Effect of highly expressed IL-35 on Th1 and Th17 plasticity in colorectal cancer

doi:10.3969/j.issn.1673-8640.2024.04.006

Abstract

Abstract:

Objective To investigate the effect of highly expressed inhibitory cytokine interleukin（IL）-35 on the plasticity of T helper cell 1（Th1） and T helper cell 17（Th17）in colorectal cancer tissues. Methods A total of 90 patients with colorectal cancer（colorectal cancer group） and 28 healthy subjects（healthy control group） were enrolled from Xinhua Hospital of Shanghai Jiao Tong University School of Medicine from March 2019 to February 2020. Flow cytometry was used to determine Th1 percentage（Th1%） and Th17 percentage（Th17%） in peripheral blood of colorectal cancer group and healthy control group. The effects of highly expressed interferon-gamma（IFN-γ） and IL-17A on overall survival and poor prognosis of colorectal cancer were analyzed based on Kaplan-Meier Plotter database. Immunohistochemistry was used to determine the expression of IL-35 in cancer tissues and adjacent tissues（2 cm from tumor margin） of colorectal cancer patients. Serum IL-35 levels in colorectal cancer and healthy control groups were determined by enzyme-linked immunosorbent assay. The effect of IL-35 on the secretion of IL-17A and IFN-γ by Th1 was observed based on the induced Th1 differentiation induction experiment in vitro. Results Compared with healthy control group，Th1% in colorectal cancer group was decreased（P=0.019），and Th17% was increased（P<0.001）. The r² values of Th1% and Th17% in colorectal cancer group（0.393 4） were different from those in healthy control group（0.041 0）（P=0.007）. Low expression of IFN-γ mRNA was a risk factor for shorter survival in colorectal cancer patients [hazard ratio（HR）=0.74，95% confidence interval（CI）0.58-0.94，P=0.013]. High expression of IL-17A mRNA was a risk factor for shorter survival in colorectal cancer patients（HR=1.26，95%CI 1.02-1.59，P=0.020）. The expression of IL-35 in colorectal cancer tissues was higher than that in adjacent tissues（P<0.001）. Serum IL-35 levels in patients with stage Ⅰ to Ⅱ（early stage） and stage Ⅲ to Ⅳ（advanced stage） colorectal cancer were higher than those in healthy control group（P<0.05）. According to the results of Th1 differentiation induction experiment in vitro，IL-35 can decrease the expression of IFN-γ and increase the expression of IL-17A in Th1. Conclusions The proportions of Th1 and Th17 in peripheral blood of patients with colorectal cancer show abnormal changes，and the expression of IL-35 increases. High expression of IL-35 can decrease the expression of Th1 IFN-γ and increase the expression of IL-17A.

Key words: Interleukin-35, Colorectal cancer, T helper cell 1, T helper cell 17, Tumor immunity

CLC Number:

R446.1

XU Yunchuan, HUANGFU Yuchan, MA Yanhui. Effect of highly expressed IL-35 on Th1 and Th17 plasticity in colorectal cancer[J]. Laboratory Medicine, 2024, 39(4): 343-350.

Figures/Tables 10

References 23

[1]	SIEGEL R L, MILLER K D, FUCHS H E, et al. Cancer statistics,2021[J]. CA Cancer J Clin, 2021, 71(1):7-33. DOI URL
[2]	ALLEMANI C, MATSUDA T, DI CARLO V, et al. Global surveillance of trends in cancer survival 2000-14(CONCORD-3):analysis of individual records for 37 513 025 patients diagnosed with one of 18 cancers from 322 population-based registries in 71 countries[J]. Lancet, 2018, 391(10125):1023-1075. DOI URL
[3]	CHEN W, ZHENG R, BAADE P D, et al. Cancer statistics in China,2015[J]. CA Cancer J Clin, 2016, 66(2):115-132. DOI URL
[4]	COLLISON L W, WORKMAN C J, KUO T T, et al. The inhibitory cytokine IL-35 contributes to regulatory T-cell function[J]. Nature, 2007, 450(7169):566-569. DOI
[5]	SHEN P, ROCH T, LAMPROPOULOU V, et al. IL-35-producing B cells are critical regulators of immunity during autoimmune and infectious diseases[J]. Nature, 2014, 507(7492):366-370. DOI
[6]	WANG Z, LIU J Q, LIU Z, et al. Tumor-derived IL-35 promotes tumor growth by enhancing myeloid cell accumulation and angiogenesis[J]. J Immunol, 2013, 190(5):2415-2423. DOI PMID
[7]	NIEDBALA W, WEI X Q, CAI B, et al. IL-35 is a novel cytokine with therapeutic effects against collagen-induced arthritis through the expansion of regulatory T cells and suppression of Th17 cells[J]. Eur J Immunol, 2007, 37(11):3021-3029. DOI PMID
[8]	ZHANG L, YU X, ZHENG L, et al. Lineage tracking reveals dynamic relationships of T cells in colorectal cancer[J]. Nature, 2018, 564(7735):268-272. DOI
[9]	ALEXANDER M, ANG Q Y, NAYAK R R, et al. Human gut bacterial metabolism drives Th17 activation and colitis[J]. Cell Host Microbe, 2022, 30(1):17-30. DOI URL
[10]	LIU H P, CAO A T, FENG T, et al. TGF-β converts Th1 cells into Th17 cells through stimulation of Runx1 expression[J]. Eur J Immunol, 2015, 45(4):1010-1018. DOI URL
[11]	MA Y, CHEN L, XIE G, et al. Elevated level of interleukin-35 in colorectal cancer induces conversion of T cells into iTr35 by activating STAT1/STAT3[J]. Oncotarget, 2016, 7(45):73003-73015. DOI PMID
[12]	MAZZONI A, MAGGI L, LIOTTA F, et al. Biological and clinical significance of T helper 17 cell plasticity[J]. Immunology, 2019, 158(4):287-295. DOI PMID
[13]	EDGE S B, COMPTON C C. The American Joint Committee on Cancer:the 7th edition of the AJCC cancer staging manual and the future of TNM[J]. Ann Surg Oncol, 2010,17:1471-1474.
[14]	LÁNCZKY A, GYÖRFFY B. Web-based survival analysis tool tailored for medical research(KMplot):development and implementation[J]. J Med Internet Res, 2021, 23(7):e27633. DOI URL
[15]	TOSOLINI M, KIRILOVSKY A, MLECNIK B, et al. Clinical impact of different classes of infiltrating T cytotoxic and helper cells(Th1,Th2,treg,Th17)in patients with colorectal cancer[J]. Cancer Res, 2011, 71(4):1263-1271. DOI URL
[16]	ZHANG L, YU X, ZHENG L, et al. Lineage tracking reveals dynamic relationships of T cells in colorectal cancer[J]. Nature, 2018, 564(7735):268-272. DOI
[17]	TERNES D, TSENKOVA M, POZDEEV V I, et al. The gut microbial metabolite formate exacerbates colorectal cancer progression[J]. Nat Metab, 2022, 4(4):458-475. DOI PMID
[18]	LI S, NA R, LI X, et al. Targeting interleukin-17 enhances tumor response to immune checkpoint inhibitors in colorectal cancer[J]. Biochim Biophys Acta Rev Cancer, 2022, 1877(4):188758. DOI URL
[19]	KRYCZEK I, BANERJEE M, CHENG P, et al. Phenotype,distribution,generation,and functional and clinical relevance of Th17 cells in the human tumor environments[J]. Blood, 2009, 114(6):1141-1149.
[20]	ZHANG J P, YAN J, XU J, et al. Increased intratumoral IL-17-producing cells correlate with poor survival in hepatocellular carcinoma patients[J]. J Hepatol, 2009, 50(5):980-989. DOI URL
[21]	MARTIN-OROZCO N, MURANSKI P, CHUNG Y, et al. T helper 17 cells promote cytotoxic T cell activation in tumor immunity[J]. Immunity, 2009, 31(5):787-798. DOI URL
[22]	PRABHALA R H, PELLURU D, FULCINITI M, et al. Elevated IL-17 produced by TH17 cells promotes myeloma cell growth and inhibits immune function in multiple myeloma[J]. Blood, 2010, 115(26):5385-5392. DOI PMID
[23]	YI P, YU W, XIONG Y, et al. IL-35:new target for immunotherapy targeting the tumor microenvironment[J]. Mol Cancer Ther, 2024, 23(2):148-158. DOI URL

TNM分期			年龄				性别				病理类型
TNM分期			≤60岁		>60岁		男		女		管状腺癌	黏液性腺癌	乳头状腺癌	未知
Ⅰ~Ⅱ期	（54例）		7（12.96）		47（87.03）		28（51.85）		26（48.15）		44（81.58）	3（5.26）	3（5.26）	4（7.89）
Ⅲ~Ⅳ期	（36例）		9（25.00）		27（75.00）		19（52.78）		17（47.22）		27（75.00）	3（8.33）	2（5.56）	4（11.11）
U值			871.00				963.00				772.50
P值			0.41				0.93				0.70
TNM分期		肿瘤部位
TNM分期		升结肠		横结肠		降结肠		乙状结肠		直肠
Ⅰ~Ⅱ期		7（12.96）		16（29.63）		4（7.41）		22（40.74）		5（9.26）
Ⅲ~Ⅳ期		6（16.67）		14（38.89）		5（13.89）		10（27.78）		1（2.78）
U值		787.50
P值		0.11

TNM分期			年龄				性别				病理类型
TNM分期			≤60岁		>60岁		男		女		管状腺癌	黏液性腺癌	乳头状腺癌	未知
Ⅰ~Ⅱ期	（54例）		7（12.96）		47（87.03）		28（51.85）		26（48.15）		44（81.58）	3（5.26）	3（5.26）	4（7.89）
Ⅲ~Ⅳ期	（36例）		9（25.00）		27（75.00）		19（52.78）		17（47.22）		27（75.00）	3（8.33）	2（5.56）	4（11.11）
U值			871.00				963.00				772.50
P值			0.41				0.93				0.70
TNM分期		肿瘤部位
TNM分期		升结肠		横结肠		降结肠		乙状结肠		直肠
Ⅰ~Ⅱ期		7（12.96）		16（29.63）		4（7.41）		22（40.74）		5（9.26）
Ⅲ~Ⅳ期		6（16.67）		14（38.89）		5（13.89）		10（27.78）		1（2.78）
U值		787.50
P值		0.11

组别	例数	Th1%/%	Th17%/%
结直肠癌组	31	10.38（4.03，15.38）	2.16（1.56，3.12）
正常对照组	28	16.75（12.37，21.89）	0.96（0.76，1.83）
U值		280	167
P值		0.019	<0.001

组别	例数	Th1%/%	Th17%/%
结直肠癌组	31	10.38（4.03，15.38）	2.16（1.56，3.12）
正常对照组	28	16.75（12.37，21.89）	0.96（0.76，1.83）
U值		280	167
P值		0.019	<0.001

临床病理特征	例数	癌组织			癌旁组织			临床病理特征	例数	癌组织			癌旁组织
临床病理特征	例数	低表达	高表达	缺失	低表达	高表达	缺失	临床病理特征	例数	低表达	高表达	缺失	低表达	高表达	缺失
年龄								T分期
≤60岁	34	15	18	1	30	2	2	T1/T2期	9	6	3	0	8	1	0
>60岁	54	36	18	0	49	3	2	T3/T4期	79	46	32	1	72	4	3
缺失	2	1	1	0	2	0	0	缺失	2	0	2	0	1	0	1
P值			0.053			0.928		P值			0.685			0.483
性别								淋巴转移
男	47	27	20	0	43	2	2	N0	56	33	23	0	51	2	3
女	43	25	17	0	38	3	2	N1/N2	34	19	14	1	30	3	1
P值			0.844			0.572		P值			0.901			0.308
肿瘤大小								远隔转移
≤5 cm	46	25	20	1	40	4	2	M0	87	51	35	1	79	4	4
>5 cm	42	16	26	9	39	1	2	M1	2	1	1	0	1	1	0
缺失	2	1	1	0	2	0	0	缺失	1	0	1	0	1	0	0
P值			0.550			0.205		P值			0.793			0.008
病理分级								TNM分期
Ⅰ级	5	5	0	0	5	0	0	Ⅰ~Ⅱ期	54	33	21	0	50	2	2
Ⅱ~Ⅲ级	85	47	37	1	76	5	4	Ⅲ~Ⅳ期	34	19	14	1	30	3	1
P值			0.054			0.569		缺失	2	0	2	0	1	0	1
								P值			0.746			0.319