The CC chemokine receptor type-2 (CCR2) is one of the members of the G protein-coupled receptor superfamily, which are expressed on the cell surface of immune and tumor cells. CCR2 binds to the C-C motif chemokine ligand 2 (CCL2)/monocyte chemoattractant protein-1 (MCP-1), which is produced by various cells, including tumor and immune-related cells. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CCR2 has been desired for treatment and diagnosis. In this study, we established a specific antihuman CCR2 (hCCR2) mAb, C2Mab-9 (mouse IgG1, kappa), using the synthetic peptide immunization method. Flow cytometric and immunocytochemical results showed that C2Mab-9 reacted with hCCR2-expressing U937 (human histiocytic lymphoma) and natural killer cells. Furthermore, C2Mab-9 showed the moderate binding affinity for both cells. Conclusively, C2Mab-9 can be a useful tool for analyzing hCCR2-related biological responses.
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References
1.
HughesCE, and NibbsRJB: A guide to chemokines and their receptors. FEBS J, 2018; 285:2944–2971.
2.
DoHTT, LeeCH, and ChoJ: Chemokines and their receptors: multifaceted roles in cancer progression and potential value as cancer prognostic markers. Cancers (Basel), 2020; 12:287.
3.
SokolCL, and LusterAD: The chemokine system in innate immunity. Cold Spring Harb Perspect Biol, 2015; 7:a016303.
4.
LutherSA, and CysterJG: Chemokines as regulators of T cell differentiation. Nat Immunol, 2001; 2:102–107.
5.
UrbantatRM, VajkoczyP, and BrandenburgS: Advances in chemokine signaling pathways as therapeutic targets in glioblastoma. Cancers (Basel), 2021; 13:2983.
6.
BianconiV, SahebkarA, AtkinSL, and PirroM: The regulation and importance of monocyte chemoattractant protein-1. Curr Opin Hematol, 2018; 25:44–51.
7.
LimSY, YuzhalinAE, Gordon-WeeksAN, and MuschelRJ: Targeting the CCL2-CCR2 signaling axis in cancer metastasis. Oncotarget, 2016; 7:28697–28710.
8.
O'ConnorT, and HeikenwalderM: CCL2 in the tumor microenvironment. Adv Exp Med Biol, 2021; 1302:1–14.
9.
XuM, WangY, XiaR, WeiY, and WeiX: Role of the CCL2-CCR2 signalling axis in cancer: mechanisms and therapeutic targeting. Cell Prolif, 2021; 54:e13115.
10.
KadomotoS, IzumiK, and MizokamiA: Roles of CCL2-CCR2 axis in the tumor microenvironment. Int J Mol Sci, 2021; 22:8530.
HaoQ, VadgamaJV, and WangP: CCL2/CCR2 signaling in cancer pathogenesis. Cell Commun Signal, 2020; 18:82.
13.
TalbotJ, BianchiniFJ, NascimentoDC, OliveiraRD, SoutoFO, PintoLG, PeresRS, SilvaJR, AlmeidaSC, Louzada-JuniorP, CunhaTM, CunhaFQ, and Alves-FilhoJC: CCR2 expression in neutrophils plays a critical role in their migration into the joints in rheumatoid arthritis. Arthritis Rheumatol, 2015; 67:1751–1759.
14.
Midavaine É, CôtéJ, and SarretP: The multifaceted roles of the chemokines CCL2 and CXCL12 in osteophilic metastatic cancers. Cancer Metastasis Rev, 2021; 40:427–445.
15.
LobergRD, DayLL, HarwoodJ, YingC, St JohnLN, GilesR, NeeleyCK, and PientaKJ: CCL2 is a potent regulator of prostate cancer cell migration and proliferation. Neoplasia, 2006; 8:578–586.
16.
RogicA, PantI, GrumolatoL, Fernandez-RodriguezR, EdwardsA, DasS, SunA, YaoS, QiaoR, JafferS, SachidanandamR, AkturkG, KarlicR, SkobeM, and AaronsonSA: High endogenous CCL2 expression promotes the aggressive phenotype of human inflammatory breast cancer. Nat Commun, 2021; 12:6889.
17.
FangWB, Sofia AcevedoD, SmartC, ZindaB, AlissaN, WarrenK, FragaG, HuangLC, ShyrY, LiW, XieL, StaggsV, HongY, BehbodF, and ChengN: Expression of CCL2/CCR2 signaling proteins in breast carcinoma cells is associated with invasive progression. Sci Rep, 2021; 11:8708.
18.
AsanoT, OhishiT, TakeiJ, NakamuraT, NanamiyaR, HosonoH, TanakaT, SanoM, HaradaH, KawadaM, KanekoMK, and KatoY: Anti‑HER3 monoclonal antibody exerts antitumor activity in a mouse model of colorectal adenocarcinoma. Oncol Rep, 2021; 46:173.
19.
HosonoH, OhishiT, TakeiJ, AsanoT, SayamaY, KawadaM, KanekoMK, and KatoY: The anti-epithelial cell adhesion molecule (EpCAM) monoclonal antibody EpMab-16 exerts antitumor activity in a mouse model of colorectal adenocarcinoma. Oncol Lett, 2020; 20:383.
20.
AsanoT, NanamiyaR, TanakaT, KanekoMK, and KatoY: Development of antihuman killer cell lectin-like receptor subfamily G member 1 monoclonal antibodies for flow cytometry. Monoclon Antib Immunodiagn Immunother, 2021; 40:76–80.
21.
TakeiJ, AsanoT, NanamiyaR, NakamuraT, YanakaM, HosonoH, TanakaT, SanoM, KanekoMK, HaradaH, and KatoY: Development of anti-human T cell immunoreceptor with Ig and ITIM domains (TIGIT) monoclonal antibodies for flow cytometry. Monoclon Antib Immunodiagn Immunother, 2021; 40:71–75.
22.
SayamaY, KanekoMK, and KatoY: Development and characterization of TrMab‑6, a novel anti‑TROP2 monoclonal antibody for antigen detection in breast cancer. Mol Med Rep, 2021; 23:92.
23.
SayamaY, KanekoMK, TakeiJ, HosonoH, SanoM, AsanoT, and KatoY: Establishment of a novel anti-TROP2 monoclonal antibody TrMab-29 for immunohistochemical analysis. Biochem Biophys Rep, 2021; 25:100902.
24.
YamadaS, ItaiS, NakamuraT, YanakaM, ChangYW, SuzukiH, KanekoMK, and KatoY: Monoclonal antibody L(1)Mab-13 detected human PD-L1 in lung cancers. Monoclon Antib Immunodiagn Immunother, 2018; 37:110–115.
25.
YamadaS, KanekoMK, SayamaY, AsanoT, SanoM, YanakaM, NakamuraT, OkamotoS, HandaS, KomatsuY, NakamuraY, FurusawaY, TakeiJ, and KatoY: Development of novel mouse monoclonal antibodies against human CD19. Monoclon Antib Immunodiagn Immunother, 2020; 39:45–50.
26.
FurusawaY, KanekoMK, and KatoY: Establishment of C(20)Mab-11, a novel anti-CD20 monoclonal antibody, for the detection of B cells. Oncol Lett, 2020; 20:1961–1967.
27.
FurusawaY, KanekoMK, and KatoY: Establishment of an anti-CD20 monoclonal antibody (C(20)Mab-60) for immunohistochemical analyses. Monoclon Antib Immunodiagn Immunother, 2020; 39:112–116.
28.
YamadaS, ItaiS, NakamuraT, YanakaM, KanekoMK, and KatoY: Detection of high CD44 expression in oral cancers using the novel monoclonal antibody, C(44)Mab-5. Biochem Biophys Rep, 2018; 14:64–68.
29.
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.
30.
FurusawaY, TakeiJ, SayamaY, YamadaS, KanekoMK, and KatoY: Development of an anti-bear podoplanin monoclonal antibody PMab-247 for immunohistochemical analysis. Biochem Biophys Rep, 2019; 18:100644.
31.
HosonoH, AsanoT, TakeiJ, SanoM, TanakaT, KanekoMK, and KatoY: Development of an anti-elephant podoplanin monoclonal antibody PMab-265 for flow cytometry. Monoclon Antib Immunodiagn Immunother, 2021; 40:141–145.
32.
KatoY, FurusawaY, SanoM, TakeiJ, NakamuraT, YanakaM, OkamotoS, HandaS, KomatsuY, AsanoT, SayamaY, and KanekoMK: Development of an anti-sheep podoplanin monoclonal antibody PMab-256 for immunohistochemical analysis of lymphatic endothelial cells. Monoclon Antib Immunodiagn Immunother, 2020; 39:82–90.
33.
OkiH, HonmaR, OgasawaraS, FujiiY, LiuX, TakagiM, KanekoMK, and KatoY: Development of sensitive monoclonal antibody PMab-2 against rat podoplanin. Monoclon Antib Immunodiagn Immunother, 2015; 34:396–403.
34.
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.
35.
YamadaS, OgasawaraS, KanekoMK, and KatoY: LpMab-23: a cancer-specific monoclonal antibody against human podoplanin. Monoclon Antib Immunodiagn Immunother, 2017; 36:72–76.
36.
HonmaR, FujiiY, OgasawaraS, OkiH, LiuX, NakamuraT, KanekoMK, TakagiM, and KatoY: Establishment of novel monoclonal antibody PMab-32 against rabbit podoplanin. Monoclon Antib Immunodiagn Immunother, 2016; 35:41–47.
37.
YamadaS, ItaiS, NakamuraT, YanakaM, SaidohN, ChangYW, HandaS, HaradaH, KagawaY, IchiiO, KonnaiS, KanekoMK, and KatoY: PMab-52: specific and sensitive monoclonal antibody against cat podoplanin for immunohistochemistry. Monoclon Antib Immunodiagn Immunother, 2017; 36:224–230.
38.
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.
39.
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.
40.
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.
41.
SchönebergT, and LiebscherI: Mutations in G protein-coupled receptors: mechanisms, pathophysiology and potential therapeutic approaches. Pharmacol Rev, 2021; 73:89–119.
YangH, ZhangQ, XuM, WangL, ChenX, FengY, LiY, ZhangX, CuiW, and JiaX: CCL2-CCR2 axis recruits tumor associated macrophages to induce immune evasion through PD-1 signaling in esophageal carcinogenesis. Mol Cancer, 2020; 19:41.
44.
PanekWK, PituchKC, MiskaJ, KimJW, RashidiA, KanojiaD, Lopez-RosasA, HanY, YuD, ChangCL, KaneJR, ZhangP, CorderoA, and LesniakMS: Local application of autologous platelet-rich fibrin patch (PRF-P) suppresses regulatory T cell recruitment in a murine glioma model. Mol Neurobiol, 2019; 56:5032–5040.
45.
YeXZ, XuSL, XinYH, YuSC, PingYF, ChenL, XiaoHL, WangB, YiL, WangQL, JiangXF, YangL, ZhangP, QianC, CuiYH, ZhangX, and BianXW: Tumor-associated microglia/macrophages enhance the invasion of glioma stem-like cells via TGF-β1 signaling pathway. J Immunol, 2012; 189:444–453.
46.
CraneCA, AhnBJ, HanSJ, and ParsaAT: Soluble factors secreted by glioblastoma cell lines facilitate recruitment, survival, and expansion of regulatory T cells: implications for immunotherapy. Neuro Oncol, 2012; 14:584–595.
47.
HuangB, LeiZ, ZhaoJ, GongW, LiuJ, ChenZ, LiuY, LiD, YuanY, ZhangGM, and FengZH: CCL2/CCR2 pathway mediates recruitment of myeloid suppressor cells to cancers. Cancer Lett, 2007; 252:86–92.
Flores-ToroJA, LuoD, GopinathA, SarkisianMR, CampbellJJ, CharoIF, SinghR, SchallTJ, DattaM, JainRK, MitchellDA, and HarrisonJK: CCR2 inhibition reduces tumor myeloid cells and unmasks a checkpoint inhibitor effect to slow progression of resistant murine gliomas. Proc Natl Acad Sci U S A, 2020; 117:1129–1138.
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