It has been widely accepted that monoclonal antibody (mAb) is an effective tool for cancer immunotherapy. The C-C motif chemokine receptor 8 (CCR8) is highly expressed in regulatory T cells and many cancers and is associated with the progression of the cancers. However, its role in cancer progression remains unclear. Thus, the development of mAbs for CCR8 leads to cancer immunotherapy and elucidation of unknown mechanisms of CCR8-dependent cancer progression. In this study, we have developed an anti-mouse CCR8 (mCCR8) mAb (clone C8Mab-1, rat IgG2a, kappa) using the Cell-Based Immunization and Screening (CBIS) method. We showed that C8Mab-1 and its recombinant antibody (recC8Mab-1) bind to mCCR8-overexpressed Chinese hamster ovary (CHO)-K1 cells (CHO/mCCR8), but not to the parental CHO-K1 cells, in flow cytometry and immunofluorescence. Moreover, C8Mab-1 and recC8Mab-1 specifically reacted to P388 (a mouse lymphocyte-like cells) and J774-1 (a mouse macrophage-like cells), which express endogenous mCCR8, in both applications. These results suggest that C8Mab-1, developed using the CBIS method, is useful for flow cytometry and immunocytochemistry against exogenous and endogenous mCCR8.
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References
1.
van den BulkJ, VerdegaalEM, and de MirandaNF: Cancer immunotherapy: Broadening the scope of targetable tumours. Open Biol, 2018; 8:180037.
2.
Sasidharan NairV, and ElkordE: Immune checkpoint inhibitors in cancer therapy: A focus on T-regulatory cells. Immunol Cell Biol, 2018; 96:21–33.
3.
LiC, JiangP, WeiS, XuX, and WangJ: Regulatory T cells in tumor microenvironment: New mechanisms, potential therapeutic strategies and future prospects. Mol Cancer, 2020; 19:116.
4.
De SimoneM, ArrigoniA, RossettiG, GruarinP, RanzaniV, PolitanoC, BonnalRJP, ProvasiE, SarnicolaML, PanzeriI, MoroM, CrostiM, MazzaraS, VairaV, BosariS, PalleschiA, SantambrogioL, BovoG, ZucchiniN, TotisM, GianottiL, CesanaG, PeregoRA, MaroniN, Pisani CerettiA, OpocherE, De FrancescoR, GeginatJ, StunnenbergHG, AbrignaniS, and PaganiM: Transcriptional landscape of human tissue lymphocytes unveils uniqueness of tumor-infiltrating T regulatory cells. Immunity, 2016; 45:1135–1147.
5.
TiffanyHL, LautensLL, GaoJL, PeaseJ, LocatiM, CombadiereC, ModiW, BonnerTI, and MurphyPM: Identification of CCR8: A human monocyte and thymus receptor for the CC chemokine I-309. J Exp Med, 1997; 186:165–170.
6.
IslamSA, LingMF, LeungJ, ShrefflerWG, and LusterAD: Identification of human CCR8 as a CCL18 receptor. J Exp Med, 2013; 210:1889–1898.
7.
IslamSA, ChangDS, ColvinRA, ByrneMH, McCullyML, MoserB, LiraSA, CharoIF, and LusterAD: Mouse CCL8, a CCR8 agonist, promotes atopic dermatitis by recruiting IL-5+ T(H)2 cells. Nat Immunol, 2011; 12:167–177.
8.
HowardOM, DongHF, ShirakawaAK, and OppenheimJJ: LEC induces chemotaxis and adhesion by interacting with CCR1 and CCR8. Blood, 2000; 96:840–845.
9.
PlitasG, KonopackiC, WuK, BosPD, MorrowM, PutintsevaEV, ChudakovDM, and RudenskyAY: Regulatory T cells exhibit distinct features in human breast cancer. Immunity, 2016; 45:1122–1134.
10.
FragaM, YanezM, ShermanM, LlerenaF, HernandezM, NourdinG, AlvarezF, UrrizolaJ, RiveraC, LampertiL, NovaL, CastroS, ZambranoO, CifuentesA, CamposL, MoyaS, PastorJ, NunezM, GaticaJ, FigueroaJ, ZunigaF, SalomonC, CerdaG, PuentesR, LabarcaG, VidalM, McGregorR, and Nova-LampertiE: Immunomodulation of T helper cells by tumor microenvironment in oral cancer is associated with CCR8 expression and rapid membrane vitamin D signaling pathway. Front Immunol, 2021; 12:643298.
11.
LiuX, XuX, DengW, HuangM, WuY, ZhouZ, ZhuK, WangY, ChengX, ZhouX, ChenL, LiY, WangG, and FuB: CCL18 enhances migration, invasion and EMT by binding CCR8 in bladder cancer cells. Mol Med Rep, 2019; 19:1678–1686.
12.
IellemA, MarianiM, LangR, RecaldeH, Panina-BordignonP, SinigagliaF, and D'AmbrosioD: Unique chemotactic response profile and specific expression of chemokine receptors CCR4 and CCR8 by CD4+CD25+ regulatory T cells. J Exp Med, 2001; 194:847–853.
13.
SpinettiG, BernardiniG, CamardaG, MangoniA, SantoniA, CapogrossiMC, and NapolitanoM: The chemokine receptor CCR8 mediates rescue from dexamethasone-induced apoptosis via an ERK-dependent pathway. J Leukoc Biol, 2003; 73:201–207.
14.
DenisC, DeiterenK, MortierA, TounsiA, FransenE, ProostP, RenauldJC, and LambeirAM: C-terminal clipping of chemokine CCL1/I-309 enhances CCR8-mediated intracellular calcium release and anti-apoptotic activity. PLoS One, 2012; 7:e34199.
AsanoT, NanamiyaR, TakeiJ, NakamuraT, YanakaM, HosonoH, TanakaT, SanoM, KanekoMK, and KatoY: Development of anti-mouse CC chemokine receptor 3 monoclonal antibodies for flow cytometry. Monoclon Antib Immunodiagn Immunother, 2021; 40:107–112.
17.
NanamiyaR, TakeiJ, AsanoT, TanakaT, SanoM, NakamuraT, YanakaM, HosonoH, KanekoMK, and KatoY: Development of anti-human CC chemokine receptor 9 monoclonal antibodies for flow cytometry. Monoclon Antib Immunodiagn Immunother, 2021; 40:101–106.
18.
TanakaT, NanamiyaR, TakeiJ, NakamuraT, YanakaM, HosonoH, SanoM, AsanoT, KanekoMK, and KatoY: Development of anti-mouse CC chemokine receptor 8 monoclonal antibodies for flow cytometry. Monoclon Antib Immunodiagn Immunother, 2021; 40:65–70.
19.
YamadaS, ItaiS, NakamuraT, YanakaM, KanekoMK, and KatoY: Detection of high CD44 expression in oral cancers using the novel monoclonal antibody, C44Mab-5. Biochem Biophys Rep, 2018; 14:64–68.
20.
YamadaS, ItaiS, NakamuraT, YanakaM, ChangYW, SuzukiH, KanekoMK, and KatoY: Monoclonal antibody L1Mab-13 detected human PD-L1 in lung cancers. Monoclon Antib Immunodiagn Immunother, 2018; 37:110–115.
21.
TanakaT, AsanoT, SanoM, TakeiJ, HosonoH, NanamiyaR, NakamuraT, YanakaM, HaradaH, FukuiM, SuzukiH, UchidaK, NakagawaT, KatoY, and KanekoMK: Development of monoclonal antibody PMab-269 against California Sea Lion podoplanin. Monoclon Antib Immunodiagn Immunother, 2021; 40:124–133.
22.
KanekoMK, SanoM, TakeiJ, AsanoT, SayamaY, HosonoH, KobayashiA, KonnaiS, and KatoY: Development and characterization of anti-sheep podoplanin monoclonal antibodies PMab-253 and PMab-260. Monoclon Antib Immunodiagn Immunother, 2020; 39:144–155.
23.
FurusawaY, KanekoMK, and KatoY: Establishment of C20Mab-11, a novel anti-CD20 monoclonal antibody, for the detection of B cells. Oncol Lett, 2020; 20:1961–1967.
24.
FurusawaY, KanekoMK, and KatoY: Establishment of an anti-CD20 monoclonal antibody (C20Mab-60) for immunohistochemical analyses. Monoclon Antib Immunodiagn Immunother, 2020; 39:112–116.
25.
ItaiS, FujiiY, NakamuraT, ChangYW, YanakaM, SaidohN, HandaS, SuzukiH, HaradaH, YamadaS, KanekoMK, and KatoY: Establishment of CMab-43, a sensitive and specific anti-CD133 monoclonal antibody, for immunohistochemistry. Monoclon Antib Immunodiagn Immunother, 2017; 36:231–235.
26.
VillarrealDO, L'HuillierA, ArmingtonS, MottersheadC, FilippovaEV, CoderBD, PetitRG, and PrinciottaMF: Targeting CCR8 induces protective antitumor immunity and enhances vaccine-induced responses in colon cancer. Cancer Res, 2018; 78:5340–5348.
27.
WangT, ZhouQ, ZengH, ZhangH, LiuZ, ShaoJ, WangZ, XiongY, WangJ, BaiQ, XiaY, WangY, LiuL, ZhuY, XuL, DaiB, GuoJ, ChangY, WangX, and XuJ: CCR8 blockade primes anti-tumor immunity through intratumoral regulatory T cells destabilization in muscle-invasive bladder cancer. Cancer Immunol Immunother, 2020; 69:1855–1867.
28.
Van DammeH, DombrechtB, KissM, RooseH, AllenE, Van OvermeireE, KanchevaD, MartensL, MurgaskiA, BardetPMR, BlanckeG, JansM, BolliE, MartinsMS, ElkrimY, DooleyJ, BoonL, SchwarzeJK, TackeF, MovahediK, VandammeN, NeynsB, OcakS, ScheyltjensI, VereeckeL, NanaFA, MerchiersP, LaouiD, and Van GinderachterJA: Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy. J Immunother Cancer, 2021; 9:e001749.
29.
CampbellJR, McDonaldBR, MeskoPB, SiemersNO, SinghPB, SelbyM, SproulTW, KormanAJ, VlachLM, HouserJ, SambanthamoorthyS, LuK, HatcherSV, LohreJ, JainR, and LanRY: Fc-optimized anti-CCR8 antibody depletes regulatory T cells in human tumor models. Cancer Res, 2021; 81:2983–2994.
30.
SolerD, ChapmanTR, PoissonLR, WangL, Cote-SierraJ, RyanM, McDonaldA, BadolaS, FedykE, CoyleAJ, HodgeMR, and KolbeckR: CCR8 expression identifies CD4 memory T cells enriched for FOXP3+ regulatory and Th2 effector lymphocytes. J Immunol, 2006; 177:6940–6951.
31.
MikhakZ, FukuiM, FarsidjaniA, MedoffBD, TagerAM, and LusterAD: Contribution of CCR4 and CCR8 to antigen-specific TH2 cell trafficking in allergic pulmonary inflammation. J Allergy Clin Immunol, 2009; 123:67–73.e63.
32.
BarsheshetY, WildbaumG, LevyE, VitenshteinA, AkinseyeC, GriggsJ, LiraSA, and KarinN: CCR8+FOXp3+ Treg cells as master drivers of immune regulation. Proc Natl Acad Sci U S A, 2017; 114:6086–6091.
33.
Blanco-PerezF, KatoY, Gonzalez-MenendezI, LainoJ, OhbayashiM, BurggrafM, KrauseM, KirbergJ, IwakuraY, MartellaM, Quintanilla-MartinezL, ShibataN, ViethsS, ScheurerS, and TodaM: CCR8 leads to eosinophil migration and regulates neutrophil migration in murine allergic enteritis. Sci Rep, 2019; 9:9608.
34.
ZychowskaM, RojewskaE, PiotrowskaA, KreinerG, NalepaI, and MikaJ: Spinal CCL1/CCR8 signaling interplay as a potential therapeutic target—Evidence from a mouse diabetic neuropathy model. Int Immunopharmacol, 2017; 52:261–271.
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