Zahra Faghih; Saeideh Sadat Shobeiri; Ali Ariafar; Mohsen Sarkarian; Shahryar Zeighami; Nazanin Nazari; Saeed Abbasi-Sarvak; Nasrollah Erfani
Volume 13, Issue 4 , December 2016, , Pages 237-248
Abstract
Background: Cytotoxic CD8+ T cells, as essential parts of the adaptive immune
system, play pivotal roles in anti-tumor immune responses. It is well documented that
cytokine expression profiles and activation status of these cells during anti-tumor
immune responses affect the outcome of host-tumor ...
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Background: Cytotoxic CD8+ T cells, as essential parts of the adaptive immune
system, play pivotal roles in anti-tumor immune responses. It is well documented that
cytokine expression profiles and activation status of these cells during anti-tumor
immune responses affect the outcome of host-tumor interaction. Objective: To
investigate the percentages of CD8+ lymphocytes and their subsets in tumor draining
lymph nodes of patients with bladder cancer. Methods: Forty-five patients with bladder
cancer, candidate for radical cystectomy, were recruited. Mononuclear cells were
isolated from draining lymph nodes using Ficoll-Hypaque gradient centrifugation, and
were activated by PMA/Ionomycin in the presence of Golgi inhibitors. The cells were
then permeabilized and stained with appropriate flourochrome conjugated antibodies
against CD3, CD8, IFN-γ, IL-17 and IL-4 molecules. Data were collected on a fourcolor
flow cytometer and analyzed by CellQuestPro software. Results: Despite no
difference in the frequency of IL-17 producing CD8+ (Tc17) lymphocytes, the mean
expression of IL-17 in this subset was significantly elevated in high-grade patients
(p=0.011). The percentage of double positive IFN-γ/IL-17 CD8+ lymphocytes was also
significantly increased in node positive patients compared to node negative ones
(p=0.046). Our results also demonstrated that the percentage of IFN-γ producing CD8+
(Tc1) lymphocytes was significantly increased in the patients with higher histological
grade compared to those with lower ones (p=0.038). Conclusion: IFN-γ and IL-17
producing CD8+ T cells may increase in advanced stages of bladder cancer, but their
correlation with tumor prognosis remains to be investigated.
Nazanin Nazari; Shirin Farjadian
Volume 13, Issue 3 , September 2016, , Pages 178-185
Abstract
Background: HLA-G is a nonclassical HLA class I molecule which, when elevated in tumor cells, is one of the main factors involved in tumor evasion of immune responses including NK and T cells. Objective: To evaluate the effect of HLA-G downregulation on NK cell cytotoxicity in tumor cell lines. Methods: ...
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Background: HLA-G is a nonclassical HLA class I molecule which, when elevated in tumor cells, is one of the main factors involved in tumor evasion of immune responses including NK and T cells. Objective: To evaluate the effect of HLA-G downregulation on NK cell cytotoxicity in tumor cell lines. Methods: The expression level of HLA-G was measured by real-time PCR and flowcytometry after transfection of SKOV3 by shRNA.1, which targets specific sequences in exon 4, or shRNA.2, which targets both exons 4 and 6. NK-92MI cell cytotoxicity against transfected or untransfected target cell lines was measured with the lactate dehydrogenase (LDH) release assay. The Jeg-3 cell line was used as a positive control. Results: Membrane-bound HLA-G expression levels decreased significantly in both cell lines after transfection with both shRNAs compared to their corresponding untransfected cells (p<0.05). Jeg-3 cells were better lysed than SKOV3 cells by NK cells during the first 48 h after transfection with both shRNAs. Compared to untransfected cells, shRNA.1-transfected SKOV3 cells were significantly more lysed by NK cells 24 h post-transfection (p=0.043). Conclusion: As a clinical approach, HLA-G downregulation with shRNA may be effective in cancer therapy by improving immune cell activation.