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Objective To investigate the expressions of CD28 and CD117 in patients with newly diagnosed multiple myeloma (MM) and their clinical significances. Methods The clinical data of 115 newly diagnosed MM patients in the First Affiliated Hospital of Zhengzhou University from May 2015 to December 2017 were retrospectively analyzed. The expressions of CD28 and CD117 were detected by using multiparameter flow cytometry. The relationship between the expressions of CD28 and CD117 and MM staging and clinical parameters was analyzed. The staging was performed according to the International Staging System (ISS). Results Among these 115 patients, there were 15 patients with CD117 positive and 30 patients with CD28 positive. Erythrocyte sedimentation rate (r ＝ -0.481, P ＝ 0.039), Cˉreactive protein level (r ＝ -0.314, P＝0.015), the proportion of plasma cells detected by bone marrow cytology (r＝-0.027, P＝0.001) were negatively correlated with CD117 positive expressions. CD28 positive expression was positively correlated with lactate dehydrogenase level (r ＝ 0.249, P ＝ 0.033) and ISS stage (r ＝ 0.319, P ＝ 0.017), while it was negatively correlated with hemoglobin level (r ＝ -0.372, P ＝ 0.026). CD28 positive was associated with light chain type, and nonˉsecretory type mostly occurred (P ＝ 0.016). The incidence of osteolytic lesions in CD28 positive group and CD117 positive group was high, but there was no statistical difference between CD28 positive group, CD117 positive group and CD28 negative group, CD117 negative group (P ＝ 0.052, P＝0.479). Conclusions The positive expression of CD117 in the early stage of MM patients is higher than that in the advanced stage, and the expression of CD28 positive in the advanced stage of MM patients is higher than that in the early stage. CD28 and CD117 can be used as indicators of prognosis stratification in the patients with newly diagnosed MM.
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Objective To explore the expression and clinical significance of CD 8 +CD28 -T lymphocytes in the patients with endometriosis ( EMT) complicating with pelvic fluid .Methods Eighty patients with endometriosis complicating with pelvic fluid ( ca-ses of ⅠtoⅣwere 15, 12, 22, and 31, respectively) were enrolled.The peripheral blood as well as the pelvic fluid were collected and flow cytometry was applied to detect the frequencies of CD 8 +CD28 -T lymphocytes and their intracellular cytokines , including transforming growth factor β1 (TGF-β1) and interleukin 10 (IL-10).Twenty health women with the same range of age were enrolled as the control group .Results Compared with the control ones , the frequencies of peripheral blood CD 8 +CD28 -T lymphocytes , TGF-β1, and IL-10 of the EMT subjects were increased significantly (all P Ⅲ>Ⅱ>Ⅰand the difference between every two of the Ⅰ~Ⅳgroups was significant (all P Ⅲ>Ⅱ>Ⅰ.Moreover, the difference of pelvic fluid TGF-β1 between any two groups was more apparent compared with CD 8 +CD28 -T lymphocytes and IL-10 ( P <0.05 ) .Spearman correlation analysis showed that any one of CD8 +CD28 -T lymphocytes, TGF-β1, and IL-10 positively correlated with any stage of ⅠtoⅣ( rs =0.791, 0.753,0.726 and all P <0.05).Conclusions CD8 +CD28 -T lymphocytes, TGF-β1, and IL-10 were closely related to the stages and could be the negative roles in the pathogenesis of EMT .TGF-β1 played a vital role in the formation of pelvic fluid .
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Objective To investigate the significance of changes in expression of co-stimulatory molecules on T lymphocytes in patients with chronic hepatitis B (CHB) infection.Methods In a casecontrol study,a total of 82 CHB cases including 50 male cases and 32 female cases (the mean age was 42.32 ±3.74) were enrolled in the First Affiliated Hospital to Soochow University from October 2012 to November 2013,together with 30 cases health control (15 male cases and 15 female cases,and the mean age was 42.32 ± 3.74).Patients were divided into three groups:40 cases of non-treated,30 cases of effective-treated and 12 cases of ineffective-treated with anti-viral drugs and immune-related therapy.The expression levels of CD28,CTLA-4,PD-1,Tim-3 on T cells subset from peripheral blood were determined by Flow Cytometry.Serum load of HBV DNA was detected by Real-time PCR and the serology markers such as HBeAg and ALT were detected by conventional methods The Kruskal-Wallis test was used to analysis groups comparison.Independent samples t-test and Mann-Whitney U test were used to two sample comparison.Spearman's rank correlation test was used to analyze the correlation.Results The expression levels of CD28,CTLA-4 and PD-1 on CD4 in health control group was the highest compared to ineffectivetreated group [404.65 (331.65-536.09) vs 277.15 (249.90-344.25) (H=29.81,P＜0.001);32.89 (29.69-39.69) vs 19.26 (11.90-20.56) (H =43.13,P ＜0.001),respectively].The expression level of PD-1 on CD8 in pre-treated group was the highest,while expression level in health control group was the lowest [15.47 (12.50-17.78) vs 3.05 (1.41-3.97) (H=56.60,P＜0.001)].Similarly,the expression level of Tim-3 on CD4 in ineffective-treated group was the highest,while Tim expression on CD4 in health control group was the lowest [199.62 (55.61-239.45) vs 70.62 (53.88-112.32) (H =41.03,P ＜ 0.001)].The expression level of Tim-3 on CD8 in pre-treated group was the highest,while it was the lowest in health control group [82.50 (78.69-84.58) vs 3.07 (1.56-6.87) (H=74.84,P ＜0.001)].Compared to the group with low viral load,the expression level of PD-1 on CD8 both in the pre-treated group with high viral load and the post-treated group was significantly increased [17.87 (13.38-20.94) (U=25.00,P＜0.001); 16.95 (5.39-18.27) (U=63.50,P＜0.001),respectively].Importantly,in the post-treated group,the PD-1 expression on CD4 T (r =0.689,P ＜0.001) and on CD8 T (r =0.751,P ＜ 0.001) was also positively correlated with ALT.Conclusions The abnormal expression of these co-stimulatory molecules may be present in the antiviral treatment process of CHB.It may provide new clues for the reasonable treatment of CHB.
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ObjectiveTo investigate the expression and significance of CD28- cells, CD4+ and CD8+T lymphocytes in the peripheral blood and synovial fluid in patients with rheumatoid arthritis ( RA ). Methods The expression of CD28, CD4+, CD8+ T lymphocytes and inducible co-stimulator(ICOS) in the peripheral blood and synovial fluid in 45 patients with RA were detected by three-color flow cytometry. Independent sample's t test was used for statistical analysis between the two groups. ResultsSynovial fluid CD4+CD28+ICOS+, CD4+CD28- ICOS+ , CD8 + CD28 + , CD8 + CD28 + 1COS+ T lymphocytes were significantly increased than the peripheral blood in RA patients[(36±19)％ vs (15±8)％, t=-4.234, P＜0.01; (2.1±2.2)％ vs (0.6±1.4)％, t=-3.143, P＜0.01; (62±15)％ vs (47±18)％, t=-2.885, P＜0.01; (9±9)％ vs (3±3)％,t=-2.131, P＜0.05], Synovial fluid CD8+CD28-T lymphoc-ytes were significantly reduced than the peripheral blood[(38±15)％ vs (54±18)％, t=2.975, P＜0.01], Synovial fluid CD8+ CD28-ICOS+, CD4+CD28+and CD4+ CD28- T lymphocytes had no significant difference than the peripheral blood (P＞0.05). Compared with peripheral blood in the same patients with RA, CD4+CD28+ ICOS+, CD8+ CD28+ T lymphocyteswere significantly increased[(38±18)％ vs (16±10)％, t=-4.065, P＜0.01 ; (61±16)％ vs (41±21)％, t=-4.065,P＜0.01], CD8+CD28-T was significantly reduced[(39±16)％ vs (59±21)％, t=2.949, P＜0.01]. The level of CD4+ CD28-, CD8+ CD28-, CD28-ICOS+ T lymphocytes in the active and remission patients with RA was not significantly different (P＞0.05). ConclusionSynovial fluid CD28T lymphocyte subsets disturbance and the abnormal expression of ICOS in patients with RA may play important roles in the mechanism of joint damage.
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Objective To study the expression of CD27 and CD28 in antigen-specific CD4~+T cells in patients with pulmonary tuberculosis and healthy people, and understand the role of differentiated stages of CD4~+T cells in the pathogenesis of tuberculosis. Methods The expression of CD27 and CD28 was analyzed by CD4, CD154, CD27 and CD28 staining and flow cytometry. The distributions of CD27 and CD28 in antigen-specific CD4~+T cells were compared between patients with pulmonary tuberculosis and healthy controls. Results In patients of pulmonary tuberculosis, the frequencies of CD27 + CD28 + (early differentiated stage), CD27~- CD28~+ and CD27~+ CD28~- (intermediate differentiated stage), CD27~- CD28~-(fully differentiated stage) T cell subsets in antigen specific CD4~+T cells were (49. 55 ±6. 15)%, (26. 85 ±3. 87)% ,(7. 2 ± 1.37)% and ( 16. 35 ±3.97)%, respectively. In healthy controls, the frequencies of the four subsets in antigen-specific CD4~+T cells were ( 51.81 ± 4. 94 ) %, ( 29. 83 ± 5.33 ) %, ( 12. 65 ±4. 48)% and (5.7±2)%, respectively. The early differentiated CD4~+T cell was the major subset both in patients and healthy people, however, which had significant difference compared with the fully differentiated subset ( t = 2. 26, P < 0. 05 ). Conclusion The population frequency of the fully differentiated CD4~+T cells in patients with pulmonary tuberculosis was significantly higher than that in healthy people. This suggested that the differentiation degree of the antigen-specific CD4~+T cell might be related with pulmonary tuberculosis.
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Objective To explore the expression of membrane form of CD28 (mCD28) on T lymphoeytes and the serum level of soluble CD28 (sCD28) in elderly patients with primary non-small cell lung cancer (NSCLC) in order to investigate the relationship between age-related changes of CD28 and the development of NSCLC in elderly patients. Methods 63 elderly patients with NSCLC, 35 elderly patients with lung benign lesion, 30 elderly healthy donors, 30 young healthy donors, 20 young patients with lung benign lesion and 20 young patients with NSCLC were enrolled in this study. The mCD28 on T cells and the serum level of sCD28 were measured by four-color flow eytometric assay and enzyme linked immunosorbent respectively, and the relationship between CD28 and clinical characteristics of NSCLC was analysed Results The expression of mCD28 was decreased and the serum level of sCD28 was increased in elderly patients with NSCLC compared with the other groups (F= 184.25, P<0. 01 ; F= 365.40, P<0.01). The expression of mCD28 was significantly lower and the level of sCD28 was significantly higher in elderly healthy donors than those in young healthy donors and young patients with lung benign lesion (P<0. 05). There were no significantly statistical differences in expression of mCD28 and level of sCD28 between elderly healthy donors and elderly patients with lung benign lesion [(42.84±5.82)% vs. (46.09±-7.34)%, (39.38±6.02)μg/L vs. (35.84±5.02)μg/L, P>0. 05]. Logistic regression analysis showed that aging (OR=2. 432), down-regulation of mCD28 expression (OR=0. 876) and up-regulation of sCD28 level (OR= 1. 113) were the risk factors for lung cancer. In the elderly patients with NSCLC, there were significant differences in mCD28 expression and sCD28 level between stages Ⅲ-Ⅳ and stages Ⅰ-Ⅱ [(16. 51± 5.64)% vs. (24.41±8.24)%, (75.03±5.98) μg/L vs. (66.73±7.52)μg/L; t=4.497,4.794, both P <0. 01]. However, there were no significantly statistical differences among different pathological types (F=0. 609, 0. 302, both P > 0. 05). Conclusions The down-regulation of mCD28 expression and up-regulation of sCD28 level with advancing age play an important role in the oncogenesis and development of primary non-small cell lung cancer in the elderly patients.
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Objective To express human inducible eostimulator (ICOS) extracellular region and IgG Fc fusion protein, and analyze their function in allogenie lymphocyte proliferation in vitro and in vivo. Methods Human ICOS extraeellular region and IgG Fc fragment were cloned into a soluble expression vector. ICOS-Ig fusion protein was expressed and purified in CHO cells. To monitor primary MLR, Balb/c spleen T cells were isolated as responder cells, and irradiated C57BL/6 spleen cells as stimulator cells. 50 μg/ml ICOS-Ig or IgG was added to primary MLR cultures. The cells responsive rates were detected by 3 H-TdR methods. ELISA tested supernatants for eytokines (IL-2,IL-4, IL-10 and IFN-γ). T cells of each group in primary MLR were cultured as responder cells for secondary MLR, and irradiated C57BL/6 (donor) or C3H (third party) spleen cells as stimulator cells. Similar indexes were detected in secondary MLR. Then vital dye CFSE was used to study alloreactive T cell proliferation in vivo. CFSE-labeled C57BL/6 spleen cells were transferred to irradiated Balb/c mice. Mice were then intraperitoneally injected with 0. 2 mg IgG, ICOS-Ig or CsA each day.At the 3rd day after transfection, the spleen cells of the mice were harvested to detect CD4+ CFSE+ and CD8+ CFSE+ by FACS. Results In primary MLR, ICOS-Ig inhibited allogenic T-cell proliferation with inhibition rate being (58 ± 8)% in 50 μg/ml, and increased IFN-γ secretion. In secondary MLR, ICOS-Ig specifically inhibited the proliferation of donor spleen cells with inhibition rate being (42±8)%, and in ICOS-Ig group the levels of IL-4 and IL-10 were lower and the level of IFN-γ higher than in IgG group. However, ICOS-Ig didn't inhibit the proliferation of third-party spleen cells. In the CFSE dye assay, CFSE intensity of CD4+ and CD8+ T cells in ICOS-Ig and CsA groups was stronger than that in control group (P < 0. 05), while CFSE intensity in combined treatment group were even stronger than that in ICOS-Ig and CsA groups (P<0. 05). Conclusion ICOS-Ig could inhibit allo-reactive T cell proliferation in vitro and in vivo, and induce donor-specific T cell hyporesponsiveness specifically.
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Objective To investigate.the co-stimulatory pathway of gastric cancer patients by examining the expression of co-stimulatory molecules CD28,CTLA.4(CDl 52)and B7 molecules on peripheral blood lymphocytes(PBLC)in patients with gastric cancer,and to discuss the role of costimulatory pathway in the pathogenesis of gastric cancer.Methods The expression of CD28,CD152,CD3,CD4,CD8 and B7 molecules on peripheral blood lymphocytes and T cell subpopulation were measured by flow cytometry in 56 patients with gastric cancer,and the data was compared and analyzed with that in gastric benign tumor group and healthy control group.Results The expression levels of CD3+,CD4+,CD3+ CD28+,B7-1(CD80),B7-2(CD86)and CD41/CD8+on PBLC in patients with gastric cancer was significantly lower than that of benign stomach tumor group and healthy control group.The expression levels of CD3+ CDl52+ and CD8+ on PBLC in patients with gastric cancer were significantly higher than those of gastric benign tumor group and healthy control group.Conclusion Peripheral lymphocytcs in gastric cancer patients express low level of CD28 and B7 molecules and high level of CDI 52 molecules,which results in dysfunction of B7:CD28/CTLA-4 co-stimulation pathway and in turn affects the capacity of T cell to eliminate tumor cells,which makes tumor cells evading the immune surveillance and metastasizing.
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Objective To explore the possible role of CD28 +/CD152 +:B7 eostimulators in immune pathophysiology of severe pneumonia.Methods 22 severe pneumonia peripheral blood sample were used to analyze the expression of CD3+ T cell CD28+,CD152+,CD14++ on mononuelear cell CD86+,and HLA - DR + by FACS expression.The relationship between CD28+,CTLA4,CD86+ and the HLA-DR +,and the relationship between APACHE Ⅱ Grading,CD28+,CD152+,CD86+ and HLA-DR + were analyzed.Results Compared with the control group,the expression of CD3 + T cell,CD86+ and HLA - DR + were remarkably reduced while the expression of CD28+ and CD152+ were markedly increased in patients with severe pneumonia who were hospitalized in 24 h(P＜0.05).However,T cells with positive CD8+ CD3+ and CD4+ CD3+ had no significant change between two groups(P＞0.05).For patients with severe pneumonia who survived,the APACHE Ⅱ scores were significantly reduced while the expression of CD28+,CD152+,CD86+,HLA-DR + and CD3+ + cells were significantly increased after 10 days from admission(P＜0.05).By contrast,T cells with positive CD8+ CD3+ and CD4+CD3+ had no significant change between two groups(P＞0.05).There were no relation between costimulators CD28+ and HLA - DR + (r=-0.12,P=0.54)and APACHE Ⅱ scores(r=-0.30,P=0.19) in control group.CD86+ and HLA - DR + showed positive correlation(r=0.65,P=0.00).CD86+ and APACHE Ⅱ scores had no correlation(r=-0.38,P=0.09).Conclusion The costimulators expressed abnormally in circumference blood of patients with severe pneumonia,CD86+ decreased,but CD28+,CD152+ increased.T cell of circumference blood was at the condition of "anergy".The increase of CD28+,CD86+,CD86+ and HLA - DR + during convalescence stages in patient with severe pneumonia showed that spocific immunity was advantageous for restoration in these patients.The relationship among CD86+,CTLA4 and HLA - DR + indicated that CD28+/CD152+:B7 play an role in the occurrence and development of severe pneumonia.
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Objective To explored the role of CD40/CD40L and B7-1/CD28 interaction in the immunopathologic process of folic acid-induced nephropathy(FAN) and investigate the intervention effect of the anti-B7-1 monoclone antibody(B7-1mAb). Methods GD1 mice were administrated by i. p with FA only or co-treated with B7-1 mAb for the intervention study. Kidneys were harvested for immunohistochemical, Western blot assays and serum for renal function evaluation at day 1, 3,5, 7, 14 and 21. Results Continuous expression of CD40, B7-1 from day 1 to day 21 on the renal tubular epithelial cells was detected in this model. CD40 was up-regulated more than five times in the kidney at every test time point (P
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Objective To investigate the role of costimulatory molecules CD28/B7 in the pathogenesis of psoriasis. Methods The expression of CD28, CD80 and CD86 was detected in the lesional skin of 22 cases by immunohistochemical technique (ABC), and in the peripheral blood lymphocytes of 17 cases by flow cytometry respectively. As control, normal skin and lymphocytes from peripheral blood of healthy volunteers were also tested. Results The immunohistochemistry study showed that the expression of CD28, CD80 and CD86 in the psoriatic lesions was significantly higher than that in the normal controls (P .05). Conclusion CD28, CD80 and CD86 might play a certain role in the pathogenesis in psoriasis.
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Objective To investigate the significance of B7-CD28/CTLA-4 (CD152),co-stimulatory molecules of T lymphocytes,in the onset,development and prognosis of genital congdyloma acuminatum (CA) in females.Methods Flow cytometry was utilized to detect the expression levels of CD80/CD86 in peripheral blood lymphocytes and of CD28/CD152 (CTLA-4) in CD4~+/CD8~+ T lymphocytes from 30 CA patients (17 primary CA,13 recurrent CA,15 at recovery stage of CA) and 15 healthy volunteers as con- trols.Results No significant difference was found for the frequencies of CD80~+,CD86~+,CD4~+/CD28~+ and CD8~+/CD28~+ lymphocytes between the primary or recurrent group and the control group.The frequencies of CD8~+/CD28~+,CD4~+/CD28~+ and CD80~+ lymphocytes were significantly higher in the recovery group than those in the recurrent group (P0.05).Conclusions There is an abnormal expression of co-stimulatory molecules B7-CD28/CTLA-4 (CD152) in periphoral blood lym- phocytes,CD4~+ and CD8~+ T lymphocytes in female patients with genital CA,and the expression abnormaility is closely linked with different disease stages of CA.
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Objective To investigate the significance of measurement of the expression CD28 on tumor infiltrating lymphocytes in patients with stomach cancer.Methods Subjects for this investigation include the tumor tissues and its nearby normal tissues from 48 patients with stomach cancer.The levels of the expression of CD28, CD8 on tumor infiltrating lymphocytes were analyzed by flow cytometry.Results First, the levels of the expression of CD8+CD28+ and CD8-CD28+ on tumor infiltrating lymphocytes from the tumor tissues were remarkably lower than from its nearby normal tissues (P
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Objective To investigate the expression of inducible co-stimulator (ICOS) on T lymphocytes in peripheral blood from the patients with systemic lupus erythematosus (SLE). Methods Two-color immunofluorescent staining and flow cytometric assay were used to analyze the level of ICOS expression on T lymphocytes in peripheral blood from 33 SLE patients and 16 healthy volunteers. The data of systemic lupus erythematosus diseases activity index (SLEDAI) in SLE patients were also collected. Results The level of ICOS expression on T cells from active SLE group was markedly higher than that in the control group and the inactive SLE group (P 0.05). However, a positive correlation between the expression level of ICOS and the SLEDAI score was found in both active SLE group and inactive SLE group(r = 0.711,P = 0.001?r = 0.561,P = 0.03). Conclusions The expression of ICOS on T cell in peripheral blood from active SLE is increased, which might play a role in the pathogenesis of SLE.
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AIM: The expression of CD28, CD56 and CD57 on CD8+T cells in the peripheral blood of young (age range 20-35) and elderly(age range 60-75) healthy donors were compared to explore the change of the cellular immune function with aging.METHODS: Three-color fluorescent flow cytometry was performed to analyze the differences in percentage of CD8+CD28+, CD8+CD56+ and CD8+CD57+T cells in the peripheral blood between the young and elderly groups.RESULTS: CD8+CD28+T cells in the peripheral blood of the elderly group was significantly lower than those in the young group, with percentage of 34.07?5.28 and 49.84?7.43,respectively (P