Comparison of osteogenic and chondrogenic differentiation ability of buccal fat pad derived mesenchymal stem cells and gingival derived cells

Statement of the Problem: One major goal of tissue engineering and regenerative medicine is to find an appropriate source of mesenchymal stem cells [MSCs] with higher differentiation ability

Purpose: In this experimental study, the osteogenic and chondrogenic differentiation ability of buccal fat pad derived MSCs [BFP-MSCs] with gingival derived cells [GDCs] were compared

Materials and Method: BFP-MSCs and GDCs were cultured enzymatically and expanded. The expanded cells were analyzed for membrane-associated markers, using flow cytometry. Then the ability of these cells to differentiate into osteocyte and chondrocyte was assessed morphologically and by mRNA expression of collagen I [COLL], BGLA and bone morphogenetic protein 2 [BMP2] using qRT-PCR

Results: Flow cytometry analysis showed that both BFP-MSCs and GDCs expressed the characteristic stem cell markers such as CD73, CD44, and CD90, whereas they did not express hematopoietic markers. Mineralized calcium deposition was observed apparently in BFP-MSCs cultured in osteogenic medium but GDCs showed fewer mineralized nodules. The mRNA expression levels of BGLA and BMP2 showed 7×105 and 733-fold more mRNA expression in BFP-MSCs treated with differentiation media compared to the control group. In chondrogenic differentiation, BFP-MSCs transformed from a spindle to a cuboidal shape while GDCs showed only a slight transformation. In addition, mRNA expression of COLL showed 282-fold higher expression in BFP-MSCs in comparison to the control group. Such significant difference in mRNA expression of BGLA, BMP2, and COLL was not observed in GDCs compared to their corresponding controls

Conclusion: Based on the present results, BFP yields a greater proportion of stem cells compared to gingiva. Therefore, this tissue can be introduced as an easily available source for the treatment of periodontal defects and other maxillofacial injuries

Comparative proteomic analysis of tumor mesenchymal-like stem cells derived from high grade versus low grade gliomas

Objective: Gliomas are the most common primary brain tumors, and have been ranked as the fourth leading cause of cancer death. Tumor mesenchymal-like stem cells [tMSCs] contribute to the aggressive behavior of glial tumors by providing a favorable microenvironment for the malignant cells. The aim of our study was to identify differential proteome of tMSCs derived from low vs. high grade glioma tumors

Materials and Methods: Patients with newly diagnosed low and high grade gliomas were included in this case control study. tMSCs were isolated from tumors using enzymatic digestion validated by flow cytometer analysis after sub-culturing. Differential proteomic analysis of tMSCs derived from low and high grade tumors was performed by two-dimensional gel electrophoresis and mass spectrometry. Protein spots with more than two fold differences and P values below 0.05 were considered as differentially expressed ones

Results: In tMSCs isolated from low and high grade gliomas, different isoforms of mesenchymal- related proteins vimentin and transgelin were differentially expressed. Overexpressed proteins in tMSCs isolated from low grade gliomas were mitochondrial manganese-containing superoxide dismutase [Mn-SOD], 40S ribosomal protein SA, and GTP-binding nuclear protein, while in tMSCs isolated from high grade gliomas, cathepsin B, endoplasmin, ezrin, peroxiredoxin1, and pyruvate kinase [PK] were found to be significantly overexpressed

Conclusion: For the first time, we analyzed the differential proteomic profiles of tMSCs isolated from glioma tumors with different grades. It was found that molecules related to mesenchymal cells [vimentin and transglin], in addition to those related to tumor aggressiveness with potential secretory behavior [e.g. cathepsin B] were among differentially expressed proteins

Adipose derived stem cells exert immunomodulatory effects on natural killer cells in breast cancer

Objective: Adipose derived stem cells [ASCs], as one of the important stromal cells in the tumor microenvironment, are determined with immunomodulatory effects. The principle aim of this study was to evaluate the immunosuppressive effects of ASCs on natural killer [NK] cells

Materials and Methods: In this experimental study, we assessed the expressions of indolamine 2, 3-dioxygenase [IDO1], IDO2 and human leukocyte antigen-G5 [HLA-G5] in ASCs isolated from breast cancer patients with different stages as well as normal individuals, using quantitative reverse transcriptase-polymerase chain reaction [qRT-PCR]. Immunomodulatory effects of ASCs on the expression of CD16, CD56, CD69, NKG2D, NKp30, NKG2A and NKp44 was also assessed in peripheral blood lymphocytes [PBLs] by flow-cytometry

Results: Our result showed that IDO1, IDO2 and HLA-G5 had higher mRNA expressions in ASCs isolated from breast cancer patients than those from normal individuals [P>0.05]. mRNA expression of these molecules were higher in ASCs isolated from breast cancer patients with stage III tumors than those with stage II. The indirect culture of ASCs isolated from breast cancer patients and normal individuals with activated PBLs significantly reduced NKG2D+ and CD69+ NK cells [P<0.05]

Conclusion: Results of the present study suggest more evidences for the immunosuppression of ASCs on NK cells, providing conditions in favor of tumor immune evasion

Construction of CTLA-4-Ig fusion gene in pBudCE4.1 expression vector

CTLA-4 inhibitory signals prevent cell cycle progression and IL-2 production, leading to a halt on an ongoing immune response. CTLA4-Ig fusion proteins con-tain the extracellular domain of CTLA-4 and Fc fragment of human IgG antibody. In this study we aimed to fuse the ctla-4 gene encoding the extracellular domain of CTLA-4 molecule with igg1 gene encoding Fc region of human IgG. After primer design, PCR reaction was performed using pfu polymerase en-zyme and specific primers. PCR amplified fragment was ligated into the vector con-taining the human igg1 gene. The resulting fusion fragment of ctla-4 and human igg1 genes was ligated to pBudCE4.1 expression vector. Extracellular domain of ctla-4 gene was ligated to igg1 gene and then ctla4-ig fragment was cloned into pBudCE4.1 vector. Construction of the expression vector was confirmed by restriction pattern analysis and sequencing. By confirming the construct, in the next step, the recombinant DNA will be used to produce CTLA4-Ig recombinant protein for the clinical uses

Downregulation of MMP2 and Bcl-2 in adipose derived stem cells [ASCs] following transfection with IP-10 gene

Mesenchymal Stem Cells [MSCs] are recently introduced as novel immunological gene carriers for treatment of cancer. It is believed that balance between the expression of angiogenic and anti-angiogenic factors, such as SDF-1 and IP-10, may regulate neovascularization within the tumor. In this study, we compared the expression of important tumor promoting mediators in IP-10-transfected Adipose Derived Stem Cells [ASCs] to those transfected with SDF-1. ASCs were isolated from adipose tissue of a normal subject undergoing cosmetic mamoplasty surgery using collagenase. ASCs were transfected with IP-10 or SDF-1 propagated plasmids by electroporation method and Lipofectamin 2000. Expressions of SDF-1, CXCR4, IP-10, Bcl-2, MMP2, IL-10, IGF-1, and VEGF were detected in transfected ASCs using quantitative Real-Time Polymerase Chain Reaction [qRT-PCR]. Results showed that the expressions of SDF-1, CXCR4, Bcl-2, MMP2, IL-10, IGF-1, and VEGF were upregulated in SDF-1-transfected ASCs. In contrast, Bcl-2 and MMP2 transcripts showed 45×103 and 10 fold lower expression in ASCs transfected with IP-10 compared to non-transfected cells. Anti-angiogenic chemokines such as IP-10 may modulate tumor promoting properties of ASCs and would be introduced as novel candidates for tumor immunotherapy; however, further studies are needed to be conducted

Association between angiotensinogen M235T polymorphism and preeclampsia in Iranian pregnant women

To determine the possible association between the M235T variant of angiotensinogen gene and preeclampsia in Iranian preeclamtic women with hypertension during pregnancy. During a case control study, we used polymerase chain reaction-based restriction fragment length polymorphism [PCR-RFLP] analysis to investigate the association between M235T polymorphism in preeclamtic women compared to normotensive controls. The M235T polymorphism was significantly associated with increased preeclampsia risk in the studied population as supported by a p value of 0.017 and chi-square value of 8.12. The frequency of mutated allele and genotype distribution showed a significant difference between preeclamtic women and control groups. The result indicates that the AGT M235T polymorphism plays a significant role in preeclampsia observed in selected Iranian preeclamtic women, and it can be considered as a major risk factor for preeclampsia

IL-23/IL-27 ratio in peripheral blood of patients with breast cancer

Background: Interleukin [IL]-23 and IL-27 are two IL-12-related cytokines which their function may dramatically influence the inflammatory response to tumor development. IL-12 and IL-27 seem to have antagonistic roles with IL-23 in tumor site. In this study, IL-23 and IL-27 mRNA expressions were analyzed in peripheral blood of patients with breast cancer and healthy volunteers using quantitative real-time PCR

Methods: Peripheral blood samples were collected from 50 women with breast cancer and 50 healthy ones. The total RNA was extracted from peripheral blood after lysis with ammonium chloride and TRizol reagent and the cDNA was synthesized. The expression of IL-23 and IL-27 gene transcripts was determined with real-time polymerase chain reaction [qRT-PCR] using Syber Green PCR Master Mix

Results: It is found that IL-23 and IL-27 transcripts had significantly higher expression in peripheral blood of patients compared with the healthy controls. The ratio of IL-23 transcript expression to IL-27 was 3.4 fold lower in the studied patients compared with the normal individuals

Conclusion: It is concluded that the over expression of IL-23 and IL-27 gene transcript in peripheral blood of breast cancer patients may be an immune response against tumor development and the inflammatory response plays a critical role in tumor development via up regulating the corresponding cytokines. However, the IL-23/IL-27 ratio may play an important role in cytokine-based immunotherapy against cancer. Further research should be carried out to assess these cytokines in a larger sample size

IL-17 and IL-4 producing CD8+ T cells in tumor draining lymph nodes of breast cancer patients: positive association with tumor progression

CD8+ cytotoxic T lymphocytes have been recently divided based on their cytokine expression profile. To evaluate the percentages of CD8+lymphocytes and their effector subsets including Tc1, Tc2 and Tc17 in the tumor draining lymph nodes [TDLNs] of patients with breast cancer. Single cell suspensions were obtained from TDLNs of 42 patients with breast cancer. Staining of the cell surface markers and intracellular cytokines was performed using appropriate fluorochrome-conjugated antibodies. The data was acquired on a four-color flow cytometer and was analyzed by CellQuestPro software package. The percentages of different CD8+ cell subtypes [Tc1, Tc2 and Tc17] were quantified in CD8+T lymphocytes. The comparison was made between LN+ versus LN- patients, as well as patients in different clinico-pathological status. The percentage of Tc1, Tc2 and Tc17 subsets were not significantly different between LN+ and LN- patients. Despite no difference in the percentages of Tc1 cells in LN+ patients with infiltrative ductal carcinoma [IDC], the mean expression of IFN-gamma by Tc1 cells decreased significantly in comparison to LN- patients. On the other hand, the percentages of Tc2 and Tc17 effector subsets were increased in advanced stages [p=0.018 and p=0.009, respectively]. As the first study to investigate various effector subtypes of CD8+ lymphocytes in TDLNs of patients with breast cancer, our data collectively suggests a positive association between IL-17- and IL-4-producing CD8+ T cell percentages [Tc2 and Tc17] in TDLNs with breast cancer progression. Although the number of Tc1 cells seems not to be affected by cancer progression, down-regulation of IFN-gamma by these cells seems to be associated with tumor metastasis to TDLNs. These findings may have implications in cancer immunotherapy based on CD8+ effector subsets.

Polymorphism in exon 2 of CD1 genes in southwest of Iran

The CD1 family is less variable transmembrane antigen presenting molecules related to the MHC molecules. CD1a and CD1e genes are the most polymorphic ones associated with autoimmune diseases. The aim was to better clarify the map of CD1 genes in Southwest Iranian normal population for implications in vaccine design. In this study we investigated the polymorphism of CD1a, CD1d and CD1e in 311 healthy individuals from Fars Province in Southwest of Iran by PCR-SSP method. Six of individuals had homozygote CD1a 01/01 genotype and 248 had homozygote CD1a 02/02 genotype. CD1d was found to be monomorphic with all tested individuals showing CD1d 01/01 genotype. Hundred and eleven individuals had homozygote CD1e 01/01 genotype and 48 had homozygote CD1e 02/02 genotype. The frequencies of CD1a 01 and CD1a 02 alleles were 11% and 89% while the frequencies of CD1e 01 and CD1e 02 alleles were 60.1% and 39.9%, respectively. Consistent with previous reports on other genes, a high degree of similarity in CD1a and CD1e allelic distribution was observed between Southwest Iranians and other Indo-European populations. However, the allelic frequency of the CD1a and CD1e alleles showed a significant difference from those of Chinese Han and She populations. These data are notable in the light of relatively recent genetic admixture along the Silk Road. Considering the significance of CD1 alleles in some autoimmune and infectious diseases and with the admixed nature of Iranian population, mapping the distribution of CD1e alleles in different regions of Iran can be useful in future designing of preventive and therapeutic vaccines

Non-viral transfection methods optimized for gene delivery to a lung cancer cell line

Mehr-80 is a newly established adherent human large cell lung cancer cell line that has not been transfected until now. This study aims to define the optimal transfection conditions and effects of some critical elements for enhancing gene delivery to this cell line by utilizing different non-viral transfection Procedures. In the current study, calcium phosphate [CaP], DEAE-dextran, superfect, electroporation and lipofection transfection methods were used to optimize delivery of a plasmid construct that expressed Green Fluorescent Protein [GFP]. Transgene expression was detected by fluorescent microscopy and flow cytometry. Toxicities of the methods were estimated by trypan blue staining. In order to evaluate the density of the transfected gene, we used a plasmid construct that expressed the Stromal cell-Derived Factor-1 [SDF-1] gene and measured its expression by real-time PCR. Mean levels of GFP-expressing cells 48 hr after transfection were 8.4% [CaP], 8.2% [DEAE-dextran], 4.9% [superfect], 34.1% [electroporation], and 40.1% [lipofection]. Lipofection had the highest intense SDF-1 expression of the analyzed methods. This study has shown that the lipofection and electroporation methods were more efficient at gene delivery to Mehr-80 cells. The quantity of DNA per transfection, reagent concentration, and incubation time were identified as essential factors for successful transfection in all of the studied methods

Strong association of ctla-4 variation [ct60a/g] and ctla-4 haplotypes with predisposition of Iranians to head and neck cancer

Variations in Cytotoxic T Lymphocyte Antigen-4 [CTLA-4] affect the expression and function of this protein. We aimed to investigate the association of +49 A/G [rs231775], +1822 C/T [rs231779] and +6230 A/G [CT60, rs3087243] genetic variations, as well as the merged haplotypes in CTLA-4 gene with susceptibility to, or progression of head and neck cancer. Eighty patients with confirmed head and neck [HN] cancer [age 54.9 +/- 16.1 years] and 85 healthy age/sexmatched controls [age 56.3 +/- 12.4 years] were enrolled in the study. Genotypes were investigated by the PCR-RFLP method. Arlequin software package was used to check for Hardy-Weinberg equilibration, and to estimate the haplotypes. At position +6230 A/G [CT60], AA genotype, as well as A allele was significantly decreased in patients with HN cancers than controls [18.8% vs. 40.7%, p=0.004; odds ratio=0.34, and 46.3% vs. 61.7, p=0.007; odds ratio=0.53%, respectively]. Nearly the same results were obtained when we compared the subgroup of patients with squamous cell carcinoma of the HN [SCC-HN] with control subjects. The frequencies of genotypes and alleles at other positions were not significantly different between patients and controls, however ACG, GTA and GCA haplotypes emerged from three investigated loci occurred with significantly more frequencies in patients [p<0.0001], while ACA and GTG haplotypes were more frequent among controls [p<0.0001]. Significant differences of haplotypes, genotypes and alleles frequencies resisted the Bonferroni correction. Our results suggest that CT60 A allele, as well as ACA and GTG haplotypes in CTLA-4 gene may have protective roles against HN cancer in Iranian population, while ACG, GTA and specially GCA haplotypes may render susceptibility.

CCL22 16C/A genetic variation is not associated with breast carcinoma in southern Iranian population

CCL22/MDC is a CC chemokine with a critical role in regulation of the immune balance in physiological condition. CCL22/CCR-4 ligation has been documented to participate in the migration of regulatory T [Treg] cells and Th2 lymphocytes to the site of breast tumors; circumstances that are known to be associated with poor prognosis. To investigate the association of a single nucleotide polymorphism [SNP] in CCL22 gene; 16C/A [rs4359426; Asp2Ala], with susceptibility to breast cancer in a sample of Iranian population. Methods: 161 patients with pathologically confirmed breast carcinoma [mean age 49.3 +/- 11.5 yrs] and 178 agematched healthy women [mean age: 49.3 +/- 12.9 yrs] were studied. CCL22 genotypes were investigated by the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism [PCR-RFLP] method. Data was verified by direct automated sequencing. Arlequin analysis showed no deviation from Hardy-Weinberg equilibrium. The most frequent genotype in both patient and control groups was wild type CC genotype with frequency of 146 out of 161 [90.7%] among patients and 153 out of 178 [86.0%] in control group [p=0.24]. The frequency of CA genotype was 15 [9.3%] and 23 [12.9%] in patients and controls, respectively [p=0.38]. No AA genotype was observed among patients but this genotype was observed with the frequency of 2 out of 178 [1.1%] in control subjects. The minor allele frequency [MAF] was 0.07 in the population. No correlation was found between the investigated genotypes and clinicopathological characteristics of the patients. Conclusively, results of this investigation do not support the association of 16C/A SNP [rs4359426; Asp2Ala] in CCL22 gene with susceptibility to, and progression of, breast cancer in Iranian population

Lack of association of mouse mammary tumor virus-like sequences in Iranian breast cancer patients

The present study aimed to detect mouse mammary tumor virus [MMTV]-like sequences in Iranian breast cancer patients in the city of Shiraz, located in southwest Iran. We searched for two MMTV genetic regions in the peripheral blood leukocytes of 300 women with breast cancer, 300 age-matched healthy control subjects, and 50 breast tumor tissues. Two regions of MMTV, 660 bp and 250 bp, were searched by nested polymerase chain reactions. None of the above two regions were detected. There were no differences between the control group and the breast cancer group. Our findings did not show any association of MMTV-like sequences with breast cancer development in Iranian patients in Shiraz

Prognostic significance of MMP2 and MMP9 functional promoter single nucleotide polymorphisms in head and neck squamous cell carcinoma

Matrix metalloproteinases comprise a family of enzyme that is able to degrade components of extra cellular matrix. There are single nucleotide polymorphisms in the promoter regions of several genes with ability to influence cancer susceptibility. The aim of this study was to analyses association between MMP2 and MMP9 promoter polymorphisms and head and neck squamous cell carcinoma occurrence and progression. A case- control study was performed including 80 head and neck squamous cell carcinoma patients and healthy controls for MMP2 and 86 head and neck squamous cell carcinoma patients and 72 healthy controls for MMAP9. Blood samples were genotyped for MMP2 and MMP9 using polymerization chain reaction-restriction fragment length polymorphism method [PCR-RFLP]. Statistical analysis was performed using SPSS 12.0 software. Our results showed that distribution of MMP2 genotype between controls and patients was significantly different [Chi[2] = 10.3, P= 0.005]. Comparison between CC genotype in HNSCC patients and controls showed that C allele modified the risk of HNSCC progression [OR= 2.6, 95% CI, 1.0046-6.729]. The MMP9 genotype distribution among HNSCC patients was significantly different [Chi[2] = 14.56, P= 0.0007]. The frequency of TT genotype in HNSCC patients was different from healthy controls and was more common genotype in HNSCC cases [OR= 2.18, 95% CI, 0.7052-6.7854]. Our results suggested an association of the MMP2 and MMP9SNP with the development of HNSCC. Also, our results showed that MMP, MMP9 genotypes and smoking were related to HNSCC progression

CTLA4 gene variants in autoimmunity and cancer: a comparative review

Gene association studies are less appealing in cancer compared to autoimmune diseases. Complexity, heterogeneity, variation in histological types, age at onset, short survival, and acute versus chronic conditions are cancer related factors which are different from an organ specific autoimmune disease, such as Grave's disease, on which a large body of multicentre data is accumulated. For years the focus of attention was on diversity and polymorphism of major histocompatibility complex in respect to human diseases specially the autoimmune diseases, but in recent years, access to other human gene sequences prompted investigators to focus on genes encoding the immune regulatory proteins such as the co-stimulatory, adhesion molecules, cytokines and chemokines and their receptors. Among them, CTLA4 [CD152] has been in the centre of attention for its pivotal role in autoimmunity and cancer. Although not fully understood, CTLA4 with no doubt plays an important role in the maintenance of the immune response by its expression on activated and regulatory T cells. CTLA4 [Gene ID:1493, MIM number:123890] has many variants and polymorphic forms, some present in regulatory positions, some in 3' UTR and the most important one in the leader sequence [+49 A/G]. As a pivotal regulatory element of the immune responses magnitude, CTLA4 could be considered as a two-blade knife, for which only the optimal expression ensures an effective, but at the same time, safe immune response. It can accordingly be speculated that CTLA4 alleles associated with extraordinary expression could make a person more susceptible to tumor growth and/or progression. On the other hand, alleles associated with a compromised CTLA4 expression/function may accelerate the formation and/or manifestation of inflammatory autoimmune disorder. I hypothesized a spectrum of the functional dichotomy of CTLA4 SNPs diverging from autoimmunity to cancer. To examine these hypotheses, results from previously published investigations on CTLA4 polymorphisms together with the work done by our own group are discussed in details. Because the most published data are about the polymorphism at position +49, I concentrated on this position; however the data regarding other SNPs are also included for comparison. To support the significance of CTLA4 gene variation in these two major human diseases evidences from organ transplantation are also included. As will be discussed in the manuscript, our work and reports by others from a normal population perspective support the hypothesis that individuals inheriting a GG genotype at position +49, for which lower CTLA4 expression has been extensively suggested, are more susceptible for developing autoimmune disorders and those with AA genotype, with an existence of a state of self-tolerance, may have a higher chance of developing cancer. CTLA4 SNPs may accordingly be considered as a crucial element, along with other known or yet unknown mechanisms, in keeping the immune balance in predisposed individuals to cancer and autoimmunity. Although an spectrum line can be drawn between autoimmunity and cancer by considering published data regarding CTLA4 +49 polymorphism, the extreme functional dichotomy of this SNP appears to be more complex and difficult to understand, but there is no doubt that the future investigations will resolve most ambiguities

Serum HER2 level in epithelial ovarian cancer

The proto-oncogene HER2 plays a key role in the control of cellular proliferation. Its overexpression has been reported to be associated with a poor prognosis in cancer, particularly in breast cancer. In the present study, serum HER2 levels were investigated in patients diagnosed with epithelial ovarian cancer. Serum HER2 levels were detected by an ELISA commercial kit in 51 patients and 33 healthy individuals. The mean serum HER2 level was found to be significantly higher in patients than healthy controls [P=0.005]. In 29% of patients, serum HER2 levels were higher than the cut-off value. HER2 serum level was not associated with tumor stage at diagnosis. Elevation of HER2 in a high proportion of patients with epithelial ovarian cancer further strengthens the importance of this molecule in the pathogenesis of ovarian cancer

CTLA-4 Exon one +49 A/G gene variants in patients with superficial and invasive bladder cancer: a study in southern Iran

Cytotoxic T-cell lymphocyte antigen 4 [CTLA-4] is a member of the superfamily of immunoglobulins that are mainly expressed by activated T cells. It is established that blockade of CTLA-4 receptors leads to the enhancement of an immune response. Different polymorphisms of the CTLA-4 gene have been described which cause increased susceptibility to various malignancies such as lymphoma or multiple myeloma. Considering that bladder cancer is one of the most prevalent cancers worldwide, we have evaluated the role of CTLA-4 gene polymorphism at position +49 A/G in the formation or progression of bladder cancer in southern Iran. A total of 226 individuals between February 2005 and June 2006 were included and placed into two subgroups: patients diagnosed with bladder cancer and a control group. Demographic data and risk factors were collected from both groups. The DNA of all subjects was extracted from their blood samples. Different genotypes of the CTLA-4 gene were determined using the restriction fragment length polymorphism [RFLP] technique and data were compared in both groups by using Pearson's chi-square test. The prevalence of AA, AG and GG genotypes at position 49, according to the PCR-RFLP method, were 57.5%, 37.2% and 5.3% in the control group, respectively. In the patient group, the prevalence of these genotypes was: AA in 57.5%, AG in 32.7% and GG in 9.8%. Statistical analysis of data showed no significant difference in both groups [P value=0.40]. Also there was no correlation between different genotypes of the CTLA-4 gene and invasiveness of the disease in our cases. Although polymorphism of the CTLA-4 gene at position 49 of exon-1 increases susceptibility to several malignancies, our study showed no relationship between polymorphism at this position and genetic susceptibility to the development of bladder cancer, nor was there any association with disease progression

Mesenchymal stem cells do not suppress lymphoblastic leukemic cell line proliferation

Several studies have demonstrated the immunosuppresive effects of mesenchymal stem cells [MSCs] in allogeneic or mitogenic interactions. Cell-cell contact inhibition and secretion of suppressive soluble factors have been suggested in this regard. To investigate if adipose derived MSCs could inhibit Jurkat lymphoblastic leukemia T cell proliferation during coculture. Adherent cells with the ability of cellular growth were isolated from normal adipose tissues. Initial characterization of growing cells by flow cytometry suggested their mesenchymal stem cell characteristics. Cells were maintained in culture and used during third to fifth culture passages. Jurkat or allogeneic peripheral blood mononuclear cells [PBMCs] were labeled with carboxy fluorescein diacetate succinimidyl ester and cocultured with increasing doses of MSCs or MSC culture supernatant. Proliferation of PBMCs or Jurkat cells under these conditions was assessed by flow cytometry after 2 and 3 days of coculture, respectively. Results showed the expression of CD105, CD166 and CD44, and the absence of CD45, CD34 and CD14 on the surface of MSC like cells. Moreover, initial differentiation studies showed the potential of cell differentiation into hepatocytes. Comparison of Jurkat cell proliferation in the presence and absence of MSCs showed no significant difference, with 70% of cells displaying signs of at least one cell division. Similarly, the highest concentration of MSC culture supernatant [50% vol/vol] had no significant effect on Jurkat cell proliferation [p>0.6]. The same MSC lots significantly suppressed the allogeneic PHA activated PBMCs under similar culture conditions. Using Jurkat cells as a model of leukemia T cells, our results indicated an uncertainty about the suppressive effect of MSCs and their inhibitory metabolites on tumor or leukemia cell proliferation. Additional systematic studies with MSCs of different sources are needed to fully characterize the immunological properties of MSCs be-fore planning clinical applications

Analysis of human leukocyte antigen class II gene polymorphism in Iranian patients with Papillon-Lefevre syndrome: a family study

Papillon-Lefevre syndrome [PLS] is a rare autosomal recessive disorder characterized by palmoplantar hyperkeratosis and early development of aggressive periodontitis. Although cathepsin C [CTSC] gene mutations have been established in about 70-80% of PLS patients, it is assumed that the patients may have dysfunctioning of immune defense mechanisms. To assess the association of HLA class II genes and PLS. HLA class II genes were typed in nine Iranian PLS patients and their family members and the results were compared to 816 Iranian healthy subjects. The results of this study revealed that DRB1*0101 and DRB1*0301 alleles were more frequent in PLS patients than in normal controls. However, there was no significant difference between PLS patients and normal controls. Moreover, the same haplotypes and genotype combinations were also observed in some patients and their healthy siblings. The results of this study showed no strong association between HLA class II alleles and PLS

Production and characterization of a monoclonal antibody against an antigen on the surface of non-small cell carcinoma of the lung

Lung carcinoma is a multiple type cancer comprising of small cell and non-small cell carcinomas [NSCLC]. For therapeutic and diagnostic purposes, serum monoclonal antibodies have been produced against lung cancer. To characterize a murine monoclonal antibody [ME3D11] reactive with human NSCLC. A murine monoclonal antibody [ME3D11] reactive with human NSCLC was selected after immunization of BALB/c mice with a human large cell carcinoma with neuroendocrine differentiation, and was tested by immunofloursence staining and Western blot analysis. Our study showed that the antigen recognized by ME3D11 antibody was a cell surface antigen of 170kDa. This antigen is expressed on the cell surface of all NSCLC and a few carcinoma cell lines. In contrast, this antigen is neither expressed on the cell surface of human sarcoma, nor on the hematopoietic and normal cell lines. This antibody had no effect on spontaneous proliferation of Mehr-80 cell line in vitro. High degree of binding of this monoclonal antibody to NSCLC and some other carcinoma cells warrants further studies on its potential use in diagnosis and therapy of cancer by conjugation to drugs, toxins or radionuclides