References
1. Barber GN. STING: infection, inflammation and cancer. Nat Rev
Immunol. 2015;15(12):760-70.
2. Gall A, Treuting P, Elkon KB, Loo YM, Gale M, Jr., Barber GN, et al.
Autoimmunity initiates in nonhematopoietic cells and progresses via
lymphocytes in an interferon-dependent autoimmune disease. Immunity.
2012;36(1):120-31.
3. Hotchkiss RS, Swanson PE, Freeman BD, Tinsley KW, Cobb JP, Matuschak
GM, et al. Apoptotic cell death in patients with sepsis, shock, and
multiple organ dysfunction. Crit Care Med. 1999;27(7):1230-51.
4. Denny MF, Yalavarthi S, Zhao W, Thacker SG, Anderson M, Sandy AR, et
al. A distinct subset of proinflammatory neutrophils isolated from
patients with systemic lupus erythematosus induces vascular damage and
synthesizes type I IFNs. J Immunol. 2010;184(6):3284-97.
5. Cekic C, Sag D, Day YJ, Linden J. Extracellular adenosine regulates
naive T cell development and peripheral maintenance. J Exp Med.
2013;210(12):2693-706.
6. El-Darahali A, Fawcett H, Mader JS, Conrad DM, Hoskin DW.
Adenosine-induced apoptosis in EL-4 thymoma cells is caspase-independent
and mediated through a non-classical adenosine receptor. Exp Mol Pathol.
2005;79(3):249-58.
7. Takahashi HK, Iwagaki H, Hamano R, Kanke T, Liu K, Sadamori H, et al.
Effect of adenosine receptor subtypes stimulation on mixed lymphocyte
reaction. Eur J Pharmacol. 2007;564(1-3):204-10.
8. Cronstein BN, Levin RI, Philips M, Hirschhorn R, Abramson SB,
Weissmann G. Neutrophil adherence to endothelium is enhanced via
adenosine A1 receptors and inhibited via adenosine A2 receptors. J
Immunol. 1992;148(7):2201-6.
9. Gu C, Ma YC, Benjamin J, Littman D, Chao MV, Huang XY. Apoptotic
signaling through the beta -adrenergic receptor. A new Gs effector
pathway. J Biol Chem. 2000;275(27):20726-33.
10. Salmon JE, Cronstein BN. Fc gamma receptor-mediated functions in
neutrophils are modulated by adenosine receptor occupancy. A1 receptors
are stimulatory and A2 receptors are inhibitory. J Immunol.
1990;145(7):2235-40.
11. Barletta KE, Ley K, Mehrad B. Regulation of neutrophil function by
adenosine. Arterioscler Thromb Vasc Biol. 2012;32(4):856-64.
12. Ohta A, Sitkovsky M. Role of G-protein-coupled adenosine receptors
in downregulation of inflammation and protection from tissue damage.
Nature. 2001;414(6866):916-20.
13. Riff R, Cohen Y, Eini-Rider H, Naamani O, Mazar J, Haviv YS, et al.
Systemic inflammatory response syndrome-related lymphopenia is
associated with adenosine A1 receptor dysfunction. J Leukoc Biol.
2017;102(1):95-103.
14. Leiss H, Niederreiter B, Bandur T, Schwarzecker B, Bluml S, Steiner
G, et al. Pristane-induced lupus as a model of human lupus arthritis:
evolvement of autoantibodies, internal organ and joint inflammation.
Lupus. 2013;22(8):778-92.
15. Hinson RM, Williams JA, Shacter E. Elevated interleukin 6 is induced
by prostaglandin E2 in a murine model of inflammation: possible role of
cyclooxygenase-2. Proc Natl Acad Sci U S A. 1996;93(10):4885-90.
16. Nordan RP, Potter M. A macrophage-derived factor required by
plasmacytomas for survival and proliferation in vitro. Science.
1986;233(4763):566-9.
17. Surawut S, Makjaroen J, Thim-Uam A, Wongphoom J, Palaga T, Pisitkun
P, et al. Increased susceptibility against Cryptococcus neoformans of
lupus mouse models (pristane-induction and FcGRIIb deficiency) is
associated with activated macrophage, regardless of genetic background.
J Microbiol. 2018.
18. Bortoluzzi A, Vincenzi F, Govoni M, Padovan M, Ravani A, Borea PA,
et al. A2A adenosine receptor upregulation correlates with disease
activity in patients with systemic lupus erythematosus. Arthritis Res
Ther. 2016;18:192.
19. Goldshtein H, Hausmann MJ, Douvdevani A. A rapid direct fluorescent
assay for cell-free DNA quantification in biological fluids. Ann Clin
Biochem. 2009;46(Pt 6):488-94.
20. Lenzen S. The mechanisms of alloxan- and streptozotocin-induced
diabetes. Diabetologia. 2008;51(2):216-26.
21. Gao F, Zheng ZM. Animal models of diabetic neuropathic pain. Exp
Clin Endocrinol Diabetes. 2014;122(2):100-6.
22. Agarwal N, Helmstaedter JP, Rangel Roja D, Kumar Bali K, Gangadharan
V, Kuner R. [EXPRESS] Evoked hypoalgesia is accompanied by tonic
pain and immune cell infiltration in the dorsal root ganglia at late
stages of diabetic neuropathy in mice. Mol Pain. 2018:1744806918817975.
23. Huang AC, Hu L, Kauffman SA, Zhang W, Shmulevich I. Using cell fate
attractors to uncover transcriptional regulation of HL60 neutrophil
differentiation. BMC Syst Biol. 2009;3:20.
24. Swamydas M, Luo Y, Dorf ME, Lionakis MS. Isolation of Mouse
Neutrophils. Curr Protoc Immunol. 2015;110:3 20 1-3 15.
25. Remijsen Q, Vanden Berghe T, Wirawan E, Asselbergh B, Parthoens E,
De Rycke R, et al. Neutrophil extracellular trap cell death requires
both autophagy and superoxide generation. Cell Res. 2011;21(2):290-304.
26. Nadesalingam A, Chen JHK, Farahvash A, Khan MA. Hypertonic Saline
Suppresses NADPH Oxidase-Dependent Neutrophil Extracellular Trap
Formation and Promotes Apoptosis. Front Immunol. 2018;9:359.
27. Naamani O, Riff R, Chaimovitz C, Mazar J, Douvdevani A.
Pharmacological preconditioning with adenosine A1 receptor agonist
induces immunosuppression and improves graft survival in novel
allogeneic transplantation models. Sci Rep. 2020;10(1):4464.
28. Guo XX, Wang Y, Wang K, Ji BP, Zhou F. Stability of a type 2
diabetes rat model induced by high-fat diet feeding with low-dose
streptozotocin injection. Journal of Zhejiang University Science B.
2018;19(7):559-69.
29. Inaba M, Nishizawa Y, Song K, Tanishita H, Okuno S, Miki T, et al.
Partial protection of 1 alpha-hydroxyvitamin D3 against the development
of diabetes induced by multiple low-dose streptozotocin injection in
CD-1 mice. Metabolism: clinical and experimental. 1992;41(6):631-5.
30. Tsutsui S, Schnermann J, Noorbakhsh F, Henry S, Yong VW, Winston BW,
et al. A1 adenosine receptor upregulation and activation attenuates
neuroinflammation and demyelination in a model of multiple sclerosis. J
Neurosci. 2004;24(6):1521-9.
31. Sitkovsky MV, Lukashev D, Apasov S, Kojima H, Koshiba M, Caldwell C,
et al. Physiological control of immune response and inflammatory tissue
damage by hypoxia-inducible factors and adenosine A2A receptors. Annu
Rev Immunol. 2004;22:657-82.
32. Jacobson KA, Gao ZG. Adenosine receptors as therapeutic targets. Nat
Rev Drug Discov. 2006;5(3):247-64.
33. Hasko G, Cronstein BN. Adenosine: an endogenous regulator of innate
immunity. Trends Immunol. 2004;25(1):33-9.
34. Rogachev B, Ziv NY, Mazar J, Nakav S, Chaimovitz C, Zlotnik M, et
al. Adenosine is upregulated during peritonitis and is involved in
downregulation of inflammation. Kidney Int. 2006;70(4):675-81.
35. Reeves WH, Lee PY, Weinstein JS, Satoh M, Lu L. Induction of
autoimmunity by pristane and other naturally occurring hydrocarbons.
Trends Immunol. 2009;30(9):455-64.
36. Armstrong JM, Chen JF, Schwarzschild MA, Apasov S, Smith PT,
Caldwell C, et al. Gene dose effect reveals no Gs-coupled A2A adenosine
receptor reserve in murine T-lymphocytes: studies of cells from
A2A-receptor-gene-deficient mice. Biochem J. 2001;354(Pt 1):123-30.
37. Teramachi J, Kukita A, Li YJ, Ushijima Y, Ohkuma H, Wada N, et al.
Adenosine abolishes MTX-induced suppression of osteoclastogenesis and
inflammatory bone destruction in adjuvant-induced arthritis. Lab Invest.
2011;91(5):719-31.
38. Nakav S, Chaimovitz C, Sufaro Y, Lewis EC, Shaked G, Czeiger D, et
al. Anti-inflammatory preconditioning by agonists of adenosine A1
receptor. PLoS One. 2008;3(5):e2107.
39. Cristovao-Ferreira S, Navarro G, Brugarolas M, Perez-Capote K, Vaz
SH, Fattorini G, et al. A1R-A2AR heteromers coupled to Gs and G i/0
proteins modulate GABA transport into astrocytes. Purinergic Signal.
2013;9(3):433-49.
40. Merayo-Chalico J, Rajme-Lopez S, Barrera-Vargas A, Alcocer-Varela J,
Diaz-Zamudio M, Gomez-Martin D. Lymphopenia and autoimmunity: A
double-edged sword. Human immunology. 2016;77(10):921-9.
41. Vila LM, Alarcon GS, McGwin G, Jr., Bastian HM, Fessler BJ, Reveille
JD, et al. Systemic lupus erythematosus in a multiethnic US cohort,
XXXVII: association of lymphopenia with clinical manifestations,
serologic abnormalities, disease activity, and damage accrual. Arthritis
and rheumatism. 2006;55(5):799-806.
42. Mirzayan MJ, Schmidt RE, Witte T. Prognostic parameters for flare in
systemic lupus erythematosus. Rheumatology. 2000;39(12):1316-9.
43. Min B, Yamane H, Hu-Li J, Paul WE. Spontaneous and homeostatic
proliferation of CD4 T cells are regulated by different mechanisms.
Journal of immunology. 2005;174(10):6039-44.
44. Baccala R, Theofilopoulos AN. The new paradigm of T-cell homeostatic
proliferation-induced autoimmunity. Trends in immunology.
2005;26(1):5-8.
45. Deaglio S, Dwyer KM, Gao W, Friedman D, Usheva A, Erat A, et al.
Adenosine generation catalyzed by CD39 and CD73 expressed on regulatory
T cells mediates immune suppression. J Exp Med. 2007;204(6):1257-65.
46. Zhang L, Yang N, Wang S, Huang B, Li F, Tan H, et al. Adenosine 2A
receptor is protective against renal injury in MRL/lpr mice. Lupus.
2011;20(7):667-77.
47. Boomer JS, To K, Chang KC, Takasu O, Osborne DF, Walton AH, et al.
Immunosuppression in patients who die of sepsis and multiple organ
failure. JAMA. 2011;306(23):2594-605.
48. Wu J, Sun L, Chen X, Du F, Shi H, Chen C, et al. Cyclic GMP-AMP is
an endogenous second messenger in innate immune signaling by cytosolic
DNA. Science. 2013;339(6121):826-30.
49. Liu YW, Yang T, Zhao L, Ni Z, Yang N, He F, et al. Activation of
Adenosine 2A receptor inhibits neutrophil apoptosis in an
autophagy-dependent manner in mice with systemic inflammatory response
syndrome. Sci Rep. 2016;6:33614.
50. Ali RA, Gandhi AA, Meng H, Yalavarthi S, Vreede AP, Estes SK, et al.
Adenosine receptor agonism protects against NETosis and thrombosis in
antiphospholipid syndrome. Nat Commun. 2019;10(1):1916.
51. Gupta S, Kaplan MJ. The role of neutrophils and NETosis in
autoimmune and renal diseases. Nat Rev Nephrol. 2016;12(7):402-13.
Figure 1. Susceptibility of A1R-KO mice to PIL.C57BL/6 WT and A1R-KO mice were injected with pristane
and monitored for alopecia, chronic wounds, or death for 36 weeks. (A)
Rate of disease appearance * p <0.05 (n=8-14).
Mantel-Cox test sighs of disease graphs. After 36 weeks, mice were
sacrificed and analyzed for (B) anti-dsDNA levels in serum of surviving
mice by ELISA, and (C) spleen size of surviving mice was measured. *p <0.05, ** p <0.01 (n=3-8). Values are
mean ± SE.
Figure 2. Lymphopenia following pristane injection. Following
pristane injection to C57BL/6, WT mice, and A1R-KO mice
(n=3-6). Blood counts were performed at the indicated time points. (A)
WBC, (B) lymphocyte, (C) neutrophils, and (D) monocytes. *p <0.05, ** p <0.01, compared to control
(WT at T=0). Values are mean ± SE.
Figure 3. A1R and A2AR mRNA
levels in PIL. Pristane was injected into C57BL/6 WT and
A1R-KO mice. To examine the dynamic expression of the
two high-affinity adenosine receptors, A1R and
A2AR, the spleen was removed at indicated time points
(6h, 24h, 48h, and 10 days). A1R and
A2AR mRNA levels in adherent splenocytes were analyzed
by real-time PCR and normalized by housekeeping RPL-12 levels. Results
are median + interquartile range, (n=3-6) * p <0.05,
between expression levels of each receptor to expression at time 0.
least two independent experiments performed
Figure 4. Adenosine receptors and susceptibility to induced
autoimmune type-1 diabetes. Mice were injected with low dose
streptozotocin (STZ, 50mg/kg) for five consecutive days. Mice were
considered diabetic when glucose remained above 200mg/dl. The experiment
was ended when, in one of the groups, all animals were sick. (A) C57BL/6
WT and A1R-KO mice, (B) Balb/C WT and
A2AR-KO mice. Mantel-Cox test sighs of disease graphs.
(n=5-7).
Figure 5. cfDNA in autoimmune diseases. Peritoneal lavage was
collected to examine the levels of cfDNA in (A) PIL model at several
time points after pristane injection, and in (B) T1D model at day 10
after first STZ injection. Analyzed for cfDNA levels by a direct rapid
fluorometric assay with the fluorochrome SYBR Gold (lower panel).
Results are median + interquartile range, (n=3-6) *p <0.05, ** p <0.01, between expression
levels of each receptor to expression at time 0. ^^p <0.01, ^^^ p <0.001 compared
to WT at the same time point (n=5-7). At least two independent
experiments performed
Figure 6. Adenosine receptors regulate NET production. (A)
HL-60 cells differentiated by retinoic acid (RA) to neutrophil-like
cells (2x105 cells/well) in triplicates were
pre-exposed to A1 adenosine receptor agonist (CCPA, 1nM)
or A2A adenosine receptor agonist (GCS, 30nM), and then
cells were stimulated with PMA (200nM) and
H202 (0.03%) in the presence of DNA
fluorescent dye (5 µM SYTOX Green) for 3h. Relative NETs production was
measured by fluorescence in 96-well plates and normalized to NETs
production by cells treated with only
H2O2+PMA. (B) Neutrophils
(2x105 cells/well) in triplicates were isolated from
bone marrow of WT and A1R-KO mice (C57BL/6 background)
and WT and A2AR-KO mice (BALB/c). Relative NETs
production was measured by fluorescence and normalized to NETs
production by WT neutrophils treated with
H2O2+PMA.*p < 0.05; **p
< 0.01. Values are mean ± SE. At least two independent
experiments performed