ABSTRACT
Objective The aim of this study was to investigate the effect of flow cytometry on peripheral blood T cell subsets in patients with malignant lymphoma and its relationship with clinicopathological and tumor types.Methods Ninety-eight patients with malignant lymphoma treated in our hospital from August 2014 to September 2016 were selected as the study group.Ninety-eight healthy subjects were selected as the control group.The peripheral blood T cell subsets(CD3 +,CD4 +,CD8 +,CD4 +/CD8 +)were detected in patients and healthy controls by flow cytometry.Results The levels of CD3 +and CD4 +/CD8 +in the study group were (55.63±11.25)and(1.32±0.62),respectively,which were significantly lower than those(68.96±12.63)and (1.59±0.59)of the control group(P<0.05).The levels of CD4 +and CD8 +were(33.67±8.14)and(26.02±4.67),respectively in the study group,were no difference from the control group(34.12±8.33)and(25.67±4.53)(P>0.05).The levels of CD3+and CD4 +/CD8 +in patients with Hodgkin's lymphoma were(54.63±11.36),(1.22±0.65),respectively,and(55.52±12.02),(1.34±0.71)for non-hodgkin lymphoma.They were significantly decreoseg in the control group(68.96±12.63 for CD3 +and 1.59±0.59 for CD4 +/CD8 +) (P<0.05).The level of CD4 +and CD8 +were no difference amoupst Hodgkin's lymphoma(33.78±8.23 for CD4 +and 25.74±4.88 for CD8 +),non-Hodgkin's lymphoma(25.74±4.88 for CD4 +and 33.62±8.74 for CD8 +)and control group(34.12±8.33 for CD4 +and 25.67±4.53 for CD8 +)(P>0.05).The levels of CD3 +,CD4+and CD4 +/CD8 +in patients with Ⅲ ~Ⅳ stage malignant lymphoma were(52.66±12.47), (28.25±6.32)and(1.30±0.62),respectively,which were significantly lower than those(68.96±12.63), (34.12±8.33)and(1.59±0.59)in the control group(P<0.05).The level of CD3 +in patients with phase Ⅰ-Ⅱ malignant lymphoma(58.63±11.85)was significantly lower than that in the control group(68.96±12.63)(P<0.05).The level of CD8 +in patients with phase Ⅰ-Ⅱ malignant lymphoma(29.63±3.57)was significantly higher than that in the control group(25.67±4.53)(P<0.05).Conclusion The detection of peripheral blood T cell subsets by flow cytometry can be used as an important methods to diagnose the disease,staging and immune status of patients with malignant lymphoma,which has high application value.
ABSTRACT
Objective The aim of this study was to investigate the effect of flow cytometry on peripheral blood T cell subsets in patients with malignant lymphoma and its relationship with clinicopathological and tumor types.Methods Ninety-eight patients with malignant lymphoma treated in our hospital from August 2014 to September 2016 were selected as the study group.Ninety-eight healthy subjects were selected as the control group.The peripheral blood T cell subsets(CD3 +,CD4 +,CD8 +,CD4 +/CD8 +)were detected in patients and healthy controls by flow cytometry.Results The levels of CD3 +and CD4 +/CD8 +in the study group were (55.63±11.25)and(1.32±0.62),respectively,which were significantly lower than those(68.96±12.63)and (1.59±0.59)of the control group(P<0.05).The levels of CD4 +and CD8 +were(33.67±8.14)and(26.02±4.67),respectively in the study group,were no difference from the control group(34.12±8.33)and(25.67±4.53)(P>0.05).The levels of CD3+and CD4 +/CD8 +in patients with Hodgkin's lymphoma were(54.63±11.36),(1.22±0.65),respectively,and(55.52±12.02),(1.34±0.71)for non-hodgkin lymphoma.They were significantly decreoseg in the control group(68.96±12.63 for CD3 +and 1.59±0.59 for CD4 +/CD8 +) (P<0.05).The level of CD4 +and CD8 +were no difference amoupst Hodgkin's lymphoma(33.78±8.23 for CD4 +and 25.74±4.88 for CD8 +),non-Hodgkin's lymphoma(25.74±4.88 for CD4 +and 33.62±8.74 for CD8 +)and control group(34.12±8.33 for CD4 +and 25.67±4.53 for CD8 +)(P>0.05).The levels of CD3 +,CD4+and CD4 +/CD8 +in patients with Ⅲ ~Ⅳ stage malignant lymphoma were(52.66±12.47), (28.25±6.32)and(1.30±0.62),respectively,which were significantly lower than those(68.96±12.63), (34.12±8.33)and(1.59±0.59)in the control group(P<0.05).The level of CD3 +in patients with phase Ⅰ-Ⅱ malignant lymphoma(58.63±11.85)was significantly lower than that in the control group(68.96±12.63)(P<0.05).The level of CD8 +in patients with phase Ⅰ-Ⅱ malignant lymphoma(29.63±3.57)was significantly higher than that in the control group(25.67±4.53)(P<0.05).Conclusion The detection of peripheral blood T cell subsets by flow cytometry can be used as an important methods to diagnose the disease,staging and immune status of patients with malignant lymphoma,which has high application value.
ABSTRACT
Objective To express soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal carcinomas in E. coli HB2151 and to purify the soluble ScFv and identify its antigen binding activities to find new target vectors for the diagnosis and therapy of colorectal carcinomas. Methods The phage clones displaying ScFv fragment of the monoclonal antibody MC3 were used to infect E. coli HB2151 to express soluble antibodies. The soluble ScFvs were identified by Dot blot and Western blot and their antigen binding activities were determined by ELISA. The VH and VL DNAs of the ScFv DNA derived were sequenced based on the dideoxy method. Results The soluble MC3 ScFvs were expressed successfully. The expression products with a proximate MW of 32?10 3 were mainly secreted into the periplasm. The soluble ScFv containing periplasmatic extracts derived from three clones could inhibit the binding of MC3 with its antigen, and the inhibition rates were 41.19%, 36.89% and 33.77% respectively. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup and ? type. Conclusion Generation of E. coli HB2151 expressed ScFv of monoclonal antibody MC3 paves the way for further use of the antibody.
ABSTRACT
Objective To clone the cDNA encoding human HMGB1, express it in E. coli, and identify its biological activity. Methods Human HMGB1 cDNA was amplified by RT-PCR and cloned into vector pUC19. After sequence analysis, the cDNA was ligated into prokaryotic expression vector pQE-80L and induced by IPTG to express HMGB1. The protein was purified with Ni~(2+)-NTA chromatography and polymyxin B affinity column. To identify the function of purified protein, the product was co-cultured with THP1 cells. Results Recombinant expression plasmid pQE-80L/HMGB1 was constructed successfully. After purification, the protein purity reached 96%. The recombinant HMGB1 stimulated THP1 to secrete TNF-? . Conclusion The highly purified HMGB1 was obtained successfully, which showed biological activity. These results lay the foundation for further research on the function of human HMGB1.