Immune Netw.  2017 Apr;17(2):116-120. 10.4110/in.2017.17.2.116.

IL-32-induced Inflammatory Cytokines Are Selectively Suppressed by α1-antitrypsin in Mouse Bone Marrow Cells

Affiliations
  • 1Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology, Konkuk University, Seoul 05029, Korea. soohyun@konkuk.ac.kr
  • 2YbdYbiotech Research Center, Seoul 08589, Korea.
  • 3Department of Molecular Science and Technology, Ajou University, Suwon 16499, Korea.
  • 4College of Veterinary Medicine, Konkuk University, Seoul 05029, Korea.
  • 5Division of Pulmonology, Department of Internal Medicine, School of Medicine, Konkuk University, Seoul 05029, Korea.
  • 6Department of Medicine, Pusan Paik Hospital, College of Medicine, Inje University, Busan 47392, Korea.
  • 7Korea Food Research Institute, Seongnam 13539, Korea.
  • 8Departments of Medicine and Academic Affairs, National Jewish Health, Denver, CO 80206, USA.
  • 9Division of Nephrology, Department of Internal Medicine, Jeju National University School of Medicine, Jeju 63243, Korea.
  • 10College of Veterinary Medicine and Veterinary Science Research Institute, Konkuk University, Seoul 05029, Korea.

Abstract

The induction of interleukin (IL)-32 in bone marrow (BM) inflammation is crucial in graft versus host disease (GvHD) that is a common side effect of allogeneic BM transplantation. Clinical trials on α-1 antitrypsin (AAT) in patients with GvHD are based on the preliminary human and mouse studies on AAT reducing the severity of GvHD. Proteinase 3 (PR3) is an IL-32-binding protein that was isolated from human urine. IL-32 primarily induces inflammatory cytokines in myeloid cells, probably due to PR3 expression on the membrane of the myeloid lineage cells. The inhibitory activity of AAT on serine proteinases may explain the anti-inflammatory effect of AAT on GvHD. However, the anti-inflammatory activity of AAT on BM cells remains unclear. Mouse BM cells were treated with IL-32γ and different inflammatory stimuli to investigate the anti-inflammatory activity of AAT. Recombinant AAT-Fc fusion protein inhibited IL-32γ-induced IL-6 expression in BM cells, but failed to suppress that induced by other stimuli. In addition, the binding of IL-32γ to PR3 was abrogated by AAT-Fc. The data suggest that the specific anti-inflammatory effect of AAT in mouse BM cells is due to the blocking of IL-32 binding to membrane PR3.

Keyword

Interleukin-32; Mouse bone marrow cell; Proteinase 3; Interleukin-6; Recombinant AAT-Fc

MeSH Terms

Animals
Bone Marrow Cells*
Bone Marrow*
Cytokines*
Graft vs Host Disease
Humans
Inflammation
Interleukin-6
Interleukins
Membranes
Mice*
Myeloblastin
Myeloid Cells
Serine Proteases
Cytokines
Interleukin-6
Interleukins
Myeloblastin
Serine Proteases

Figure

  • Figure 1 The anti-inflammatory effect of recombinant AAT-Fc in mouse BM cells. (A) IL-6 level in the supernatant of mouse BM cells was measured by sandwich ELISA (R&D Systems, Minneapolis MN). (B) The induction of IL-6 by different stimuli was measured without AAT-Fc (open bar) or with pre-incubation of AAT-Fc (closed bar, 2 µg/mL) for 3 h. (C) LPS-, (D) TNFα-, (E) IL-32γ-, and (F) IL-1α-mediated IL-6 level in the supernatant of BM cells isolated from the mice treated with AAT-Fc for 48 h was compared to that in the supernatant of BM cells isolated from the untreated control mice. Data in A are comparisons between untreated control and stimulated mice. Data in B are comparisons between RPMI control and AAT-Fc pre-incubated cells. Data in E are comparisons between BM cells isolated from untreated mice and BM cells isolated from mice treated with AAT-Fc (2 mg/kg) for 48 h. Mean±SEM; *p<0.05; #p<0.001 from three replicates. Representative data from 1 of 3 independent experiments are shown.

  • Figure 2 Suppression of inflammatory signaling in recombinant AAT-Fc-pretreated mouse BM cells, AAT-Fc competing with IL-32γ binding to PR3-Fc, and the dogma of AAT anti-inflammatory activity on IL-32-induced inflammation in BM transplantation. (A) The phosphorylation of NF-κB and p38MAPK by IL-32γ in BM cells from 6-week-old C57BL/6 mice was assessed using anti-phospho antibodies. (B) Similar experiments were performed by injecting AAT-Fc (2 mg/kg) into mice for 48 h prior to isolating BM cells. (C) Recombinant PR3-Fc was used for IL-32γ binding assay. IL-32γ was added at the concentration indicated on the bottom, and IL-32γ binds to PR3-Fc in a dose dependent manner. (D) The binding of IL-32γ was blocked with PR3-Fc in a dose dependent manner. Data in C are comparisons between control blank and IL-32γ-treated cells. Data in D are comparisons between control without AAT-Fc and with AAT-Fc. Mean±SEM; *p<0.05; **p<0.01; #p<0.001 from three replicates. Representative data from 1 of 4 independent experiments are shown. (C) Schematic representation of AAT-Fc suppression on IL-32-mediated inflammation in BM transplantation. Allogeneic BM transplantation induces different pro-inflammatory cytokines, such as IL-32, TNFα, IL-1α, and IL-6, resulting in immune reaction causing GvHD and death, while the specific IL-32 blockade AAT-Fc reduces immune reaction increasing survival.


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Reference

1. Schwartz RH, Van Ess JD, Johnstone DE, Dreyfuss EM, Abrishami MA, Chai H. Alpha-1 antitrypsin in childhood asthma. J Allergy Clin Immunol. 1977; 59:31–34.
Article
2. Becker K, Frieling T, Haussinger D. Quantification of fecal alpha 1-antitrypsin excretion for assessment of inflammatory bowel diseases. Eur J Med Res. 1998; 3:65–70.
3. Campbell EJ, Campbell MA, Boukedes SS, Owen CA. Quantum proteolysis by neutrophils: implications for pulmonary emphysema in alpha 1-antitrypsin deficiency. J Clin Invest. 1999; 104:337–344.
Article
4. Elzouki AN, Eriksson S, Lofberg R, Nassberger L, Wieslander J, Lindgren S. The prevalence and clinical significance of alpha 1-antitrypsin deficiency (PiZ) and ANCA specificities (proteinase 3, BPI) in patients with ulcerative colitis. Inflamm Bowel Dis. 1999; 5:246–252.
Article
5. Malerba M, Radaeli A, Ceriani L, Tantucci C, Grassi V. Airway hyperresponsiveness in a large group of subjects with alpha1-antitrypsin deficiency: a cross-sectional controlled study. J Intern Med. 2003; 253:351–358.
Article
6. Aldonyte R, Eriksson S, Piitulainen E, Wallmark A, Janciauskiene S. Analysis of systemic biomarkers in COPD patients. COPD. 2004; 1:155–164.
Article
7. Griese M, Latzin P, Kappler M, Weckerle K, Heinzlmaier T, Bernhardt T, Hartl D. alpha1-Antitrypsin inhalation reduces airway inflammation in cystic fibrosis patients. Eur Respir J. 2007; 29:240–250.
Article
8. Kwak A, Lee Y, Kim H, Kim S. Intracellular interleukin (IL)-1 family cytokine processing enzyme. Arch Pharm Res. 2016; 39:1556–1564.
Article
9. Novick D, Rubinstein M, Azam T, Rabinkov A, Dinarello CA, Kim SH. Proteinase 3 is an IL-32 binding protein. Proc Natl Acad Sci U S A. 2006; 103:3316–3321.
Article
10. Kim S, Lee S, Her E, Bae S, Choi J, Hong J, Jaekal J, Yoon D, Azam T, Dinarello CA, Kim S. Proteinase 3-processed form of the recombinant IL-32 separate domain. BMB Rep. 2008; 41:814–819.
Article
11. Bae S, Kang T, Hong J, Lee S, Choi J, Jhun H, Kwak A, Hong K, Kim E, Jo S, Kim S. Contradictory functions (activation/termination) of neutrophil proteinase 3 enzyme (PR3) in interleukin-33 biological activity. J Biol Chem. 2012; 287:8205–8213.
Article
12. Kim SJ, Lee S, Kwak A, Kim E, Jo S, Bae S, Lee Y, Ryoo S, Choi J, Kim S. Interleukin-32gamma transgenic mice resist LPS-mediated septic shock. J Microbiol Biotechnol. 2014; 24:1133–1142.
13. Kim S. Interleukin-32 in inflammatory autoimmune diseases. Immune Netw. 2014; 14:123–127.
Article
14. Jhun H, Choi J, Hong J, Lee S, Kwak A, Kim E, Jo S, Ryoo S, Lim Y, Yoon DY, Hong JT, Kim TS, Lee Y, Song K, Kim S. IL-32gamma overexpression accelerates streptozotocin (STZ)-induced type 1 diabetes. Cytokine. 2014; 69:1–5.
Article
15. Bae S, Kim YG, Choi J, Hong J, Lee S, Kang T, Jeon H, Hong K, Kim E, Kwak A, Lee CK, Yoo B, Park YB, Song EY, Kim S. Elevated interleukin-32 expression in granulomatosis with polyangiitis. Rheumatology (Oxford). 2012; 51:1979–1988.
Article
16. Jaekal J, Jhun H, Hong J, Park S, Lee J, Yoon D, Lee S, Her E, Yang Y, G Rho, Kim S. Cloning and characterization of bovine interleukin-32 beta isoform. Vet Immunol Immunopathol. 2010; 137:166–171.
Article
17. Hong J, Bae S, Kang Y, Yoon D, Bai X, Chan ED, Azam T, Dinarello CA, Lee S, Her E, Rho G, Kim S. Suppressing IL-32 in monocytes impairs the induction of the proinflammatory cytokines TNFalpha and IL-1beta. Cytokine. 2010; 49:171–176.
Article
18. Kim SH, Han SY, Azam T, Yoon DY, Dinarello CA. Interleukin-32: a cytokine and inducer of TNFalpha. Immunity. 2005; 22:131–142.
19. Marcondes AM, Li X, Tabellini L, Bartenstein M, Kabacka J, Sale GE, Hansen JA, Dinarello CA, Deeg HJ. Inhibition of IL-32 activation by alpha-1 antitrypsin suppresses alloreactivity and increases survival in an allogeneic murine marrow transplantation model. Blood. 2011; 118:5031–5039.
Article
20. Lee S, Kim E, Jhun H, Hong J, Kwak A, Jo S, Bae S, Lee J, Kim B, Lee J, Youn S, Kim S, Kim M, Kim H, Lee Y, Choi DK, Kim YS, Kim S. Proinsulin shares a motif with interleukin-1alpha (IL-1alpha) and induces inflammatory cytokine via interleukin-1 receptor 1. J Biol Chem. 2016; 291:14620–14627.
Article
21. Lee S, Lee Y, Hong K, Hong J, Bae S, Choi J, Jhun H, Kwak A, Kim E, Jo S, Dinarello CA. Effect of recombinant alpha1-antitrypsin Fc-fused (AAT-Fc) protein on the inhibition of inflammatory cytokine production and streptozotocin-induced diabetes. Mol Med. 2013; 19:65–71.
Article
22. Choi J, Bae S, Hong J, Ryoo S, Jhun H, Hong K, Yoon D, Lee S, Her E, Choi W, Kim J, Azam T, Dinarello CA, Kim S. Paradoxical effects of constitutive human IL-32{gamma} in transgenic mice during experimental colitis. Proc Natl Acad Sci U S A. 2010; 107:21082–21086.
Article
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