Can immune parameters be used as predictors to distinguish between pulmonary multidrug-resistant and drug-sensitive tuberculosis?

Introduction: Despite the development and wide implementation of Directly Observed Therapy Strategies (DOTS), multidrug-resistant tuberculosis (MDR-TB) remains a serious global health threat. In this study, the role of host immune response in patients with MDR-TB is investigated and compared with that of patients with smear-positive drug-sensitive tuberculosis (SP-TB

Combination of Early and Late Growth Factors to Enrich Transplantable Cord Blood CD34+Cells in Short Term Cultures

Objective: Although umbilical cord blood (UCB) is a useful source of stem and progenitor cells for bone marrow transplantation, it is not possible to obtain sufficient number of transplantable cells from a single human umbilical cord for transplantation in adult patients. In this study, we combined early and late growth factors in short term cultures to enrich cord blood CD34+ hematopoietic progenitor cell count for adult patients. Material and Methods: Cord blood mononuclear cells (CBMC) harvested from 15 healthy volunteers, delivering in term, incubated in culture medium supplemented with interleukin-3 (IL-3), IL-6, IL-11, stem cell factor, flt3/flk2 ligand and thrombopoietin for 14 days. Starting cells and cultured cells (day 7 and 14 cells) were evaluated for; CD4, CD8, CD33, CD34, CD38 and HLA-DR by flow cytometry and seeded in semisolid methylcellulose culture for determining BFU-E, CFU-GM and CFU-GEMM colony forming abilities. Results: In the short-term cultures, total CBMC count significantly increased (p= 0.0001). Compared to starting cells, the expressions of CD34 (p= 0.002), CD33 (p= 0.0001), HLA-DR (p= 0.0001) and CD8 (p= 0.002) increased, but those of CD38 (p= 0.057) and CD4 (p= 0.0001) were suppressed on day 14 of cultures. There were an increase in generation of CFU-GM (p= 0.131) and CFU-GEMM (p= 0.134) colonies and a significant decrease in generation of BFU-E colonies (p= 0.0001). Conclusion: Ex vivo expansion of CBMC, under combination of the growth factors, contribute to CD34+ progenitor cell count and may help to reach sufficient amount of transplantable cells for adult patients. Suspension cultures may lead to elimination of graft-versus-host disease risk and protection of graft-versus-leukemia effect, through the decrease of CD4+ T cells and maintenance of CD8+ T lymphocytes, respectively

Antiproliferative effects of fluorine substitute 3,5-di-tert-butylphenol bearing Schiff bases using CFSE-based cell proliferation assay

The determination of antiproliferative properties of compounds on tumour cells is important for assessment of their efficacy in cancer treatment. CFSE-labelled K562 cells were incubated with doxorubicin and ortho-or para-fluorosubstitute Schiff bases (compounds 1 and 2 respectively). CFSE intensities were analysed using flow cytometry. K562 cells treated with doxorubicin resulted in homogeneous high intensity fluorescence after 96 h of incubation. Schiff bases exhibited antiproliferative effects, but lower than doxorubicin. Our results reveal that CFSE assay can be used for determining in vitro antiproliferative features of anticancer drugs and/or compounds from herbal or chemical sources

Concanavalin A and Phytohaemagglutinin Stimulated Lymphoproliferative Responses in Cord Blood Mononuclear Cells

Experimental and clinical evidences have demonstrated that umbilical cord blood is an efficient source of transplantable haematopoietic stem and progenitor cells. After transplantation, initial exposure to alloantigen-presenting cells (allo-APC) induced lymphoproliferative responses in cord blood T cells. However, this response is lower than that by adult T cells. The aim of this study was to investigate and compare lymphoprolifertive responses of cord blood mononuclear cells (CBMC) and adult peripheral blood mononuclear cells (PBMC). CBMC and PBMC immunoproliferative responses were measured by MTT assay after stimulation of respective primary culture cells with phytohaemagglutinin (PHA) or Concanavalin A (Con A) for 76 hours. Lymphoproliferative response of CBMC was significantly lower compared to that of PBMC

Lymphocyte Markers and Proliferative Responses to Microbial Antigens in Patients with Allergic Rhinitis

Objective: Atopy is a condition of predisposition to allergic reaction to environmental allergens, and T cells have a critical role in initiating and ending allergic responses. This study was conducted to evaluate the T cell responses of atopic patients with allergic rhinitis who have allergen-hyperreactive memory CD4 T cells in vitro. Material and Methods: Cell surface markers (CD3, CD4, CD8, CD19, CD28, CD45RA, CD45RO, CD95, HLA-DR) were analyzed for T and B lymphocytes by flow cytometry using fluorescein isothiocyanate (FITC) or phycoerythrin (PE) labeled monoclonal antibodies. T cell proliferative response assessing pokeweed mitogen (PWM), tetanus toxoid (TT), purified protein derivative of mycobacterium (PPD) and cytomegalovirus antigen (CMV) were examined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction assay. Results: Immune profile and lymphoproliferative responses of 19 allergic rhinitis (AR) patients with positive prick skin test and nasal blockage, discharge, sneeze, nasal or ocular itching (mean age 33.2 +/- 8.4), and 10 healthy controls (mean age 31.6 +/- 9.1) were evaluated. CD3 and CD4 expression was higher in AR patients than in healthy controls. Memory (CD45RO) and activated (CD28) T cell levels were higher, but lymphoproliferation to PWM, TT, PPD, and CWV was decreased in AR patients. Conclusion: The high CD28 and CD45RO expression associated with atopy symptoms indicated that immune reactions in AR patients tended to show an undesirable shift toward Th2 skewed with high levels of allergen-reactive memory T cells. Consequently, the reduced lymphoproliferation to non-allergenic stimulants such as mitogens, bacterial and viral antigens in AR patients may lead to reduced immune response capability to infectious agents

A Pilot Study on the Isolation of Dental Pulp Stem Cells, Potential of Forming Colonies and Defining the Content of Stem Cells

Objective: Dental pulp has been used as a source in various stem cells experiments in the recent years. The aim of this study was to isolate dental pulp stem cells in sterilized conditions, to define the content level of stem cells and to optimize the methods that were used in obtaining single cell suspension for cell culture experiments. Material and Methods: In this study, a third molar tooth was used. Dental pulp tissue was digested with collagenase Type 1 (3 mg/mL) in alpha-MEM containing 10% fetal calf serum to obtain single cell suspension. The cells were incubated in 6-well-plates with culture medium at 5% CO(2) for 72 hours at moist conditions. After 72 hours, the cells were examined by inverted microscopy for cell morphology and development of colonies and the ratio of CD34 positive cells were defined with flow cytometry. Results: The development of the cell colonies was observed in culture. Three different cell populations were defined from the culture and the ratios of these populations that expressed CD34 were 10.99%, 12.99% and 2.99%. Conclusion: Considering the high rate of CD34 positive cells and the capability of forming colonies in in vitro culture, dental pulp is a good stem cell source

In Vitro Cytotoxic and Proapoptotic Activities of Anatolian Macrovipera Lebetina Obtusa (Dwigubski, 1832) Crude Venom on Cultured K562 Human Chronic Myelogenous Leukemia Cells

WOS: 000373662500005In the context of searching for anticancer compounds in natural products, snake venom is one of the important sources for peptide/protein based bioactive molecules. Proteins and peptides with anticancer activity were purified and identified from snake venoms. The aim of the present study was to determine the in vitro cytotoxicity of Macrovipera lebetina obtusa (Blunt-Nosed Viper) crude venom from southeastern Anatolia against K562 human chronic myelogenous leukemia (CML) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Adenosine tripohsphate (ATP) assays. Additionally, the apoptosis induction was assessed by morphological evaluation and immunohistochemical analysis for activated caspase-3. For histopahtological evaluation, haematoxylineosin, giemsa and papanicolau stains were used in combination. M. L obtusa venom showed dose-dependent toxicity against K562 cells after 72 h treatment with different concentrations of crude venom. 1050 values were 0.45 and 0.37 mu g/mL for MTT and ATP assays, respectively. Nuclear fragmentation and condensation, apoptotic bodies and activation of caspase-3, as an induction of apoptosis were also observed in K562 cells. Since apoptosis-inducing compounds are important for the treatment of cancer, further studies on Anatolian M. I. obtusa venom could result in the purification and identification of new proteins and peptides, which might have therapeutic value for the treatment of CML