Adenosine A<sub>3</sub> agonists reverse neuropathic pain via T cell–mediated production of IL-10

Mariaconcetta Durante; Silvia Squillace; Filomena Lauro; Luigino Antonio Giancotti; Elisabetta Coppi; Federica Cherchi; Lorenzo Di Cesare Mannelli; Carla Ghelardini; Grant Kolar; Carrie Wahlman; Adeleye Opejin; Cuiying Xiao; Marc L. Reitman; Dilip K. Tosh; Daniel Hawiger; Kenneth A. Jacobson; Daniela Salvemini

doi:10.1172/JCI139299

Adenosine A₃ agonists reverse neuropathic pain via T cell–mediated production of IL-10

Mariaconcetta Durante,^1,2 Silvia Squillace,^1,3,4 Filomena Lauro,^1,3,5 Luigino Antonio Giancotti,^1,3 Elisabetta Coppi,² Federica Cherchi,² Lorenzo Di Cesare Mannelli,² Carla Ghelardini,² Grant Kolar,^3,6 Carrie Wahlman,¹ Adeleye Opejin,⁷ Cuiying Xiao,⁸ Marc L. Reitman,⁸ Dilip K. Tosh,⁸ Daniel Hawiger,⁷ Kenneth A. Jacobson,⁸ and Daniela Salvemini^1,3

Authorship note: MD, SS, and FL are co–first authors.

Published February 23, 2021 - More info

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Abstract

The A₃ adenosine receptor (A₃AR) has emerged as a therapeutic target with A₃AR agonists to tackle the global challenge of neuropathic pain, and investigation into its mode of action is essential for ongoing clinical development. Immune cell A₃ARs, and their activation during pathology, modulate cytokine release. Thus, the use of immune cells as a cellular substrate for the pharmacological action of A₃AR agonists is enticing, but unknown. The present study discovered that Rag-KO mice lacking T and B cells, as compared with WT mice, are insensitive to the anti-allodynic effects of A₃AR agonists. Similar findings were observed in interleukin-10 and interleukin-10 receptor knockout mice. Adoptive transfer of CD4⁺ T cells from WT mice infiltrated the dorsal root ganglion (DRG) and restored A₃AR agonist-mediated anti-allodynia in Rag-KO mice. CD4⁺ T cells from Adora3-KO or Il10-KO mice did not. Transfer of CD4⁺ T cells from WT mice, but not Il10-KO mice, into Il10-KO mice or Adora3-KO mice fully reinstated the anti-allodynic effects of A₃AR activation. Notably, A₃AR agonism reduced DRG neuron excitability when cocultured with CD4⁺ T cells in an IL-10–dependent manner. A₃AR action on CD4⁺ T cells infiltrated in the DRG decreased phosphorylation of GluN2B-containing N-methyl-D-aspartate receptors at Tyr1472, a modification associated with regulating neuronal hypersensitivity. Our findings establish that activation of A₃AR on CD4⁺ T cells to release IL-10 is required and sufficient evidence for the use of A₃AR agonists as therapeutics.

Introduction

Chronic neuropathic pain constitutes a large unmet medical need affecting 15–30 million people in the United States (1), and the annual economic burden cannot be underscored (2). Neuropathic pain arises when peripheral nerves are injured by trauma, disease, or toxins. Neuropathic pains are chronic, severe, debilitating, and exceedingly difficult to treat with currently available analgesics (3). Novel nonnarcotic analgesics are needed. Recently, the Gi-coupled A₃ adenosine receptor (A₃AR) was identified as a novel target for therapeutic intervention with selective A₃AR agonists (4–6). Continued investigation into their mode of action is essential, as these are in clinical development. Human and rodent immune cells, and in particular T cells (including CD4⁺ and CD8⁺), express high A₃AR levels (7), but whether these receptors play a role in the beneficial agonist effects in neuropathic pain is unknown. Interestingly, A₃AR activation on circulating immune cells harvested from animal models of autoimmune disorders blocks the formation of neuroexcitatory/inflammatory cytokines such as TNF and interleukin 1β and enhances interleukin-10 (IL-10) release (8). Similar findings were obtained with immune cells harvested from patients with autoimmune disorders validating the target in humans (9, 10). IL-10 is a potent anti-inflammatory and neuroprotective cytokine with documented positive effects in mitigating neuropathic pain (11, 12). These data, in parallel fields of studies, point to a potential link between immune cells and IL-10 in A₃AR agonist action. Using behavioral, genetic, pharmacological, and electrophysiological approaches, the present study explores the contribution of T cells to the pharmacological actions of A₃AR agonists in traumatic nerve injury–induced neuropathic pain.

Results and Discussion

Mouse sciatic nerve chronic constriction injury (CCI) leads to neuropathic pain (mechano-allodynia) that is maximal by day 7 (D7) and maintained for several weeks after injury (13). Intraperitoneal injection of highly selective A₃AR agonist MRS5980 at time of peak neuropathic pain reverses mechano-allodynia in both female and male mice (Figure 1A and Supplemental Figure 1; supplemental material available online with this article; https://doi.org/10.1172/JCI139299DS1), with effects lost in mice deficient in T and B cells (Rag-KO mice) (Figure 1A and Supplemental Figure 4). No significant difference in mechano-allodynia between WT and Rag-KO mice after nerve injury was observed, confirming previous studies (14). A₃AR agonist doses were chosen from our previous studies to cause a near-to-maximal reversal of mechano-allodynia in this model (15). Adoptive transfer (D7 after CCI) of CD3⁺ T cells from WT mice restored the A₃AR agonist effects in Rag-KO mice (Figure 1B and Supplemental Figure 1). CD8⁺ T cell adoptive transfer from WT mice did not restore A₃AR agonist anti-allodynic effects in Rag-KO mice. In contrast, adoptive transfer of CD4⁺ T cells fully reinstated anti-allodynic effects in both male and female Rag-KO mice (Figure 1, C and D and Supplemental Figure 1). CD4⁺ T cell adoptive transfer from A₃AR knockout (Adora3-KO) mice failed to restore the anti-allodynic effects of A₃AR agonists in Rag-KO mice, indicating that A₃AR activation on CD4⁺ T cells is required for A₃AR agonist anti-allodynic activity (Figure 1E and Supplemental Figure 1). The anti-allodynic responses to morphine were unaltered in Rag-KO mice compared with WT mice (Supplemental Figure 2), confirming that a lack of anti-allodynic responses is not a general, nonspecific response.

Anti-allodynic effects of A3AR agonists require CD4+ T cells.

Figure 1

Anti-allodynic effects of A₃AR agonists require CD4⁺ T cells. (A) Injection of MRS5980 (1 mg/kg, i.p.) given at time of peak neuropathic pain reversed allodynia in male and female WT mice (n = 7 males and n = 5 females) but not Rag-KO mice (n = 6 males and n = 5 females). Adoptive transfer of CD3⁺ (B, n = 5) or CD4⁺ (D, n = 15 males and n = 5 females) T cells but not CD8⁺ T cells (C, n = 9) from WT mice into Rag-KO mice restored the anti-allodynic effects of MRS5980. Adoptive transfer of CD4⁺ T cells from Adora3-KO mice (E, n = 4) failed to restore the anti-allodynic effect of MRS5980. Injection of MRS5980 (1 mg/kg, i.p.), ipsilateral to nerve injury during peak mechano-allodynia reversed allodynia in WT mice (n = 5) but not in Il10-KO (n = 5) or Il10r-KO (n = 6) mice (F). The anti-allodynic effect of MRS5980 lost in both male and female Rag-KO mice after adoptive transfer of CD4⁺ T cells from Il10-KO mice (G, n = 6 males and n = 5 females) was restored after adoptive transfer of CD4⁺ T cells from Il10r-KO mice (H, n = 9). Data are mean ± SD (A–H). *P < 0.05 vs. D0; ^†P < 0.05 vs D8/BL by 2-way repeated measures ANOVA (A and F) or 1-way ANOVA with Dunnett’s pair-wise comparisons (B–E, G, and H). ^#P < 0.05 vs. WT by 2-way repeated measures ANOVA with Sidak (A) or Tukey’s (F) pair-wise comparisons.

These results suggest that, following A₃AR activation, CD4⁺ T cells release mediators that rapidly reverse allodynia. Therefore, we focused on IL-10, which can be released by T cells (16) and is able to reverse neuropathic pain states (17, 18). Moreover, neurons as well as both CD4⁺ T cells and CD8⁺ T cells express A₃AR, IL-10, and IL-10R (19, 20). The anti-allodynic effects exerted by A₃AR agonists were lost in Il10-KO and in IL-10 receptor (Il10r-KO) mice (Figure 1F and Supplemental Figure 1). Thus, an intact IL-10/IL-10R system is required for A₃AR agonist effect (5). In order to test whether CD4⁺ T cells are a source of IL-10, we examined A₃AR agonist responses in Rag-KO mice that were adoptively transferred with CD4⁺ T cells from Il10-KO mice. In both male and female Rag-KO mice repopulated with CD4⁺ T cells from Il10-KO mice, A₃AR agonists failed to reverse mechano-allodynia (Figure 1G and Supplemental Figure 1), establishing CD4⁺ T cells as the predominant IL-10 source. In contrast, the A₃AR agonist anti-allodynic effects were uncompromised in Rag-KO mice reconstituted with CD4⁺ T cells from Il10r-KO mice (Figure 1H and Supplemental Figure 1). Collectively, the data suggest that CD4⁺ T cell–derived IL-10, but not the presence of IL-10 receptor on the CD4⁺ cells, is necessary for the effects of A₃AR agonists.

In Il10-KO mice, adoptive transfer of CD4⁺ T cells from WT but not Il10-KO mice restored the anti-allodynic effects of A₃AR agonists (Figure 2, A–C and Supplemental Figure 1). This information supports similar findings in Rag-KO mice and the premise that CD4⁺ T cell–derived IL-10 is necessary for A₃AR agonist effects.

Activation of A3AR expressed on CD4+ T cells is required in the anti-allody

Figure 2

Activation of A₃AR expressed on CD4⁺ T cells is required in the anti-allodynic effects of A₃AR agonist; role of IL-10. The anti-allodynic effects of MRS5980 were lost in Il10-KO (A, n = 6) and Il10r-KO (D, n = 6) mice. CD4⁺ T cell adoptive transfer from WT mice (B, n = 6) but not from Il10-KO mice (C, n = 6) restored the anti-allodynic effects of MRS5980 in Il10-KO mice. Conversely, adoptive transfer of CD4⁺ T cells from WT mice (E, n = 6) did not restore the anti-allodynic effects of MRS5980 in Il10r-KO mice. The anti-allodynic effects of MRS5980 were lost in Adora3-KO mice (F, n = 4). Adoptive transfer of CD4⁺ T cells from WT mice (G, n = 6) but not from Il10-KO mice (H, n = 4) restored the anti-allodynic effects of MRS5980 in Adora3-KO mice. Data are mean ± SD. *P < 0.05 vs. D0; ^†P < 0.05 vs. D8/BL by 1-way ANOVA with Dunnett’s pair-wise comparisons.

A₃AR effects lost in Il10r-KO mice are not restored by adoptive transfer of WT CD4⁺ T cells (Figure 2, D and E and Supplemental Figure 1), reinforcing the notion that CD4⁺ T cell–derived IL-10 is essential in A₃AR agonists’ mode of action.

To determine whether A₃AR activation on CD4⁺ T cells is required and sufficient for the IL-10 response, behavioral outcomes in Adora3-KO mice were investigated. A₃AR agonists did not reverse mechano-allodynia in Adora3-KO mice (Figure 2F and Supplemental Figure 1). However, adoptive transfer of CD4⁺ T cells from WT donors but not from IL-10–KO mice into Adora3-KO mice completely restored the agonists’ anti-allodynic effects (Figure 2, G and H and Supplemental Figure 1). These results establish that A₃AR activation on CD4⁺ T cells drives the IL-10 response. As previously described (5, 21, 22), we observed no reduction of mechano-allodynia in Il10-KO and Il10r-KO mice compared with WT mice. Moreover, the anti-allodynic responses to morphine were not altered in Il10-KO (23) and Adora3-KO (24) mice compared with WT mice. No changes in contralateral paws were observed in any study (Supplemental Figures 3 and 4).

The hypersensitivity of primary sensory neurons that develops in the DRG is critically important in neuropathic pain development (25), and increased phosphorylation of GluN2B-containing N-methyl-D-aspartate receptors (NMDARs) at Tyr1472 [GluN2B(Tyr1472)] contributes to this increase (26, 27). Our data suggest that CD4⁺ T cell infiltration in the DRG attenuates neuronal excitability following A₃AR activation. A- and C-type DRG neurons express the IL-10 receptor (alpha subunit, IL-10RA) (28). IL-10 can block phosphorylation of NMDARs by attenuating NMDA-induced intracellular calcium concentration increases (29), inhibiting protein kinases and phosphatases known to regulate NMDAR channel activity (30), inhibiting DRG neuronal firing (28, 31), and reducing neuronal firing indirectly by have effects on nonneuronal cells (11). Consistently, application of IL-10 to DRG neurons isolated from naive mice prevented action potential (AP) initiation (Figure 3, A–C, Supplemental Figure 5). Of note, DRG neurons exposed to IL-10 were still able to respond to the transient receptor potential vanilloid 1 (TPV1) agonist capsaicin (Supplemental Figure 6).

Figure 3

Functional effects of IL-10 on cell firing in DRG neurons, and CD4⁺ T cell infiltration in mouse DRG neurons. (A) Original current-clamp traces recorded by whole-cell patch-clamp technique in a typical naive mouse DRG neuron where IL-10 (0.5 μg/mL) reversibly inhibits AP firing evoked by a depolarizing ramp current injection (1 second; 30 pA; lower inset) once every 30 seconds. Dotted lines indicate the 0 mV level. The number of APs elicited by the current ramp was plotted as a function of time in the same cell (B) or was expressed as pooled data (mean ± SEM) in the bar graph (C, n = 6). *P = 0.0018, paired Student’s t test; scale bars: 300 ms; 50 mV (C). CD4⁺ T cells (arrow) (magnification ×40) are present in the ipsilateral DRG of the Rag-KO mice reconstituted with CD4⁺ T cells from WT GFP mice (green, GFP; blue, DAPI) (D, E; n = 7). MRS5980 reduced Tyr1472 phosphorylation of GluN2B in the DRG of Rag-KO mice after adoptive transfer of CD4⁺ T cells from WT mice (F, n = 9). Density of each p-Tyr₁₄₇₂GluN2B band was calculated relative to α-tubulin. Data are mean ± SEM (E) or mean ± SD (F). *P < 0.05. WT+veh or ipsilateral; ^†P < 0.05 vs. Rag-KO+veh by 2-tailed Student’s t test (E) or 1-way ANOVA (F) with Dunnett’s pair-wise comparisons.

Our study next examined whether A₃AR agonism leads to inhibition of DRG neuronal excitability via IL-10 release from CD4⁺ T cells. Immunofluorescence analysis of DRGs harvested from Rag-KO mice following adoptive transfer of CD4⁺ T cells from WT mice expressing enhanced green fluorescence protein (GFP) showed increased CD4⁺ T cell numbers in DRG ipsilateral to nerve injury compared with those that were contralateral (Figure 3, D and E). The A_2AAR receptor subtype, not the A₃AR, seems to have the predominant role in lymphocyte migration (32, 33). Furthermore, in C57BL/6 mice, it has been reported that the absence of IL-10 receptor on the CD4⁺ T cell surface does not impair trafficking in inflammatory conditions, suggesting the IL-10/IL-10R system is nonessential for T cell migration (34). So, although A₃AR activation of IL-10 inhibition may affect T cell migration, we consider this to be unlikely. Intraperitoneal injection of MRS5980 caused a significant decrease in GluN2B(Tyr1472) phosphorylation in DRG ipsilateral to nerve injury in Rag-KO mice after CD4⁺ T cell adoptive transfer from WT mice compared with Rag-KO mice with no adoptive transfer (Figure 3F).

To explore potential cross-talk between CD4⁺ T cells and neurons in the DRG, we performed an in vitro study, coculturing primary mouse DRG neurons with primary mouse CD4⁺ T cells—both cell types isolated from naive animals. A₃AR agonist MRS5980 significantly decreased the number of APs evoked by a 30 pA ramp current in DRG neurons when cocultured with CD4⁺ T cells (Figure 4, A–C). Concurrently, a marked increase in current threshold (Supplemental Table 1) was detected. These effects were prevented by an anti-IL-10 antibody (Figure 4, D–F) but not by a control IgG isotype (Supplemental Figure 7) and were not observed when DRG neurons were cocultured with CD8⁺ T cells (Figure 4, G–I). MRS5980 did not alter cell excitability when CD4⁺ T cells were absent in the DRG culture (Figure 4, J–L). This result is at variance with findings that we recently published, in which A₃AR activation reduced neuronal firing in rat DRG neurons (35). This difference is possibly due to the reported lack of A₃AR expression in mouse DRG neurons (36, 37). When mouse DRG neurons were cultured in the absence of CD4⁺ T cells (Figure 4L), cocultured with CD4⁺ T cells (Figure 4C), cocultured with CD4⁺ T cells with anti-IL-10 antibody present (Figure 4F), or cocultured with CD8⁺ T cells (Figure 4I), the number of APs elicited by the current ramp in control conditions (before MRS5980 application) was similar among the groups. Results were replicated in DRG and CD4⁺ T cells isolated from CCI animals on D7 (Supplemental Figure 8 and Supplemental Table 1). Of note, DRG neurons isolated from CCI mice presented a markedly smaller current threshold to first AP (Supplemental Table 1), so ramp current injection was lowered to 15 pA to avoid signal saturation (Supplemental Figure 8).

IL-10 released by CD4+ T cells is required for A3AR agonist–mediated inhibi

Figure 4

IL-10 released by CD4⁺ T cells is required for A₃AR agonist–mediated inhibition of AP firing in cocultured mouse DRG neurons isolated from naive mice. Original current-clamp traces recorded by whole-cell patch-clamp technique in typical mouse DRG neurons. AP firing was evoked by a depolarizing ramp current injection (1 second; 30 pA; lower inset) once every 30 seconds. The A₃AR agonist MRS5980 (300 nM) was applied in (A) DRG-CD4⁺ T cell cocultures, (D) DRG-CD4⁺ T cell cocultures in the presence of anti-IL-10 antibody (Ab IL-10; 0.5 μg/ml), (G) DRG-CD8⁺ T cell cocultures, and (J) DRG cultures. The number of APs elicited by the current ramp was plotted as a function of time in 4 different representative cells (B, E, H, and K) or was expressed as pooled data (mean ± SEM) in the bar graphs (L, n = 6; C, n = 10; F, n = 11; I, n = 7). Dotted gray lines indicate the 0 mV level. *P = 0.0120, paired Student’s t test. The number of APs elicited before MRS5980 application (with bars, ctrl) was not different in DRG neurons cultured alone (L), DRG neurons cocultured with CD4⁺ T cells (C), DRG neurons cocultured with CD4⁺ T cells in the presence of anti-IL-10 antibody (F), or DRG neurons cocultured with CD8⁺ T cells (I). One-way ANOVA with Bonferroni comparison: L vs C: P = 0.3981; C vs. F: P = 0.1034; L vs. F: P > 0.9999; L vs. I: P > 0.9999. Scale bars: 300 ms; 50 mV.

Collectively, these results suggested a model whereby A₃AR agonists reverse established hypersensitivity by activating A₃AR expressed on CD4⁺ T cells to release IL-10, reducing neuronal DRG excitability (Graphical Abstract).

Author contributions

DS conceived and designed the studies. EC designed the electrophysiology studies. MD, FL, SS, LDCM, CG, GK, CX, CW, LAG, FC, and EC performed the experiment and analysis. DKT, MLR, and KAJ provided key reagents. AO and DH provided technical input. DS, MD, SS, and FL prepared the manuscript with input from all authors. The order of the first co–authors was determined by degree of involvement with the project; MD and SS were involved during the project’s pilot phase; FL joined during the project’s maturation.

References

NIH. Peripheral neuropathy fact sheet. https://www.ninds.nih.gov/Disorders/Patient-Caregiver-Education/Fact-Sheets/Peripheral-Neuropathy-Fact-Sheet Updated March 16, 2020. Accessed December 7, 2017.

Institute of Medicine (US) Committee on Advancing Pain Research, Care, Education. Relieving Pain in America: A Blueprint for Transforming Prevention, Care, Education, and Research. Nation Academies Press; 2011.

Finnerup NB, et al. Pharmacotherapy for neuropathic pain in adults: a systematic review and meta-analysis. Lancet Neurol. 2015;14(2):162–173.
View this article via: PubMed CrossRef Google Scholar

Janes K, et al. Spinal neuroimmune activation is independent of T-cell infiltration and attenuated by A3 adenosine receptor agonists in a model of oxaliplatin-induced peripheral neuropathy. Brain Behav Immun. 2015;44:91–99.
View this article via: PubMed CrossRef Google Scholar

Wahlman C, et al. Chemotherapy-induced pain is promoted by enhanced spinal adenosine kinase levels through astrocyte-dependent mechanisms. Pain. 2018;159(6):1025–1034.
View this article via: PubMed CrossRef Google Scholar

Jacobson KA, et al. Treatment of chronic neuropathic pain: purine receptor modulation. Pain. 2020;161(7):1425–1441.
View this article via: PubMed CrossRef Google Scholar

Borea PA, et al. The A3 adenosine receptor: history and perspectives. Pharmacol Rev. 2015;67(1):74–102.
View this article via: PubMed CrossRef Google Scholar

Bar-Yehuda S, et al. Inhibition of experimental auto-immune uveitis by the A3 adenosine receptor agonist CF101. Int J Mol Med. 2011;28(5):727–731.
View this article via: PubMed Google Scholar

Ravani A, et al. Role and function of A2A and A(3) adenosine receptors in patients with ankylosing spondylitis, psoriatic arthritis and rheumatoid arthritis. Int J Mol Sci. 2017;18(4):697.
View this article via: CrossRef Google Scholar

Varani K, et al. A2A and A3 adenosine receptor expression in rheumatoid arthritis: upregulation, inverse correlation with disease activity score and suppression of inflammatory cytokine and metalloproteinase release. Arthritis Res Ther. 2011;13(6):R197.
View this article via: PubMed CrossRef Google Scholar

Burmeister AR, Marriott I. The interleukin-10 family of cytokines and their role in the CNS. Front Cell Neurosci. 2018;12:458.
View this article via: PubMed Google Scholar

Vanderwall AG, et al. Effects of spinal non-viral interleukin-10 gene therapy formulated with d-mannose in neuropathic interleukin-10 deficient mice: behavioral characterization, mRNA and protein analysis in pain relevant tissues. Brain Behav Immun. 2018;69:91–112.
View this article via: PubMed CrossRef Google Scholar

Bennett GJ, Xie YK. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain. 1988;33(1):87–107.
View this article via: PubMed CrossRef Google Scholar

Sorge RE, et al. Different immune cells mediate mechanical pain hypersensitivity in male and female mice. Nat Neurosci. 2015;18(8):1081–1083.
View this article via: PubMed CrossRef Google Scholar

Little JW, et al. Endogenous adenosine A3 receptor activation selectively alleviates persistent pain states. Brain. 2015;138(Pt 1):28–35.
View this article via: PubMed Google Scholar

Trinchieri G. Interleukin-10 production by effector T cells: Th1 cells show self control. J Exp Med. 2007;204(2):239–243.
View this article via: PubMed CrossRef Google Scholar

Khan J, et al. Interleukin-10 levels in rat models of nerve damage and neuropathic pain. Neurosci Lett. 2015;592:99–106.
View this article via: PubMed CrossRef Google Scholar

Davoli-Ferreira M, et al. Regulatory T cells counteract neuropathic pain through inhibition of the Th1 response at the site of peripheral nerve injury. Pain. 2020;161(8):1730–1743.
View this article via: PubMed CrossRef Google Scholar

Stubbington MJ, et al. An atlas of mouse CD4(+) T cell transcriptomes. Biol Direct. 2015;10:14.
View this article via: PubMed Google Scholar

Li Q, et al. Developmental heterogeneity of microglia and brain myeloid cells revealed by deep single-cell RNA sequencing. Neuron. 2019;101(2):207–223.
View this article via: PubMed CrossRef Google Scholar

McKelvey R, et al. Neuropathic pain is constitutively suppressed in early life by anti-inflammatory neuroimmune regulation. J Neurosci. 2015;35(2):457–466.
View this article via: PubMed CrossRef Google Scholar

Alvarez P, et al. Nociceptor interleukin 10 receptor 1 is critical for muscle analgesia induced by repeated bouts of eccentric exercise in the rat. Pain. 2017;158(8):1481–1488.
View this article via: PubMed CrossRef Google Scholar

Chen Z, et al. Sphingosine-1-phosphate receptor 1 activation in astrocytes contributes to neuropathic pain. Proc Natl Acad Sci U S A. 2019;116(21):10557–10562.
View this article via: PubMed CrossRef Google Scholar

Doyle TM, et al. Chronic morphine-induced changes in signaling at the A₃ adenosine receptor contribute to morphine-induced hyperalgesia, tolerance, and withdrawal. J Pharmacol Exp Ther. 2020;374(2):331–341.
View this article via: PubMed CrossRef Google Scholar

Stemkowski PL, et al. Increased excitability of medium-sized dorsal root ganglion neurons by prolonged interleukin-1β exposure is K(+) channel dependent and reversible. J Physiol. 2015;593(16):3739–3755.
View this article via: PubMed CrossRef Google Scholar

Li J, et al. Electrophysiological characterization of N-methyl-D-aspartate receptors in rat dorsal root ganglia neurons. Pain. 2004;109(3):443–452.
View this article via: PubMed CrossRef Google Scholar

Chen W, et al. BDNF released during neuropathic pain potentiates NMDA receptors in primary afferent terminals. Eur J Neurosci. 2014;39(9):1439–1454.
View this article via: PubMed CrossRef Google Scholar

Shen KF, et al. Interleukin-10 down-regulates voltage gated sodium channels in rat dorsal root ganglion neurons. Exp Neurol. 2013;247:466–475.
View this article via: PubMed CrossRef Google Scholar

Turovskaya MV, et al. Interleukin-10 modulates [Ca2+]i response induced by repeated NMDA receptor activation with brief hypoxia through inhibition of InsP(3)-sensitive internal stores in hippocampal neurons. Neurosci Lett. 2012;516(1):151–155.
View this article via: PubMed CrossRef Google Scholar

Wang YT, Salter MW. Regulation of NMDA receptors by tyrosine kinases and phosphatases. Nature. 1994;369(6477):233–235.
View this article via: PubMed CrossRef Google Scholar

Krukowski K, et al. CD8+ T cells and endogenous IL-10 are required for resolution of chemotherapy-induced neuropathic pain. J Neurosci. 2016;36(43):11074–11083.
View this article via: PubMed CrossRef Google Scholar

Hoskin DW, et al. Inhibition of T cell and natural killer cell function by adenosine and its contribution to immune evasion by tumor cells (Review). Int J Oncol. 2008;32(3):527–535.
View this article via: PubMed Google Scholar

Linden J, Cekic C. Regulation of lymphocyte function by adenosine. Arterioscler Thromb Vasc Biol. 2012;32(9):2097–2103.
View this article via: PubMed CrossRef Google Scholar

Diefenhardt P, et al. IL-10 receptor signaling empowers regulatory T cells to control Th17 responses and protect from GN. J Am Soc Nephrol. 2018;29(7):1825–1837.
View this article via: PubMed CrossRef Google Scholar

Coppi E, et al. Adenosine A3 receptor activation inhibits pronociceptive N-type Ca2+ currents and cell excitability in dorsal root ganglion neurons. Pain. 2019;160(5):1103–1118.
View this article via: PubMed CrossRef Google Scholar

Ray P, et al. Comparative transcriptome profiling of the human and mouse dorsal root ganglia: an RNA-seq-based resource for pain and sensory neuroscience research. Pain. 2018;159(7):1325–1345.
View this article via: PubMed CrossRef Google Scholar

Usoskin D, et al. Unbiased classification of sensory neuron types by large-scale single-cell RNA sequencing. Nat Neurosci. 2015;18(1):145–153.
View this article via: PubMed CrossRef Google Scholar

[1] NIH. Peripheral neuropathy fact sheet. https://www.ninds.nih.gov/Disorders/Patient-Caregiver-Education/Fact-Sheets/Peripheral-Neuropathy-Fact-Sheet Updated March 16, 2020. Accessed December 7, 2017.

[2] Institute of Medicine (US) Committee on Advancing Pain Research, Care, Education. Relieving Pain in America: A Blueprint for Transforming Prevention, Care, Education, and Research. Nation Academies Press; 2011.

[3] Finnerup NB, et al. Pharmacotherapy for neuropathic pain in adults: a systematic review and meta-analysis. Lancet Neurol. 2015;14(2):162–173.
View this article via: PubMed CrossRef Google Scholar

[4] Janes K, et al. Spinal neuroimmune activation is independent of T-cell infiltration and attenuated by A3 adenosine receptor agonists in a model of oxaliplatin-induced peripheral neuropathy. Brain Behav Immun. 2015;44:91–99.
View this article via: PubMed CrossRef Google Scholar

[5] Wahlman C, et al. Chemotherapy-induced pain is promoted by enhanced spinal adenosine kinase levels through astrocyte-dependent mechanisms. Pain. 2018;159(6):1025–1034.
View this article via: PubMed CrossRef Google Scholar

[6] Jacobson KA, et al. Treatment of chronic neuropathic pain: purine receptor modulation. Pain. 2020;161(7):1425–1441.
View this article via: PubMed CrossRef Google Scholar

[7] Borea PA, et al. The A3 adenosine receptor: history and perspectives. Pharmacol Rev. 2015;67(1):74–102.
View this article via: PubMed CrossRef Google Scholar

[8] Bar-Yehuda S, et al. Inhibition of experimental auto-immune uveitis by the A3 adenosine receptor agonist CF101. Int J Mol Med. 2011;28(5):727–731.
View this article via: PubMed Google Scholar

[9] Ravani A, et al. Role and function of A2A and A(3) adenosine receptors in patients with ankylosing spondylitis, psoriatic arthritis and rheumatoid arthritis. Int J Mol Sci. 2017;18(4):697.
View this article via: CrossRef Google Scholar

[10] Varani K, et al. A2A and A3 adenosine receptor expression in rheumatoid arthritis: upregulation, inverse correlation with disease activity score and suppression of inflammatory cytokine and metalloproteinase release. Arthritis Res Ther. 2011;13(6):R197.
View this article via: PubMed CrossRef Google Scholar

[11] Burmeister AR, Marriott I. The interleukin-10 family of cytokines and their role in the CNS. Front Cell Neurosci. 2018;12:458.
View this article via: PubMed Google Scholar

[12] Vanderwall AG, et al. Effects of spinal non-viral interleukin-10 gene therapy formulated with d-mannose in neuropathic interleukin-10 deficient mice: behavioral characterization, mRNA and protein analysis in pain relevant tissues. Brain Behav Immun. 2018;69:91–112.
View this article via: PubMed CrossRef Google Scholar

[13] Bennett GJ, Xie YK. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain. 1988;33(1):87–107.
View this article via: PubMed CrossRef Google Scholar

[14] Sorge RE, et al. Different immune cells mediate mechanical pain hypersensitivity in male and female mice. Nat Neurosci. 2015;18(8):1081–1083.
View this article via: PubMed CrossRef Google Scholar

[15] Little JW, et al. Endogenous adenosine A3 receptor activation selectively alleviates persistent pain states. Brain. 2015;138(Pt 1):28–35.
View this article via: PubMed Google Scholar

[16] Trinchieri G. Interleukin-10 production by effector T cells: Th1 cells show self control. J Exp Med. 2007;204(2):239–243.
View this article via: PubMed CrossRef Google Scholar

[17] Khan J, et al. Interleukin-10 levels in rat models of nerve damage and neuropathic pain. Neurosci Lett. 2015;592:99–106.
View this article via: PubMed CrossRef Google Scholar

[18] Davoli-Ferreira M, et al. Regulatory T cells counteract neuropathic pain through inhibition of the Th1 response at the site of peripheral nerve injury. Pain. 2020;161(8):1730–1743.
View this article via: PubMed CrossRef Google Scholar

[19] Stubbington MJ, et al. An atlas of mouse CD4(+) T cell transcriptomes. Biol Direct. 2015;10:14.
View this article via: PubMed Google Scholar

[20] Li Q, et al. Developmental heterogeneity of microglia and brain myeloid cells revealed by deep single-cell RNA sequencing. Neuron. 2019;101(2):207–223.
View this article via: PubMed CrossRef Google Scholar

[21] McKelvey R, et al. Neuropathic pain is constitutively suppressed in early life by anti-inflammatory neuroimmune regulation. J Neurosci. 2015;35(2):457–466.
View this article via: PubMed CrossRef Google Scholar

[22] Alvarez P, et al. Nociceptor interleukin 10 receptor 1 is critical for muscle analgesia induced by repeated bouts of eccentric exercise in the rat. Pain. 2017;158(8):1481–1488.
View this article via: PubMed CrossRef Google Scholar

[23] Chen Z, et al. Sphingosine-1-phosphate receptor 1 activation in astrocytes contributes to neuropathic pain. Proc Natl Acad Sci U S A. 2019;116(21):10557–10562.
View this article via: PubMed CrossRef Google Scholar

[24] Doyle TM, et al. Chronic morphine-induced changes in signaling at the A₃ adenosine receptor contribute to morphine-induced hyperalgesia, tolerance, and withdrawal. J Pharmacol Exp Ther. 2020;374(2):331–341.
View this article via: PubMed CrossRef Google Scholar

[25] Stemkowski PL, et al. Increased excitability of medium-sized dorsal root ganglion neurons by prolonged interleukin-1β exposure is K(+) channel dependent and reversible. J Physiol. 2015;593(16):3739–3755.
View this article via: PubMed CrossRef Google Scholar

[26] Li J, et al. Electrophysiological characterization of N-methyl-D-aspartate receptors in rat dorsal root ganglia neurons. Pain. 2004;109(3):443–452.
View this article via: PubMed CrossRef Google Scholar

[27] Chen W, et al. BDNF released during neuropathic pain potentiates NMDA receptors in primary afferent terminals. Eur J Neurosci. 2014;39(9):1439–1454.
View this article via: PubMed CrossRef Google Scholar

[28] Shen KF, et al. Interleukin-10 down-regulates voltage gated sodium channels in rat dorsal root ganglion neurons. Exp Neurol. 2013;247:466–475.
View this article via: PubMed CrossRef Google Scholar

[29] Turovskaya MV, et al. Interleukin-10 modulates [Ca2+]i response induced by repeated NMDA receptor activation with brief hypoxia through inhibition of InsP(3)-sensitive internal stores in hippocampal neurons. Neurosci Lett. 2012;516(1):151–155.
View this article via: PubMed CrossRef Google Scholar

[30] Wang YT, Salter MW. Regulation of NMDA receptors by tyrosine kinases and phosphatases. Nature. 1994;369(6477):233–235.
View this article via: PubMed CrossRef Google Scholar

[31] Krukowski K, et al. CD8+ T cells and endogenous IL-10 are required for resolution of chemotherapy-induced neuropathic pain. J Neurosci. 2016;36(43):11074–11083.
View this article via: PubMed CrossRef Google Scholar

[32] Hoskin DW, et al. Inhibition of T cell and natural killer cell function by adenosine and its contribution to immune evasion by tumor cells (Review). Int J Oncol. 2008;32(3):527–535.
View this article via: PubMed Google Scholar

[33] Linden J, Cekic C. Regulation of lymphocyte function by adenosine. Arterioscler Thromb Vasc Biol. 2012;32(9):2097–2103.
View this article via: PubMed CrossRef Google Scholar

[34] Diefenhardt P, et al. IL-10 receptor signaling empowers regulatory T cells to control Th17 responses and protect from GN. J Am Soc Nephrol. 2018;29(7):1825–1837.
View this article via: PubMed CrossRef Google Scholar

[35] Coppi E, et al. Adenosine A3 receptor activation inhibits pronociceptive N-type Ca2+ currents and cell excitability in dorsal root ganglion neurons. Pain. 2019;160(5):1103–1118.
View this article via: PubMed CrossRef Google Scholar

[36] Ray P, et al. Comparative transcriptome profiling of the human and mouse dorsal root ganglia: an RNA-seq-based resource for pain and sensory neuroscience research. Pain. 2018;159(7):1325–1345.
View this article via: PubMed CrossRef Google Scholar

[37] Usoskin D, et al. Unbiased classification of sensory neuron types by large-scale single-cell RNA sequencing. Nat Neurosci. 2015;18(1):145–153.
View this article via: PubMed CrossRef Google Scholar

Adenosine A₃ agonists reverse neuropathic pain via T cell–mediated production of IL-10

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Adenosine A₃ agonists reverse neuropathic pain via T cell–mediated production of IL-10