The Study Of The Relation Between The Phenotypes Of Macrophages In Milky Spots In Human Greater Omentum And Gastric Carcinomas With Different Clinical Stages

Objective:To explore the relation between the phenotypes of macrophages in milky spots in the greater omentum and gastric carcinomas with different stages.Methods:Specimens of human great omentum of gastric carcinomas with different stages(50cases of30in stage T1-3and20in stage T4)and normal people(10cases)were collected in Qingdao municipal hospital from March2011to December2011.Using a modified three-step incubation method and Ficoll density gradient centrifugation to isolate and purify macrophages.The cells were identified by morphological observation and immunofluorescence staining.Flow Cytometry(FCM)was used to determine the expression of CD68+/CD206-and CD206+macrophages.Enzyme linked immunosorbent assay(ELISA)was used to determine the secretion of IL-10and IL-12in different time points.And the relation between the phenotypes of macrophages in milky spots in the greater omentum and the pathological features of gastric carcinomas was analyzed.Results:The obtained cells were and were confirmed to be CD68+macrophages with a positive rate of69.9%by immunofluorescence staining.By FCM the number of CD68+/CD206-Mφ in milky spots in greater omentum in normal controls,T1-3group and T4group were(8.2959±0.0546)×105,(7.4535±0.0546)×105,(5.9144±0.2958)×105,respectively.The number of CD68+/CD206-Mφ in normal controls and T1-3group was larger than that in in T4group(F=3418.713、6305.821,P<0.01).There were significant difference among the there groups(χ2=49.186,P<0.01).The number of CD206+M2in normal controls,T1-3group and T4group were(0.1695±0.0242)×105,(1.0068±0.1213)×105,(5.9144±0.2958)×105,respectively.There were significant difference among the there groups(x2=49.194,P<0.01).The number of CD206+M2in T4group was larger than that in normal controls and T1-3group(Z=-4.399s-5.942,P<0.01).The number of CD68+/CD206’Mcp was negative correlated with that of CD206+M2and T stages of gastric carcinomas(r=-0.586^-0.832,P<0.01).But The number of CD206+M2was positive correlated with T stages of gastric carcinomas(r=0.826,P<0.01).By ELISA The average secretion of IL-10in Normal controls and T1-3group were lower than T4group(P<0.05).The average secretion of IL-12in T4group were lower than Normal controls and T1-3group(P0.05).The secretion of IL-12was negative correlated with that of IL-10and T stages of gastric carcinomas(r=-0.194^-0.476,P<0.01).But the secretion of IL-10was positive correlated with T stages of gastric carcinomas(r=0.552,P<0.01).There was a close correlation between the expression of CD68+/CD206"and CD206+M2and whether invading serosa,lymphnode metastasis and clinical staging in patients of gastric carcinomas(P<0.05).Conclusion:(1)The influencing factor of the phenotypes of macrophages in milky spots in the greater omentum include whether invading serosa and lymphnode metastasis;(2)Milky spots express small amount of M2type macrophages with a low expression of CD206but high secretion of IL-12of patient with gastric carcinomas in stage T1-3and are still in immune defenses period;(3)When tumor invades adjacent structures in the T4stages,Milky spots mainly express M2type macrophages with a high expression of CD206and high secretion of IL-10.