macrophage-differentiation-polarization-into-an-m2-like-phenotype

Macrophage Differentiation and Polarization into an M2-Like Phenotype using a Human Monocyte-Like THP-1 Leukemia Cell Line

Tumor-associated macrophages (TAM) can switch their expression and cytokine profile according to external stimuli. This remarkable plasticity enables TAM ...

University of Louisville

Recent clinical trials investigating the role of hyperoxia in decreasing surgical site infection have reported conflicting results. Immunologic mechanisms through which supplemental oxygen could act have not been elucidated fully. The authors sought to investigate the effects of hyperoxia on previously tested and prognostically significant innate immune parameters to uncover the potential effects of hyperoxia at the cellular level.

Oxygen and surgical site infection: a study of underlying immunologic mechanisms.

The tolerance of mouse strains to cecal ligation and puncture (CLP), a clinically relevant model of sepsis, can vary greatly. We compared the immune response and bacterial eradication during CLP in two mouse strains with different susceptibilities to the lethal effects in an effort to understand alterations in tolerance.

Genetic background determines susceptibility during murine septic peritonitis.

Natural killer (NK) cells are major cytokine producers during bacterial sepsis, but their precise role is undefined. This study investigates the effect of NK cell depletion with and without prior activation on macrophage function and bacterial clearance during cecal ligation and puncture. Two different NK cell-depleting antibodies were used: anti-asialo-GM1 (GM1), a nonactivating antibody, and anti-NK1.1 (NK1.1), an NK cell-activating antibody. C57BL/6 mice were NK depleted with either GM1 or NK1.1 by intraperitoneal injection 7 and 3 days before experimentation. Control animals received isotype immunoglobulin G. Depletion was confirmed by flow cytometry. Bacterial levels in peritoneal washout, blood, and liver were determined 4 hours after cecal ligation and puncture. Macrophage activation was measured by phagocytosis ability and by production of nitric oxide and interleukin-6. Depletion with GM1 resulted in significantly higher bacterial levels at 4 hours, whereas depletion with NK1.1 had the opposite effect of significantly decreasing bacterial levels. Macrophage phagocytosis ability was significantly increased in mice depleted with NK1.1 compared with those mice depleted with GM1. We conclude that activation of NK cells improves bacterial clearance by priming macrophages to help clear a subsequent bacterial challenge. Macrophages are less able to clear bacteria when NK cells are depleted without activation. NK cells are therefore important in bacterial clearance through interactions with macrophages.

Natural killer cell activation primes macrophages to clear bacterial infection.

&#x6211;&#x4EEC;&#x4EE5;&#x524D;&#x6240;&#x6307;&#x51FA;&#x4E0D;&#x8DB3;&#x4E2D;&#x6027;&#x7C92;&#x7EC6;&#x80DE;&#x805A;&#x96C6;&#xFF0C;&#x76F4;&#x5230;&#x7EC6;&#x83CC;&#x611F;&#x67D3;&#x90E8;&#x4F4D;&#x7684;&#x80BF;&#x7624;&#x574F;&#x6B7B;&#x56E0;&#x5B50;-&alpha; &#x53CD;&#x5E94;&#x7F3A;&#x4E4F;&#x6570;&#x5B57; &gt; 10(5) &#x83CC;&#x843D;&#x5F62;&#x6210;&#x5355;&#x4F4D;&#x6162;&#x6027;&#x5C0F;&#x9F20;&#x8179;&#x819C;&#x708E;&#x3001; &#x540E;&#x7EC6;&#x83CC;&#x6311;&#x6218;&#x4E0E;&#x514B;&#x96F7;&#x4F2F;&#x6746;&#x83CC;&#x80BA;&#x708E; (&#x514B;&#x96F7;&#x4F2F;&#x83CC;&#xFF09; &#x6697;&#x793A;&#x6709;&#x7F3A;&#x9677;&#x4E3B;&#x673A;&#x54CD;&#x5E94;&#x7684;&#x6A21;&#x5F0F;&#x3002;&#x8FD9;&#x9879;&#x7814;&#x7A76;&#x7684;&#x76EE;&#x7684;&#x662F;&#x786E;&#x5B9A;&#x6F5C;&#x5728;&#x514D;&#x75AB;&#x6291;&#x5236; IL-10 &#x7684;&#x5F71;&#x54CD;&#x5728;&#x6B64;&#x6A21;&#x578B;&#x4E2D;&#x7684;&#x6162;&#x6027;&#x8179;&#x819C;&#x708E;&#x3002;Balb/c &#x52A8;&#x7269;&#x662F;&#x8179;&#x8154;&#x6CE8;&#x5C04; 10 &#xFF08;3&#xFF09; &#x83CC;&#x843D;&#x5F62;&#x6210;&#x5355;&#x4F4D;&#x514B;&#x96F7;&#x4F2F;&#x83CC;&#x3002;&#x5E86;&#x5927;&#x9709;&#x7D20; (5 &#x6BEB;&#x514B;/&#x5343;&#x514B;/&#x5929;&#x53EB;&#xFF09; &#x76AE;&#x4E0B;&#x7ED9;&#x4E88; (s.c.) &#x4E3A;&#x671F;&#x4E24;&#x5929;&#xFF0C;&#x7136;&#x540E;&#x64A4;&#x56DE;&#x3002;&#x52A8;&#x7269;&#x6CBB;&#x7597;&#x6297;-IL-10 &#x6297;&#x4F53;&#x6216; igg &#x6297;&#x4F53;&#x540C;&#x79CD;&#x63A7;&#x5236; (s.c.) &#x4E4B;&#x524D;&#x6216;&#x4E4B;&#x540E;&#x514B;&#x96F7;&#x4F2F;&#x83CC;&#x7BA1;&#x7406;&#x3002;&#x751F;&#x5B58;&#x786E;&#x5B9A; 14 &#x5929;&#x4EE5;&#x4E0A;&#x3002;&#x540C;&#x6837;&#x5904;&#x7406;&#x7684;&#x52A8;&#x7269;&#x6536;&#x83B7;&#x540E; 48 h &#x83B7;&#x5F97;&#x809D;&#x810F;&#x3001; &#x8179;&#x8154;&#x6DB2;&#x548C;&#x8840;&#x6DB2;&#x3002;&#x7EC6;&#x83CC;&#x548C;&#x4E2D;&#x6027;&#x7C92;&#x7EC6;&#x80DE;&#x8BA1;&#x6570;&#x786E;&#x5B9A;&#x3002;&#x80BF;&#x7624;&#x574F;&#x6B7B;&#x56E0;&#x5B50;-&alpha; &#x548C;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-10 &#x7684;&#x9176;&#x8054;&#x514D;&#x75AB;&#x5438;&#x9644;&#x6CD5;&#x6D4B;&#x91CF;&#x3002;&#x6297;-IL-10 &#x6297;&#x4F53;&#x663E;&#x8457;&#x589E;&#x52A0;&#x751F;&#x5B58;&#x548C;&#x89C2;&#x5BDF;&#x5230;&#x7684;&#x8F66;&#x53A2;&#x5185;&#x7684;&#x7EC6;&#x83CC;&#x6E05;&#x9664;&#x3002;&#x6297;-IL-10 &#x7BA1;&#x7406;&#x5E76;&#x4E0D;&#x5BFC;&#x81F4;&#x589E;&#x52A0;&#x80BF;&#x7624;&#x574F;&#x6B7B;&#x56E0;&#x5B50;-&alpha; &#x6D53;&#x5EA6;&#x6216;&#x4E2D;&#x6027;&#x7C92;&#x7EC6;&#x80DE;&#x79EF;&#x7D2F;&#x5728;&#x7AD9;&#x70B9;&#x7684;&#x8F83;&#x4F4E;&#x7EA7;&#x522B;&#x7684;&#x514B;&#x96F7;&#x4F2F;&#x83CC;&#x611F;&#x67D3;&#x3002;&#x6211;&#x4EEC;&#x5F97;&#x51FA;&#x7ED3;&#x8BBA;&#xFF1A; &#x5185;&#x6E90;&#x6027;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-10 &#x5728;&#x6B64;&#x6A21;&#x578B;&#x4E2D;&#x7684;&#x6162;&#x6027;&#x8179;&#x819C;&#x708E;&#x751F;&#x5B58;&#x548C;&#x7EC6;&#x83CC;&#x6E05;&#x9664;&#x6709;&#x5BB3;&#x3002;

&#x5185;&#x6E90;&#x6027;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-10 &#x5BFC;&#x81F4;&#x53D7;&#x635F;&#x7EC6;&#x83CC;&#x6E05;&#x9664;&#xFF0C;&#x5E76;&#x51CF;&#x5C11;&#x6162;&#x6027;&#x8179;&#x819C;&#x708E;&#x5C0F;&#x9F20;&#x6A21;&#x578B;&#x4E2D;&#x7684;&#x751F;&#x5B58;&#x3002;

We previously observed insufficient neutrophil accumulation and a lack of TNF-alpha response at the site of infection until bacteria numbers >10(5) colony forming units in our model of chronic murine peritonitis, suggesting a defective host response after bacterial challenge with Klebsiella pneumoniae (Klebsiella). The aim of this study was to determine a potentially immunosuppressive effect of IL-10 in this model of chronic peritonitis. Balb/c animals were injected with 10(3) colony forming units Klebsiella intraperitoneally. Gentamicin (5 mg/kg/day BID) was given subcutaneously (s.c.) for two days and then withdrawn. Animals were treated with anti-IL-10 antibody or IgG isotype control (s.c.) before or after Klebsiella administration. Survival was determined over 14 days. Similarly treated animals were harvested after 48 h to obtain liver tissue, peritoneal fluid and blood. Bacteria and neutrophil counts were determined. TNF-alpha and IL-10 were measured by ELISA. Anti-IL-10 antibody significantly increased survival and bacterial clearance in the observed compartments. Anti-IL-10 administration did not lead to an increase in TNF-alpha concentrations or neutrophil accumulation at the site of infection at lower levels of Klebsiella. We conclude that endogenous IL-10 is detrimental for survival and bacterial clearance in this model of chronic peritonitis.

Endogenous IL-10 leads to impaired bacterial clearance and reduced survival in a murine model of chronic peritonitis.

Nous avons observ&eacute; pr&eacute;c&eacute;demment accumulation de neutrophile insuffisante et l&#39;absence de r&eacute;ponse de TNF-alpha sur le site de l&#39;infection &agrave; bact&eacute;ries num&eacute;ros &gt; colonie de 10 5 unit&eacute;s formant dans notre mod&egrave;le de chronique p&eacute;ritonite murine, sugg&eacute;rant une r&eacute;ponse de l&#39;h&ocirc;te d&eacute;fectueux apr&egrave;s d&eacute;fi bact&eacute;rien avec Klebsiella pneumoniae (Klebsiella). Le but de cette &eacute;tude &eacute;tait de d&eacute;terminer un effet potentiellement immunosuppresseur d&#39;IL-10 dans ce mod&egrave;le de p&eacute;ritonite chronique. Animaux BALB/c ont &eacute;t&eacute; inject&eacute;s par voie intrap&eacute;riton&eacute;ale avec 10 3 unit&eacute;s formant des colonies Klebsiella. Gentamicine (5 mg/kg/jour BID) a &eacute;t&eacute; donn&eacute; par voie sous-cutan&eacute;e (s.c.) pendant deux jours et ensuite retir&eacute;. Animaux ont &eacute;t&eacute; trait&eacute;s avec des anticorps anti-IL-10 ou contr&ocirc;le d&#39;isotype IgG (c.s.) avant ou apr&egrave;s l&#39;administration de Klebsiella. La survie a &eacute;t&eacute; d&eacute;termin&eacute;e sur une p&eacute;riode de 14 jours. De m&ecirc;me, les animaux trait&eacute;s ont &eacute;t&eacute; r&eacute;colt&eacute;s apr&egrave;s 48 h d&#39;obtenir des tissus h&eacute;patiques, liquide p&eacute;riton&eacute;al et le sang. On a d&eacute;termin&eacute; des bact&eacute;ries et des neutrophiles. TNF-alpha et IL-10 ont &eacute;t&eacute; mesur&eacute;s par ELISA. Anticorps anti-IL-10 a consid&eacute;rablement augment&eacute; la survie et la clairance bact&eacute;rienne dans les compartiments observ&eacute;es. Anti-IL-10 administration n&#39;a pas entra&icirc;n&eacute; une augmentation des concentrations de TNF-alpha ou neutrophile accumulation sur le site de l&#39;infection &agrave; des niveaux inf&eacute;rieurs de Klebsiella. Nous concluons qu&#39;endog&egrave;ne IL-10 est n&eacute;faste pour la clairance bact&eacute;rienne et de survie dans ce mod&egrave;le de p&eacute;ritonite chronique.

Endog&egrave;ne de IL-10 conduit &agrave; la clairance bact&eacute;rienne avec facult&eacute;s affaiblies et r&eacute;duit la survie dans un mod&egrave;le murin de p&eacute;ritonite chronique.

Wir zuvor beobachteten unzureichende neutrophil Anh&auml;ufung und Mangel an TNF-Alpha-Antwort am Ort der Infektion bis Bakterien Zahlen &gt; 10 Absatz 5 Kolonie bildende Einheiten in unserem Modell chronischer murinen Bauchfellentz&uuml;ndung, eine defekte Wirtsantwort nach bakteriellen Challenge mit Klebsiella Pneumoniae (Klebsiella) vorschlagen. Ziel dieser Studie war eine potenziell immunsuppressive Wirkung von IL-10 in diesem Modell chronische Bauchfellentz&uuml;ndung zu bestimmen. Balb/c Tieren wurden mit 10 Absatz 3 Kolonie bilden Einheiten Klebsiella intraperitoneal verabreicht. Gentamicin (5 mg/kg/Tag BID) erhielt subkutan (s.c.) f&uuml;r zwei Tage und dann zur&uuml;ckgezogen. Die Tiere wurden mit Anti-IL-10 Antik&ouml;rper oder IgG Isotype Kontrolle (s.c.) vor oder nach der Verabreichung von Klebsiella behandelt. &Uuml;berleben war &uuml;ber 14 Tage bestimmt. In &auml;hnlicher Weise wurden behandelte Tieren geerntet, nach 48 h Lebergewebe, peritonealen Fl&uuml;ssigkeit und Blut zu erhalten. Bakterien und neutrophil Grafen wurden ermittelt. TNF-Alpha und IL-10 wurden von ELISA gemessen. Anti-IL-10 Antik&ouml;rper deutlich erh&ouml;ht, &Uuml;berleben und bakterielle Spiel in die beobachteten F&auml;cher. Anti-IL-10 Verwaltung f&uuml;hrten nicht zu einem Anstieg der Konzentrationen von TNF-Alpha oder neutrophil Anh&auml;ufung am Standort der Infektion auf den unteren Ebenen von Klebsiella. Wir schlie&szlig;en, dass endogene IL-10 f&uuml;r &Uuml;berleben und bakteriellen Clearance in diesem Modell chronische Bauchfellentz&uuml;ndung abtr&auml;glich ist.

Endogenen IL-10 f&uuml;hrt zu gest&ouml;rter bakterieller Freiraum und &Uuml;berleben in einem murinen Modell chronische Bauchfellentz&uuml;ndung reduziert.

Abbiamo precedentemente osservato accumulo di neutrofili insufficiente e una mancanza di risposta di TNF-alfa nel sito di infezione fino a batteri numeri &amp;gt; 5 Colonia formando unit&agrave; nel nostro modello di peritonite cronica di murino, suggerendo una risposta ospite difettoso dopo sfida batterica con Klebsiella pneumoniae (Klebsiella). Lo scopo di questo studio era di determinare un effetto potenzialmente immunosoppressori di IL-10 in questo modello di peritonite cronica. Animali Balb/c sono stati iniettati con 10 unit&agrave; formanti colonie Klebsiella per via intraperitoneale. Gentamicina (5 mg/kg/giorno di offerta) &egrave; stato dato per via sottocutanea (s.c.) per due giorni e poi ritirato. Gli animali sono stati trattati con anticorpo anti-IL-10 o controllo di isotipo IgG (s.c.) prima o dopo la somministrazione di Klebsiella. Sopravvivenza &egrave; stato determinato per 14 giorni. Allo stesso modo degli animali trattati sono state raccolte dopo 48 h per ottenere tessuto epatico, liquido peritoneale e sangue. La conta dei neutrofili e batteri sono stata determinata. TNF-alfa e IL-10 sono stati misurati con ELISA. Anticorpo anti-IL-10 aumentato significativamente la sopravvivenza e la clearance batterica ribaltine osservati. Somministrazione di anti-IL-10 non ha condotto ad un aumento della concentrazione di TNF-alfa o accumulo di neutrofili presso il sito di infezione ai livelli inferiori della Klebsiella. Concludiamo che IL endogeno-10 &egrave; dannosa per la sopravvivenza e batterica clearance in questo modello di peritonite cronica.

Endogena di IL-10 conduce alla clearance batterica alterata e ridotta sopravvivenza in un modello murino di peritonite cronica.

&#x6211;&#x3005; &#x306F;&#x4EE5;&#x524D;&#x756A;&#x53F7;&#x304C;&#x4E0D;&#x8DB3;&#x3057;&#x3066;&#x3044;&#x308B;&#x597D;&#x4E2D;&#x7403;&#x96C6;&#x7A4D;&#x304A;&#x3088;&#x3073; TNF-&alpha; &#x306E;&#x5FDC;&#x7B54;&#x307E;&#x3067;&#x306E;&#x7D30;&#x83CC;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x3067;&#x306E;&#x6B20;&#x5982;&#x3092;&#x89B3;&#x5BDF;&#x76EE; 10 (5) &#x30B3;&#x30ED;&#x30CB;&#x30FC;&#x306E;&#x6162;&#x6027;&#x306E;&#x30DE;&#x30A6;&#x30B9;&#x8179;&#x819C;&#x708E;, &#x80BA;&#x708E;&#x687F;&#x83CC; (&#x30AF;&#x30EC;&#x30D6;&#x30B7;&#x30A8;&#x30E9;) &#x7D30;&#x83CC;&#x6311;&#x6226;&#x5F8C;&#x4E0D;&#x826F;&#x30DB;&#x30B9;&#x30C8;&#x5FDC;&#x7B54;&#x3092;&#x793A;&#x5506;&#x306E;&#x6211;&#x3005; &#x306E;&#x30E2;&#x30C7;&#x30EB;&#x306E;&#x5358;&#x4F4D;&#x3092;&#x5F62;&#x6210;&#x3057;&#x307E;&#x3059;&#x3002;&#x6162;&#x6027;&#x6027;&#x8179;&#x819C;&#x708E;&#x306E;&#x3053;&#x306E;&#x30E2;&#x30C7;&#x30EB;&#x306B;&#x304A;&#x3051;&#x308B; IL-10 &#x306E;&#x6F5C;&#x5728;&#x7684;&#x514D;&#x75AB;&#x6291;&#x5236;&#x52B9;&#x679C;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x305F;.Balb/c &#x52D5;&#x7269; 10 (3) &#x306F;&#x3001;&#x30B3;&#x30ED;&#x30CB;&#x30FC;&#x5F62;&#x6210;&#x5358;&#x4F4D;&#x30AF;&#x30EC;&#x30D6;&#x30B7;&#x30A8;&#x30E9;&#x8179;&#x8154;&#x5185;&#x6CE8;&#x5165;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x30B2;&#x30F3;&#x30BF;&#x30DE;&#x30A4;&#x30B7;&#x30F3; (5 mg/kg/&#x65E5;&#x306E;&#x5165;&#x672D;) &#x76AE;&#x4E0B;&#x3092;&#x4E0E;&#x3048;&#x3089;&#x308C;&#x305F; (&#x30B5;&#x30A6;&#x30B9;&#x30AB;&#x30ED;&#x30E9;&#x30A4;&#x30CA;&#x5DDE;) &#x306E; 2 &#x65E5;&#x9593;&#x3068;&#x305D;&#x306E;&#x5F8C;&#x64A4;&#x9000;&#x3002;&#x52D5;&#x7269;&#x30A2;&#x30F3;&#x30C1;-IL-10 &#x6297;&#x4F53;&#x307E;&#x305F;&#x306F; IgG &#x30A2;&#x30A4;&#x30BD;&#x30BF;&#x30A4;&#x30D7; &#x30B3;&#x30F3;&#x30C8;&#x30ED;&#x30FC;&#x30EB; (&#x30B5;&#x30A6;&#x30B9;&#x30AB;&#x30ED;&#x30E9;&#x30A4;&#x30CA;&#x5DDE;) &#x306E;&#x524D;&#x306B;&#x3001;&#x307E;&#x305F;&#x306F;&#x30AF;&#x30EC;&#x30D6;&#x30B7;&#x30A8;&#x30E9;&#x6295;&#x4E0E;&#x5F8C;&#x306B;&#x6271;&#x308F;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x751F;&#x5B58;&#x7387;&#x306F; 14 &#x65E5;&#x9593;&#x3092;&#x6C7A;&#x5B9A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x540C;&#x69D8;&#x306B;&#x6271;&#x308F;&#x308C;&#x305F;&#x52D5;&#x7269;&#x809D;&#x7D44;&#x7E54;&#x3001;&#x8179;&#x8154;&#x6DB2;&#x3001;&#x8840;&#x6DB2;&#x3092;&#x5165;&#x624B;&#x3059;&#x308B; 48 &#x6642;&#x9593;&#x5F8C;&#x306B;&#x53CE;&#x7A6B;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x7D30;&#x83CC;&#x304A;&#x3088;&#x3073;&#x597D;&#x4E2D;&#x7403;&#x6570;&#x3092;&#x6C7A;&#x5B9A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;TNF &alpha; &#x3068; IL-10 elisa &#x6CD5;&#x306B;&#x3088;&#x308A;&#x6E2C;&#x5B9A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x30A2;&#x30F3;&#x30C1;-IL-10 &#x6297;&#x4F53;&#x306F;&#x751F;&#x5B58;&#x3068;&#x7D30;&#x83CC;&#x30AF;&#x30EA;&#x30A2;&#x30E9;&#x30F3;&#x30B9;&#x89B3;&#x6E2C;&#x306E;&#x30B3;&#x30F3;&#x30D1;&#x30FC;&#x30C8;&#x30E1;&#x30F3;&#x30C8;&#x306B;&#x5927;&#x5E45;&#x5897;&#x52A0;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x30A2;&#x30F3;&#x30C1;-IL-10 &#x7BA1;&#x7406; TNF-&alpha; &#x6FC3;&#x5EA6;&#x307E;&#x305F;&#x306F;&#x611F;&#x67D3;&#x306E;&#x687F;&#x83CC;&#x306E;&#x4E0B;&#x4F4D;&#x30EC;&#x30D9;&#x30EB;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x3067;&#x597D;&#x4E2D;&#x7403;&#x96C6;&#x7A4D;&#x306E;&#x5897;&#x52A0;&#x306B;&#x306F;&#x81F3;&#x308A;&#x307E;&#x305B;&#x3093;&#x3067;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x306F;&#x5185;&#x56E0;&#x6027; IL-10 &#x304C;&#x6162;&#x6027;&#x6027;&#x8179;&#x819C;&#x708E;&#x306E;&#x3053;&#x306E;&#x30E2;&#x30C7;&#x30EB;&#x306E;&#x751F;&#x5B58;&#x3068;&#x7D30;&#x83CC;&#x30AF;&#x30EA;&#x30A2;&#x30E9;&#x30F3;&#x30B9;&#x306E;&#x305F;&#x3081;&#x6709;&#x5BB3;&#x3067;&#x3042;&#x308B;&#x3068;&#x7D50;&#x8AD6;&#x3057;&#x307E;&#x3059;&#x3002;

&#x5185;&#x56E0;&#x6027; IL-10 &#x969C;&#x5BB3;&#x306E;&#x7D30;&#x83CC;&#x30AF;&#x30EA;&#x30A2;&#x30E9;&#x30F3;&#x30B9;&#x306B;&#x3064;&#x306A;&#x304C;&#x308B;&#x3057;&#x3001;&#x6162;&#x6027;&#x6027;&#x8179;&#x819C;&#x708E;&#x306E;&#x30DE;&#x30A6;&#x30B9;&#x30E2;&#x30C7;&#x30EB;&#x3067;&#x306E;&#x751F;&#x5B58;&#x3092;&#x6E1B;&#x5C11;&#x3057;&#x307E;&#x3059;&#x3002;

&#xC6B0;&#xB9AC;&#xB294; &#xC774;&#xC804;&#xC5D0; &#xAD00;&#xCC30; &#xBD80;&#xC871; &#xD55C; &#xD638; &#xCD95;&#xC801;&#xACFC; &#xBC15;&#xD14C;&#xB9AC;&#xC544;&#xAE4C;&#xC9C0; &#xAC10;&#xC5FC;&#xC758; &#xC0AC;&#xC774;&#xD2B8;&#xC5D0; TNF-&#xC54C;&#xD30C; &#xC751;&#xB2F5;&#xC758; &#xBD80;&#xC871; &#xC22B;&#xC790; &amp;gt; 10(5) &#xC2DD;&#xBBFC;&#xC9C0; Klebsiella &#xADE0; (Klebsiella)&#xC640; &#xC138;&#xADE0; &#xB3C4;&#xC804; &#xD6C4; &#xACB0;&#xD568;&#xC774; &#xC788;&#xB294; &#xD638;&#xC2A4;&#xD2B8; &#xC751;&#xB2F5; &#xC81C;&#xC548; &#xB9CC;&#xC131; murine &#xBCF5; &#xB9C9; &#xC5FC;&#xC758; &#xC6B0;&#xB9AC;&#xC758; &#xBAA8;&#xB378;&#xC758; &#xB2E8;&#xC704;&#xB97C; &#xD615;&#xC131; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC758; &#xBAA9;&#xD45C;&#xB294; &#xB9CC;&#xC131; &#xBCF5; &#xB9C9; &#xC5FC;&#xC774;&#xC774; &#xBAA8;&#xB378;&#xC5D0;&#xC11C; IL-10&#xC758; &#xC7A0;&#xC7AC;&#xC801;&#xC73C;&#xB85C; &#xBA74;&#xC5ED; &#xD6A8;&#xACFC; &#xACB0;&#xC815; &#xD588;&#xB2E4;. Balb/c &#xB3D9;&#xBB3C; intraperitoneally 10 (3) &#xCF5C;&#xB85C; &#xB2C8; &#xD615;&#xC131; &#xB2E8;&#xC704; Klebsiella &#xC8FC;&#xC785; &#xD588;&#xB2E4;. &#xACE0; (5 mg/kg/&#xC77C; &#xC785;&#xCC30;) subcutaneously &#xC8FC;&#xC5B4;&#xC84C;&#xB2E4; (&#xC0AC;&#xC6B0;&#xC2A4; &#xCE90;&#xB864;&#xB77C;&#xC774;&#xB098;) &#xC774;&#xD2C0; &#xB3D9;&#xC548; &#xD558; &#xACE0; &#xB2E4;&#xC74C; &#xCCA0;&#xD68C;. &#xB3D9;&#xBB3C; Klebsiella &#xAD00;&#xB9AC; &#xC804;&#xD6C4; &#xC548;&#xD2F0;-IL-10 &#xD56D; &#xCCB4; &#xB610;&#xB294; IgG isotype &#xCEE8;&#xD2B8;&#xB864; (&#xC0AC;&#xC6B0;&#xC2A4; &#xCE90;&#xB864;&#xB77C;&#xC774;&#xB098;)&#xB85C; &#xCE58;&#xB8CC; &#xD588;&#xB2E4;. &#xC0DD;&#xC874;&#xC740; 14 &#xC77C; &#xB3D9;&#xC548;&#xC744; &#xACB0;&#xC815; &#xD588;&#xB2E4;. &#xB9C8;&#xCC2C;&#xAC00;&#xC9C0;&#xB85C; &#xCE58;&#xB8CC; &#xB3D9;&#xBB3C; &#xAC04; &#xC870;&#xC9C1;, &#xBCF5; &#xB9C9; &#xC561;&#xACFC; &#xD608;&#xC561; 48 h &#xD6C4; &#xC218;&#xD655; &#xD588;&#xB2E4;. &#xBC15;&#xD14C;&#xB9AC;&#xC544;&#xC640; &#xD638; &#xAC74;&#xC758; &#xACB0;&#xC815; &#xD588;&#xB2E4;. TNF-&#xC54C;&#xD30C;&#xC640; &#xC77C;&#xB9AC;&#xB178;&#xC774; 10 &#xC77C; &#xB77C;&#xC5D0; &#xC758;&#xD574; &#xCE21;&#xC815; &#xB418;&#xC5C8;&#xB2E4;. &#xC548;&#xD2F0;-IL-10 &#xD56D; &#xCCB4; &#xC0DD;&#xC874;&#xACFC; &#xAD00;&#xCC30; &#xB41C; &#xAD6C;&#xD68D;&#xC5D0;&#xC11C; &#xC138;&#xADE0;&#xC131; &#xD074;&#xB9AC;&#xC5B4;&#xB7F0;&#xC2A4;&#xC5D0; &#xD06C;&#xAC8C; &#xC99D;&#xAC00; &#xD588;&#xB2E4;. &#xC548;&#xD2F0;-IL-10 &#xAD00;&#xB9AC; TNF-&#xC54C;&#xD30C; &#xB18D;&#xB3C4; &#xB610;&#xB294; Klebsiella&#xC758; &#xD558;&#xC704; &#xC218;&#xC900;&#xC5D0;&#xC11C; &#xAC10;&#xC5FC;&#xC758; &#xC0AC;&#xC774;&#xD2B8;&#xC5D0; &#xD638; &#xCD95;&#xC801; &#xC99D;&#xAC00; &#xC774;&#xC5B4;&#xC9C8; &#xD558;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xB0B4; IL-10&#xC740; &#xD574;&#xB85C;&#xC6B4; &#xB9CC;&#xC131; &#xBCF5; &#xB9C9; &#xC5FC;&#xC774;&#xC774; &#xBAA8;&#xB378;&#xC5D0;&#xC11C; &#xC0DD;&#xC874; &#xBC0F; &#xC138;&#xADE0;&#xC131; &#xD5C8;&#xAC00; &#xB300; &#xD55C; &#xACB0;&#xB860;.

&#xB0B4; IL-10&#xACFC; &#xC7A5;&#xC560;&#xC778;&#xB41C; &#xC138;&#xADE0; &#xD074;&#xB9AC;&#xC5B4;&#xB7F0;&#xC2A4; &#xB9AC;&#xB4DC; &#xB9CC;&#xC131; &#xBCF5; &#xB9C9; &#xC5FC;&#xC758; murine &#xBAA8;&#xB378;&#xC5D0;&#xC11C; &#xC0DD;&#xC874;&#xC744; &#xAC10;&#xC18C;.

Anteriormente observamos insuficiente acumulaci&oacute;n de neutr&oacute;fila y la falta de respuesta de TNF-alfa en el sitio de la infecci&oacute;n hasta bacterias n&uacute;meros &gt; 5 10 formadoras unidades en nuestro modelo de la peritonitis cr&oacute;nica murina, sugiriendo una respuesta defectuosa del anfitri&oacute;n despu&eacute;s de desaf&iacute;o bacteriana con Klebsiella pneumoniae (Klebsiella). El objetivo de este estudio fue determinar un efecto potencialmente inmunosupresor de IL-10 en este modelo de peritonitis cr&oacute;nica. Animales BALB/c se inyectan por v&iacute;a intraperitoneal con 10 (3) unidades formadoras de colonias Klebsiella. Gentamicina (5 mg/kg/d&iacute;a BID) fue dado por v&iacute;a subcut&aacute;nea (s.c.) durante dos d&iacute;as y luego retirado. Animales fueron tratados con el anticuerpo anti-IL-10 o control de isotipo IgG (s.c.) antes o despu&eacute;s de la administraci&oacute;n de la Klebsiella. Supervivencia se determin&oacute; durante 14 d&iacute;as. Del mismo modo los animales tratados fueron cosechados despu&eacute;s de 48 h para obtener tejido del h&iacute;gado, sangre y l&iacute;quido peritoneal. Se determinaron las bacterias y los recuentos de neutr&oacute;filos. TNF-alfa e IL-10 fueron medidos por ELISA. Anticuerpo anti-IL-10 aument&oacute; significativamente la supervivencia y la separaci&oacute;n bacteriana en los compartimientos del observado. Administraci&oacute;n de anti-IL-10 no condujo a un aumento en las concentraciones de TNF-alfa o acumulaci&oacute;n de neutr&oacute;fila en el sitio de la infecci&oacute;n en los niveles inferiores de Klebsiella. Concluimos que Il end&oacute;geno-10 es perjudicial para el despacho de supervivencia y bacteriana en este modelo de peritonitis cr&oacute;nica.

Il end&oacute;geno-10 conduce a la separaci&oacute;n bacteriana deteriorada y reduce la supervivencia en un modelo murino de peritonitis cr&oacute;nica.

To examine cellular and immunologic mechanisms by which intraoperative hypothermia affects surgical patients.

Hypothermia and surgery: immunologic mechanisms for current practice.

Early clinical trials investigating the role of tightly controlled glucose levels showed marked benefit in survival of critically ill patients. However, a recent meta-analysis and large randomized controlled trial have failed to reproduce the benefit, showing instead substantially increased risk of dangerous hypoglycemia. We sought to investigate the effects of varying glucose concentrations on previously tested, prognostically significant, innate immune parameters, to define any potential effects of glucose at the cellular level.

Glucose and surgical sepsis: a study of underlying immunologic mechanisms.

To investigate liver damage and abscess formation in murine, secondary peritonitis.

Liver injury and abscess formation in secondary murine peritonitis.

Unintentional hypothermia is a well-described risk factor for death and complications after elective and emergency surgery. The molecular mechanisms by which hypothermia exerts its detrimental effects are not well understood. Differences in cytokine production and the overall cell function have been reported under hypothermic conditions. We investigated the effect of a range of clinically relevant temperatures on cytokine production and microRNA (miRNA) expression in a whole-blood model. We found that there was a wide variation in tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-10 production among different subjects, ranging from low to high TNF-α producers. The intersubject variation can also be found on the transcriptional level: high producers had higher upregulation of TNF-α messenger RNA than intermediate and low producers. This variation in TNF-α was reproducible in each individual. Temperature seems to modulate TNF-α production among these different groups. miRNA expression was modulated by temperature. miRNA-181a might control, or be a part of the mechanism which controls, TNF-α production. However, an analysis of whole-leukocyte RNA does not allow the investigation of mechanisms in a specific leukocyte subpopulation such as monocytes, because these changes may be concealed by miRNA expression changes in the other leukocyte subsets. In conclusion, TNF-α, IL-6, and IL-10 production is highly variable among different persons, but temperature affects the expression of miRNAs, which may consequently alter the production of TNF-α.

Does clinically relevant temperature change miRNA and cytokine expression in whole blood?

Therapeutic hypothermia is commonly used to improve neurological outcomes in patients after cardiac arrest. However, therapeutic hypothermia increases sepsis risk and unintentional hypothermia in surgical patients increases infectious complications. Nonetheless, the molecular mechanisms by which hypothermia dysregulates innate immunity are incompletely understood. We found that exposure of human monocytes to cold (32°C) potentiated LPS-induced production of TNF and IL-6, while blunting IL-10 production. This dysregulation was associated with increased expression of microRNA-155 (miR-155), which potentiates Toll-like receptor (TLR) signaling by negatively regulating Ship1 and Socs1. Indeed, Ship1 and Socs1 were suppressed at 32°C and miR-155 antagomirs increased Ship1 and Socs1 and reversed the alterations in cytokine production in cold-exposed monocytes. In contrast, miR-155 mimics phenocopied the effects of cold exposure, reducing Ship1 and Socs1 and altering TNF and IL-10 production. In a murine model of LPS-induced peritonitis, cold exposure potentiated hypothermia and decreased survival (10 vs. 50%; P < 0.05), effects that were associated with increased miR-155, suppression of Ship1 and Socs1, and alterations in TNF and IL-10. Importantly, miR-155-deficiency reduced hypothermia and improved survival (78 vs. 32%, P < 0.05), which was associated with increased Ship1, Socs1, and IL-10. These results establish a causal role of miR-155 in the dysregulation of the inflammatory response to hypothermia.

MicroRNA-155 potentiates the inflammatory response in hypothermia by suppressing IL-10 production.

To investigate whether warming to normal body temperature or to febrile range temperature (39°C) is able to reverse the detrimental effects of hypothermia.

Warming to 39°C but Not to 37°C Ameliorates the Effects on the Monocyte Response by Hypothermia.

Hypothermia is a well-known risk factor for postoperative complications because it prolongs the monocyte inflammatory response. The purpose of this study was to investigate whether temperature-activated ion channels (transient receptor protein channels [TRP] A1 and V1) mediate the effects of temperature on monocytes.

TRPA1 mediates the effects of hypothermia on the monocyte inflammatory response.

We have reported a new phenomenon in acute wound healing following the use of intracellular ATP delivery-extremely rapid tissue regeneration, which starts less than 24 h after surgery, and is accompanied by massive macrophage trafficking, in situ proliferation, and direct collagen production. This unusual process bypasses the formation of the traditional provisional extracellular matrix and significantly shortens the wound healing process. Although macrophages/monocytes are known to play a critical role in the initiation and progression of wound healing, their in situ proliferation and direct collagen production in wound healing have never been reported previously. We have explored these two very specific pathways during wound healing, while excluding confounding factors in the in vivo environment by analyzing wound samples and performing in vitro studies. The use of immunohistochemical studies enabled the detection of in situ macrophage proliferation in ATP-vesicle treated wounds. Primary human macrophages and Raw 264.7 cells were used for an in vitro study involving treatment with ATP vesicles, free Mg-ATP alone, lipid vesicles alone, Regranex, or culture medium. Collagen type 1α 1, MCP-1, IL-6, and IL-10 levels were determined by ELISA of the culture supernatant. The intracellular collagen type 1α1 localization was determined with immunocytochemistry. ATP-vesicle treated wounds showed high immunoreactivity towards BrdU and PCNA antigens, indicating in situ proliferation. Most of the cultured macrophages treated with ATP-vesicles maintained their classic phenotype and expressed high levels of collagen type 1α1 for a longer duration than was observed with cells treated with Regranex. These studies provide the first clear evidence of in situ macrophage proliferation and direct collagen production during wound healing. These findings provide part of the explanation for the extremely rapid tissue regeneration, and this treatment may hold promise for acute and chronic wound care.

Rapid tissue regeneration induced by intracellular ATP delivery-A preliminary mechanistic study.

This study focuses on impaired monocyte function, which occurs in some patients after trauma, major elective surgery, or sepsis. This monocyte impairment increases the risk of secondary infection and death. We aimed to determine the influence IκK-16 had on monocytes using an ex-vivo model of human monocyte impairment. We included the effects of the well-studied comparators interferon-gamma (IFN-γ) and granulocyte-macrophage colony-stimulating factor (GM-CSF) on impaired monocytes. Primary human monocytes were stimulated with 10ng/mL of lipopolysaccharide (LPS) for 16h and then challenged with 100ng/mL LPS to assess the monocyte inflammatory response. Treatment regimens, consisting of either IκK-16, IFN-γ, or GM-CSF, were administered to impaired monocytes near the time of initial LPS stimulation. Stimulation with 10ng/mL LPS initially promoted a pro-inflammatory response but subsequently impaired production of both tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) and decreased HLA-DR expression. IκK-16 treatment attenuated TNF-α production and programmed death-ligand 1 (PD-L1) expression and increased IL-10 and CD14 expression. IFN-γ treatment increased TNF-α production as well as PD-L1 and HLA-DR expression. In conclusion, limiting early inflammation with IκK-16 suppresses TNF-α production and PD-L1 expression but enhances IL-10 production and preserves CD14 expression for potential future exposure to infective stimuli.

The effect of IκK-16 on lipopolysaccharide-induced impaired monocytes.

Severe injury can lead to immune dysfunction and predispose patients to infection and death. Micro-RNAs regulate gene expression and may act as biomarkers for susceptibility to infection. The aim of this study was to examine the temporal and differential expression of previously identified dysregulated micro-RNAs in patients with severe injury.

Temporal expression of circulating miRNA after severe injury.

Following major trauma, sepsis or surgery, some patients exhibit an impaired monocyte inflammatory response that is characterized by a decreased response to a subsequent bacterial challenge. To investigate this poorly understood phenomenon, we adopted an in-vitro model of endotoxin tolerance utilising primary human CD14 + monocytes to focus on the effect of impairment on IκKα/β, a critical part of the NFκB pathway. Impaired monocytes had decreased IκKα mRNA and protein expression and decreased phosphorylation of the IκKα/β complex. The impaired monocyte secretome demonstrated a distinct cytokine/chemokine footprint from the naïve monocyte, and that TNF-α was the most sensitive cytokine or chemokine in this setting of impairment. Inhibition of IκKα/β with a novel selective inhibitor reproduced the impaired monocyte phenotype with decreased production of TNF-α, IL-6, IL-12p70, IL-10, GM-CSF, VEGF, MIP-1β, TNF-β, IFN-α2 and IL-7 in response to an LPS challenge. Surgical patients with infection also exhibited an impaired monocyte phenotype and had decreased SITPEC, TAK1 and MEKK gene expression, which are important for IκKα/β activation. Our results emphasize that impaired monocyte function is, at least in part, related to dysregulated IκKα/β activation, and that IκKα/β is likely involved in mounting a sufficient monocyte inflammatory response. Future studies may wish to focus on adjuvant therapies that augment IκKα/β function to restore monocyte function in this clinically important problem.

The role and function of IκKα/β in monocyte impairment.

Immune dysfunction can occur during sepsis or following major trauma. Decreased monocyte HLA-DR expression and cytokine responses are associated with mortality. Recent studies have shown that adaptive immune system defects can also occur in such patients, characterised by increased PD-L1 expression and associated T-cell anergy. The aim of this study was to determine the effects of an immune adjuvant, interferon-gamma, on monocyte PD-L1 expression and T-cell activation in an ex-vivo human whole blood model of infection. We found that with interferon-gamma treatment, monocytes had increased HLA-DR expression and augmented TNF-α production in response to LPS stimulation, with a decrease in IL-10 levels. Both LPS and interferon-gamma increased the level of monocyte PD-L1 expression, and that a combination of both agents synergistically stimulated a further increase in PD-L1 levels as measured by flow cytometry. However, despite elevated PD-L1 expression, both CD4 and CD8 T-cell activation was not diminished by the addition of interferon-gamma treatment. These findings suggest that PD-L1 may not be a reliable marker for T-cell anergy, and that interferon-gamma remains an adjuvant of interest that can improve the monocyte inflammatory response while preserving T-cell activation.

Article	Year
Oxygen and surgical site infection: a study of underlying immunologic mechanisms. Anesthesiology\| PubMed ID: 20613473	2010
Genetic background determines susceptibility during murine septic peritonitis. The Journal of surgical research\| PubMed ID: 11792151	2002
Natural killer cell activation primes macrophages to clear bacterial infection. The American surgeon\| PubMed ID: 12953826	2003
Endogenous IL-10 leads to impaired bacterial clearance and reduced survival in a murine model of chronic peritonitis. Cytokine\| PubMed ID: 18053738	2007
Hypothermia and surgery: immunologic mechanisms for current practice. Annals of surgery\| PubMed ID: 19561472	2009
Glucose and surgical sepsis: a study of underlying immunologic mechanisms. Journal of the American College of Surgeons\| PubMed ID: 20510806	2010
Liver injury and abscess formation in secondary murine peritonitis. Inflammation research : official journal of the European Histamine Research Society ... [et al.]\| PubMed ID: 20976524	2011
Does clinically relevant temperature change miRNA and cytokine expression in whole blood? Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research\| PubMed ID: 22909186	2012
MicroRNA-155 potentiates the inflammatory response in hypothermia by suppressing IL-10 production. FASEB journal : official publication of the Federation of American Societies for Experimental Biology\| PubMed ID: 25231976	2014
Warming to 39°C but Not to 37°C Ameliorates the Effects on the Monocyte Response by Hypothermia. Annals of surgery\| PubMed ID: 25719808	2016
TRPA1 mediates the effects of hypothermia on the monocyte inflammatory response. Surgery\| PubMed ID: 26054320	2015
Rapid tissue regeneration induced by intracellular ATP delivery-A preliminary mechanistic study. PloS one\| PubMed ID: 28380006	2017
The effect of IκK-16 on lipopolysaccharide-induced impaired monocytes. Immunobiology\| PubMed ID: 29126656	2018
Temporal expression of circulating miRNA after severe injury. Surgery\| PubMed ID: 30077391	2018
The role and function of IκKα/β in monocyte impairment. Scientific reports\| PubMed ID: 32699255	2020
Interferon-gamma increases monocyte PD-L1 but does not diminish T-cell activation. Cellular immunology\| PubMed ID: 32891037	2020

Sarah A. Gardner

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