NK cells recognize MHC class I molecules and inhibit the killing of target cells that have self MHC class I molecules through their inhibitory receptors. Therefore inhibitory NK receptor negatively regulate NK cell functions, including target cell lysis, through their binding to MHC class I molecules. The development of leukemia and malignant tumors might be induced as a consequence of the failure of immune surveillance. Leukemic cells and tumor cells can escape from immune recognition by cytotoxic T cells because of the low expression level of tumor antigen and/or HLA class I molecules. In such cases, non-tumor antigen-restricted cytotoxic cells such as NK cells and NK receptor-expressing cells may be important to overcome tolerance in the tumorbearing host. Donor allogeneic NK cells can attack patient's leukemic cell resulting in the enhancement of graft-versus-leukemia（GVL）effect and at the same time attack host antigen-presenting cells resulting in the suppression of graft-versus-host disease（GVHD）in allogeneic stem cell transplantation with Killer cell immunoglobulin-like recepto（r KIR）-ligand mismatch donor. Therefore, NK cell and KIR incompatibility may have an important role in the clinical outcome of immunological cell therapy such as allogeneic stem cell transplantation for not only hematological malignancies but also solid cancers.
Tumor cells release many kinds of immunosuppressive and pro-inflammatory factors that facilitate tumor immune escape and tumor growth. In addition, many kinds of immunosuppressive cells accumulate in the tumor microenvironment. These factors are ultimately responsible for orchestrating immunosuppressive network at the tumor site. CD1d-restricted invariant natural killer T (iNKT) cells play crucial roles in both the innate and acquired immune responses. Activation of iNKT cells leads to downstream activation of other cell types in the immune system, which is crucial to the protective anti-tumor immune responses. In this review, we discuss the contribution of iNKT-dendritic cell (DC) cross-talk governing T helper (Th) balance and the possible mechanisms by which NKT cells regulate the tumor microenvironment to enhance the anti-tumor immune responses.
Colorectal cancer (CRC) is the third most common nd cancers in the world with a very high mortality rate. In many countries like Canada and Japan it is the 2nd after lung cancer. Most CRC are sporadic, and others are familial or genetic. Sequence-time Fishbone of multistage colorectal cancer is associated between macro scopic developing [from adenomatous polyp, severe dysplasia, carcinoma, to metastasis] and up-regulated follow by mutation of several gene expression [APC, K-ras, p53, DCC] has been figured. The dancing of CD4- CD8 occur during the evolution of adenomatous polyp, severe dysplasia to carcinoma. Sequence APC, K-ras,p53, DCC gene mutation in the Fishbone has been transformed to a Firecrackers (up-regulated then down regulated APC gene will up-regulated the next gene K-ras, and p53. When K-ras and 53 is down-regulated DCC gene is up-regulated, when DCC gene is mutated metastasis happens). Non genetic, non gender, famil ial, geographical dependency (sporadic), also in young people, increase 2 to about 200 times in last decade ask many scientist who is the firelighter of the firecrackers gene mutation? The aim of this study was to identi fy the firelighter of the colorectal cancer development and progression and find that U-238 which emits alpha and beta ray during their decay is the agent.
An elevated CEA, confirmed by retesting, needs further evaluation for metastatic disease, but does not justify the institution of adjuvant therapy or systemic therapy for presumed metastatic disease. It is because of CEA has only moderate 70% sensitivity and specificity. Decreasing CD4, CD8 and ratio of CD4/CD8 increase the specificity of this CEA function. This study figured a fishbone and then transformed to firecrackers of APC, K-ras, p53, DCC mutation, microsatellite instability which has commonly occured during the evolution of adenomatous polyp, severe dysplasia to carcinoma. The position of the dancing of CD4 and CD8-PE is put on this firecrackers corridor. Using the dancing of CD4 and CD8-PE is along with T-reg (T4reg and T8reg) in the time sequence firecrackers with upregulated-downregulated gen sequences. Up and down of CD4 and CD8 react in between each gene mutation stage., increase the sensitivity and specificity of CEA (CD66e) for CRC prognosis. Quasi experimental got ten patients pre- and post-operative CRC. Analyzing were done with Fisher exact test for categorical determination. Decreasing CD8+ and CD4/CD8 has increased the sensitivity and specificity of CEA to 100%.
Cancer immunotherapy has been used in clinical trials and shows a powerful life force. But if it is benefit for the treatment of gastric cancer is still not well studied. Present study discussed the antitumor efficiency of individualized immunotherapy induced by dendritic cells transfected with total antigen of autologouse gastric cancer cells and reported the initial results of our pilot clinical trail.
Autologous DCs were isolated from PBMC and proliferated in supplement with GM-CSF and IL-4. Resulting immature DCs were loaded with autologous tumor antigen (tumor lysate or total RNA) and matured in supplement with TNF-α (tu-DC). Autologous T lymphocytes were isolated from PBMC and cocultured with tu-DC to prepare antigen-specific CTLs. Autologous primary cancer cells were obtained from resected specimens of the patients with gastric cancer and cultured as target cells. Phenotypic characters of DCs were detected and evaluated by FCM. The efficiency of cytotoxicity and cytokine release was measured and estimated by using assays of Cytotoxicity and ELISA.
Humanized immune system was reconstructed on 4-week-old nude mice by intraperitoneal (i.p.) injection of human PBMC with cell number of 3×107 each. Prophylactic assay and therapeutic assay were performed by subcutaneously challenge of primary cancer cells after or before autologous DC vaccination, respectively, and the tumor burden were observed.
Nine patients with gastric cancer were subjected to a pilot clinical trail. Two patients with stage Ib-IIIa underwent a radical D2 resection, 5 patients with stage IIIb-IV underwent palliative resection, and 2 patients with stage IV underwent an open-and-close operation. Autologous tumor lysate were prepared from fresh specimens of the patients. DC vaccines and tumor specific CTLs were prepared by using the methods above mentioned. To suppress Treg and to promote calreticulin (CRT) exposure, CTX 1000 mg and ADM 60 mg were intravenously infused a day before immunotherapy. Tumor-specific CTLs were therapied by intravenous infusing with one day interval, and DC vaccines were inoculated to the axillary lymph nodes with two therapies interval. Thymosin alpha 1 (Tα1) was injected once a day subcutaneously during the therapy. The levels of Treg were measured before and after chemotherapy by FCM, and the Incidence of recurrence, Karnofsky performance scale, tumor burden, and side effects were observed and evaluated.
Purpose: Some studies suggest that the Cluster of Differentiation 8 (CD8+ T cells) is down-regulated and this reduction may be involved in immune evasion in cancer patients. The present study was designed to investigate CD8+ T lympho cytes and its relationship to Carcinoembryonic Antigen (CEA) in colorectal cancer patients. CEA is correlated with volume tumor with response therapy anti tumor and associated with remnant tumor after resection (sensitivity and specificity 70-80%). High CEA level in preoperative figured probability of post operative recurrent. CEA is not useful for screening general population or early colon Ca but adjunct toward the diagnostic of patient with Colon Carcinoma (Association between CEA level and the stage of malignancy). CEA should not be used to rule out the presence of a malignancy. The best clinical use is in assessing the prognosis of patients with colorectal carcinoma. Plus the drop of CD8, the sensitivity and specificity of recurrent become forward 100%. Experimental Design: CD8+ T cells was evaluated by multicolor flow cytometry. CD8-PE was count with the background of CD3-FITC and CD45-PerCP. CEA in the sera was quantitated by ELISA. Ca Colorecti were included from Top Referral Hospital of Indonesia: Cipto Mangunkusumo Hospital. CEA> 5 ng/ml as D+ and Ratio CD4/CD8 range 1.5-2.2 and increasing CD8 as TResult: Sensitivity become 80% and specificity 100% Conclusions: CD8-PE has an important role to increase sensitivity and specificity of CEA in colorectal carcinoma prognosis.
Although many genomic/epigenomic alterations involved in pathogenesis of gastrointestinal tumors have been reported, their precise molecular mechanisms remain unclear. Using those alterations as landmarks, genes, which are involved in the cancer development/progression and useful for diagnostic markers as well as therapeutic targets in personalized medicine, can be identified.
We have focused on genomic copy-number aberrations, especially the remarkanle aberrations, i.e., local high-level amplifications and homozygous deletions, and tried to identify novel tumor-related genes as targets for those alterations in various types of neoplasms including gastrointestinal cancers. We applied conventional and in-house bacterial artificial chromosome (BAC)-array-based comparative genomic hybridization (CGH). As amplification targets, many genes were identified, and some of them, such as cIAP1 and ZASC1 identified from esophageal cancer and CCND3 from colon cancer, seem to be important as prognosticators as well as therapeutic targets. From homozygously deleted regions detected in esophageal and gastric cancer, several genes were also identified as tumor suppressors frequently silenced through homozygous loss (genetic) or promoter hypermethylation (epigenetic). In addition, BAC array-based methylated CpG island amplification (BAMCA), a newly developed method to screen abnormally methylated sequences using BAC-array, accelerated the identification of tumor suppressor genes whose expression was altered by DNA methylation.
Genome-wide surveys of genetic and epigenetic abnormalities interpreting them in the context of broader knowledge of cancers facilitates the identification of crucial genes and pathways involved in the gastrointestinal carcinogenesis, resulting in the development of new diagnostic methods and molecular targeting therapies of gastrointestinal cancers.
Cancer stem-like cell subpopulations, referred to as “side-population” SP cells have been identified in several tumors based on their ability to efflux the fluorescent dye Hoechst 33342. While SP cells have been identified in the normal human endometrium, they have not been characterized in endometrial cancer. In this study, we isolated and characterized the SP cells in an endometrial cancer cell line and in rat endometrial cells expressing oncogenic [12Val] human K-Ras protein. These SP cells showed 1) reductions in the expression levels of differentiation markers 2) long-term repopulating properties 3) self-renewal capacity 4) enhancement of migration and podia formation 5) enhancement of tumorigenicity 6) bipotent developmental potential (tumor cells and stroma-like cells). In nude mice, SP cells formed large, invasive tumors, which were composed of both tumor and extracellular matrix enriched stromal-like cells. The expression levels of vimentin,αsmooth muscle actin and collagen III were enhanced in SP-tumors compared with the levels in non-SP-tumors. Analysis of microdissected samples and fluorescence in situ hybridization showed that the stromal -like cells with enriched ECM contained human DNA, confirming that the stromal-like cells were derived from the inoculated cells. In a matrigel assay, SP cells differentiated into _-SMA -expressing cells. These findings demonstrate that SP cells have cancer stem like cell features including the potential to differentiate into the mesenchymal cell lineage.
We have previously reported that acupuncture may regulate the immune system and activate cellular and humoral immunity. In this study, we examined the effects of acupuncture treatment on lymphocyte subsets, intracellular cytokine production, and the catecholamine levels in human peripheral blood in order to elucidate the relationships between the immune system, the autonomic nervous system, and the endocrine system. We sampled peripheral blood from 10 healthy volunteers（age, 20.6 ± 1.6 years）before and 1 day after acupuncture, at the same time on each day, and determined the total and differential leukocyte counts and the levels of 3 catecholamines （adrenaline, noradrenaline, and dopamine）. Further, using flow cytometry, we analyzed the lymphocyte subsets and cells producing the intracellular cytokines interferon（IFN）-γ, interleukin（IL）-4, and IL-1β.
The leukocyte and granulocyte counts significantly decreased after acupuncture（p < 0.05）. The number of CD4＋ cells also significantly decreased（p < 0.01）, whereas that of CD8＋ cells significantly increased（p < 0.01）; the CD56＋/CD3− natural killer（NK）cell and CD11b＋ cell counts also tended to increase. Moreover, the number of IFN-γ-, IL-4-, and IL-1β-producing cells significantly increased（p < 0.01）, and the cell count of IFN-γ-producing cells was the highest. The levels of the 3 catecholamines, in particular adrenaline and noradrenaline, tended to decrease after acupuncture. Thus, acupuncture induced a decrease in the granulocyte counts and adrenaline levels as well as an increase in the number of CD8＋ and IFN-γ-producing cells. These results indicate that the signals produced from acupuncture treatment are transmitted from the peripheral nerve to the central nervous system; this transmission may suppress the secretion of adrenaline via suppression of the sympathetic nerve, and enhance the activity of IFN-γ-producing CD8＋ T cells and NK cells.
It was first pointed out by Inokuchi et al. that ploidy differences in superficially spreading and penetrating gastric cancers may reflect different genetic constitution. Further studies, however, revealed that such a correlation is typically seen only in the undifferentiated-type and that ploidy can alter during tumor progression. DNA ploidy alteration is now considered as an aspect of chromosomal instability. By mapping cytometrically determined DNA ploidy of metaphase cells in individual cancers, we found that (1) DNA-aneuploidy was rare in early cancers and accounted for 70% of advanced cancers, and was almost always accompanied by DNA-diploid cancer cells in the mucosa, suggesting that advanced cancers with aneuploidy may be preceded by DNAdiploid early cancers. (2) The incidence of cycling polyploid cells accompanied in the stemline may be more important than the ploidy mode, itself, for an assessment of chromosomal instability. In DNA-diploid mucosal and extramucosal parts of advanced cancers, the incidences of cycling polyploid cells were as low as that in DNA-diploid early cancers and as high as that in DNA-aneuploid part, respectively. To demonstrate that cycling polypoid cells precede DNA-aneuploidy, we examined the distribution of the cells with duplicated chromosomespecific FISH signals in tissue sections by confocal microscopy. It was found that the cells with duplicated signals were distributed either sporadically or in small clusters. The latter pattern may represent cycling polyploid populations, from which a DNA-aneuploid subclone with growth advantages may arise through further alterations of modal chromosome number. Our studies suggest that the time when DNA-aneuploidy occurs in the tumor and the degree of chromosomal instability (incidence of cycling polyploidy) may be more important clinically than the presence or absence of DNA-aneuploidy.
DNA histograms on log and linear channels were compared for peak height, half width of peak, and distance between G1 and G2/M peaks. Cell cycle analyses of DNA histograms for log and linear channels were also compared. Advantages of log channel DNA histograms are described.
High-sensitivity analysis of erythrocytes using flow cytometry (FCM) is performed for a definitive diagnosis of subclinical paroxysmal nocturnal hemoglobinuria (PNH).
In this study, we conducted line-specific analyses of CD55/59 antigens using FCM by CD235a/CD11b gating of erythrocytes/granulocytes, compared the results between these two cell fractions, and evaluated the causes of differences.
Peripheral blood samples collected in EDTA from 88 subjects were employed. Three, 43, 24, and 18 of them had PNH (including 1 case of PNH/AA), aplastic anemia, myelodysplastic syndrome (MDS), and other disorders, respectively.
Analyses of erythrocytes and granulocytes showed that the rate of concordance was 72%. All subjects with PNH, 47% of AA cases, and 4% of MDS cases were PNH-cell-positive in both erythrocytes and granulocytes. Differences in the results between the two cell fractions were observed in 22 cases (25%), and, in most cases, the PNH cells were positive only in erythrocytes. This discrepancy was noted in 23, 25, and 33% of AA, MDS, and other disorders, respectively, showing similar rates in all disorders. This occurred around the regions of cells showing a low expression of CD235a in scattergrams.
Since the involvement of small-sized erythrocytes with a smaller surface area of the cell membrane was suspected as the cause for the difference, CD55/59 fluorescence intensity depending on the size of erythrocytes was compared.
It was revealed that the expression of CD55/59 decreased with the size of erythrocytes, and that most PNH cells were small erythrocytes.
On the other hand, when only larger-sized erythrocytes were analyzed, the concordance rate with granulocytes rose to 88%.
With high-sensitivity analysis of PNH erythrocytes, small-sized erythrocytes with a low expression of CD55/59 are likely to be recognized as PNH blood cells, which causes false-positive results.