Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Veterans Affairs Palo Alto Health Care System

Stanford University School of Medicine

To study the implication of prokineticin 1 (PK1/EG-VEGF) and prokineticin 2 (PK2/Bv8) in hepatocellular carcinoma angiogenesis.

Prokineticin 2/Bv8 is expressed in Kupffer cells in liver and is down regulated in human hepatocellular carcinoma.

Prokineticins are a novel family of secreted peptides with diverse regulatory roles, one of which is their capacity to modulate immunity in humans and in other species. Prokineticins are small peptides of 8 kDa that mediate their biological activities by signaling through two homologous G-protein-coupled receptors (prokineticin receptor 1 and prokineticin receptor 2). This family of peptides is characterized by a completely conserved N-terminal hexapeptide crucial for their bioactivities and a unique structural motif comprising five disulfide bonds. Prokineticins and their receptors are highly expressed in bone marrow, in peripheral circulating leukocytes, in inflamed tissues and in resident organ immune cells. Their structure, size, signaling and biological activities are reminiscent of the chemokine superfamily. In this review, emphasis is placed on the properties of prokineticins as cytokines and their role in the immune system.

Cytokine properties of prokineticins.

Prokineticin 1 and 2 (PROK1 and PROK2) are two small proteins largely expressed in inflammatory tissues and involved in monocyte activation and differentiation. The focus of this study was to evaluate whether PROK1 was able to induce chemokine secretion in human monocytes, in monocyte-derived macrophages and in monocyte-derived dendritic cells, an aspect not addressed thus far. Here, we show for the first time, using flow cytometry, that PROK receptors 1 and 2 are present on the surface of human monocytes. Subsequently, monocytes were selected to investigate the chemokine response after stimulation by PROK1. Our results show that only three chemokines (CCL4, CXCL1 and CXCL8) were significantly induced at both the transcript and protein level, and that PROK1 induces most potently CXCL8, in a dose-dependent manner. From a mechanistic point of view, by blocking independently Galphai protein or intracellular calcium, monocytes lose the ability to secrete CXCL8 in response to PROK1. Finally, we observed that CCL4, CXCL1 and CXCL8 secretion, following PROK1 induction, is only observed in monocytes and not in monocyte-derived macrophages and dendritic cells. Our results demonstrate that, in vitro, the differentiation status of monocytes influences chemokine production after stimulation by PROK1, and that this chemokine production is geared toward a pro-inflammatory response. This could represent a novel amplification loop of leukocyte recruitment, extravasation and tissue invasion.

Prokineticin 1 induces CCL4, CXCL1 and CXCL8 in human monocytes but not in macrophages and dendritic cells.

This review will examine the roles of prokineticins in the different neoplasms that have been investigated and will discuss how and why the prokineticin family presents such interesting prospects in the research against cancer. Prokineticins belong to a family of highly conserved small peptides (8kDa) discovered a decade ago in frog skin secretions and snake venom. The mammalian orthologs consist of two prokineticin peptides, prokineticin 1 (PROK1) and prokineticin 2 (PROK2) that signal through two G-protein coupled receptors: prokineticin receptor 1 (PROKR1) and prokineticin receptor 2 (PROKR2). Over the last decade of research, the PROK/PROKR system has been associated with a considerable number of physiological and pathological functions. Due to this wide spectrum of functions, notably potent angiogenic and immunoregulatory activities, numerous investigators have researched the PROK/PROKR system's role in cancer development in a variety of tissues.

Prokineticins in angiogenesis and cancer.

Development of hepatocellular carcinoma (HCC) is a complex and progressive disease that involves cycles of liver cell death, inflammation, and tissue regeneration/remodelling. Chemokines and chemokine receptors play numerous and integral roles in the disease progression of HCC. Here we investigated the novel chemokine receptor CXCR7/RDC1 in HCC progression, its two known ligands CXCL12 and CXCL11, as well as the other CXCL12 receptor, CXCR4. Our results show that in a cohort of 408 human HCCs, CXCR7 and CXCL11 were significantly higher in tumours compared to normal liver controls (5- and 10-fold, respectively). Immunohistochemical (IHC) staining on human HCC sections confirmed that both CXCL11 and CXCR7 were much higher in cancer tissues. Furthermore, IHC staining revealed that CXCR7 protein was only expressed in endothelial cells whereas CXCL11 exhibited a much broader tissue expression. At the cellular level we observed that in vitro, human microvascular endothelial cells (HMEC-1) up-regulated CXCR7 under hypoxic and acidic pH conditions, which are well known characteristics of the HCC tumour micro-environment. As for its ligand, we observed that IFNγ robustly induced CXCL11 in hepatic stellate cells, hepatocytes, and HMEC-1s. In addition, in the mouse Diethylnitrosamine model of hepatocarcinogenesis we observed a very strong induction of CXCR7 and CXCL11 transcripts, confirming that CXCR7/CXCL11 up-regulation is conserved between human and mice liver cancer. Altogether, our results strongly support the hypothesis that the CXCL11/CXCR7 pathway is involved HCC progression.

CXCR7 is up-regulated in human and murine hepatocellular carcinoma and is specifically expressed by endothelial cells.

(&#x6284;&#x9001;&#x6BCD;&#x9898;) &#x8D8B;&#x5316;&#x56E0;&#x5B50;&#x53D7;&#x4F53;&#x6837; 2 (CCRL2) &#x7ED1;&#x5B9A;&#x767D;&#x7EC6;&#x80DE;&#x8D8B;&#x5316; chemerin &#x548C;&#x53EF;&#x8C03;&#x8282;&#x5730;&#x65B9;&#x5404;&#x7EA7;&#x7684;&#x5F15;&#x8BF1;&#x5242;&#xFF0C;&#x4F46;&#x672C;&#x8EAB;&#x4E0D;&#x652F;&#x6301;&#x7EC6;&#x80DE;&#x8FC1;&#x79FB;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x8868;&#x660E; CCRL2 &#x548C;&#x8840;&#x7BA1;&#x7EC6;&#x80DE;&#x9ECF;&#x9644;&#x5206;&#x5B50;-1 &#x5BF9;&#x57F9;&#x517B;&#x7684;&#x4EBA;&#x548C;&#x5C0F;&#x9F20;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE; (EC) &#x548C;&#x708E;&#x75C7;&#x523A;&#x6FC0;&#x7EC6;&#x80DE;&#x682A;&#x589E;&#x9AD8;&#x3002;CCRL2 &#x611F;&#x5E94;&#x662F;&#x4F9D;&#x8D56; NF-&kappa; B &#x548C; JAK/STAT &#x4FE1;&#x53F7;&#x901A;&#x8DEF;&#xFF0C;&#x5E76;&#x6FC0;&#x6D3B;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x4E13;&#x95E8;&#x7ED1;&#x5B9A; chemerin&#x3002;&#x5728;&#x4F53;&#x5185;&#xFF0C;CCRL2 &#x4E00;&#x5BF9;&#x8868;&#x793A;&#x662F;&#x7531;&#x80BA;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x9AD8;&#x7EA7;&#x522B;&#x548C;&#x4F4E;&#x7EA7;&#x522B;&#x7531;&#x809D;&#x5185;&#x76AE;&#x7EC6;&#x80DE; &#xFF1B;&#x809D;&#x4F46;&#x4E0D;&#x662F;&#x80BA;&#x6B27;&#x5171;&#x4F53;&#x901A;&#x8FC7;&#x548C;&#x54CD;&#x5E94;&#x7CFB;&#x7EDF;&#x6027;&#x8102;&#x591A;&#x7CD6;&#x6CE8;&#x5C04;&#x6DB2;&#x8FDB;&#x4E00;&#x6B65;&#x4E0A;&#x8C03;&#x7684;&#x53D7;&#x4F53;&#x3002;&#x603B; chemerin &#x7684;&#x8840;&#x6D46;&#x6C34;&#x5E73;&#x5347;&#x9AD8; CCRL2(-/-) &#x5C0F;&#x9F20;&#x548C;&#x540E; CCRL2(-/-) &#x5C0F;&#x9F20;&#x7CFB;&#x7EDF;&#x6027; LPS &#x6CBB;&#x7597;&#x4E0E;&#x91CE;&#x751F;&#x578B;&#x5C0F;&#x9F20;&#x76F8;&#x6BD4;&#x90FD;&#x5927;&#x5927;&#x589E;&#x5F3A;&#x3002;&#x540E;&#x4F53;&#x5185;&#xFF0C;&#x7C7B;&#x4F3C;&#x7684;&#x8D8B;&#x5316;&#x56E0;&#x5B50;&#x53D7;&#x4F53; 1 &#x7684;&#x5185;&#x6BD2;&#x7D20;&#x7684;&#x6025;&#x6027;&#x80BA;&#x90E8;&#x708E;&#x75C7; (CMKLR1)(+) NK &#x7EC6;&#x80DE;&#x62DB;&#x8058;&#x7ED9;&#x822A;&#x7A7A;&#x516C;&#x53F8;&#x5927;&#x5927;&#x53D7;&#x635F; CCRL2(-/-) &#x5C0F;&#x9F20;&#x4E0E;&#x91CE;&#x751F;&#x578B;&#x5C0F;&#x9F20;&#x76F8;&#x6BD4;&#x3002;&#x5728;&#x8BD5;&#x7BA1;&#x91CC;&#xFF0C;chemerin &#x7ED1;&#x5B9A;&#x5230; CCRL2 &#x5BF9;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x901A;&#x8FC7; &alpha;(4)&beta;(1) &#x6574;&#x5408;&#x7D20;/&#x8840;&#x7BA1;&#x7EC6;&#x80DE;&#x9ECF;&#x9644;&#x5206;&#x5B50;-1 &#x4F9D;&#x8D56;&#x7684;&#x673A;&#x5236;&#x89E6;&#x53D1; CMKLR1(+) &#x6DCB;&#x5DF4;&#x7EC4;&#x7EC7;&#x7EC6;&#x80DE;&#x7684;&#x9C81;&#x68D2;&#x7C98;&#x9644;&#x3002;&#x603B;&#x4E4B;&#xFF0C;CCRL2 &#x8868;&#x793A;&#x6559;&#x7EDF;&#x4F1A;&#x7684;&#x7EC4;&#x7EC7;&#x548C;&#x6FC0;&#x6D3B;&#x4F9D;&#x8D56;&#x7684;&#x65B9;&#x5F0F;&#x3001; &#x8C03;&#x8282;&#x5FAA;&#x73AF; chemerin &#x7EA7;&#x522B;&#x548C;&#x5176;&#x751F;&#x7269;&#x6D3B;&#x6027;&#xFF0C;&#x5E76;&#x589E;&#x5F3A;&#x4E86; chemerin &#x548C; CMKLR1 &#x4F9D;&#x8D56;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;/EC &#x7C98;&#x9644;&#x4F53;&#x5916;&#x548C;&#x4F53;&#x5185;&#x53D1;&#x708E;&#x822A;&#x7A7A;&#x516C;&#x53F8;&#x62DB;&#x8058;&#x3002;&#x5176;&#x8868;&#x8FBE;&#x548C;/&#x6216;&#x7531;&#x708E;&#x75C7;&#x523A;&#x6FC0;&#x8BF1;&#x5BFC;&#x5BF9;&#x6B27;&#x5171;&#x4F53;&#x63D0;&#x4F9B;&#x5C0F;&#x8BF4;&#x548C;&#x5F53;&#x5730;&#x4E30;&#x5BCC;&#x7684;&#x89C4;&#x7BA1;&#x62DB;&#x8058; CMKLR1(+) &#x7EC6;&#x80DE;&#x7684;&#x708E;&#x75C7;&#x5730;&#x76D8; chemerin &#x7684;&#x5177;&#x4F53;&#x673A;&#x5236;&#x3002;

&#x8868;&#x8FBE;&#x3001; &#x8C03;&#x63A7;&#xFF0C;&#x975E;&#x5178;&#x578B;&#x7684; Chemerin &#x53D7;&#x4F53; CCRL2 &#x5BF9;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x7684;&#x529F;&#x80FD;&#x3002;

Chemokine (CC motif) receptor-like 2 (CCRL2) binds leukocyte chemoattractant chemerin and can regulate local levels of the attractant, but does not itself support cell migration. In this study, we show that CCRL2 and VCAM-1 are upregulated on cultured human and mouse vascular endothelial cells (EC) and cell lines by proinflammatory stimuli. CCRL2 induction is dependent on NF-ÎºB and JAK/STAT signaling pathways, and activated endothelial cells specifically bind chemerin. In vivo, CCRL2 is constitutively expressed at high levels by lung endothelial cells and at lower levels by liver endothelium; and liver but not lung EC respond to systemic LPS injection by further upregulation of the receptor. Plasma levels of total chemerin are elevated in CCRL2(-/-) mice and are significantly enhanced after systemic LPS treatment in CCRL2(-/-) mice compared with wild-type mice. Following acute LPS-induced pulmonary inflammation in vivo, chemokine-like receptor 1 (CMKLR1)(+) NK cell recruitment to the airways is significantly impaired in CCRL2(-/-) mice compared with wild-type mice. In vitro, chemerin binding to CCRL2 on endothelial cells triggers robust adhesion of CMKLR1(+) lymphoid cells through an Î±(4)Î²(1) integrin/VCAM-1-dependent mechanism. In conclusion, CCRL2 is expressed by EC in a tissue- and activation-dependent fashion, regulates circulating chemerin levels and its bioactivity, and enhances chemerin- and CMKLR1-dependent lymphocyte/EC adhesion in vitro and recruitment to inflamed airways in vivo. Its expression and/or induction on EC by proinflammatory stimuli provide a novel and specific mechanism for the local enrichment of chemerin at inflammatory sites, regulating the recruitment of CMKLR1(+) cells.

Expression, Regulation, and Function of Atypical Chemerin Receptor CCRL2 on Endothelial Cells.

Chemokine (CC motif) receptor type 2 (CCRL2) lie les leucocytes chemoattractant chemerin et peut r&eacute;guler les niveaux locaux de la r&eacute;pulsion, mais pas lui-m&ecirc;me ne prend en charge la migration cellulaire. Dans cette &eacute;tude, nous montrons que les CCRL2 et VCAM-1 sont surexprim&eacute;s homme cultiv&eacute; et des cellules endoth&eacute;liales vasculaires de souris (EC) et des lign&eacute;es cellulaires par des stimuli pro-inflammatoires. CCRL2 induction d&eacute;pend de NF-&kappa;B et JAK/STAT, voies de signalisation, et les cellules endoth&eacute;liales activ&eacute;es lient sp&eacute;cifiquement chemerin. In vivo, CCRL2 est constitutivement exprim&eacute; par les cellules endoth&eacute;liales pulmonaires &agrave; des niveaux &eacute;lev&eacute;s et &agrave; des niveaux inf&eacute;rieurs de l&#39;endoth&eacute;lium du foie ; et foie mais pas le poumon EC r&eacute;pond &agrave; injection syst&eacute;mique de LPS en augmentation suppl&eacute;mentaire du r&eacute;cepteur. Les taux plasmatiques de chemerin total sont &eacute;lev&eacute;s chez la souris CCRL2(-/-) et sont sensiblement am&eacute;lior&eacute;es apr&egrave;s que traitement syst&eacute;mique de LPS chez la souris CCRL2(-/-) par rapport aux souris de type sauvage. Suite &agrave; une inflammation pulmonaire induite par LPS aigu&euml; in vivo, r&eacute;cepteur de chimiokine-like 1 (recrutement de cellules NK CMKLR1)(+) pour les voies respiratoires est significativement r&eacute;duite chez les souris CCRL2(-/-) par rapport aux souris de type sauvage. In vitro, la liaison de chemerin &agrave; CCRL2 sur les cellules endoth&eacute;liales d&eacute;clenche robuste adh&eacute;rence de cellules lympho&iuml;des CMKLR1(+) gr&acirc;ce &agrave; un m&eacute;canisme d&#39;int&eacute;grine/VCAM-1-dependent &alpha;(4)&beta;(1). En conclusion, les CCRL2 s&#39;exprime par EC de fa&ccedil;on d&eacute;pendante du tissu et activation, r&eacute;gule en circulation chemerin niveaux et sa bioactivit&eacute; et am&eacute;liore l&#39;adh&eacute;rence de lymphocytes/EC-chemerin - et CMKLR1-dependent in vitro et le recrutement &agrave; enflamm&eacute;e airways in vivo. Son expression et/ou induction sur EC par des stimuli pro-inflammatoires fournit un roman et un m&eacute;canisme sp&eacute;cifique pour l&#39;enrichissement local de chemerin dans les sites inflammatoires, r&eacute;gissant le recrutement des cellules CMKLR1(+).

Expression, r&egrave;glement et la fonction de CCRL2 atypique Chemerin r&eacute;cepteurs sur les cellules endoth&eacute;liales.

Chemokine (CC-Motiv) Rezeptor-wie 2 (CCRL2) bindet der Leukozyte Chemoattractant Chemerin und lokaler Ebene von den Lockstoff regulieren kann, aber nicht selbst unterst&uuml;tzt Zellmigration. In dieser Studie zeigen wir, dass CCRL2 und VCAM-1 herraufreguliert auf kultivierten Menschen und Maus vaskul&auml;re Endothelzellen (EG) und Zelllinien durch Proinflammatory Stimuli. CCRL2 Induktion ist abh&auml;ngig von NF-&kappa;B und JAK/STAT-Signalisierung Wege und aktivierte endotheliale Zellen spezifisch binden Chemerin. In vivo wird CCRL2 konstitutiv auf hohem Niveau von Endothelzellen der Lunge und auf den unteren Ebenen von Leber Endothel ausgedr&uuml;ckt; und Leber, aber nicht die Lunge EG reagieren auf systemische LPS-Injektion durch weitere Hochregulation des Rezeptors. Plasmaspiegel von total Chemerin sind erh&ouml;ht bei CCRL2(-/-) M&auml;usen und sind erheblich verbessert worden, nachdem systemische LPS-Behandlung bei CCRL2(-/-) M&auml;usen mit Wildtyp M&auml;usen verglichen. Nach akuten LPS-induzierte pulmonale Entz&uuml;ndung in Vivo Chemokin-Like-Rezeptor 1 (CMKLR1)(+) NK Zelle Anwerbung f&uuml;r die Atemwege ist bei CCRL2(-/-) M&auml;usen im Vergleich zu Wildtyp M&auml;usen erheblich beeintr&auml;chtigt. In-vitro-Chemerin Bindung an CCRL2 in Endothelzellen l&ouml;st robuste Adh&auml;sion der CMKLR1(+) lymphoide Zellen durch einen &alpha;(4)&beta;(1) Integrin/VCAM-1-abh&auml;ngigen Mechanismus. Abschlie&szlig;end CCRL2 wird durch die EG in Gewebe und Aktivierung-abh&auml;ngige Weise ausgedr&uuml;ckt, regelt die zirkulierenden Chemerin Ebenen und seiner Bioaktivit&auml;t, und verbessert die Chemerin und CMKLR1-abh&auml;ngigen Lymphozyten/EG Adh&auml;sion in-vitro- und die Rekrutierung von in-vivo entz&uuml;ndeten Airways. Seinen Ausdruck und/oder Induktion auf EG von Proinflammatory Reize bieten eine neuartige und spezifischen Mechanismus f&uuml;r die lokale Anreicherung von Chemerin an entz&uuml;ndlichen Standorten, die Rekrutierung von CMKLR1(+) Zellen regulieren.

Ausdruck, Regulierung und Funktion der atypischen Chemerin Rezeptor CCRL2 in Endothelzellen.

Chemochina (motivo CC) receptor-like 2 (CCRL2) associa leucocita chemioattrattanti chemerin e possibile regolare i livelli locali dell&#39;esca, ma non supporta la migrazione cellulare. In questo studio, ci mostrano che CCRL2 e VCAM-1 sono sovraregolati uomo colto e cellule endoteliali vascolari del mouse (CE) e linee cellulari da stimoli proinfiammatori. CCRL2 induzione &egrave; dipendente da NF-&kappa;B e JAK/STAT vie di segnalazione, e cellule endoteliali attivate legano specificamente chemerin. In vivo, CCRL2 &egrave; costitutivamente espresso a livelli elevati di cellule endoteliali polmonare e a livelli inferiori di fegato endotelio; e fegato ma non polmone CE risposta a iniezione sistemica di LPS da ulteriore sovraregolazione del recettore. Livelli plasmatici di chemerin totale sono elevati nei topi CCRL2(-/-) e sono significativamente migliorati dopo trattamento sistemico LPS in topi CCRL2(-/-) rispetto ai topi wild-type. Seguito di acuta infiammazione polmonare indotta da LPS in vivo, simil-chemokine receptor 1 (reclutamento delle cellule NK CMKLR1)(+) alle vie aeree &egrave; significativamente alterata nei topi CCRL2(-/-) rispetto ai topi wild-type. In vitro, associazione chemerin a CCRL2 sulle cellule endoteliali innesca robusto adesione delle cellule linfoidi CMKLR1(+) attraverso un meccanismo di integrina/VCAM-1-dipendente &alpha;(4)&beta;(1). In conclusione, CCRL2 &egrave; espressa dalla CE in una moda tessuto e attivazione dipendente, regola la circolazione chemerin livelli e la sua bioattivit&agrave; e migliora - chemerin - e CMKLR1-dipendente dei linfociti/CE adesione in vitro e reclutamento per vie aeree infiammate in vivo. Sua espressione e/o induzione su CE da stimoli proinfiammatori fornisce un romanzo e un meccanismo specifico per l&#39;arricchimento locale di chemerin nei siti infiammatori, che regolano il reclutamento delle cellule CMKLR1(+).

Espressione, regolazione e funzione di atipico CCRL2 Chemerin recettore sulle cellule endoteliali.

&#x30B1;&#x30E2;&#x30AB;&#x30A4;&#x30F3; &#xFF08;CC &#x30E2;&#x30C1;&#x30FC;&#x30D5;&#xFF09; &#x53D7;&#x5BB9;&#x4F53;&#x69D8; 2 (CCRL2) &#x304C;&#x767D;&#x8840;&#x7403;&#x904A;&#x8D70; chemerin &#x3092;&#x30D0;&#x30A4;&#x30F3;&#x30C9;&#x3057;&#x3001;&#x8A98;&#x5F15;&#x306E;&#x30ED;&#x30FC;&#x30AB;&#x30EB; &#x30EC;&#x30D9;&#x30EB;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x304C;&#x306A;&#x3044;&#x81EA;&#x4F53;&#x7D30;&#x80DE;&#x306E;&#x79FB;&#x52D5;&#x3092;&#x30B5;&#x30DD;&#x30FC;&#x30C8;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;, CCRL2 &#x3068; VCAM 1 &#x767A;&#x73FE;&#x4EA2;&#x9032;&#x3072;&#x3068;&#x57F9;&#x990A;&#x3068;&#x30DE;&#x30A6;&#x30B9;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7D30;&#x80DE; (EC) &#x304A;&#x3088;&#x3073;&#x708E;&#x75C7;&#x523A;&#x6FC0;&#x306B;&#x3088;&#x308B;&#x7D30;&#x80DE;&#x682A;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;CCRL2 &#x8A98;&#x5C0E;&#x306F; NF-&kappa; &#x3068; JAK/STAT &#x30B7;&#x30B0;&#x30CA;&#x30EA;&#x30F3;&#x30B0;&#x7D4C;&#x8DEF;&#x306B;&#x4F9D;&#x5B58;&#x3059;&#x308B;&#x3057;&#x3001;&#x6D3B;&#x6027;&#x5316;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x306F;&#x5177;&#x4F53;&#x7684;&#x306B; chemerin &#x3092;&#x30D0;&#x30A4;&#x30F3;&#x30C9;&#x3057;&#x307E;&#x3059;&#x3002;In vivo CCRL2 &#x80BA;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x306B;&#x3088;&#x308B;&#x9AD8;&#x30EC;&#x30D9;&#x30EB;&#x304A;&#x3088;&#x3073;&#x305D;&#x306E;&#x4E0B;&#x4F4D;&#x306E;&#x30EC;&#x30D9;&#x30EB;&#x306B;&#x3088;&#x3063;&#x3066;&#x809D;&#x81D3;&#x5185;&#x76AE;&#x767A;&#x73FE;&#x3067;&#x3059;;&#x809D;&#x81D3;&#x304C;&#x306A;&#x3044;&#x80BA; EC &#x306B;&#x5168;&#x8EAB; lps &#x306E;&#x53D7;&#x5BB9;&#x4F53;&#x306E;&#x3055;&#x3089;&#x306E;&#x30A2;&#x30C3;&#x30D7;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306B;&#x3088;&#x3063;&#x3066;&#x5FDC;&#x7B54;&#x3057;&#x307E;&#x3059;&#x3002;&#x8840;&#x6F3F;&#x7DCF; chemerin &#x306E; CCRL2(-/-) &#x30DE;&#x30A6;&#x30B9;&#x306B;&#x304A;&#x3051;&#x308B;&#x9AD8;&#x67B6;&#x3055;&#x308C;&#x5168;&#x8EAB;&#x306E; LPS &#x51E6;&#x7406; CCRL2(-/-) &#x30DE;&#x30A6;&#x30B9;&#x306B;&#x304A;&#x3051;&#x308B;&#x91CE;&#x751F;&#x578B;&#x30DE;&#x30A6;&#x30B9;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x5F8C;&#x3092;&#x5927;&#x5E45;&#x306B;&#x5F37;&#x5316;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x6B21;&#x306E;&#x6025;&#x6027;&#x306E; LPS &#x8A98;&#x767A;&#x80BA;&#x708E;&#x75C7; in vivo&#x3001;&#x30B1;&#x30E2;&#x30AB;&#x30A4;&#x30F3;&#x69D8;&#x53D7;&#x5BB9;&#x4F53; 1 &#xFF08;CMKLR1)(+) NK &#x7D30;&#x80DE;&#x306E;&#x52D5;&#x54E1;&#x3001;&#x6C17;&#x9053;&#x306B;&#x306F;&#x8457;&#x3057;&#x304F;&#x640D;&#x306A;&#x308F;&#x308C;&#x305F;&#x91CE;&#x751F;&#x578B;&#x30DE;&#x30A6;&#x30B9;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066; CCRL2(-/-) &#x30DE;&#x30A6;&#x30B9;&#x3067;&#x3002;In vitro CCRL2 &#x5185;&#x76AE;&#x7D30;&#x80DE;&#x3078;&#x306E; chemerin &#x30D0;&#x30A4;&#x30F3;&#x30C9; CMKLR1(+) &#x30EA;&#x30F3;&#x30D1;&#x7D30;&#x80DE;&#x306E;&#x5805;&#x7262;&#x306A;&#x63A5;&#x7740;&#x306F; &alpha;(4)&beta;(1) &#x30A4;&#x30F3;&#x30C6;&#x30B0;&#x30EA;&#x30F3;/VCAM-1 &#x4F9D;&#x5B58;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306B;&#x3088;&#x3063;&#x3066;&#x30C8;&#x30EA;&#x30AC;&#x30FC;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x7D50;&#x8AD6;&#x3068;&#x3057;&#x3066;&#x3001;CCRL2 EC &#x306B;&#x3088;&#x308B;&#x7D44;&#x7E54;&#x3068;&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D9;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x4F9D;&#x5B58;&#x30D5;&#x30A1;&#x30C3;&#x30B7;&#x30E7;&#x30F3;&#x3067;&#x8868;&#x3055;&#x308C;&#x308B;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x5FAA;&#x74B0; chemerin &#x30EC;&#x30D9;&#x30EB;&#x3068;&#x305D;&#x306E;&#x751F;&#x7269;&#x6D3B;&#x6027;&#x3068; chemerin &#x3068; CMKLR1 &#x306B;&#x4F9D;&#x5B58;&#x3059;&#x308B;&#x30EA;&#x30F3;&#x30D1;&#x7403;/EC &#x63A5;&#x7740;&#x4F53;&#x5916;&#x3068;&#x708E;&#x75C7;&#x6C17;&#x9053; in vivo &#x3067;&#x306E;&#x52DF;&#x96C6;&#x3092;&#x5F37;&#x5316;&#x3057;&#x307E;&#x3059;&#x3002;&#x305D;&#x306E;&#x5F0F;&#x304A;&#x3088;&#x3073;/&#x307E;&#x305F;&#x306F;&#x708E;&#x75C7;&#x6027;&#x523A;&#x6FC0;&#x306B;&#x3088;&#x308B; ec &#x8A98;&#x5C0E;&#x5C0F;&#x8AAC;&#x3068; chemerin CMKLR1(+) &#x7D30;&#x80DE;&#x306E;&#x52DF;&#x96C6;&#x3092;&#x898F;&#x5236;&#x3059;&#x308B;&#x708E;&#x75C7;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x3067;&#x306E;&#x30ED;&#x30FC;&#x30AB;&#x30EB;&#x306E;&#x6FC3;&#x7E2E;&#x306E;&#x305F;&#x3081;&#x306E;&#x56FA;&#x6709;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;

&#x5F0F;&#x3001;&#x8ABF;&#x7BC0;&#x3001;&#x304A;&#x3088;&#x3073;&#x975E;&#x5B9A;&#x578B;&#x306E; Chemerin &#x53D7;&#x5BB9;&#x4F53; CCRL2 &#x5185;&#x76AE;&#x7D30;&#x80DE;&#x306E;&#x6A5F;&#x80FD;&#x3002;

(CC &#xBAA8;&#xD2F0;&#xBE0C;) Chemokine &#xC218;&#xC6A9; &#xCCB4; &#xAC19;&#xC740; 2 (CCRL2) &#xBC31;&#xD608;&#xAD6C; chemoattractant chemerin&#xC5D0; &#xBC14;&#xC778;&#xB529;&#xD558;&#xACE0; &#xC720;&#xC778;, &#xB85C;&#xCEEC; &#xB808;&#xBCA8;&#xC744; &#xC870;&#xC808;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xD558;&#xC9C0;&#xB9CC; &#xC140; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158;&#xC744; &#xC9C0;&#xC6D0; &#xC790;&#xCCB4;&#xAC00; &#xB418;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xB294; ccrl2&#xC640; VCAM-1 &#xAD50;&#xC591; &#xC778;&#xAC04;&#xACFC; &#xB9C8;&#xC6B0;&#xC2A4; &#xD608;&#xAD00; &#xB0B4; &#xD53C; &#xC138;&#xD3EC; (EC)&#xC640; proinflammatory &#xC790;&#xADF9;&#xC5D0; &#xC758;&#xD574; &#xC138;&#xD3EC; &#xB77C;&#xC778;&#xC5D0; upregulated&#xB97C; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. CCRL2 &#xC720;&#xB3C4; NF &kappa;B JAK/STAT &#xC2E0;&#xD638; &#xACBD;&#xB85C;&#xC5D0; &#xB530;&#xB77C; &#xB2EC;&#xB77C; &#xC9D1;&#xB2C8;&#xB2E4; &#xBC0F; &#xD65C;&#xC131;&#xD654; &#xB41C; &#xB0B4; &#xD53C; &#xC138;&#xD3EC; &#xD2B9;&#xD788; chemerin&#xC5D0; &#xBC14;&#xC778;&#xB529;&#xD569;&#xB2C8;&#xB2E4;. Vivo&#xC5D0;&#xC11C; CCRL2 constitutively &#xD45C;&#xD604; &#xD3D0; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC5D0; &#xC758;&#xD574; &#xC0C1;&#xBD80;&#xC5D0; &#xACE0; &#xAC04; endothelium;&#xC5D0; &#xC758;&#xD574; &#xD558;&#xC704; &#xC218;&#xC900;&#xC5D0;&#xC11C; &#xADF8;&#xB9AC;&#xACE0; &#xAC04;&#xB9CC; &#xD3D0; &#xC544;&#xB2C8; EC &#xC218;&#xC6A9; &#xCCB4;&#xC758; &#xCD94;&#xAC00; upregulation &#xD558; &#xC5EC; &#xC870;&#xC9C1;&#xC758; LPS &#xC8FC;&#xC785;&#xC5D0; &#xC751;&#xB2F5;. &#xCD1D; chemerin &#xD50C;&#xB77C;&#xC988;&#xB9C8; &#xC218;&#xC900;&#xC740; CCRL2(-/-) &#xB9C8;&#xC6B0;&#xC2A4;&#xC5D0; &#xB192;&#xC740; CCRL2(-/-) &#xB9C8;&#xC6B0;&#xC2A4;&#xC5D0; &#xC870;&#xC9C1;&#xC758; LPS &#xCE58;&#xB8CC; &#xC57C;&#xC0DD;-&#xD0C0;&#xC785; &#xB9C8;&#xC6B0;&#xC2A4;&#xC640; &#xBE44;&#xAD50; &#xD6C4; &#xD06C;&#xAC8C; &#xD5A5;&#xC0C1; &#xB41C;. &#xBE44;&#xBCF4;, chemokine &#xAC19;&#xC740; &#xC218;&#xC6A9; &#xCCB4; 1&#xC5D0;&#xC5D0;&#xC11C; &#xAE09;&#xC131; LPS-&#xC720;&#xBC1C; &#xD3D0; &#xC5FC;&#xC99D;&#xC5D0; &#xB530;&#xB77C; (&#xAE30;&#xB3C4; CMKLR1)(+) NK &#xC140; &#xCC44;&#xC6A9;&#xC740; &#xC57C;&#xC0DD;-&#xD0C0;&#xC785; &#xB9C8;&#xC6B0;&#xC2A4;&#xC5D0; &#xBE44;&#xD574; CCRL2(-/-) &#xB9C8;&#xC6B0;&#xC2A4;&#xC5D0; &#xD06C;&#xAC8C; &#xC7A5;&#xC560;&#xAC00;. &#xCCB4; &#xC678;, &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC5D0; CCRL2 chemerin &#xBC14;&#xC778;&#xB529; &alpha;(4)&beta;(1) integrin/VCAM 1 &#xC885;&#xC18D; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC744; &#xD1B5;&#xD574; CMKLR1(+) &#xB9BC;&#xD504; &#xC138;&#xD3EC;&#xC758; &#xAC15;&#xB825;&#xD55C; &#xC811;&#xCC29;&#xC744; &#xD2B8;&#xB9AC;&#xAC70;&#xD569;&#xB2C8;&#xB2E4;. &#xB05D;&#xC73C;&#xB85C;, CCRL2 &#xC870;&#xC9C1; &#xD65C;&#xC131;&#xD654;&#xC5D0; &#xC885;&#xC18D; &#xD328;&#xC158; &#xB2E8;&#xC704;&#xB85C; EC&#xC5D0; &#xC758;&#xD574;, &#xADDC;&#xC81C; &#xC21C;&#xD658; chemerin &#xC218;&#xC900; &#xBC0F; &#xADF8;&#xAC83;&#xC758; bioactivity &#xBC0F; chemerin &#xBC0F; CMKLR1 &#xC885;&#xC18D; &#xB9BC;&#xD504; &#xAD6C;/EC &#xC811;&#xCC29; &#xCCB4; &#xC678; &#xBC0F; &#xC5FC;&#xC99D;&#xC774; &#xD56D;&#xACF5; vivo&#xC5D0;&#xC11C; &#xCC44;&#xC6A9;&#xC744; &#xD5A5;&#xC0C1; &#xC2DC;&#xD0B5;&#xB2C8;&#xB2E4;. &#xC2DD; &#xB610;&#xB294; proinflammatory &#xC790;&#xADF9;&#xC5D0; &#xC758;&#xD574; EC&#xC5D0; &#xC720;&#xB3C4; &#xC18C;&#xC124; &#xBC0F; CMKLR1(+) &#xC140;&#xC758; &#xBAA8;&#xC9D1;&#xC744; &#xADDC;&#xC81C; &#xD558;&#xB294; &#xC5FC;&#xC99D; &#xC131; &#xC0AC;&#xC774;&#xD2B8;&#xC5D0; chemerin&#xC758; &#xB85C;&#xCEEC; &#xB18D;&#xCD95;&#xC5D0; &#xB300; &#xD55C; &#xAD6C;&#xCCB4;&#xC801;&#xC778; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC744; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;.

&#xC2DD;, &#xADDC;&#xC81C;, &#xADF8;&#xB9AC;&#xACE0; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC5D0; &#xD754;&#xD558;&#xC9C0; &#xC54A;&#xC740; Chemerin &#xC218;&#xC6A9; &#xCCB4; ccrl2&#xC758; &#xAE30;&#xB2A5;.

Chemokine (CC Adorno) receptor tipo 2 (CCRL2) se une del leucocito chemoattractant chemerin y puede regular los niveles locales del atrayente, pero no soporta migraci&oacute;n celular. En este estudio demostramos que la CCRL2 y VCAM-1 son upregulated humanos cultivados y c&eacute;lulas endoteliales vasculares de rat&oacute;n (EC) y l&iacute;neas celulares por est&iacute;mulos proinflamatorios. CCRL2 inducci&oacute;n depende de NF-&kappa;B y v&iacute;as de se&ntilde;alizaci&oacute;n JAK/STAT, y las c&eacute;lulas endoteliales activadas unen espec&iacute;ficamente chemerin. En vivo, CCRL2 es constitutivamente en altos niveles por las c&eacute;lulas endoteliales de pulm&oacute;n y en los niveles inferiores por el endotelio del h&iacute;gado; y h&iacute;gado pero no pulm&oacute;n CE responde a inyecci&oacute;n sist&eacute;mica de LPS m&aacute;s upregulation del receptor. Los niveles plasm&aacute;ticos de chemerin total son elevados en ratones CCRL2(-/-) y se han mejorado significativamente despu&eacute;s del tratamiento sist&eacute;mico LPS en ratones CCRL2(-/-) en comparaci&oacute;n con ratones de tipo salvaje. Tras la inflamaci&oacute;n pulmonar aguda inducida por LPS en vivo, como chemokine receptor 1 (reclutamiento de c&eacute;lulas NK CMKLR1)(+) a las v&iacute;as respiratorias se deteriora significativamente en ratones CCRL2(-/-) en comparaci&oacute;n con los ratones de tipo salvaje. In vitro, Uni&oacute;n de chemerin a CCRL2 en las c&eacute;lulas endoteliales provoca fuerte adhesi&oacute;n de c&eacute;lulas linfoides CMKLR1(+) a trav&eacute;s de un mecanismo de integrina/VCAM-1-dependiente &alpha;(4)&beta;(1). En conclusi&oacute;n, el CCRL2 se expresa por la CE de manera dependiente del tejido y activaci&oacute;n, regula niveles de chemerin y su bioactividad de la circulaci&oacute;n y mejora la adhesi&oacute;n de linfocitos/CE-chemerin - y CMKLR1-dependent in vitro y reclutamiento a v&iacute;as respiratorias inflamadas en vivo. Su expresi&oacute;n ni inducci&oacute;n en EC por est&iacute;mulos proinflamatorios ofrecen una novela y un mecanismo espec&iacute;fico para el enriquecimiento local de chemerin en sitios inflamatorios, regular el reclutamiento de c&eacute;lulas CMKLR1(+).

Expresi&oacute;n, regulaci&oacute;n y funci&oacute;n de CCRL2 Chemerin Receptor anormal en las c&eacute;lulas endoteliales.

Infiltration of specialized immune cells regulates the growth and survival of neoplasia. Here, in a survey of public whole genome expression datasets we found that the gene for chemerin, a widely expressed endogenous chemoattractant protein, is down-regulated in melanoma as well as other human tumors. Moreover, high chemerin messenger RNA expression in tumors correlated with improved outcome in human melanoma. In experiments using the B16 transplantable mouse melanoma, tumor-expressed chemerin inhibited in vivo tumor growth without altering in vitro proliferation. Growth inhibition was associated with an altered profile of tumor-infiltrating cells with an increase in natural killer (NK) cells and a relative reduction in myeloid-derived suppressor cells and putative immune inhibitory plasmacytoid dendritic cells. Tumor inhibition required host expression of CMKLR1 (chemokine-like receptor 1), the chemoattractant receptor for chemerin, and was abrogated by NK cell depletion. Intratumoral injection of chemerin also inhibited tumor growth, suggesting the potential for therapeutic application. These results show that chemerin, whether expressed by tumor cells or within the tumor environment, can recruit host immune defenses that inhibit tumorigenesis and suggest that down-regulation of chemerin may be an important mechanism of tumor immune evasion.

The chemoattractant chemerin suppresses melanoma by recruiting natural killer cell antitumor defenses.

Despite circumstantial evidence postulating a protective role for NK cells in many fibrotic conditions, their contribution to the development of pulmonary fibrosis has yet to be tested. Lung-migrating NK cells are thought to attenuate the development of bleomycin induced pulmonary fibrosis (BIPF) by providing anti-fibrotic mediators and cytokines, such as IFN-γ. If true, we reasoned that depletion of NK cells during experimentally-induced fibrotic disease would lead to exacerbated fibrosis. To test this, we treated mice with NK cell-depleting antisera (anti-asialo GM1) and evaluated lung inflammation and fibrosis in the BIPF model. While NK cell infiltration into the airways was maximal at day 10 after bleomycin injection, NK cells represented a minor portion (1-3%) of the total leukocytes in BAL fluid. Anti-asialo GM1 significantly abrogated NK cell numbers over the course of the disease. Depletion of NK cells with anti-asialo GM1 before and throughout the BIPF model, or during just the fibrotic phase did not alter fibrosis development or affect the levels of any of the pro-inflammatory/pro-fibrotic cytokines measured (IL-1β, IL-17, IFN-γ, TGF-β and TNF-α). In addition, adoptively transferred NK cells, which were detectable systemically and in the airways throughout BIPF, failed to impact lung fibrosis. These findings indicate that NK cells likely do not play an essential protective role in controlling pulmonary fibrosis development.

Justin Monnier

Palo Alto Institute for Research and Education

Veterans Affairs Palo Alto Health Care System

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Justin Monnier

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Palo Alto Institute for Research and Education

Veterans Affairs Palo Alto Health Care System

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Palo Alto Institute for Research and Education