OBJECTIVETo investigate the effect of FCGR3A polymorphisms on the antibody-dependent cell-mediated cytotoxicity (ADCC) activity induced by cetuximab against A549 cells.

METHODSA549 cell line was used as target cells and NKTm cells as effector cells. FCGR3A polymorphisms were detected by direct sequencing. The ADCC activity mediated by cetuximab was assessed by CCK-8 assay.

RESULTSThree genotypes of FCGR3A were detected:V/V,V/F,and F/F. The ADCC activity of NKTm cells with these three different genotypes mediated by cetuximab were significantly different (P=0.0015). NKTm cells with FCGR3A-158V/V genotypes had significantly higher ADCC activity than FCGR3A-V/F or F/F genotypes (P<0.01),whereas the ADCC activity between V/F and F/F genotype showed no statistical significance(P>0.05).

CONCLUSIONFCGR3A polymorphisms have an impact on ADCC activity mediated by cetuximab in NKTm cells.

PURPOSE: During differentiation of HL-60 cells by all-trans retinoic acid(ATRA), we analyzed the expression of Fcr receptors and Mac-1 by molecules of equivalent soluble fluorochromes(MESF) and functional studies. METHODS: HL-60 cells were induced to differentiate by adding 1micrometer ATRA. On the 4th and 7th day after stimulation as well as before stimulation, the cells were analyzed for phenotypic and functional differentiation. Phenotypic analysis was performed by flow cytometry after staining the cells with PE-conjugated anti-human CD64(FcrRI), CD32(FcrRII), CD16(FcrRIII), CD11b, CD18. The measured fluorescent intensity was transformed into MESF. Phagocytic activity was measured by flow cytometry after incubation of the cells with fluorochrome-conjugated beads. Respiratory burst was measured by chemiluminescence assay. ADCC was measured by hemoglobin release assay. Opsonophagocytic activity was measured by fungicidal assay. Correlation between MESF of FcrR and Mac-1 and function of HL-60 was measured. RESULTS: Percent positive cells and MESF of CD11b and FcrRI increased on the 4th day and decreased on the 7th day. Percent positive cells of CD18 was 99% regardless of differentiation. But MESF of CD18 was increased on the 4th day and decreased on the 7th day. Percent positive cells of FcrRII were above 90% regardless of differentiation. MESF of FcrRII showed no significant change. FcrRIII expression was not induced. Phagocytic activity of HL-60 cells was increased twofold. Chemiluminescence of HL-60 cells was increased up to 60-fold on the 7th day. ADCC of HL-60 cells was incerased up to 2.5-fold on the 7th day. Opsonophagocytic activity increased twice on the 4th day. ADCC and opsonophagocytic activity correlates with the expression of CD11b/CD18 and FcrRII. CONCLUSION: Differentiation of HL-60 cells with ATRA induces several functional maturations until 7 days with expression of FcrR and Mac-1.
Antibody-Dependent Cell Cytotoxicity ; Flow Cytometry ; HL-60 Cells* ; Humans ; Luminescence ; Respiratory Burst ; Tretinoin*

BACKGROUND: Zinc which is widely used to treat Behcet's disease, is known to be an important modulator in various aspects of immunity including cell mediated immunity (CMI). CMI is suspected of playing a major role in the pathogenesis of Behçet's disease. OBJECTIVE: This study was done to clarify the relationship of CMI and zinc in Behçet's disease. METHODS: Serum zinc level, NK cell activity, and ADCC were measured in 83 patients with Behçet's diseade. The results were analyzed using multiple regression analysis. RESULTS: ADCC and serum zinc level were found to be two significant variables that affect NK cell activity positively and negatively, respectively. CONCLUSION: Serum zinc is presumed to exert inhibitory effect on NK cell activity but does not affect ADCC in Behçet's disease patients.
Antibody-Dependent Cell Cytotoxicity ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; Zinc*

In recent years, as increasing of monoclonal antibody application in clinic, the antitumor effect of antibody dependent cell-mediated cytotoxicity (ADCC) get increasing attention. The natural killer (NK) cells are the most important effector cells mediating specific antitumor of ADCC; the phagocytes, T-cells and granulocytes have the definite effect on antitumor of ADCC. ADCC is confirmed as the important mechanism and means for clinically treating the cancers with monoclonal antibodies. The IgG antibody firstly combines with target cells (tumor cells) through antigen-binding sites, and then FcγR on effector cells identifies its Fc fragment and mediates ADCC. Today many kinds of monoclonal antibodies have been put into clinical application such as rituximab and other new anti-CD20 monoclonal antibodies including trastuzumab, erbitux, cetuximab, edrecolomab, nimotuzumab, gemtuzumab ozogamicin and so on, which all can mediate ADCC. The antitumor effects of ADCC mediated by monoclonal antibody can be influenced by IgG Fc receptor gene polymorphism, tumor cell antigen, serum antibody levels, cytokines and drugs etc. As to peripheral blood mononuclear cells, ADCC efficacies of FcγRIIIa-158V/V and FcγRIIa-131H/H are higher than that of other genotypes, while increasing the level of tumor antigen and decreasing the level of serum antibody or adding some cytokines (IL-2, IL-21, IL-15, etc) may elevate the ADCC effect mediated by monoclonal antibodies. Avoiding use of certain drugs (dexamethasone, TNF antagonist) or appropriately using of ondansetron and clemastine also can enhance the anti-tumor effect of ADCC mediated by monoclonal antibodies. In short, ADCC is very important in clinical application for anti-tumor treatment, but its efficacy may be impacted by multiple factors.In this article, the killing mechanisms of ADCC, the clinical use of monoclonal antibodies with antitumor effect of ADCC, the factors influencing anti-tumor efficacy of ADCC, and the antitumor effects of ADCC by other cells are reviewed.
Antibodies, Monoclonal ; therapeutic use ; Antibody-Dependent Cell Cytotoxicity ; Humans ; Immunotherapy ; Killer Cells, Natural ; Neoplasms ; therapy

OBJECTIVE: To investigate the injury effect of anti-donor specific antibody (DSA) on human umbilical vein enolothelial cells (HUVEC) in NK-mediated antibody-dependent cell cytotoxity (ADCC). METHODS: The peripheral blood of 10 healthy donors was colleced for allo-HSCT of AML patients diagnosed in Department of Hemology of the Tumor Hospital affiliated to Shanxi Medical University, then the peripheral blood NK cells were isolated and used as the effector cells; the HUVEC of passages 9-6 were selected and co-cultured with DSA, then the DSA-binding HUVEC were used as the target cells (CDH group), while the DSA-unbinding HUVEC were used as negative control (UDH group). After co-culture of effecor cells with target cells, the expression of IFN-γ was detected by flow cytometry and the HUVEC activity was detected by using MTT method, so as to indirectily reflect the injury effect of DSA-mediated ADCC on endothelial cells. RESULTS: With the increase of effector-target (E:T) ratio, the activity of HUVEC decreased, the expression level of IFN-γ increased. Under the some effector-target ratio (1∶1, 10∶1, 20∶1), the activity of HUVEC in CDH group was significantly lower than that of UDH group, and the expression of IFN-γ was significantly higher than that of the UDH group (P<0.05). CONCLUSION DSA can damage vascular endothelial cells through the ADCC effect mediated by NK cells.
Antibodies, Monoclonal ; Antibody-Dependent Cell Cytotoxicity ; Endothelial Cells ; Flow Cytometry ; Humans ; Killer Cells, Natural

The effects of chimeric monoclonal antibodies (cMAbs), prostaglandin E, (PGE,), and indomethacin (INDO) on antibody-dependent cellular cytotoxicity (ADCC) against human squamous cell carcinoma of head and neck (SCCHN) cell line were examined. Using the PCI-50 SCCHN cell line as target and normal human peripheral blood mononuclear cells as effector, ADCC was enhanced by the treatment of cMAbs (1.25 p,g/ml), but was inhibited by exogenous PGE (5 X 10' M). The effects of cMAb and PGE were dose-dependent. Maximal suppression of activity occured when PGE was present during the entire 4-hr 'Cr-release assay period, whereas pretreatment of effector cells with PGE had minimal inhibitory effect after washing. These results indicate that decreased ADCC seen with SCCHN targets treated with PGE is related to post-binding events, such as binding of effector and target cells. Pre-treatment of effector cells with INDO (1 ug/ml) resulted in restoration of NK activity which was inhibited by PGE. Our in vitro results suggest that INDO can increase tumor cell killing by the reversal of the suppression for many imrnune functions by PGE.
Antibodies, Monoclonal ; Antibody-Dependent Cell Cytotoxicity ; Carcinoma, Squamous Cell* ; Cell Line ; Head* ; Homicide ; Humans* ; Indomethacin ; Neck* ; Prostaglandins E

Autoimmune postpartum thyroiditis (PPT) has been thought of as one of the organ-specific autoimmune diseases. The present study was designed to investigate whether the immunological changes during the postpartum period might induce this disease, by comparing the circulating lymphocyte subsets and antibody-dependent cell-mediated cytotoxicity (ADCC) between normal postpartum women and PPT patients. The results were as follows: 1) No significant differences in the circulating total T lymphocyte population, or suppressor T lymphocyte subsets, or in Th/Ts ratio were found among 25 PPT patients, 11 normal postpartum women and 11 normal non-pregnant women. 2) In PPT patients, helper T lymphocyte subsets were fewer in proportion than those of normal postpartum or non-pregnant women. However, B lymphocyte population (19.7 +/- 7.8%) and ADCC activity (.41 +/- 13) in PPT patients were comparable to those in normal postpartum women (18.3 +/- 4.8%, .42 +/- .11), although they were significantly greater than those in normal normal non-pregnant women (13.3 +/- 5.9%, .29 +/- .07). In conclusion, the enhancement of immune activities observed in PPT patients was comparable to that in normal postpartum women, suggesting that some other causative or triggering factors might be responsible for the occurrence of this disease.
Adult ; Antibody-Dependent Cell Cytotoxicity ; B-Lymphocytes/immunology ; Female ; Human ; Pregnancy ; Puerperal Disorders/immunology* ; T-Lymphocytes/immunology ; Thyroiditis, Autoimmune/immunology*

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.
Antibodies, Monoclonal, Murine-Derived ; pharmacology ; Antibody-Dependent Cell Cytotoxicity ; Antigens, CD20 ; immunology ; Genes, Reporter ; Humans ; Reproducibility of Results ; Rituximab

The NK activity and ADCC of peripheral blood mononuclear cell were examined to evaluate the contribution of ADCC and NK activity to host immune response against lung cancer. The NK activity and ADCC were examined in 58 patients with primary lung cancer and 40 healthy volunteers as normal controls. The NK activity of patients with lung cancer was significantly subnormal, but ADCC was at a normal level. The NK activity was decreased in non-small cell lung cancer (NSCLC), but not in small cell lung cancer (SCLC) compared to normal controls. According to stage, the NK activity in stage II, III-M0 and III-M1 NSCLC showed low levels compared to that of stage I NSCLC, but there was no difference of NK activity in patients with SCLC. The NK activity was not affected by performance status. There was no significant difference of ADCC in patients with lung cancer according to cell type, stage and performance compared with that of normal controls. The NK activity and ADCC were not changed after chemotherapy and operation respectively.
*Antibody-Dependent Cell Cytotoxicity ; Human ; Killer Cells, Natural/*immunology ; Lung Neoplasms/*immunology ; Neoplasm Staging ; Support, Non-U.S. Gov't

PURPOSE: The Mesima-Ex is a kind of biologic response modifier, which is extracted from a mushroom called Phellinus linteus. Mesima-Ex consists of various chemical compounds which include protein bound polysaccharide, mucoprotein, triterpenoid, and quinones. Mesima-Ex exerts its antitumor effects by augmenting host immune response without any toxic side effects. In vitro study, Mesima-Ex seems to potentiates antibody dependent cell mediated cytotoxicity (ADCC) and cell mediated cytotoxicity (CMI) against tumor cells. We initiated this study to verify antitumor effects of Mesima-Ex as an antineoplastic agent. MATERIALS AND METHOD: Gastric cancer patients who underwent curative resection with normal hepatic and renal function were eligible. They were divided into two groups by random number table. One group (N=30: Mesima-Ex group) received postoperative adjuvant chemotherapy with 5-FU (500 mg/m2 weekly), adriamycin (40 mg/m2 every 3 weeks) and Mesima-Ex (6 cap daily per Os). Another group (N=37: control group) received 5-FU and adriamycin only without Mesima-Ex. NK (natural killer cell) activity, ADCC (antibody dependent cell mediated cytotoxicity), CD4 , and CD8 cells were measured and an analysis of disease free survival rate of the two study groups was performed. RESULTS: Sixty seven patients were enrolled in this study. Their median age was 55 years old. NK activity (basal activity: 25%) was enhanced significantly at the 2nd, and 4th months in the Mesima-Ex group (28.9%, 43.4%, p<0.05). ADCC was also enhanced from 37% to 42.1% at the 2nd month in the Mesima-Ex group (p<0.05). The control group did not show any significant change in NK activity or ADCC. The CD4 cell ratio was increased from 37% to 42.1% at the 2nd months in the Mesima-Ex group but not in the control group (p<0.05). There was no significant change in CD8 subsets (p>0.05). There were no toxic side effects more than grade III from Mesima-Ex administration. The two year disease free survival rate was higher in the Mesima-Ex group than that of the control group (77% vs 58%, p<0.05). CONCLUSION: Mesima-Ex can be used safely as an immunomodulator with standard chemotherapeutic agents for purpose of adjuvant chemotherapy. Mesima-Ex was effective in augmenting host immune response in vitro. The Mesima-Ex group showed a higher two year disease free survival rate than that of the control group.
Agaricales ; Antibody-Dependent Cell Cytotoxicity ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Doxorubicin ; Fluorouracil ; Humans ; Middle Aged ; Quinones ; Stomach Neoplasms*