Lentivirus-mediated overexpression of suppressor of cytokine signaling-3 reduces neutrophilic airway inflammation 
by suppressing T-helper 17 responses in mice with chronic Pseudomonas aeruginosa lung infections

Ding,Feng‑Ming; Zhang,Xing‑Yi; Chen,Yu‑Qing; Liao,Ruo‑Min; Xie,Guo‑Gang; Zhang,Peng‑Yu; Shao,Ping; Zhang,Min

doi:10.3892/ijmm.2018.3417

Lentivirus-mediated overexpression of suppressor of cytokine signaling-3 reduces neutrophilic airway inflammation by suppressing T-helper 17 responses in mice with chronic Pseudomonas aeruginosa lung infections

Authors:
- Feng‑Ming Ding
- Xing‑Yi Zhang
- Yu‑Qing Chen
- Ruo‑Min Liao
- Guo‑Gang Xie
- Peng‑Yu Zhang
- Ping Shao
- Min Zhang
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Affiliations: Department of Respiratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China
Published online on: January 23, 2018 https://doi.org/10.3892/ijmm.2018.3417
Pages: 2193-2200

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Abstract

The aim of the present study was to explore the effect of overexpressed suppressor of cytokine signaling‑3 (SOCS3) on T-helper (Th)17 cell responses and neutrophilic airway inflammation in mice with chronic Pseudomonas aeruginosa (PA) infections. SOCS3 expression was enhanced via the administration of tail vein injections of therapeutic lentivirus in mice with chronic PA lung infections. SOCS3 expression in the blood and lung tissue was assessed using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis. Total and differential cell numbers and myeloperoxidase levels in the bronchoalveolar lavage (BAL) fluid were assessed, as well as the number of bacterial colonies in the lungs. Histological analysis of lung tissue was performed using hematoxylin and eosin staining and phosphorylated‑signal transducer and activator of transcription‑3 (p‑STAT3) expression was measured by western blot analysis and immunohistochemistry. The expression of STAT3 mRNA and retinoid‑related orphan receptor (ROR)γt were measured by RT‑qPCR. The percentage of interleukin (IL)‑17+ cells among cluster of differentiation (CD)4+ cells was calculated using flow cytometry and levels of IL‑17A and IL‑6 were assessed by ELISA. The expression of SOCS3 was significantly increased in CD4+ T cells following lentivirus injection and the inflammation of neutrophilic airways was notably ameliorated. Enhanced SOCS3 expression was associated with a significant decrease in the expression of p‑STAT3 and RORγt in CD4+ T cells. Additionally, the percentage of IL‑17+ cells among CD4+ T cells and the IL‑17 contents in the BAL fluid were significantly decreased. Lentivirus‑mediated overexpression of SOCS3 was revealed to ameliorate neutrophilic airway inflammation by inhibiting pulmonary Th17 responses in mice with chronic PA lung infections.

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1	Hengzhuang W, Song Z, Ciofu O, Onsøyen E, Rye PD and Høiby N: OligoG CF-5/20 disruption of mucoid Pseudomonas aeruginosa biofilm in a murine lung infection model. Antimicrob Agents Chemother. 60:2620–2626. 2016. View Article : Google Scholar : PubMed/NCBI
2	Mayer-Hamblett N, Kloster M, Rosenfeld M, Gibson RL, Retsch-Bogart GZ, Emerson J, Thompson V and Ramsey BW: Impact of sustained eradication of new pseudomonas aeruginosa infection on long-term outcomes in cystic fibrosis. Clin Infect Dis. 61:707–715. 2015. View Article : Google Scholar : PubMed/NCBI
3	Sharma G, Rao S, Bansal A, Dang S, Gupta S and Gabrani R: Pseudomonas aeruginosa biofilm: Potential therapeutic targets. Biologicals. 42:1–7. 2014. View Article : Google Scholar
4	Darch SE, McNally A, Harrison F, Corander J, Barr HL, Paszkiewicz K, Holden S, Fogarty A, Crusz SA and Diggle SP: Recombination is a key driver of genomic and phenotypic diversity in a Pseudomonas aeruginosa population during cystic fibrosis infection. Sci Rep. 5:76492015. View Article : Google Scholar : PubMed/NCBI
5	Armstrong DS, Hook SM, Jamsen KM, Nixon GM, Carzino R, Carlin JB, Robertson CF and Grimwood K: Lower airway inflammation in infants with cystic fibrosis detected by newborn screening. Pediatr Pulmonol. 40:500–510. 2005. View Article : Google Scholar : PubMed/NCBI
6	Regamey N, Tsartsali L, Hilliard TN, Fuchs O, Tan HL, Zhu J, Qiu YS, Alton EW, Jeffery PK, Bush A and Davies JC: Distinct patterns of inflammation in the airway lumen and bronchial mucosa of children with cystic fibrosis. Thorax. 67:164–170. 2012. View Article : Google Scholar
7	Kolls JK: CD4⁺ T-cell subsets and host defense in the lung. Immunol Rev. 252:156–163. 2013. View Article : Google Scholar : PubMed/NCBI
8	Veldhoen M, Hocking RJ, Atkins CJ, Locksley RM and Stockinger B: TGFbeta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-producing T cells. Immunity. 24:179–189. 2006. View Article : Google Scholar : PubMed/NCBI
9	Mangan PR, Harrington LE, O'Quinn DB, Helms WS, Bullard DC, Elson CO, Hatton RD, Wahl SM, Schoeb TR and Weaver CT: Transforming growth factor-β induces development of the T_H17 lineage. Nature. 441:231–234. 2006. View Article : Google Scholar : PubMed/NCBI
10	Yang XO, Pappu BP, Nurieva R, Akimzhanov A, Kang HS, Chung Y, Li M, Shah B, Panopoulos AD, Schluns KS, et al: T helper 17 lineage differentiation is programmed by orphan nuclear receptors ROR alpha and ROR gamma. Immunity. 28:29–39. 2008. View Article : Google Scholar : PubMed/NCBI
11	Ivanov II, Mckenzie BS, Liang Z, Tadokoro CE, Lepelley A, Lafaille JJ, Cua DJ and Littman DR: The orphan nuclear receptor RORgammat directs the differentiation program of proinflammatory IL-17⁺ T helper cells. Cell. 126:1121–1133. 2006. View Article : Google Scholar : PubMed/NCBI
12	Kolls JK and Lindén A: Interleukin-17 family members and inflammation. Immunity. 21:467–476. 2004. View Article : Google Scholar : PubMed/NCBI
13	Lubberts E: The role of IL-17 and family members in the pathogenesis of arthritis. Curr Opin Investig Drugs. 4:572–577. 2003.PubMed/NCBI
14	Dubin PJ and Kolls JK: IL-23 mediates inflammatory responses to mucoid Pseudomonas aeruginosa lung infection in mice. Am J Physiol Lung Cell Mol Physiol. 292:L519–L528. 2007. View Article : Google Scholar
15	Linossi EM, Babon JJ, Hilton DJ and Nicholson SE: Suppression of cytokine signaling: The SOCS prespective. Cytokine Growth Factor Rev. 24:241–248. 2013. View Article : Google Scholar : PubMed/NCBI
16	Yoshimura A and Yasukawa H: JAK's SOCS: A mechanism of inhibition. Immunity. 36:157–159. 2012. View Article : Google Scholar : PubMed/NCBI
17	Liu X, Ren S, Qu X, Ge C, Cheng K and Zhao RC: Mesenchymal stem cells inhibit Th17 cells differentiation via IFN-γ-mediated SOCS3 activation. Immunol Res. 61:219–229. 2015. View Article : Google Scholar : PubMed/NCBI
18	Tanaka K, Ichiyama K, Hashimoto M, Yoshida H, Takimoto T, Takaesu G, Torisu T, Hanada T, Yasukawa H, Fukuyama S, et al: Loss of suppressor of cytokine signaling 1 in helper T cells leads to defective Th17 differentiation by enhancing antagonistic effects of IFN-gamma on STAT3 and Smads. J Immunol. 180:3746–3756. 2008. View Article : Google Scholar : PubMed/NCBI
19	Chen Z, Laurence A, Kanno Y, Pacher-Zavisin M, Zhu BM, Tato C, Yoshimura A, Hennighausen L and O'Shea JJ: Selective regulatory function of Socs3 in the formation of IL-17-secreting T cells. Proc Natl Acad Sci USA. 103:8137–8142. 2006. View Article : Google Scholar : PubMed/NCBI
20	van Heeckeren AM and Schluchter MD: Murine models of chronic Pseudomonas aeruginosa lung infection. Lab Anim. 36:291–312. 2002. View Article : Google Scholar : PubMed/NCBI
21	Brown BD, Venneri MA, Zingale A, Sergi Sergi L and Naldini L: Endogenous microRNA regulation suppresses transgene expression in hematopoietic lineages and enables stable gene transfer. Nat Med. 12:585–591. 2006. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔ^C T method. Methods. 25:402–408. 2011. View Article : Google Scholar
23	Dalpke A and Heeg K: Suppressors of cytokine signaling proteins in innate and adaptive immune responses. Arch Immunol Ther Exp. 51:91–103. 2003.
24	Carow B and Rottenberg ME: SOCS3, a major regulator of infection and inflammation. Front Immunol. 5:582014. View Article : Google Scholar : PubMed/NCBI
25	Khader SA, Pearl JE, Sakamoto K, Gilmartin L, Bell GK, Jelley-Gibbs DM, Ghilardi N, deSauvage F and Cooper AM: IL-23 compensates for the absence of IL-12p70 and is essential for the IL-17 response during tuberculosis but is dispensable for protection and antigen-specific IFN-gamma responses if IL-12p70 is available. J Immunol. 175:788–795. 2005. View Article : Google Scholar : PubMed/NCBI
26	Nguyen-Jackson H, Panopoulos AD, Zhang H, Li HS and Watowich SS: STAT3 controls the neutrophil migratory response to CXCR2 ligands by direct activation of G-CSF-induced CXCR2 expression and via modulation of CXCR2 signal transduction. Blood. 115:3354–3363. 2010. View Article : Google Scholar : PubMed/NCBI
27	Xu D and Qu CK: Protein tyrosine phosphatases in the JAK/STAT pathway. Front Biosci. 13:4925–4932. 2008. View Article : Google Scholar : PubMed/NCBI
28	Shuai K and Liu B: Regulation of JAK-STAT signalling in the immune system. Nat Rev Immunol. 3:900–911. 2003. View Article : Google Scholar : PubMed/NCBI
29	Shouda T, Yoshida T, Hanada T, Wakioka T, Oishi M, Miyoshi K, Komiya S, Kosai K, Hanakawa Y, Hashimoto K, et al: Induction of the cytokine signal regulator SOCS3/CIS3 as a therapeutic strategy for treating inflammatory arthritis. J Clin Invest. 108:1781–1788. 2001. View Article : Google Scholar : PubMed/NCBI
30	Suzuki A, Hanada T, Mitsuyama K, Yoshida T, Kamizono S, Hoshino T, Kubo M, Yamashita A, Okabe M, Takeda K, et al: Cis3/Socs3/Ssi3 plays a negative regulatory role in STAT3 activation and intestinal inflammation. J Exp Med. 193:471–481. 2001. View Article : Google Scholar : PubMed/NCBI
31	Harris TJ, Grosso JF, Yen HR, Xin H, Kortylewski M, Albesiano E, Hipkiss EL, Getnet D, Goldberg MV, Maris CH, et al: Cutting edge: An in vivo requirement for STAT3 signaling in TH17 development and TH17-dependent autoimmunity. J Immunol. 179:4313–4317. 2007. View Article : Google Scholar : PubMed/NCBI
32	Ernst M, Najdovska M, Grail D, Lundgren-May T, Buchert M, Tye H, Matthews VB, Armes J, Bhathal PS, Hughes NR, et al: STAT3 and STAT1 mediate IL-11-dependent and inflammation- associated gastric tumorigenesis in gp130 receptor mutant mice. J Clin Invest. 118:1727–1738. 2008.PubMed/NCBI
33	Brender C, Tannahill GM, Jenkins BJ, Fletcher J, Columbus R, Saris CJ, Ernst M, Nicola NA, Hilton DJ, Alexander WS and Starr R: Suppressor of cytokine signaling 3 regulates CD8 T-cell proliferation by inhibition of interleukins 6 and 27. Blood. 110:2528–2536. 2007. View Article : Google Scholar : PubMed/NCBI
34	Moseley TA, Haudenschild DR, Rose L and Reddi AH: Interleukin-17 family and IL-17 receptors. Cytokine Growth Factor Rev. 14:155–174. 2003. View Article : Google Scholar : PubMed/NCBI
35	Ye P, Garvey PB, Zhang P, Nelson S, Bagby G, Summer WR, Schwarzenberger P, Shellito JE and Kolls JK: Interleukin-17 and lung host defense against Klebsiella pneumoniae infection. Am J Respir Cell Mol Biol. 25:335–340. 2001. View Article : Google Scholar : PubMed/NCBI
36	Happel KI, Zheng M, Young E, Quinton LJ, Lockhart E, Ramsay AJ, Shellito JE, Schurr JR, Bagby GJ, Nelson S and Kolls JK: Cutting edge: Roles of Toll-like receptor 4 and IL-23 in IL-17 expression in response to Klebsiella pneumoniae infection. J Immunol. 170:4432–4436. 2003. View Article : Google Scholar : PubMed/NCBI
37	Lindén A and Adachi M: Neutrophilic airway inflammation and IL-17. Allergy. 57:769–775. 2002. View Article : Google Scholar : PubMed/NCBI