This study investigated the role of the alternative receptor for complement activation fragment C5a, C5aR2, in secondary inflammatory pathology after contusive spinal cord injury (SCI) in mice. C5ar2–/– mice exhibited decreased intraparenchymal tumor necrosis factor alpha and interleukin-6 acutely post-injury, but these reductions did not translate into improved outcomes. We show that loss of C5aR2 leads to increased lesion volumes, reduced myelin sparing, and significantly worsened recovery from SCI in C5ar2–/– animals compared to wild-type (WT) controls. Loss of C5aR2 did not alter leukocyte mobilization from the bone marrow in response to SCI, and neutrophil recruitment/presence at the lesion site was also not different between genotypes. Acute treatment of SCI mice with the selective C5aR1 antagonist, PMX205, improved SCI outcomes, compared to vehicle controls, and, importantly, fully alleviated the worsened recovery of C5ar2–/– mice compared to their WT counterparts. Collectively, these findings indicate that C5aR2 is neuroprotective and a novel target to restrain injurious C5a signaling after a major neurotraumatic event.
Get full access to this article
View all access options for this article.
References
1.
QiaoF., AtkinsonC., KindyM.S., ShunmugavelA., MorganB.P., SongH., and TomlinsonS. (2010). The alternative and terminal pathways of complement mediate post-traumatic spinal cord inflammation and injury. Am. J. Pathol., 177, 3061–3070.
2.
AndersonA.J., RobertS., HuangW., YoungW., and CotmanC.W. (2004). Activation of complement pathways after contusion-induced spinal cord injury. J. Neurotrauma, 21, 1831–1846.
3.
BrennanF.H., GordonR., LaoH.W., BigginsP.J., TaylorS.M., FranklinR.J., WoodruffT.M., and RuitenbergM.J. (2015). The complement receptor C5aR controls acute inflammation and astrogliosis following spinal cord injury. J. Neurosci., 35, 6517–6531.
4.
DunkelbergerJ.R., and SongW.C. (2010). Complement and its role in innate and adaptive immune responses. Cell Res. 20, 34–50.
5.
MantheyH.D., WoodruffT.M., TaylorS.M., and MonkP.N. (2009). Complement component 5a (C5a). Int. J. Biochem. Cell Biol., 41, 2114–2117.
6.
NatafS., StahelP.F., DavoustN., and BarnumS.R. (1999). Complement anaphylatoxin receptors on neurons: new tricks for old receptors?. Trends Neurosci. 22, 397–402.
7.
FarkasI., BaranyiL., LipositsZ.S., YamamotoT., and OkadaH. (1998). Complement C5a anaphylatoxin fragment causes apoptosis in TGW neuroblastoma cells. Neuroscience, 86, 903–911.
8.
PavlovskiD., ThundyilJ., MonkP.N., WetselR.A., TaylorS.M., and WoodruffT.M. (2012). Generation of complement component C5a by ischemic neurons promotes neuronal apoptosis. FASEB J. 26, 3680–3690.
9.
LeinhaseI., HolersV.M., ThurmanJ.M., HarhausenD., SchmidtO.I., PietzckerM., TahaM.E., RittirschD., Huber-LangM., SmithW.R., WardP.A., and StahelP.F. (2006). Reduced neuronal cell death after experimental brain injury in mice lacking a functional alternative pathway of complement activation. BMC Neurosci., 7, 55.
10.
FarkasI., TakahashiM., FukudaA., YamamotoN., AkatsuH., BaranyiL., TateyamaH., YamamotoT., OkadaN., and OkadaH. (2003). Complement C5a receptor-mediated signaling may be involved in neurodegeneration in Alzheimer's disease. J. Immunology, 170, 5764–5771.
11.
OsakaH., MukherjeeP., AisenP.S., and PasinettiG.M. (1999). Complement-derived anaphylatoxin C5a protects against glutamate-mediated neurotoxicity. J. Cell. Biochem., 73, 303–311.
12.
MukherjeeP., and PasinettiG.M. (2001). Complement anaphylatoxin C5a neuroprotects through mitogen-activated protein kinase-dependent inhibition of caspase 3. J. Neurochem., 77, 43–49.
13.
MukherjeeP., ThomasS., and PasinettiG.M. (2008). Complement anaphylatoxin C5a neuroprotects through regulation of glutamate receptor subunit 2 in vitro and in vivo. J. Neuroinflammation1, 5, 5.
14.
BambergC.E., MackayC.R., LeeH., ZahraD., JacksonJ., LimY.S., WhitfeldP.L., CraigS., CorsiniE., LuB., GerardC., and GerardN.P. (2010). The C5a receptor (C5aR) C5L2 is a modulator of C5aR-mediated signal transduction. J. Biol. Chem., 285, 7633–7644.
LiG., FanR.M., ChenJ.L., WangC.M., ZengY.C., HanC., JiaoS., XiaX.P., ChenW., and YaoS.T. (2014). Neuroprotective effects of argatroban and C5a receptor antagonist (PMX53) following intracerebral haemorrhage. Clin. Exp. Immunol., 175, 285–295.
17.
Van BeekJ., BernaudinM., PetitE., GasqueP., NouvelotA., MacKenzieE.T., and FontaineM. (2000). Expression of receptors for complement anaphylatoxins C3a and C5a following permanent focal cerebral ischemia in the mouse. Exp. Neurol., 161, 373–382.
BeckK.D., NguyenH.X., GalvanM.D., SalazarD.L., WoodruffT.M., and AndersonA.J. (2010). Quantitative analysis of cellular inflammation after traumatic spinal cord injury: evidence for a multiphasic inflammatory response in the acute to chronic environment. Brain, 133, 433–447.
20.
LiL., XiongZ.Y., QianZ.M., ZhaoT.Z., FengH., HuS., HuR., KeY., and LinJ. (2014). Complement C5a is detrimental to histological and functional locomotor recovery after spinal cord injury in mice. Neurobiol. Dis., 66, 74–82.
21.
CrokerD.E., HalaiR., KaeslinG., WendeE., FehlhaberB., KlosA., MonkP.N., and CooperM.A. (2014). C5a2 can modulate ERK1/2 signaling in macrophages via heteromer formation with C5a1 and beta-arrestin recruitment. Immunol. Cell Biol., 92, 631–639.
22.
CrokerD.E., MonkP.N., HalaiR., KaeslinG., SchofieldZ., WuM.C., ClarkR.J., BlaskovichM.A., MorikisD., FloudasC.A., CooperM.A., and WoodruffT.M. (2016). Discovery of functionally selective C5aR2 ligands: novel modulators of C5a signalling. Immunol. Cell Biol., 94, 787–795.
23.
ArboreG., WestE.E., SpolskiR., RobertsonA.A., KlosA., RheinheimerC., DutowP., WoodruffT.M., YuZ.X., O'NeillL.A., CollR.C., SherA., LeonardW.J., KohlJ., MonkP., CooperM.A., ArnoM., AfzaliB., LachmannH.J., CopeA.P., Mayer-BarberK.D., and KemperC. (2016). T helper 1 immunity requires complement-driven NLRP3 inflammasome activity in CD4(+) T cells. Science, 352, aad1210.
24.
TaokaY., OkajimaK., UchibaM., MurakamiK., KushimotoS., JohnoM., NaruoM., OkabeH., and TakatsukiK. (1997). Role of neutrophils in spinal cord injury in the rat. Neuroscience, 79, 1177–1182.
25.
KigerlK.A., GenselJ.C., AnkenyD.P., AlexanderJ.K., DonnellyD.J., and PopovichP.G. (2009). Identification of two distinct macrophage subsets with divergent effects causing either neurotoxicity or regeneration in the injured mouse spinal cord. J. Neurosci., 29, 13435–13444.
26.
LeeS.M., RosenS., WeinsteinP., van RooijenN., and Noble-HaeussleinL.J. (2011). Prevention of both neutrophil and monocyte recruitment promotes recovery after spinal cord injury. J. Neurotrauma, 28, 1893–1907.
27.
PopovichP.G., GuanZ., WeiP., HuitingaI., van RooijenN., and StokesB.T. (1999). Depletion of hematogenous macrophages promotes partial hindlimb recovery and neuroanatomical repair after experimental spinal cord injury. Exp. Neurol., 158, 351–365.
28.
ChenN.J., MirtsosC., SuhD., LuY.C., LinW.J., McKerlieC., LeeT., BaribaultH., TianH., and YehW.C. (2007). C5L2 is critical for the biological activities of the anaphylatoxins C5a and C3a. Nature, 446, 203–207.
29.
KilkennyC., BrowneW.J., CuthillI.C., EmersonM., and AltmanD.G. (2010). Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol., 8, e1000412.
30.
ScheffS.W., RabchevskyA.G., FugacciaI., MainJ.A., and LumppJ.E.Jr. (2003). Experimental modeling of spinal cord injury: characterization of a force-defined injury device. J. Neurotrauma, 20, 179–193.
31.
HarrisonM., O'BrienA., AdamsL., CowinG., RuitenbergM.J., SengulG., and WatsonC. (2013). Vertebral landmarks for the identification of spinal cord segments in the mouse. Neuroimage, 68, 22–29.
32.
BassoD.M., FisherL.C., AndersonA.J., JakemanL.B., McTigueD.M., and PopovichP.G. (2006). Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains. J. Neurotrauma, 23, 635–659.
33.
BlomsterL.V., CowinG.J., KurniawanN.D., and RuitenbergM.J. (2013). Detection of endogenous iron deposits in the injured mouse spinal cord through high-resolution ex vivo and in vivo MRI. NMR Biomed. 26, 141–150.
34.
PineauI., and LacroixS. (2007). Proinflammatory cytokine synthesis in the injured mouse spinal cord: multiphasic expression pattern and identification of the cell types involved. J. Comp. Neurol., 500, 267–285.
35.
BachmaierK., GuzmanE., KawamuraT., GaoX., and MalikA.B. (2012). Sphingosine kinase 1 mediation of expression of the anaphylatoxin receptor C5L2 dampens the inflammatory response to endotoxin. PLoS One, 7, e30742.
36.
GerardN.P., LuB., LiuP., CraigS., FujiwaraY., OkinagaS., and GerardC. (2005). An anti-inflammatory function for the complement anaphylatoxin C5a-binding protein, C5L2. J. Biol. Chem., 280, 39677–39680.
37.
ZhangX., SchmuddeI., LaumonnierY., PandeyM.K., ClarkJ.R., KonigP., GerardN.P., GerardC., Wills-KarpM., and KohlJ. (2010). A critical role for C5L2 in the pathogenesis of experimental allergic asthma. J. Immunol., 185, 6741–6752.
38.
RabyA.C., HolstB., DaviesJ., ColmontC., LaumonnierY., ColesB., ShahS., HallJ., TopleyN., KohlJ., MorganB.P., and LabetaM.O. (2011). TLR activation enhances C5a-induced pro-inflammatory responses by negatively modulating the second C5a receptor, C5L2. Eur. J. Immunol., 41, 2741–2752.
39.
RittirschD., FlierlM.A., NadeauB.A., DayD.E., Huber-LangM., MackayC.R., ZetouneF.S., GerardN.P., CianfloneK., KohlJ., GerardC., SarmaJ.V., and WardP.A. (2008). Functional roles for C5a receptors in sepsis. Nat. Med., 14, 551–557.
40.
FergusonA.R., ChristensenR.N., GenselJ.C., MillerB.A., SunF., BeattieE.C., BresnahanJ.C., and BeattieM.S. (2008). Cell death after spinal cord injury is exacerbated by rapid TNF alpha-induced trafficking of GluR2-lacking AMPARs to the plasma membrane. J. Neurosci., 28, 11391–11400.
41.
OkadaS., NakamuraM., MikamiY., ShimazakiT., MiharaM., OhsugiY., IwamotoY., YoshizakiK., KishimotoT., ToyamaY., and OkanoH. (2004). Blockade of interleukin-6 receptor suppresses reactive astrogliosis and ameliorates functional recovery in experimental spinal cord injury. J. Neurosci. Res., 76, 265–276.
GuerreroA.R., UchidaK., NakajimaH., WatanabeS., NakamuraM., JohnsonW.E., and BabaH. (2012). Blockade of interleukin-6 signaling inhibits the classic pathway and promotes an alternative pathway of macrophage activation after spinal cord injury in mice. J. Neuroinflammation, 9, 40.
44.
ArimaH., HanadaM., HayasakaT., MasakiN., OmuraT., XuD., HasegawaT., TogawaD., YamatoY., KobayashiS., YasudaT., MatsuyamaY., and SetouM. (2014). Blockade of IL-6 signaling by MR16-1 inhibits reduction of docosahexaenoic acid-containing phosphatidylcholine levels in a mouse model of spinal cord injury. Neuroscience, 269, 1–10.
45.
YaguchiM., OhtaS., ToyamaY., KawakamiY., and TodaM. (2008). Functional recovery after spinal cord injury in mice through activation of microglia and dendritic cells after IL-12 administration. J. Neurosci. Res., 86, 1972–1980.
46.
WuM.C., BrennanF.H., LynchJ.P., MantovaniS., PhippsS., WetselR.A., RuitenbergM.J., TaylorS.M., and WoodruffT.M. (2013). The receptor for complement component C3a mediates protection from intestinal ischemia-reperfusion injuries by inhibiting neutrophil mobilization. Proc. Natl. Acad. Sci. U. S. A., 110, 9439–9444.
47.
ScolaA.M., JohswichK.O., MorganB.P., KlosA., and MonkP.N. (2009). The human complement fragment receptor, C5L2, is a recycling decoy receptor. Mol. Immunol., 46, 1149–1162.
48.
WoodruffT.M., PollittS., ProctorL.M., StocksS.Z., MantheyH.D., WilliamsH.M., MahadevanI.B., ShielsI.A., and TaylorS.M. (2005). Increased potency of a novel complement factor 5a receptor antagonist in a rat model of inflammatory bowel disease. J. Pharmacol. Exp. Ther., 314, 811–817.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
