The Suppressive Effect of Serum Samples from Patients with Chancroid on Human Mononuclear Cells Correlates with the Clinical Picture and is Interleukin-2-Dependent

1993 ◽  
Vol 4 (3) ◽  
pp. 171-173
Author(s):  
H C Korting ◽  
R Zaba ◽  
R C Ballard ◽  
D Abeck
Keyword(s):  
Mononuclear Cells ◽  
Cultured Cells ◽  
Human Lymphocytes ◽  
Interleukin 2 ◽  
Suppressive Effect ◽  
Serum Samples ◽  
Human Mononuclear Cells ◽  
Healthy Donors ◽  
Significant Difference ◽  
Dependent Cell

Plasma samples from patients with chancroid diagnosed both on clinical and microbiological grounds were assessed for their ability to inhibit mitogen-induced proliferation of human lymphocytes from healthy donors. All serum samples analysed suppressed phytohaemagglutinin A (PHA) blastogenic response. A significant difference in the observed extent was seen when serum samples from patients with and without associated lymphadenopathy were compared ( P < 0.05). Using an interleukin-2 (IL-2)-dependent cell line it could be demonstrated that the addition of patients' plasma to cultured cells markedly depressed mitogen-induced IL-2 synthesis. Results presented suggest that cell-mediated mechanisms play a role in the pathogenesis of infection due to Haemophilus ducreyi.

scholarly journals Optimizing interleukin-2 concentration, seeding density and bead-to-cell ratio of T-cell expansion for adoptive immunotherapy

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Sasan Ghaffari ◽  
Monireh Torabi-Rahvar ◽  
Sajjad Aghayan ◽  
Zahra Jabbarpour ◽  
Kobra Moradzadeh ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Growth Kinetics ◽  
Cell Expansion ◽  
Mononuclear Cells ◽  
Adoptive Cell Therapy ◽  
Interleukin 2 ◽  
Seeding Density ◽  
Healthy Donors ◽  
T Cell Expansion

Abstract Background The successful ex vivo expansion of T-cells in great numbers is the cornerstone of adoptive cell therapy. We aimed to achieve the most optimal T-cell expansion condition by comparing the expansion of T-cells at various seeding densities, IL-2 concentrations, and bead-to-cell ratios. we first expanded the peripheral blood mononuclear cells (PBMCs) of a healthy donor at a range of 20 to 500 IU/mL IL-2 concentrations, 125 × 103 to 1.5 × 106 cell/mL, and 1:10 to 10:1 B:C (Bead-to-cell) ratios and compared the results. We then expanded the PBMC of three healthy donors using the optimized conditions and examined the growth kinetics. On day 28, CD3, CD4, and CD8 expression of the cell populations were analyzed by flow cytometry. Results T-cells of the first donor showed greater expansion results in IL-2 concentrations higher than 50 IU/mL compared to 20 IU/mL (P = 0.02). A seeding density of 250 × 103 cell/mL was superior to higher or lower densities in expanding T-cells (P = 0.025). Also, we witnessed a direct correlation between the B:C ratio and T-cell expansion, in which, in 5:1 and 10:1 B:C ratios T-cell significantly expanded more than lower B:C ratios. The results of PBMC expansions of three healthy donors were similar in growth kinetics. In the optimized condition, 96–98% of the lymphocyte population expressed CD3. While the majority of these cells expressed CD8, the mean expression of CD4 in the donors was 19.3, 16.5, and 20.4%. Conclusions Our methodology demonstrates an optimized culture condition for the production of large quantities of polyclonal T-cells, which could be useful for future clinical and research studies.

scholarly journals Specific adsorption of HTLV-I to various target human and animal cells

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1303-1311 ◽  
Author(s):  
K Krichbaum-Stenger ◽  
BJ Poiesz ◽  
P Keller ◽  
G Ehrlich ◽  
J Gavalchin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Lymphoid Cells ◽  
Core Antigen ◽  
Target Cells ◽  
Animal Cells ◽  
Homologous Virus

In this report, we describe a flow cytometric analysis of HTLV-I specific binding to fresh and cultured cells on a single cell basis. This assay uses rhodamine hydrocarbon tagged, purified HTLV-I virions according to the procedure originally described for avian retroviruses. Successful HTLV-I transmission was detected by analysis of integrated HTLV-I DNA, virion-associated reverse transcriptase, and/or intracellular HTLV-I core antigen p19 expression. Only a specific virus- cell interaction was detected because nonrhodamine-tagged homologous virus or related HTLV-II interfered with tagged HTLV-I binding. In contrast, an unrelated, nonlabeled animal retrovirus was unable to block tagged HTLV binding. Of the cell lines tested, 2 nonlymphoid mammalian and 3 human lymphoid bound significantly high to moderate levels of HTLV-I-tagged virions. The other three human lymphocyte cell lines were insensitive to HTLV-I adsorption. A direct correlation was observed between HTLV-I binding sites and infectivity of human lymphoid cells alone and not other nonlymphoid animal cells. Fresh normal human mononuclear cells bound low levels of HTLV-I virions. As expected, T lymphocytes demonstrated more binding than did the non-T cell population. Enhancement of HTLV-I cell binding in a subpopulation of mononuclear target cells was achieved with phytohemagglutinin (PHA) activation and interleukin 2 (IL2) stimulation, which correlates well with previously published infectivity studies.

Can IL-2R Alpha be a Valuable Marker along with Ca 19–9 in the Diagnosis of Chronic Pancreatitis and Pancreatic Cancer?

2004 ◽  
Vol 19 (3) ◽  
pp. 196-202
Author(s):  
B. Kayhan ◽  
B. Kayhan ◽  
M. Akdoğ;an
Keyword(s):  
Pancreatic Cancer ◽  
Chronic Pancreatitis ◽  
Interleukin 2 ◽  
Carbohydrate Antigen ◽  
Control Group ◽  
Serum Samples ◽  
Cancer Group ◽  
Abdominal Symptoms ◽  
Significant Difference ◽  
Inflammatory Processes

Background Pancreatic cancer is characterized initially by non-specific abdominal symptoms followed by rapid tumor progression. Although chronic pancreatitis is a benign disorder, it can be one of the causative factors of pancreatic cancer. The level of the tumor marker carbohydrate antigen 19–9 (CA 19–9) in pancreatic cancer does not correlate with the stage of the neoplasm. Soluble interleukin 2 receptor (sIL-2R) is a cytokine that shows increased levels during some inflammatory processes and malignant disorders. Aim Our aim in this study was to investigate whether sIL-2Rα levels can be used in association with CA 19–9 in the early diagnosis of pancreatic cancer and chronic pancreatitis. Patients Serum samples were obtained from the blood of 21 pancreatic cancer patients without distant metastasis who were deemed inoperable, 16 chronic pancreatitis patients and 20 normal volunteers. Results We did not find any significant differences in CA 19–9 levels between normal controls and patients with chronic pancreatitis. There was a significant difference in the levels between the control group and the pancreatic cancer group (p=0.003) and between patients with chronic pancreatitis and those with pancreatic cancer (p=0.004). Although there was no significant difference in sIL-2Rα levels between the control group and the patient groups, we found a slight correlation between sIL-2Rα and CA 19–9 levels in the pancreatic cancer group (p=0.003, r=0.623) and a more marked correlation in the chronic pancreatitis group (p<0.01, r=0.751). Conclusion According to our results, sIL-2Rα alone is not a good candidate marker in the diagnosis of pancreatic cancer; it can, however, be used in association with CA 19–9 for this purpose.

scholarly journals Specific adsorption of HTLV-I to various target human and animal cells

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1303-1311 ◽  
Author(s):  
K Krichbaum-Stenger ◽  
BJ Poiesz ◽  
P Keller ◽  
G Ehrlich ◽  
J Gavalchin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Lymphoid Cells ◽  
Core Antigen ◽  
Target Cells ◽  
Animal Cells ◽  
Homologous Virus

Abstract In this report, we describe a flow cytometric analysis of HTLV-I specific binding to fresh and cultured cells on a single cell basis. This assay uses rhodamine hydrocarbon tagged, purified HTLV-I virions according to the procedure originally described for avian retroviruses. Successful HTLV-I transmission was detected by analysis of integrated HTLV-I DNA, virion-associated reverse transcriptase, and/or intracellular HTLV-I core antigen p19 expression. Only a specific virus- cell interaction was detected because nonrhodamine-tagged homologous virus or related HTLV-II interfered with tagged HTLV-I binding. In contrast, an unrelated, nonlabeled animal retrovirus was unable to block tagged HTLV binding. Of the cell lines tested, 2 nonlymphoid mammalian and 3 human lymphoid bound significantly high to moderate levels of HTLV-I-tagged virions. The other three human lymphocyte cell lines were insensitive to HTLV-I adsorption. A direct correlation was observed between HTLV-I binding sites and infectivity of human lymphoid cells alone and not other nonlymphoid animal cells. Fresh normal human mononuclear cells bound low levels of HTLV-I virions. As expected, T lymphocytes demonstrated more binding than did the non-T cell population. Enhancement of HTLV-I cell binding in a subpopulation of mononuclear target cells was achieved with phytohemagglutinin (PHA) activation and interleukin 2 (IL2) stimulation, which correlates well with previously published infectivity studies.

Vandellia Cordifolia Regulated Cell Proliferation and Cytokines Production in Human Mononuclear Cells

2000 ◽  
Vol 28 (03n04) ◽  
pp. 313-323 ◽  
Author(s):  
An-Pang Lin ◽  
Wei-Jern Tsai ◽  
Chi-Yen Fan ◽  
Ming-Jen Lee ◽  
Yuh-Chi Kuo
Keyword(s):  
Cell Cycle Progression ◽  
Mononuclear Cells ◽  
Tritiated Thymidine ◽  
Interleukin 2 ◽  
Interferon Γ ◽  
Thymidine Uptake ◽  
Dependent Manner ◽  
Human Mononuclear Cells ◽  
Tritiated Thymidine Uptake ◽  
Ifn Γ

Vandellia cordifolia (V. cordifolia) used for treatment inflammation in traditional Chinese medicine was selected for immunopharmacological activity test. The effects of V. cordifolia extracted fractions on human mononuclear cells (HMNC) proliferation were determined by tritiated thymidine uptake. The results indicated that VC-ME fraction suppressed HMNC proliferation activated with phytohemagglutinin (PHA) and stimulated cell cycle progression was arrested at the G0/G1 stage. The inhibitory mechanisms may involve the blocking of interleukin-2 (IL-2) and interferon-γ (IFN-γ) production, since VC-ME suppressed IL-2 and IFN-γ production of HMNC in a dose-dependent manner. Therefore, it is suggested that immunomodulatory agents are contained in V. cordifolia.

scholarly journals Good Manufacturing Practice-Compliant Expansion of Marrow-Derived Stem and Progenitor Cells for Cell Therapy

2007 ◽  
Vol 16 (7) ◽  
pp. 685-696 ◽  
Author(s):  
Martin H. Gastens ◽  
Kristin Goltry ◽  
Wolfgang Prohaska ◽  
Diethelm Tschöpe ◽  
Bernd Stratmann ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Progenitor Cells ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Ex Vivo ◽  
Good Manufacturing Practice ◽  
Minimal Risk ◽  
Multilineage Differentiation ◽  
Significant Difference ◽  
Stem And Progenitor Cells

Ex vivo expansion is being used to increase the number of stem and progenitor cells for autologous cell therapy. Initiation of pivotal clinical trials testing the efficacy of these cells for tissue repair has been hampered by the challenge of assuring safe and high-quality cell production. A strategy is described here for clinical-scale expansion of bone marrow (BM)-derived stem cells within a mixed cell population in a completely closed process from cell collection through postculture processing using sterile connectable devices. Human BM mononuclear cells (BMMNC) were isolated, cultured for 12 days, and washed postharvest using either standard open procedures in laminar flow hoods or using automated closed systems. Conditions for these studies were similar to long-term BM cultures in which hematopoietic and stromal components are cultured together. Expansion of marrow-derived stem and progenitor cells was then assessed. Cell yield, number of colony forming units (CFU), phenotype, stability, and multilineage differentiation capacity were compared from the single pass perfusion bioreactor and standard flask cultures. Purification of BMMNC using a closed Ficoll gradient process led to depletion of 98% erythrocytes and 87% granulocytes, compared to 100% and 70%, respectively, for manual processing. After closed system culture, mesenchymal progenitors, measured as CD105+CD166+CD14–CD45– and fibroblastic CFU, expanded 317- and 364-fold, respectively, while CD34+ hematopoietic progenitors were depleted 10-fold compared to starting BMMNC. Cultured cells exhibited multilineage differentiation by displaying adipogenic, osteogenic, and endothelial characteristics in vitro. No significant difference was observed between manual and bioreactor cultures. Automated culture and washing of the cell product resulted in 181 × 106 total cells that were viable and contained fibroblastic CFU for at least 24 h of storage. A combination of closed, automated technologies enabled production of good manufacturing practice (GMP)-compliant cell therapeutics, ready for use within a clinical setting, with minimal risk of microbial contamination.

scholarly journals Tacrolimus Pharmacodynamics and Pharmacogenetics along the Calcineurin Pathway in Human Lymphocytes

2014 ◽  
Vol 60 (10) ◽  
pp. 1336-1345 ◽  
Author(s):  
Ofelia M Noceti ◽  
Jean-Baptiste Woillard ◽  
Ahmed Boumediene ◽  
Patricia Esperón ◽  
Jean-Luc Taupin ◽  
...  
Keyword(s):  
Signal Transduction ◽  
T Cells ◽  
Mononuclear Cells ◽  
Interindividual Variability ◽  
Ex Vivo ◽  
Physiological Activity ◽  
Human Lymphocytes ◽  
Interleukin 2 ◽  
Therapeutic Drug ◽  
Calcineurin Pathway

Abstract BACKGROUND Although therapeutic drug monitoring has improved the clinical use of immunosuppressive drugs, there is still interpatient variability in efficacy and toxicity that pharmacodynamic monitoring may help to reduce. To select the best biomarkers of tacrolimus pharmacodynamics, we explored the strength and variability of signal transduction and the influence of polymorphisms along the calcineurin pathway. METHODS Peripheral blood mononuclear cells from 35 healthy volunteers were incubated with tacrolimus (0.1–50 ng/mL) and stimulated ex vivo. Inhibition of NFAT1 (nuclear factor of activated T cells 1) translocation to the nucleus and intracellular expression of interleukin-2 in CD4+ and CD8+ T cells and the surface activation marker CD25 in CD3+ cells were measured by flow cytometry. We sequenced the promoter regions of immunophilins and calcineurin subunits and characterized selected single nucleotide polymorphisms in the genes of the calcineurin pathway with allelic discrimination assays. RESULTS All responses closely fitted an I/Imax sigmoid model. Large interindividual variability (n = 30) in I0 and IC50 was found for all biomarkers. Moreover, strong and statistically significant associations were found between tacrolimus pharmacodynamic parameters and polymorphisms in the genes coding cyclophilin A, the calcineurin catalytic subunit α isoenzyme, and CD25. CONCLUSIONS This study demonstrates the consistency and large interindividual variability of signal transduction along the calcineurin pathway, as well as the strong influence of pharmacogenetic polymorphisms in the calcineurin cascade on both the physiological activity of this route and tacrolimus pharmacodynamics.

Chimeric CD19 antibody mediates cytotoxic activity against leukemic blasts with effector cells from pediatric patients who received T-cell–depleted allografts

Blood ◽  
2004 ◽  
Vol 103 (10) ◽  
pp. 3982-3985 ◽  
Author(s):  
Peter Lang ◽  
Karin Barbin ◽  
Tobias Feuchtinger ◽  
Johann Greil ◽  
Matthias Peipp ◽  
...  
Keyword(s):  
Nk Cells ◽  
Mononuclear Cells ◽  
Therapeutic Option ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Hla Class I ◽  
Post Transplantation ◽  
Effector Cells ◽  
Healthy Donors ◽  
B Lineage

Abstract Relapse is a major problem after transplantation in children with acute B-lineage leukemias, and new therapies are needed to increase graft-versus-leukemia (GvL) effects without inducing graft-versus-host disease (GvHD). Here, we studied the ability of effector cells recovered from patients after transplantation with positive-selected stem cells from alternative donors to induce antibody-dependent cellular cytotoxicity (ADCC). For this purpose, a chimeric CD19 antibody, CD19-4G7chim, was generated. This antibody efficiently mediated ADCC against primary acute lymphoblastic leukemia (ALL) blasts by using purified natural killer (NK) cells from healthy donors or mononuclear cells from patients as effector cells. Increased lysis was obtained after stimulation of effector cells with interleukin-2 (IL-2). ADCC was not prevented by inhibitory effects mediated by HLA class I. We propose that treatment with chimeric CD19 antibodies leading to ADCC by donor-derived NK cells may become a therapeutic option for the post-transplantation treatment of minimal residual B-lineage ALLs.

scholarly journals Nicotine inhibits the in vitro production of interleukin 2 and tumour necrosis factor-α by human mononuclear cells

1996 ◽  
Vol 35 (1) ◽  
pp. 47-51 ◽  
Author(s):  
G.S. Madretsma ◽  
G.J. Donze ◽  
A.P.M. van Dijk ◽  
C.J.A.M. Tak ◽  
J.H.P. Wilson ◽  
...  
Keyword(s):  
Tumour Necrosis ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Tumour Necrosis Factor Α ◽  
In Vitro Production ◽  
Human Mononuclear Cells ◽  
Factor Α ◽  
Vitro Production ◽  
Necrosis Factor

scholarly journals Expression of the interleukin-2 receptor gamma chain on cord blood mononuclear cells

Blood ◽  
1996 ◽  
Vol 87 (8) ◽  
pp. 3344-3350 ◽  
Author(s):  
S Saito ◽  
T Morii ◽  
H Umekage ◽  
K Makita ◽  
K Nishikawa ◽  
...  
Keyword(s):  
T Cells ◽  
Cord Blood ◽  
Nk Cells ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Gamma Chain ◽  
Blood Lymphocytes ◽  
Interleukin 2 Receptor ◽  
Significant Difference ◽  
Cord Blood Lymphocytes

Lymphocytes in umbilical cord blood and neonatal peripheral blood have been shown to have less ability in an immune reaction. In our present experimental approach to address this issue, we made use of the cord blood of full-term birth infants to investigate the expression of the interleukin- 2 receptor gamma (IL-2Rgamma) chain that is shared with receptors for IL-4, IL-7, IL-9, and IL-15 as well as IL-2. The gamma chain expression in cord blood lymphocytes was about one-third that in the lymphocytes of adults, whereas no significant difference between cord blood and adult monocytes was observed. A reduced expression of the gamma chain was observed in all of the CD4+ T cells, CD8+ T cells, gamma-delta T cells, B cells, CD16+ natural killer (NK) cells, and CD56(bright) NK cells of the cord blood lymphocytes. The reduced gamma chain expression reached two-thirds of that in adults after 3 days of culture in vitro and in infants 3 days after birth, thus implying that the increase in the gamma chain may significantly contribute to the prevention of neonatal infection.

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