increased IL-17A and TGF-β1 expression in patients with asthma and a strongly negative correlation between Bcl11b and these 2 cytokines in SA patients. Similar expression patterns of Bcl11b, IL-17A and TGF-β1 were also found in mice with HDM-induced allergic airway inflammation. We demonstrated further that Smad2/3 phosphorylation was increased in HDM-challenged mice and that ectopic expression of Bcl11b in HDM-stimulated primary mouse splenocytes reduced Smad2 phosphorylation and IL-17 expression. CONCLUSIONS: Our findings demonstrate a potential effect of Bc111b in controlling IL-17-mediated inflammation in asthma and suggest that Bc111b may be a useful therapeutic target for asthma.

Notch1 signaling pathway regulates Th17 cell differentiation and function in murine psoriasis model / 中国免疫学杂志

Objective:To determine the effect of Notch1 signaling pathway on the differentiation and function of Th17 cells in murine psoriasis model.Methods: BALB/c mice were randomly divided into psoriasis model group and control group.Murine psoriasis model was established by topical 5% imiquimod application in combination with intraperitoneal injection of α-2b interferon.The CD4+ T lymphocytes were isolated by magnetic activated cell sorter (MACS).Flow cytometric analysis (FCM) was performed to detect the percentage of Th17 cells.Real-time RT-PCR was employed to measure the mRNA levels of RORγt,IL-17A,Notch1 and Hes-1.The CD4+ T lymphocytes were then divided into γ-secretase inhibitor DAPT groups and control group,and the expression differences of Notch1 signaling molecule and its target gene Hes-1 mRNA levels,Th17 cell percentage,RORγt and IL-17A mRNA levels,and IL-17A concentrations in cell-free supernatant were detected.Results: The expression levels of Th17 cell percentage and RORγt,IL-17A,Notch1 and Hes-1 mRNA in CD4+ T lymphocytes of murine psoriasis model were significantly higher than control mouse[(2.97±0.86)% vs.(0.65±0.11)%,t=15.083;(5.75±0.61) vs.(1.57±0.43),t=21.630;(7.83±0.97) vs.(1.63±0.31),t=25.348;(7.10±1.37) vs.(1.47±0.34),t=17.386;(7.30±1.15) vs.(1.67±0.48),t=18.840,respectively,all P<0.01].Compared with control group,Th17 cell percentage,mRNA expression levels of Notch1,Hes-1,RORγt and IL-17A,and IL-17A concentrations in cell-free supernatant from cultured CD4+ T lymphocytes of murine psoriasis model were dramatically decreased in DAPT treated groups in a dose-dependent way (F=74.368,89.719,126.572,94.558,124.323 and 123.231 respectively,all P<0.01).Conclusion: Notch1 signaling pathway can regulate the differentiation and function of Th17 cells in murine psoriasis model,and may have potential value for the target immunotherapy of psoriasis.

Clinical Features and Expression of Cytokine in Post-infectious Irritable Bowel Syndrome Patients / 中山大学学报(医学科学版)

[Objective]To study the clinical features and expressions of IL-17A,IFN-γ,and IL-10 in serum and intestinal mucosa of patients with post-infectious irritable bowel syndrome and non post-infectious irritable bowel syndrome.[Methods]44 diar?rhea-predominate IBS patients(21 with PI-IBS,23 with NPI-IBS)and 10 healthy controls were recruited in this study. Investigation questionnaires of GSRS,SAS,SDS were carried out to evaluate the gastrointestinal function,anxiety status and depression status of IBS patients. The expressions of IL-17A,IFN-γ,and IL-10 in intestinal mucosa and serum were measured by immunohistochemis?try(IHC)and enzyme-linked immunosorbent assay(ELISA).[Results]The SDS scores of NPI-IBS patients were higher than those of controls(P 0.05).[Conclusion]PI-IBS and NPI-IBS patients existed various anxiety and depression. The levels of IL-17A and IFN-γ increased and level of IL-10 decreased in PI-IBS and NPI-IBS group. But the clinical symptoms and changes of cytokines of PI-IBS patients were more significant. There may exist other pathogenesis in PI-IBS but not in NPI-IBS.

Distribution of γδT17/Th17/Tc17 cells in lung of H1N1 infected mice and their relationship with immunologic injury of lung / 中国免疫学杂志

Objective:To investigate the distribution of γδT17,Th17 and Tc17 cells in the lung of mice severely infected by influenza A(H1N1)pdm09 virus and the relationship between these cells with lung immunopathalogical injury.Methods:Intranasal infection was used to establish mouse model of severe H1N1 infection.Flow cytometry assay was used to detect the proportion and number of γδT17 cells,Th17 cells and Tc17 cells in the lung.The concentrations of interleukin-17A(IL-17A),interleukin-1β(IL-1β)and interleukin-23(IL-23) in the bronchoalveolar lavage fluid and serum were assayed by enzyme-linked immunosorbent assay and Lu-minex assay.Results:①The model of mice severely infected by influenza A(H1N1)pdm09 virus was established successfully.②The ratio of γδT cells,but not CD4+T and CD8+T cells in total lymphocytes of the lung of infected mice significantly increased compared with uninfected control mice at the third day post infection(DPI)(P<0.01).③The proportion and number of γδT17 cells,Th17 cells and Tc17 cells in total γδT cells,Th cells and Tc cells in the lung of infected mice were significant higher than that in uninfected control mice at the first DPI,respectively.However,the absolute number of γδT17 cells was far more than Th17 and Tc17 cells(P<0.05);④The concentration of IL-17A in BALF increased significantly after infection(P<0.05),and the concentration of IL-17A in serum increased significantly at the third DPI(P<0.05).The concentrations of both IL-1β and IL-23 in BALF probably participating in the activation of γδT17 cells increased significantly after infection compared with uninfected control mice.Conclusion:The γδT17 cells could be activated and secreted IL-17A via γδTCR non-depended pathway and involved in inflammatory pathological injury of lung at the early stage of severe H1N1 infection.

Effects of IL-17A on fibrosis of skin and lung in a mouse model of systemic sclerosis / 中华微生物学和免疫学杂志

Objective To analyze the expression of interleukin ( IL)-17A in a mouse model of bleomycin ( BLM)-induced systemic sclerosis ( SSc) and to evaluate its effects on inflammation and fibrosis in skin and lung tissues. Methods Twenty-four female BALB/c mice were randomly divided into four groups:normal control group ( mice were subcutaneously injected with phosphate buffer ) , model group (subcutaneously injected with BLM), antibody group (injected with BLM + IL-17A monoclonal antibody), homotypic control group ( injected with BLM + isotype control) . Pathological changes in skin and lung tis-sues of those mice were observed;inflammatory and fibrotic scores were assessed. Immunohistochemistry and real-time fluorescent quantitative PCR ( RT-PCR) were used to detect the expression of IL-17A, TGF-β1 and typeⅠ collagen in skin and lung tissues of those mice at mRNA level. Mouse lung fibroblasts ( FB) de-rived from the mice of model group were cultured in vitro and then were cultured with IL-17A cytokines with or without the interference of monoclonal antibodies. Expression of typeⅠ collagen and TGF-β1 at mRNA level and levels of IL-6 and TGF-β1 in the culture supernatants were detected by RT-PCR and enzyme-linkedimmunosorbent assay ( ELISA) , respectively. Results Compared with the mice of model and homotypic control groups, those of the antibody group showed mild skin thickening, skin inflammation and lung inflam-mation as well as lower fibrosis scores (P<0. 05). The expression of IL-17A at both protein and mRNA lev-els and the expression of TGF-β1 and collagen typeⅠat mRNA level in skin and lung tissues of mice of the antibody group were significantly lower than those of the model and homotypic control group (P<0. 05). Re-sults of the in vitro cell culture of SSc mice-derived lung FB with IL-17A showed that the expression of TGF-β1 and typeⅠ collagen at mRNA level and the levels of IL-6 and TGF-β1 in the culture supernatants were decreased with the interference of anti-IL-17A monoclonal antibody (P<0. 05), but were still higher than those of the control group (P<0. 05). Conclusion IL-17A promotes the development of inflammation and fibrosis in skin and lung tissues in the mouse model of SSc. Blocking IL-17A might inhibit fibrosis in SSc by inhibiting the production of TGF-β1, IL-6 and typeⅠ collagen.

Mechanism of IL-17A induced the secretion of CXCL12 in non-small cell lung cancer cell line A549 / 局解手术学杂志

Objective To study the molecular mechanism of IL-17A induced the secretion of CXCL 12 in non-small cell lung cancer cell line A549.Methods Cultured non-small cell lung cancer cell line A549 in vitro with recombination cytokine IL-17A or STAT3 signal path-way inhibitor pre-incubated for 1 hour, and then the level of CXCL12 were detected by enzyme-linked immunosorbent assay .And chemotaxis assay was used to analyze the chemotactic movement of neutrophil .Results After IL-17A stimulation,the secretion of CXCL12 by non-small cell lung cancer cell line A549 was significantly increased(P<0.01),which is in a dose and time dependent manner .However,IL-17A in-duced the secretion of CXCL12 by A549 was significantly decreased after pre-incubated by the STAT3 inhibitor(P<0.01).In addition,neu-trophil could have chemotaxis by cell suspension obtained from IL-17A stimulated A549 cell line,but such chemotaxis would be declined while CXCL12 was neutralized.Conclusion IL-17A could induce the secretion of CXCL12 in non-small cell lung cancer cell line A549 through STAT3 signal pathway , so as to promote the chemotaxis of neutrophil .