| [Objective]B cells participate in the immune response by secreting antibodies and acting as antigen-presenting cells.It has recently been discovered that a new cell subgroup present in B cells,namely regulatory B cells,secretes IL-10,IL-35 and TGF-beta,exert negative immune regulatory effects and play an important role in infectious diseases,autoimmune diseases and tumors.Fc? receptors are the Fc segment receptors of the immunoglobulin(IgG)constant region.They belong to the immunoglobulin superfamily,mainly including ?,?,and?.There are many types of activating Fc?R,but there is only one type of inhibitory FcyR in humans and mice,namely Fc??Rb,which is widely expressed in B cells and myeloid cells.Various immune cell functions are regulated by it:inhibition of BCR-induced calcium flow,cell proliferation,and antibody secretion.In our previous study,We found that a group of new regulatory B cells secreting IL-10 with the phenotype of CD19hiFcyRIIbhi existed in lung cancer 3LL mice and melanoma B16F10 mice.This group of B cells can be inhibited by their highly secreted IL-10 Activate proliferation of CD4+T cells.In addition,we found that the frequency of CD19+Fc?R?bhiB cells in peripheral blood CD19+B cells of patients with Cutaneum Carcinoma showed no significant difference from that of normal controls.The frequency of CD19+Fc?R?bhiB cells in pulmonary malignant tumor patients is higher than that of normal controls.However,the proportion of CD 19+Fc?R?bhiB cells in peripheral blood PBMC of patients with skin malignant tumors was not significantly different from that of normal controls.Based on the above experiments,this study will investigate whether these CD19+Fc?R?bhiB cells in human peripheral blood can secrete IL-10 at high levels and whether it has negative immune regulation.[Methods]1.A total of 11 peripheral blood samples were collected from patients with skin malignancies diagnosed in Subei People's Hospital from September 2018 to December 2019,including 5 males and 6 females,aged 32-76 years,with a median age of 54 years;There were 7 cases of skin squamous cell carcinoma,2 cases of skin basal cell carcinoma,1 case of Bown's disease,and 1 case of malignant melanoma.A total of 15 peripheral blood samples were collected from patients with pulmonary malignancies diagnosed in Subei People's Hospital,including 9 males and 6 females;aged 56-82 years,median age 69 years;8 cases of lung adenocarcinoma and 5 cases of lung squamous cell carcinoma.There were 2 cases of small cell lung cancer;11 cases of peripheral blood samples were collected from normal subjects,including 6 males and 5 females,aged 22-42 years,with a median age of 32 years.CD19+B cells in peripheral blood PBMCs were sorted by flow cytometry.After cultured for 24 hours in vitro,the secretion level of interleukin 10(IL-10)in the culture supernatant was detected by ELISA.2.Collected peripheral blood samples from 10 patients diagnosed with skin malignancies from March 2019 to October 2019,including 4 males and 6 females,aged 36-78 years,with a median age of 57 years.There were 6 cases of skin squamous cell carcinoma,3 cases of basal cell carcinoma,and 1 case of Brown's disease.A total of 28 blood samples were collected from patients with pulmonary malignancies diagnosed in North Jiangsu People's Hospital,including 22 males and 6 females.The median age was 62.5 years;13 cases of lung adenocarcinoma,10 cases of lung squamous cell carcinoma,and 5 cases of small cell lung cancer;13 cases of peripheral blood samples were collected from normal physical examination,including 8 males and 5 females,aged 18-38 years.The median age was 28 years.In this study,the expression of IL-10 in CD19+Fc?R?bhi and CD19+Fc?R?blowB cells in peripheral blood PBMCs was detected by intracellular staining with flow cytometry.3.Collect 10 peripheral blood specimens from patients diagnosed with lung malignancies from the People's Hospital of Yangzhou University from October 2019 to January 2020,including 6 males and 4 females,aged 42-76 years,with a median age of 59 years;There were 6 cases of lung adenocarcinoma,3 cases of lung squamous cell carcinoma,and 1 case of small cell lung carcinoma.CD19+Fc?R?bhiB cells,CD19+Fc?R?blowB cells,and CD4+T cells were sorted from peripheral blood PBMCs of patients with lung malignancies by flow cytometry.T cells were activated with anti-CD3 and anti-CD28,and then co-cultured with CD19+Fc?R?bhiB cells or CD19+Fc?R?blowB for 7 days.Thenthe proportion of CD25+Foxp3+regulatory T cells(Treg)in CD4+T cells was detected by flow cytometry.[Results]1.IL-10 secretion from CD 19+B cell supernatants in peripheral blood PBMCs from patients with skin malignant tumors or lung malignant tumors is higher than that in the normal control group,and there are significant differences.2.The expressing levels of IL-10 in CD19+Fc?R?bhiB cells of normal people,patients with skin malignant tumors or patients with lung malignant tumors were higher than those of CD19+Fc?R?blowB cells.3.The proportion of Tregs induced by CD19+Fc?R?bhiB cells from patients with lung cancer is higher than that of autologous T cells alone.The ratio of Tregs induced by CD19+Fc?R?bhiB cells is higher than that induced by CD19+Fc?R?blowB cells.[Conclusion]CD19+Fc?R?bhiB cells in human peripheral blood PBMCs can secrete IL-10 at high levels;Compared with CD19+Fc?R?blowB cells,CD19+Fc?R?bhiB cells are more capable of inducing the production of autologous Treg cells. |
PDF Full Text Request