IL-4-deficient Mice Aggravate Hypersensitivity Pneumonitis

Hwang, Su-Jin,Chung, Doo-Hyun The Korean Association of Immunobiologists 2008 Immune Network Vol.8 No.3

Background: Hypersensitivity pneumonitis (HP) comprises a group of lung diseases resulting from repeated inhalation of various antigens such as Saccharopolyspora rectivirgula (SR). HP is categorized as a Th1 disease. Therefore, it has been suggested that IL-4, Th2 type cytokine, plays a protective role in the development of HP. However, the functional role of IL-4 in HP has not been extensively investigated in vivo. Therefore, we investigated the functional role of IL-4 in HP using IL-4 knockout (KO) mice. Methods: HP was induced by repeated exposure to SR in C57BL/6 (B6) and IL-4 KO (C57BL/6 background) mice. Results: IL-4 KO mice aggravated HP in terms of histological alteration, SR-specific immune responses, and inflammatory cell infiltration in the lungs compared with B6 mice. IL-4 KO mice produced high levels of IFN-${\gamma}$, TGF-${\beta}$ and TNF-${\alpha}$ in the lungs, whereas B6 mice showed the enhanced production of IL-4. Moreover, chemokines such as MIP-1${\alpha}$, MCP-1, and RANTES were highly expressed in IL-4 KO mice. IFN-${\gamma}$-secreting CD4, CD8 T cells, and neutrophils were enhanced in the bronchoalveolar lavage fluid (BALF) of IL-4 KO mice than those of B6 mice. The administration of recombinant(r) IL-4 restored these immunologic parameters in IL-4 KO mice. Conclusion: These results indicate that IL-4 plays a suppressive role in SR-induced HP by attenuating Th1-dominant immune responses.

IL-4 Receptor α 유전자다형성을 이용한 한국인알레르기비염환자에서의 감수성에 대한 연구

이재훈,최태욱,이정헌,이상헌,정하민,민정열,김정중 대한이비인후과학회 2005 대한이비인후과학회지 두경부외과학 Vol.48 No.8

Background and Objectives:The IL-4 receptor (IL-4R) gene has been suggested as a candidate gene for atopic diseases. The IL-4R consists of two subunits:the α chain (IL-4Rα), which is a high-affinity IL-4 binding site shared with the IL-13R, and the common γ chain shared with several other cytokine receptors that amplifies signalling of the α chain. A Gln551Arg polymorphism of the IL-4Rα gene was shown to be a gain-of-function mutation and was associated with atopy. We tested whether a Gln551Arg polymorphism of IL-4Rα gene is associated with allergic rhinitis, blood eosinophil counts and total serum IgE levels in the Korean population. Subjects and Method:Blood samples for genetic analysis were obtained from 192 individuals with allergic rhinitis and from 191 healthy subjects without atopic diseases. Polymerase chain reaction-based assay for IL-4Rα Gln551Arg was used for genotyping. Serum total IgE levels were determined by using the immunoassay. Eosinophil values were determined by eosinophil numbers per total cell numbers per μL . Results:There were no differences in the frequencies of the genotypes of IL-4Rα in the controls and patients (p>0.05). The frequencies of the IL-4Rα Arg551 allele were statistically different between controls and patients (p>0.05). Blood eosinophil count and total serum IgE levels were not statistically different in the genotypes of IL-4Rα Gln551Arg in allergic rhinitis (p>0.05). Conclusion:Our result suggests that the IL-4Rα Gln551Arg polymorphism might not give susceptibility to the development of allergic rhinitis in Koreans.

Interleukin-4 receptor-targeted delivery of Bcl-xL siRNA sensitizes tumors to chemotherapy and inhibits tumor growth

Guruprasath, Padmanaban,Kim, Jihoon,Gunassekaran, Gowri Rangaswamy,Chi, Lianhua,Kim, Soyoun,Park, Rang-Woon,Kim, Sang-Hyun,Baek, Moon-Chang,Bae, Sang Mun,Kim, Sang-Yeob,Kim, Dong-Kyu,Park, In-Kyu,Kim, Elsevier 2017 Biomaterials Vol.142 No.-

<P><B>Abstract</B></P> <P>IL-4 receptor (IL-4R) is commonly up-regulated on tumor cells, and interactions between the receptor and Interleukin-4 (IL-4) can induce the expression of anti-apoptotic proteins, including Bcl-xL. This contributes to tumor cell survival and their resistance to chemotherapy. In this study, we exploited IL-4R-targeted delivery of Bcl-xL siRNA to IL-4R-expressing tumor cells in order to sensitize them to chemotherapy. To target IL-4R, an IL-4R-binding peptide, IL4RPep-1, was attached to branched polyethyleneimine-superparamagnetic iron oxide nanoparticles (BPEI-SPION). These nanoparticles were then complexed with Bcl-xL-targeting siRNA. IL-4R-targeted BPEI-SPION/Bcl-xL siRNA more efficiently reduced Bcl-xL gene expression and enhanced cytotoxicity of doxorubicin in MDA-MB231 breast tumor cells compared to untargeted BPEI-SPION/Bcl-xL siRNA. The siRNA was released from the complexes after 15 h of incubation at pH 5.5 and was stable in the complexes up to 72 h in the serum. The IL-4R-targeted BPEI-SPION/siRNA was internalized by cells through IL-4R, successfully escaped the endosomes, and was dispersed into the cytoplasm. Near-infrared fluorescence and magnetic resonance imaging demonstrated that <I>in vivo</I> tumor homing and accumulation of IL-4R-targeted BPEI-SPION/siRNA were both higher than untargeted BPEI-SPION/siRNA. The IL-4R-targeted BPEI-SPION/Bcl-xL siRNA, in combination with doxorubicin, significantly inhibited tumor growth in mice compared to untargeted BPEI-SPION/Bcl-xL siRNA. These results suggest that the IL-4R-targeted delivery of Bcl-xL siRNA to IL-4R-expressing tumors can sensitize tumors to chemotherapy and enhance the efficacy of anti-tumor therapeutics.</P> <P><B>Graphical abstract</B></P> <P>Interaction between IL-4 and IL-4R provides downstream signaling for the expression of Bcl-xL and inhibits doxorubicin-induced apoptosis. IL4RPep-1-labeled BPEI-SPION/Bcl-xL siRNA selectively binds to IL-4R-expressing tumor cells and is internalized into the cells through the receptor-mediated endocytosis, which contributes to the silencing of the Bcl-xL gene expression. This sensitizes IL-4R-expressing tumor cells to doxorubicin and enhances its therapeutic efficacy.</P> <P>[DISPLAY OMISSION]</P>

Enhanced delivery of liposomes to lung tumor through targeting interleukin-4 receptor on both tumor cells and tumor endothelial cells

Chi, L.,Na, M.H.,Jung, H.K.,Vadevoo, S.M.P.,Kim, C.W.,Padmanaban, G.,Park, T.I.,Park, J.Y.,Hwang, I.,Park, K.U.,Liang, F.,Lu, M.,Park, J.,Kim, I.S.,Lee, B.H. Elsevier Science Publishers 2015 Journal of controlled release Vol.209 No.-

A growing body of evidence suggests that pathological lesions express tissue-specific molecular targets or biomarkers within the tissue. Interleukin-4 receptor (IL-4R) is overexpressed in many types of cancer cells, including lung cancer. Here we investigated the properties of IL-4R-binding peptide-1 (IL4RPep-1), a CRKRLDRNC peptide, and its ability to target the delivery of liposomes to lung tumor. IL4RPep-1 preferentially bound to H226 lung tumor cells which express higher levers of IL-4R compared to H460 lung tumor cells which express less IL-4R. Mutational analysis revealed that C1, R2, and R4 residues of IL4RPep-1 were the key binding determinants. IL4RPep-1-labeled liposomes containing doxorubicin were more efficiently internalized in H226 cells and effectively delivered doxorubicin into the cells compared to unlabeled liposomes. In vivo fluorescence imaging of nude mice subcutaneously xenotransplanted with H226 tumor cells indicated that IL4RPep-1-labeled liposomes accumulate more efficiently in the tumor and inhibit tumor growth more effectively compared to unlabeled liposomes. Interestingly, expression of IL-4R was high in vascular endothelial cells of tumor, while little was detected in vascular endothelial cells of control organs including the liver. IL-4R expression in cultured human vascular endothelial cells was also up-regulated when activated by a pro-inflammatory cytokine tumor necrosis factor-α. Moreover, the up-regulation of IL-4R expression was observed in primary human lung cancer tissues. These results indicate that IL-4R-targeting nanocarriers may be a useful strategy to enhance drug delivery through the recognition of IL-4R in both tumor cells and tumor endothelial cells.

IL-4와 다른 여러가지 cytokines이 사람 편도 단핵세포의 IgE 생산에 미치는 영향

이헌구,송원재,하대유 大韓免疫學會 1995 大韓免疫學會誌 Vol.17 No.3

The present study was desinged to compare IgE producing ability of cultured human tonsillar mononuclear cells (TMNC) stimulated by rhIL-4 between complete Iscove's modified Dulbeco's medium (C-IMDM, which was composed of IMDM enriched with bovine serum albumin, fetal calf serum, transferrin, insulin, and mixture of saturated and unsaturated fatty acids) and RPMI-1640 supplemented with FCS (RPMI-1640), as well as to investigate a single or combined effects of IL-4 and 7 different human cytokines (IL-2, IL-5, IL-6, IL-7, TNF-a, IFN- 7' and TGF-fi ) in a culture system on IgE production of human TMNC. It was found that C-IMDM was superior to RPMI 1640 as a culture medium for supporting the synthesis of IgE by TMNC stimulated with rhIL-4. In fact, RPMI-1640 failed to induce significant IgE synthesis by TMNC cultured for 21 days in the presence of rhlL-4 (100 U / ml). In addition, 7 different cytokines other than IL-4 failed to induce the significant IgE synthesis by TMNC cultured in C-IMDM when tested alone. However, IL-2 or IL-6, which was ineffective in inducing IgE production when tested alone, enhanced IL-4-mediated IgE production, indicating - that IL-2 or IL-6 acts synergistically with IL-4. In contrast, IFN- Y . or TGF-J1 demonstrated the inhibition of IL-4 dependent IgE synthesis by TMNC. Interestingly, IL-5, IL-7 and TNF-a had no modulatory effect on the IL-4 dependent IgE response in the applied experimental conditions. Taken together, the present results strongly suggested that 1) C-IMDM may be superior to RPMI-1640 as a culture _medium for IgE production, 2) Only IL-4 had the ability to stimulate in vitro with IgE synthesis by TMNC when tested alone. 3) IL-2 and IL-6 may be synergistic with IL-4 in IgE synthesis, 4) IFN- F and TGF-P may have the antagonistic effects on IL-4 dependent IgE synthesis and 5) IL-5, IL-7 and TNF-a may have no modulatory effects. This study also suggested that increased insight into various mechanisms of IgE regulation by cytokines may eventually lead to improve therapeutic strategies in the clinical management of IgE-mediated allergy.

정신분열병에서 Interleukin-4 시발자와 Interleukin-4 수용체 유전자의 다형성

최진욱,전태연,배치운,이경욱 대한신경정신의학회 2002 신경정신의학 Vol.41 No.5

연구목적: 정신분열병은 면역유전적 성향이 강한 정신 질환의 하나로, 최근에는 분자유전학적 기법을 이용하 여 유전자 다형성과 질환 사이의 연관성을 밝히는 시도가 정신분열병의 연구에서 중요한 부분을 차지 하고 있다. 이에 본 연구는 중추신경계와 면역기능의 조절에 관여하는 사이토카인 중 IL-4 시발자 및 IL-4 수용체 유전자 다형성과 정신분열병과의 관련성을 알아보기 위하여 IL-4 시발자와 IL-4 수용체의 유전자형과 대립유전자의 빈도를 환자군과 대조군 사이 그리고 편집형 환자군과 비편집형 환자군 사이에서 알아보았다. 방 법: DSM-Ⅳ에 의하여 정신분열병으로 진단된 환자 222명과 가톨릭 조혈모세포 정보은행에서 보유하고 있는 정상 한국인 165명의 자료를 대상으로 하였다. 전혈에서 DNA를 추출한 후 중합효소 연쇄반응을 이용하여 IL-4 시발자와 IL-4 수용체의 유전자 부위를 증폭하였다. 이들 증폭산물을 SSCP(single strand conformation polymorphism) 방법을 이용하여 유전자형과 대립유전자를 동정하였으며 자료 의 분석은 카이제곱 검증을 이용하였다. 결 과: 정신분열병 환자군과 정상 대조군 간에 IL-4 시발자와 IL-4 수용체의 유전자형과 대립유전자 발 현 빈도에는 유의한 차이가 없었으며, 편집형 환자군과 비편집형 환자군 간에서도 IL-4 시발자와 IL- 4 수용체의 유전자형과 대립유전자 발현 빈도에는 유의한 차이가 없었다. 결 론: 본 연구에서는 IL-4 시발자 및 IL-4 수용체 유전자 다형성과 정신분열병 사이의 면역유전학적 관련성을 확인할 수 없었다. 향후연구에서는 다양한 임상변인을 포함한 체계적인 연구가 이루어져야 할 것으로 생각된다 Objectives:Recently, the main focus of etiologic study in schizophrenia has been directed to molecular genetic approach including polymorphism analysis. Abnormal immunoreactivity to IL-4 promoter and IL-4Rα has been identified in patients with schizophrenia. This study was designed to investigate the relationship between schizophrenia and immunologic influences by analyzing the polymorphisms of IL-4 promoter and IL-4Rα gene that are involved in interaction between immunologic system and CNS. Methods:222 schizophrenic patients diagnosed by DSM-IV and data of 165 normal controls obtained from Catholic Hemopoietic Stem Cell Information Bank, College of medicine, the Catholic University of Korea, were used in this study. DNA was extracted from whole blood and the polymorphic loci of IL-4 promoter and IL-4Rαgene were amplified by polymerase chain reaction. Gene typing was performed by using SSCP and the results were assessed. The frequencies of allele and genotype were compared between patients and normal controls and between paranoid group and non-paranoid group. All data were analyzed χ2-test with two-tailed Fisher’s exact test. Results: 1) There were no significant differences in allele or genotype frequencies of IL-4 promoter and IL- 4Rα between the group of schizophrenic patients and controls. 2) There were no significant differences in allele or genotype frequencies of IL-4 promoter and of IL-4Rα between the group of paranoid schizophrenic patients and non-paranoid schizophrenic patients. Conclusion:These results suggest that polymorphisms of IL-4 promoter and IL-4Rα genes are unlikely related with the pathogenesis of schizophrenia.

IL-4-deficient Mice Aggravate Hypersensitivity Pneumonitis

황수진,정두현 대한면역학회 2008 Immune Network Vol.8 No.3

Background: Hypersensitivity pneumonitis (HP) comprises a group of lung diseases resulting from repeated inhalation of various antigens such as Saccharopolyspora rectivirgula (SR). HP is categorized as a Th1 disease. Therefore, it has been suggested that IL-4, Th2 type cytokine, plays a protective role in the development of HP. However, the functional role of IL-4 in HP has not been extensively investigated in vivo. Therefore, we investigated the functional role of IL-4 in HP using IL-4 knockout (KO) mice. Methods: HP was induced by repeated exposure to SR in C57BL/6 (B6) and IL-4 KO (C57BL/6 background) mice. Results: IL-4 KO mice aggravated HP in terms of histological alteration, SR-specific immune responses, and inflammatory cell infiltration in the lungs compared with B6 mice. IL-4 KO mice produced high levels of IFN-γ, TGF-β and TNF-α in the lungs, whereas B6 mice showed the enhanced production of IL-4. Moreover, chemokines such as MIP-1α, MCP-1, and RANTES were highly expressed in IL-4 KO mice. IFN-γ-secreting CD4, CD8 T cells, and neutrophils were enhanced in the bronchoalveolarlavage fluid (BALF) of IL-4 KO mice than those of B6 mice. The administr ation of recombinant(r) IL-4 restored these immunologic parameters in IL-4 KO mice. Conclusion: These results indicate that IL-4 plays a suppressive role in SR-induced HP by attenuating Th1-dominant immune responses. Background: Hypersensitivity pneumonitis (HP) comprises a group of lung diseases resulting from repeated inhalation of various antigens such as Saccharopolyspora rectivirgula (SR). HP is categorized as a Th1 disease. Therefore, it has been suggested that IL-4, Th2 type cytokine, plays a protective role in the development of HP. However, the functional role of IL-4 in HP has not been extensively investigated in vivo. Therefore, we investigated the functional role of IL-4 in HP using IL-4 knockout (KO) mice. Methods: HP was induced by repeated exposure to SR in C57BL/6 (B6) and IL-4 KO (C57BL/6 background) mice. Results: IL-4 KO mice aggravated HP in terms of histological alteration, SR-specific immune responses, and inflammatory cell infiltration in the lungs compared with B6 mice. IL-4 KO mice produced high levels of IFN-γ, TGF-β and TNF-α in the lungs, whereas B6 mice showed the enhanced production of IL-4. Moreover, chemokines such as MIP-1α, MCP-1, and RANTES were highly expressed in IL-4 KO mice. IFN-γ-secreting CD4, CD8 T cells, and neutrophils were enhanced in the bronchoalveolarlavage fluid (BALF) of IL-4 KO mice than those of B6 mice. The administr ation of recombinant(r) IL-4 restored these immunologic parameters in IL-4 KO mice. Conclusion: These results indicate that IL-4 plays a suppressive role in SR-induced HP by attenuating Th1-dominant immune responses.

IL-4 and IL-13 suppress prostaglandins production in human follicular dendritic cells by repressing COX-2 and mPGES-1 expression through JAK1 and STAT6

Cho, W.,Kim, Y.,Jeoung, D.I.,Kim, Y.M.,Choe, J. Pergamon Press 2011 Molecular immunology Vol.48 No.6

Originally discovered as a B cell growth and differentiation factor, IL-4 displays a variety of functions in many different cell types. Germinal center T cells are abundant producers of IL-4. In a recent report, we demonstrated that IL-4 inhibits prostaglandins (PGs) production in follicular dendritic cell (FDC)-like cells, HK. To understand the inhibitory mechanisms of IL-4, its effects on the biosynthesis of enzymes in charge of PG production were assessed in this study. Although IL-4 did not affect COX-1 expression, it specifically inhibited LPS-induced COX-2 biosynthesis at mRNA and protein levels. Protein expression of mPGES-1, a downstream enzyme of COX-2, was also markedly diminished by IL-4 but not by IL-10, maximizing the inhibitory activity. Next, we attempted to identify the early signaling molecules that led to this inhibition of COX-2 expression. Although IL-4 induced tyrosine phosphorylation of JAK1 and TYK2, RNA interference experiments revealed that only JAK1 was responsible for the IL-4-stimulated STAT6 phosphorylation. Knocking down JAK1 and STAT6 ablated the inhibitory effect of IL-4 on COX-2 expression and significantly reduced production of PGE<SUB>2</SUB> and prostacyclin. Similar results were obtained with IL-13. Pharmacologic inhibitors of ERK and p38 mitogen-activated protein kinases inhibited the COX-2 upregulation. However, IL-4 did not affect LPS-induced phosphorylation of ERK and p38. These results stress the essential roles of JAK1 and STAT6 in the early signaling pathway of IL-4 and IL-13 leading to suppression of COX-2 expression and repression of PG production by HK cells. Our results suggest that T cells via IL-4 play a regulatory role in PG generation in FDC. IL-4 therapeutics may be applied to immune disorders where normal and ectopic expression of germinal center reactions needs to be regulated.

천식유발 마우스에서의 폐 내 세포조성 변화와 IL-4 및 IL-10의 발현 양상

이수진,박세종,리천주,장양호,최농훈,Lee, Soo-Jin,Park, Se-Jong,Li, Tian-Zhu,Jang, Yang-Ho,Choe, Nong-Hoon 한국생명과학회 2006 생명과학회지 Vol.16 No.5

본 연구진은 일차로 BALB/C 마우스를 이용하여 항원으로 ovalbumin을 사용하여 천식을 유발하여 폐 내 세포구성에 미치는 영향을 다른 실험방법에 의해 만들어진 천식모델과 비교하여 살펴보았고 이차로 천식 발생에 중요한 역할을 하는 것으로 알려진 IL-4와 IL-10 knock out (KO) 마우스를 이용하여 천식모델을 구축하여 천식인자를 가진 개체에서의 반응과 천식관련인자가 결핍된 개체에서의 반응성의 차이를 살펴보았다. 천식의 유발은 실험 1일째 $20\;{\mu}g$ ovalbumin으로 감작시킨 후 실험 14일에 재감작을 시켰다. 그 후 nebulizer를 이용하여 nasal inhalation을 28일, 29일, 30일에 실시하여 천식을 유발시켰다. 천식 유발의 확인은 기관지폐포 세척술로 채취한 폐 내 세포액을 이용하여 총 세포수 및 염증세포의 증가와 폐 내 세포와 폐조직의 염색을 통해 분포율을 확인하였다. 천식의 발병 과정에서 IgE 관련 과민반응을 주도하는 IL-4와 Th2 세포의 기능을 억제한다고 알려진 IL-10의 면역화학염색을 통해 그 발현정도를 관찰하였다. BALB/c 마우스의 천식유발군의 경우 천식의 특징인 염증세포의 증가와 호산구의 증가와 세기관지 주위 염증세포의 침윤 및 기도 상피의 비후를 관찰할 수 있었다. 따라서 본 실험에 응용된 천식유발 방법은 적합하였으며 천식유발 절차가 성공적으로 수행되었음을 확인하였다. IL-4 및 IL-10 KO 마우스를 이용한 천식유발군의 경우 BALB/c 마우스를 이용한 천식유발군보다 호산구의 증가 정도가 미약하게 관찰되었고, 폐조직 내 염증세포의 침윤 정도도 감소하였다. 천식매개인자인 IL-4와 IL-10의 면역조직화학염색 결과에서도 양성반응이 거의 나타나지 않았다. 본 연구 결과, IL-10이 IL-4와 같이 천식과 관련하여 세기관지 염증 반응을 증가시키는 역할을 하는 것으로 생각된다. Asthma is a chronic inflammatory disorder of the airways, which characterized by bronchial hyperresponsiveness, reversible airflow limitation and respiratory symptoms. Internationally, the prevalence of asthma has been increased over last 3 decades. Recently, several studies of asthma have been reported with gradually increasing importance. To tesify the hypothesis that interleukin (IL)-4 and IL-10 may be an important determinant of the severity of airway inflammation, their expression was studied in mouse model of asthma. BALB/c mouse, IL-4 Knockout (KO) mouse and IL-10 KO mouse were sensitized with intraperitoneal injection of ovalbumin adsorbed to aluminum potassium sulfate, followed by challenges with intranasal ovalbumin on 3 consecutive days. The severity of pulmonary inflammation was assessed by eosinophilia in BAL fluid, number of total BAL cells, histopathological changes in lung tissues, and immunohistochemical staining against IL-4 and IL-10. In BAL fluid, the number of total cells was significantly increased in asthma induced mouse compare to the control. In asthma induced mouse, eosinophil was increased to 56% and neutrophil was 0.2%. In H &E stains, eosinophilic infiltration and epithelium hyperplasia were clearly noticed in asthma induced mouse. In immunohistochemical staining for IL-4 and IL-10, there was no positive reaction in control group. However, very strong reactions were appeared in asthma induced group. In this research, IL-4 and IL-10, which seem to play a central role in allergic asthma, KO mouse was utilized to test the causative relationship between airway inflammation and role of specific cytokine. Asthma induced IL-4 and IL-10 KO mice showed much decreased inflammatory reactions in the number of total BAL cells, in eosinophilic infiltration, and in immunohistochemical stains against diverse inflammatory proteins. These results suggest that IL-4 and IL-10 increase the asthmatic reactions in vivo mice model.

급성 골수성백혈병에서 Stem Cell Factor가 CD34⁺세포의 증식 및 IL-1β, IL-4, TNF-α의 분비에 미치는 효과

이석,민유홍,정소영,한지숙,고윤웅 大韓血液學會 1995 大韓血液學會誌 Vol.30 No.3