Correlations of immune cell infiltration characteristics with clinicopathological parameters in patients with clear cell renal cell carcinoma

doi:10.12122/j.issn.1673-4254.2025.06.17

Abstract

Abstract:

Objective To investigate the characteristics of immune cell infiltration in tumor samples from Chinese patients with clear cell renal cell carcinoma (ccRCC) and the correlation of immune cell infiltration with tumor stage and response to immunotherapy. Methods Tumor samples and clinicopathological data were collected from 154 ccRCC patients treated in Nanfang Hospital, Southern Medical University from October, 2020 to October, 2023. The immune cell types infiltrating the tumor tissues were identified using immunohistochemistry and immunofluorescence staining, and their correlations with the patients' clinicopathological characteristics were analyzed. Patient-derived tumor tissue fragment models (PDTF) models, constructed using tumor tissues from 22 patients, were treated with PD-1 monoclonal antibody, and T cell activation was detected using flow cytometry to assess the patients' responses to immunotherapy. Results In Chinese ccRCC patients included in this study, CD8⁺ T cells, CD4⁺ T cells, and CD3⁺ T cells were the most abundant in the tumor tissues. Higher infiltration levels of CD3⁺ T cells (P=0.004), PD-1⁺ T cells (P=0.020), CD68⁺ T cells (P=0.049), CD79⁺ T cells (P=0.049), and Tryptase⁺ cells (P=0.049) were all positively correlated with a larger tumor size (≥5 cm). A higher infiltration level of CD4⁺ T cells was associated with a lower tumor stage. Patients with higher International Society of Urological Pathology (ISUP) grades had higher infiltration levels of CD3⁺ T cells (P=0.023), CD8⁺ T cells (P=0.045), PD-1⁺ T cells (P=0.014), CD20⁺ B cells (P=0.020) and CD79⁺ B cells (P=0.049), and lower levels of Tryptase⁺ cells (P=0.001). Patients with abundant infiltrating immune cells tended to have better responses to immunotherapy. Conclusion The infiltrating immune cells are heterogeneous in Chinese ccRCC patients, and immune cell infiltration characteristics are closely correlated with clinicopathological parameters of the patients.

Key words: clear cell renal cell carcinoma, tumor-infiltrating lymphocytes, patient-derived tumor tissue fragment models

Huaxuan ZHAO, Guichao ZHANG, Jiarong LIU, Futian MO, Taoen LI, Chengyong LEI, Shidong LÜ. Correlations of immune cell infiltration characteristics with clinicopathological parameters in patients with clear cell renal cell carcinoma[J]. Journal of Southern Medical University, 2025, 45(6): 1280-1288.

Figures/Tables 8

References 35

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Antibody	Clone	Company	Dilution ratio
CD3	MX036	MXB Biotechnologies	Ready to use
CD4	SP35	MXB Biotechnologies	Ready to use
CD8	SP16	Amresco	Ready to use
FOXP3	236A/E7	Abcam	1:250
CD68	C68/684	Abcam	1:100
CD20	EP459Y	Abcam	1:300
CD79	EP3618	Abcam	1:250
Tryptase	EPR8476	Abcam	1:2500
Programmed Death-1	D4W2J	Cell Signaling Technology	1:200
Programmed Death-Ligand 1	EPR19759	Abcam	1:250

Antibody	Clone	Company	Dilution ratio
CD3	MX036	MXB Biotechnologies	Ready to use
CD4	SP35	MXB Biotechnologies	Ready to use
CD8	SP16	Amresco	Ready to use
FOXP3	236A/E7	Abcam	1:250
CD68	C68/684	Abcam	1:100
CD20	EP459Y	Abcam	1:300
CD79	EP3618	Abcam	1:250
Tryptase	EPR8476	Abcam	1:2500
Programmed Death-1	D4W2J	Cell Signaling Technology	1:200
Programmed Death-Ligand 1	EPR19759	Abcam	1:250

Antibody	Coupled fluorescein	Clone	Company
CD3	FITC	UCHT1	BD Biosciences
CD4	APC-CY7	RPA-T4	BD Biosciences
CD8	BV605	SK1	BD Biosciences
CD278 (ICOS)	PE	DX29	BD Biosciences
CD134 (OX40)	PE-CY7	ACT35	BD Biosciences
CD137	BV421	4B4-1	BD Biosciences
CD25	APC	M-A251	BD Biosciences

Antibody	Coupled fluorescein	Clone	Company
CD3	FITC	UCHT1	BD Biosciences
CD4	APC-CY7	RPA-T4	BD Biosciences
CD8	BV605	SK1	BD Biosciences
CD278 (ICOS)	PE	DX29	BD Biosciences
CD134 (OX40)	PE-CY7	ACT35	BD Biosciences
CD137	BV421	4B4-1	BD Biosciences
CD25	APC	M-A251	BD Biosciences

Characteristic	Immunohistochemical cohort (n=154)	PDTF cohort (n=22)	P
Gender			>0.999
Male	106 (68.8)	15 (68.2)
Female	48 (31.2)	7 (31.8)
Age (year)			0.811
<50	51 (33.1)	8 (36.4)
≥50	103 (66.9)	14 (63.6)
Tumor size (cm)			0.819
<5	90 (58.4)	12 (54.5)
≥5	64 (41.6)	10 (45.5)
Tumor stage			0.964
1a	75 (48.7)	10 (45.4)
1b	49 (31.8)	8 (36.4)
2a	16 (10.4)	2 (9.1)
3a	11 (7.1)	2 (9.1)
3b	2 (1.3)	0 (0)
4	1 (0.7)	0 (0)
ISUP grade			0.872
1	9 (5.8)	2 (9.1)
2	125 (81.2)	18 (81.8)
3	15 (9.7)	2 (9.1)
4	5 (3.3)	0 (0)
Clinical stage			>0.999
Ⅰ	122 (79.2)	18 (81.8)
Ⅱ	17 (11.0)	2 (9.1)
Ⅲ	13 (8.5)	2 (9.1)
IV	2 (1.3)	0 (0)