肝细胞癌相关三级淋巴样结构免疫细胞组成

目的 探讨肝细胞癌(hepatocellular carcinoma,HCC)相关三级淋巴样结构(tertiary lymphoid structure,TLS)的免疫细胞组成.方法 以2016年4月至2019年10月北京大学第一医院收治的112例原发性HCC患者为研究对象,收集其手术切除及HCC穿刺活检样本...

The immune cell components of hepatocellular carcinoma-related tertiary lymphoid structure

Objective To investigate the composition of tertiary lymphoid structure(TLS)of hepatocellular carcinoma(HCC).Methods A total of 112 patients with primary HCC admitted to Peking University First Hospital from April 2016 to October 2019 were selected as subjects.Surgical resection and liver cancer biopsy samples were collected.Tumor specimens of all patients were stained by hematoxylin-eosin(HE).The TLS positive specimens were stained with CD3,CD20,PD1,CD45RO,CD21 and CD68 by immunohistochemistry.IPP(Image Pro Plus)software was used to calculate the area percentage of positive cells in TLS,and the differences among patients with different HCC stages and different sites(intratumoral and paratumoral)were compared.Results There were 21 patients out of 112 HCC patients whose TLS were positive.The ratio of patients with early stage HCC(CNLCⅠ~Ⅱstage)in TLS positive group and TLS negative group were 76.2%(16/21)and 49.5%(45/91),respectively,the difference was statistically significant(χ²=4.919,P=0.027).For HCC patients in early stage with TLS positive,the percentages of CD3⁺[(69.19±16.43)%vs(53.25±14.48)%],CD20⁺[(63.63±17.72)%vs(44.44±16.63)%]and CD21⁺[(68.48±12.78)%vs(11.75±7.20)%]of intratumoral TLS were significantly higher than those of paratumoral TLS,the proportion of PD1⁺[(6.94±3.64)%vs(20.56±8.47)%]was lower than that of paratumoral TLS,the differences were statistically significant(all P<0.05).There were no statistically significant differences in the percentages of CD45RO⁺[(44.56±19.67)%vs(41.00±16.07)%]and CD68⁺[(10.25±6.13)%vs(8.13±5.78)%](all P>0.05).For the 16 patients with early HCC(CNLCⅠ~Ⅱ),compared with paraneoplastic TLS of themselves,the number of patients whose percentages of CD3⁺,CD20⁺and CD21⁺of intratumoral TLS increased were 11 cases,15 cases and 16 cases,respectively,the number of patients whose CD45RO⁺(increased:8 cases,decreased:8 cases)and CD68⁺(increased:7 cases,decreased:9 cases)ratio increased is similar to those CD45RO⁺and CD68⁺ratio decreased.The percentages of CD3⁺[(69.19±16.43)%vs(35.60±15.21)%],CD20⁺[(63.63±17.72)%vs(32.80±13.85)%]and CD21⁺[(68.48±12.78)%vs(10.00±6.71)%]of early stage HCC patients with TLS positive were significantly higher than those of advanced HCC patients with TLS positive(all P<0.05).There were no significant differences in the percentages of PD1⁺[(6.94±3.64)%vs(9.60±3.78)%],CD45RO⁺[(44.56±19.67)%vs(48.20±13.94)%]and CD68⁺[(10.25±6.13)%vs(7.40±3.65)%](all P>0.05).The percentages of CD3⁺[(53.25±14.48)%vs(19.80±5.07)%]and CD20⁺[(44.44±14.63)%vs(21.60±5.46)%]in paratumoral TLS of HCC patients in early stage with TLS positive were significantly higher than those of advanced HCC patients with TLS positive(all P<0.05).There were no significant differences in the percentages of CD21⁺[(11.75±7.20)%vs(11.80±3.90)%],PD1⁺[(20.56±8.47)%vs(22.00±9.49)%],CD45RO⁺[(41.00±16.07)%vs(38.60±9.99)%]and CD68⁺[(8.13±5.78)%vs(6.80±4.15)%](all P>0.05).Conclusions The proportion of T cells,B cells and dendritic cells of intratumoral TLS of patients with early stage HCC were higher than those of paratumoral TLS,and also higher than those of intratumoral TLS of patients with advanced HCC,and the proportion of depletion lymphocytes decreased,while the proportion of memory lymphocytes and macrophages didn’t change.It is suggested that TLS in HCC patients in early stage contains more activated lymphocytes and dendritic cells,which may be the reason for its anti-tumor immune response,delaying the progression of the disease and prolonging the survival of patients.