Effects Of Recombinant Human Interleukin Research Articles

Therapeutic effects of recombinant human interleukin 2 as adjunctive immunotherapy against tuberculosis: A systematic review and meta-analysis.

BackgroundInterleukin 2 (IL-2) is a cytokine secreted by activated T cells. Studies exploring recombinant human interleukin 2 (rhuIL-2) as an adjunctive immunotherapeutic agent to treat tuberculosis (TB) have shown variable results; however, the true therapeutic efficacy of rhuIL-2 administration in TB patients has not been determined.MethodsA systematic review to identify publications exploring the association between rhuIL-2-based immunotherapy for TB and outcomes (sputum culture conversion, sputum smear conversion, radiographic changes, and leukocyte phenotype changes) in patients with pulmonary TB published before June 8, 2018 was performed. Data were extracted and analyzed by two investigators independently.ResultsA total of 2,272 records were screened. Four randomized controlled trials (RCTs) comprising 656 pulmonary TB patients were finally included. The rhuIL-2 treatment could significantly improve the sputum culture conversion of TB (RR, 1.18; 95%CI: 1.03–1.36; I2 < 0.01; P = 0.019) after at least 3 months of anti-TB therapy and the sputum smear conversion of TB during anti-TB therapy. Treating multidrug-resistant tuberculosis (MDR-TB) with rhuIL-2 could improve the sputum culture conversion (RR, 1.28; 95%CI: 1.05–1.57; I2 < 0.01; P = 0.016) and smear conversion (RR, 1.28; 95%CI: 1.09–1.51; I2 < 0.01; P = 0.003) at the end of anti-TB treatment. Meanwhile, rhuIL-2-based adjunctive immunotherapy could expand the proliferation and conversion of CD4+ and natural killer (NK) cells. Three of the included studies suggested that radiographic changes could not be improved by the use of rhuIL-2 as adjunctive immunotherapy. Publication bias did not exist.ConclusionsBased on this first meta-analysis, rhuIL-2-based adjunctive immunotherapy appears to expand the proliferation and conversion of CD4+ and NK cells, as well as improve the sputum culture (at 3 months and later) and smear conversion of TB patients.

Effect of rhIL-11 on Th1/Th2 and T-bet/GATA-3 imbalance in idiopathic thrombocytopenic purpura

Objective To investigate the effect of recombinant human interleukin 11 (rhlL-11) in the treatment of idiopathic thrombocytopenic purpura (ITP) on the levels of Th1, Th2 and the expression of their transcription factors T-bet mRNA, GATA-3 mRNA. Methods Fifty-six cases adult ITP patients hospitalized in the department of hematology of the Second People’s Hospital of Datong from May 2015 to December 2016 were collected, including 21 males and 35 females, aged 29~73 years; 10 healthy people in the same period were enrolled as control group, 4 males and 6 females, aged 20~52 years.Th1 and Th2 cell ratio and Th1/Th2 ratio of ITP patients were detected by flow cytometry before and after treatment.The expression levels of transcription factor T-bet and GATA-3 were measured using real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) before and after treatment. Results The effective rate of rhlL-11 in ITP treatment was 76.8%(43/56). For the effective patients, the median PLT after treatment increased (25.0 (15.0, 36.0)×109/L vs.68.0 (49.0, 108.0)×109/L, Z=-5.712, P 0.05). Conclusion rhIL-11 can effectively correct the imbalance in Th1 and Th2 cells and the imbalance of T-bet and GATA-3 in ITP patients, but it has no obvious therapeutic effect on a small number of patients Key words: Idiopathic thrombocytopenic purpura; Recombinant human interleukin-1; T Helper cells 1; T helper cells 2; T-box expressed in T Cells; GATA binding protein 3

A Clinical Study on Juheli (Recombinant Human Interleukin - 11) in the Second Prevention of Chemotherapy Induced Thrombocytopenia

to investigate the effect and side effects of recombinant human interleukin - 11 (rhIL - 11, in Chinese Juheli, produced by Qi Lu Biotechnology CO., LTD) in the second prevention of chemotherapy induced thrombocytopenia (CIT). Cancer patients with CIT were recruited and were treated with rhIL - 11 (treatment phase, TP), and in the following cycle, all these patients administered with rhIL - 11 24 hours immediately after chemotherapy (preventive treatment phase, PTP). Duration and severity of thrombocytopenia between two phases were compared. for patients in TP or PTP, nadir values of platelet were (29.28±20.08)?109/L and (45.24±19.66)?109/L, duration of thrombocytopenia in TP and PTP was (11.52±4.33) and (8.20±+2.77)days, recovery time was (19.40±3.89)and (13.44±3.02)days, duration of rhIL - 11 administration was 10.68±2.46)and (6.28±1.77)days, number of patients needing platelet infusion was 16and4 respectively, all differences were statistically significant (p value were 0.007, 0.002, 0.000, 0.000, 0.034 respectively). For TP and PTP, number of patients with hemorrhage was 8 and 4, duration of bleeding was (5.00±0.82) and (4.50 ± 0.71) days respectively, with no statistically significant difference. Adverse reactions mainly included fever, edema, arrhythmia, joint pain, fatigue, skin rash, headache, dizziness, etc., all were not statistically significant between TP and PTP. rhIL - 11 could be well tolerated and is effective that could reduce the duration, severity of CIT, platelet transfusion, and incidence of bleeding, as well as shorten the recovery time, duration of rhIL - 11 administration. Thus, rhIL - 11 could be commended in the second prevention of CIT for patients with cancer.

Recombinant human interleukin 24 reverses Adriamycin resistance in a human breast cancer cell line

BackgroundThe major cause of multidrug resistance is over-expression of membrane P-glycoprotein (P-gp). We investigated the effect of recombinant human interleukin 24 (rhIL-24) on the Adriamycin (ADM)-resistant human breast cancer cell line MCF-7/ADM. MethodsThe cytotoxicity of rhIL-24 and ADM was determined by 3-[4,5-dimethylthiazol-2-yl], 5-diphenyl tetrazolium bromide (MTT) assays. The expression of P-gp was assessed by confocal microscopy and Western blot analysis. ResultsThe IC50 values for rhIL-24 in MCF-7/wild-type and MCF-7/ADM cells were 0.17 and 14.6μM, respectively. The IC50 value of Adriamycin in MCF-7/ADM cells decreased in a dose-dependent manner when rhIL-24 was used. The resistance modulating factor (RMF) was directly proportional to the dose of rhIL24. ADM accumulation increased while P-gp expression decreased at a low dose (4μM) of rhIL24 in MCF-7/ADM cells. The expression of P-gp was decreased at 4μM in confocal microscopy and western blot analysis. ConclusionsrhIL-24 circumvented the drug-resistance of MCF-7/ADM cells via activation of the transcription factor Stat 3. rhIl24 has potential to act as a P-gp inhibitor to reverse Adriamycin resistance in breast cancer.

Interleukin-33 promotes proliferation of primary human lung fibroblasts

Event Abstract Back to Event Interleukin-33 promotes proliferation of primary human lung fibroblasts Alexander Bocharov1* and Olga Kornilova1 1 National University of Pharmacy, Ukraine Interleukin (IL)-33 is a multifunctional cytokine that belongs to the IL-1 cytokine family and expressed by multiple organs and cell types in humans and mice. Recent studies have showed that IL-33 plays an etiological role in several fibrotic disorders and may be involved in the pathogenesis of chronic respiratory diseases, such as asthma, and pulmonary fibrosis. It has been reported that IL-33–induced cutaneous fibrosis is associated with increased fibroblast proliferation and altered expression of extracellular matrix-modifying genes. However, the role of IL-33 in regulating of functions of lung fibroblasts remains unclear. In the present study, we examined the effect of recombinant human interleukin (IL)-33 on proliferation of human lung fibroblasts. Five primary lines of normal adult human lung fibroblasts were cultured for 3-7 days in the presence of increasing concentrations of IL-33 (1, 2, 10, 50, 100 and 300 ng/ml). The proliferation of human lung fibroblasts was analyzed using non-radioactive cell proliferation assay (MTT). We have observed that normal human lung fibroblasts responded, in a dose-dependent manner, to treatment with recombinant human IL-33, by increasing proliferation rates 1.5- to 2.3 fold compared with non-stimulated control. The maximum effect of IL-33 on fibroblast proliferation was observed in cytokine concentrations range from 2 ng/ml to 100 ng/ml. These results suggest that IL-33 may play an important role in the regulation of human lung fibroblast proliferation. Human lung fibroblasts activated by IL-33 may act as effector cells not only in the pathogenesis of lung diseases but also in lung remodeling processes. Keywords: interleukin -33, human lung fibroblasts, proliferation, Pulmonary Fibrosis, fibrotic disorders Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Innate immunity Citation: Bocharov A and Kornilova O (2013). Interleukin-33 promotes proliferation of primary human lung fibroblasts. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00565 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 02 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Alexander Bocharov, National University of Pharmacy, Kharkov, Ukraine, a.a.botcharov@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Alexander Bocharov Olga Kornilova Google Alexander Bocharov Olga Kornilova Google Scholar Alexander Bocharov Olga Kornilova PubMed Alexander Bocharov Olga Kornilova Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Evaluation of the role of rhIL-11 on hematological recovery in lymphoma patients after autologous hematopoietic stem cell transplantation

To evaluate the effect of recombinant human interleukin 11 (rhIL-11) on hematological recovery after autologous hematopoietic stem cell transplantation (AHSCT) in patients with lymphoma. A retrospective study was carried out on 73 patients with lymphoma after AHSCT. The patients were divided into two groups. The study group (n = 35) received rhIL-11 1.5 mg daily from the fifth day after AHSCT to the day when platelets recovering to 80.0×10⁹/L. The control group (n = 38) did not receive rhIL-11 after AHSCT. All the 73 patients finished AHSCT from Mar 2003 to Dec 2008 in our department. Thirty-five patients received rhIL-11 and 38 patients did not. In the rhIL-11 group and control group, the nadir of platelet was (18.9 ± 5.0)×10⁹/L and (21.5 ± 6.0)×10⁹/L, respectively, with a significant difference (P = 0.04). The median time of platelet recovering to 50.0×10⁹/L was (14.3 ± 5.5) d and (13.2 ± 4.5) d (P = 0.37) in the two groups. There was no significant difference (P = 0.82) in the median numbers of platelet transfusion in the two groups. The curves of the mean of daily absolute platelet counts of the two groups were similar (P = 0.22). Adverse events related to rhIL-11 were not found in the rhIL-11 group. The results of this study do not show obviously accelerating effect of rhIL-11 on the platelet recovery in lymphoma patients after AHSCT and obvious increase of adverse events after rhIL-11 administration.

Effects of Recombinant Human Interleukin 7 on T-Cell Recovery and Thymic Output in HIV-Infected Patients Receiving Antiretroviral Therapy: Results of a Phase I/IIa Randomized, Placebo-Controlled, Multicenter Study

The immune deficiency of human immunodeficiency virus (HIV) infection is not fully corrected with ARV therapy. Interleukin-7 (IL-7) can boost CD4 T-cell counts, but optimal dosing and mechanisms of cellular increases need to be defined. We performed a randomized placebo-controlled dose escalation (10, 20 and 30 µg/kg) trial of 3 weekly doses of recombinant human IL-7 (rhIL-7) in ARV-treated HIV-infected persons with CD4 T-cell counts between 101 and 400 cells/µL and plasma HIV levels <50 copies/mL. Toxicity, activity and the impact of rhIL-7 on immune reconstitution were monitored. Doses of rhIL-7 up to 20 µg/kg were well tolerated. CD4 increases of predominantly naive and central memory T cells were brisk (averaging 323 cells/µL at 12 weeks) and durable (up to 1 year). Increased cell cycling and transient increased bcl-2 expression were noted. Expanded cells did not have the characteristics of regulatory or activated T cells. Transient low-level HIV viremia was seen in 6 of 26 treated patients; modest increases in total levels of intracellular HIV DNA were proportional to CD4 T-cell expansions. IL-7 seemed to increase thymic output and tended to improve the T-cell receptor (TCR) repertoire in persons with low TCR diversity. Three weekly doses of rhIL-7 at 20 µg/kg are well tolerated and lead to a dose-dependent CD4 T-cell increase and the broadening of TCR diversity in some subjects. These data suggest that this rhIL-7 dose could be advanced in future rhIL-7 clinical studies. NCT0047732.

Effects of recombinant human interleukin 11 on hematological malignancy after allogeneic hematopoietic cell transplantation

To investigate the effects of recombinant human interleukin 11 (rhIL-11) on hematological malignancy after allogeneic hematopoietic stem cell transplantation (allo-HSCT). A total of 48 patients with hematological malignancy from January 2006 to June 2010 were alternately enrolled into a prospective randomized study. And they were assigned into the control and rhIL-11 injection groups. Later the investigators compared two groups with regards to hematopoietic reconstitution, graft versus host disease (GVHD) classification, clinical recurrence rate and disease-free survival. With the therapy of rhIL-11, the absolute neutrophil counts recovering to 0.5 × 10(9)/L and platelet recovering to 20 × 10(9)/L were (15.1 ± 1.6) and (18.2 ± 3.3) days respectively. And they were significantly lower than those in control group [(16.1 ± 1.6) vs (22.4 ± 5.5) days, P = 0.032, 0.003]. The incidence of acute GVHD was surprisingly low in the study group (26.1% vs 50.0%, P = 0.048). There was no significant difference in chronic GVHD (36.8% vs 38.9%, P = 0.899) or relapse rate (5.1% vs 7.7%, P = 0.662) between two groups during a median follow-up period of 11.7 months. A trend of improved 3-year-disease-free survival was observed in the study group (65.4% vs 50.9%, P = 0.637). The application of rhIL-11 after allo-HSCT may accelerate both neutrophil and platelet engraftment and lower the occurrence of acute GVHD.

A Pilot Study of Interleukin-11 in Subjects with Chronic Hepatitis C and Advanced Liver Disease Nonresponsive to Antiviral Therapy

To evaluate the effects of recombinant human interleukin (rhIL)-11 on liver histology in patients with chronic hepatitis C virus (HCV) infection and advanced liver disease who had failed antiviral therapy. This was an open-label study of rhIL-11 (Neumega), Wyeth Laboratories, Collegeville, PA) at a dose of 5 microg/kg administered by subcutaneous injection daily for 12 wk. The primary efficacy endpoint was the change in the Knodell Histology Activity Index (HAI) between pre- and posttreatment liver biopsies. Secondary efficacy endpoints included changes in plasma alanine transaminase (ALT) concentrations and in the number of platelets. The Knodell HAI improved in 11 (55%) of the 20 subjects enrolled, with the mean score improving from 7.3 to 5.9 (p= 0.006). Eight subjects (40%) experienced significant improvement as defined by a decrease of at least two points in the HAI. IL-11 treatment was also associated with a decrease in ALT levels from a mean level of 113 IU/L at baseline to 65 IU/L at week 12 (p < 0.001). Platelet levels increased from a mean of 143 x 10(3)/microl at baseline to 198 x 10(3)/mul at week 12 of treatment (p < 0.001). Overall, rhIL-11 was well tolerated and no serious adverse events (AEs) were reported. The most common AE was edema of the lower extremities, which occurred in all subjects. The findings from this pilot study suggest that rhIL-11 may be beneficial for patients with hepatic inflammation and advanced liver disease associated with chronic HCV infection. Larger clinical trials are warranted to further evaluate the long-term antiinflammatory and antifibrotic effects of rhIL-11.

Effects of recombinant human interleukin 11 on thrombocytopenia and neutropenia in irradiated rhesus monkeys.

The effects of recombinant human interleukin 11 (rhIL11) on thrombocytopenia and neutropenia in irradiated rhesus monkeys were evaluated after administration different doses at different times. Twenty-three rhesus monkeys were exposed to a total-body irradiation (TBI) with a single dose of 3 Gy 60Co gamma rays. Either placebo, rhIL11 at a dose of 30, 60 or 120 microg/kg day(-1) on days 0-13, or rhIL11 at a dose of 60 microg/kg day(-1) on days 13-26 after TBI was administered to the animals. The results showed that the immediate treatment with rhIL11 but not treatment on days 13-26 resulted in much higher platelet nadirs than in the placebo-treated group. The accelerated recovery of platelets to normal levels after TBI was demonstrated in all groups treated with rhIL11, but the effects of rhIL11 were independent of dose. However, rhIL11 treatment could also accelerate the recovery of leukocytes to normal levels. The numbers of colony-forming bone marrow cells (CFU-E, CFU-Mix, CFU-MK and CFU-GM) in all groups treated with rhIL11 were increased 4- to 14-fold relative to those of the placebo group on day 30. We conclude that rhIL11 may directly promote megakaryocyte development and ameliorate myelosuppression in irradiated monkeys.

Interleukin-11 (IL-11) enhances clonal proliferation of acute myelogenous leukemia cells with strong expression of the IL-11 receptor alpha chain and signal transducing gp130.

We examined the effect of recombinant human interleukin (IL)-11 alone or in combination with various colony-stimulating factors (CSFs), including IL-3, granulocyte/macrophage (GM)-CSF, granulocyte (G)-CSF, stem cell factor (SCF), flt3 ligand (FL), and thrombopoietin (TPO), on colony formation by leukemic progenitor cells (L-CFU) obtained from 33 patients with acute myelogenous leukemia (AML). Leukemic colony formation was found in approximately 70 to 80% of the patients in the presence of at least one of the above CSFs. Although IL-11 alone did not support L-CFU, the growth of these progenitors in the presence of other cytokines was enhanced by IL-11 in 16 out of 33 patients and it showed a synergistic action with G-CSF in 12 of them. This synergistic action occurred in seven out of nine M5 patients (French-American-British (FAB) classification). A single cell clone-sorting experiment clearly demonstrated that this synergistic effect was operative at the single progenitor cell level. The number of leukemic cells proliferating in the presence of G-CSF+IL-11 was significantly higher than in the presence of G-CSF alone, suggesting that IL-11 recruited dormant leukemic progenitors into the cell cycle. Flow cytometric analysis revealed that all types of AML blast cells (M0 approximately M6) ubiquitously expressed gp130, although the level of expression was significantly higher in M5 cells. In contrast, expression of the IL-11 receptor alpha chain (IL-11Ralpha) varied between FAB types. Blast cells obtained from M1, M3 and M5 patients showed higher levels of expression, with M5 cells showing the strongest expression. Interestingly, the leukemic progenitor cells for which proliferation was synergistically enhanced by IL-11 had significantly higher expression of both IL-11Ralpha and gp130. These results suggest that administration of IL-11 in vivo may stimulate the proliferation of leukemic progenitor cells, particularly M5 cells, in the presence of G-CSF, and that the responsiveness of L-CFU to IL-11 may be predicted by a simple receptor assay.

Interleukin-2 inhibits HIV-1 replication in human macrophages by modulating expression of CD4 and CC-chemokine receptor-5.

To determine the effect of recombinant human interleukin (IL)-2 on HIV-1 replication and macrophage colony stimulating factor (M-CSF) production by HIV-1-infected monocyte-derived macrophages (MDM). Therapeutic use of IL-2 increases the number and function of CD4+ T cells. IL-2 also increases M-CSF production and M-CSF receptor expression by human monocytes, but the subsequent effects on HIV-1 replication in MDM have yet to be determined. MDM from HIV-1-seronegative donors were cultured in the presence and absence of IL-2 and infected with HIV-1. Harvested supernatants were monitored for reverse transcriptase activity and M-CSF production. Reverse transcriptase activity was significantly lower when MDM cultures were treated with IL-2 for 10 days prior to infection with HIV-1. IL-2 did not stimulate production of inhibitory chemokines or cytokines, but FACS analysis revealed that expression of CD4, the primary HIV-1 receptor, and CC-chemokine receptor-5, a coreceptor used by macrophage-tropic viruses, are down modulated after treatment with IL-2. IL-2 may not only be of benefit in restoring immune function in AIDS patients, but may also help to prevent the infection of healthy macrophages by decreasing their expression of HIV-1 receptors.

Interleukin 10 Inhibits Vascular Permeability Factor Release by Peripheral Blood Mononuclear Cells in Patients with Lipoid Nephrosis

The effect of recombinant human interleukin 10 (IL-10) on the in vitro secretion of vascular permeability factor (VPF) of peripheral blood mononuclear cells (PBMC) was evaluated in patients with lipoid nephrosis (LN). Significantly increased spontaneous and concanavalin A (ConA) stimulated secretion of VPF was detected in PBMC cultures of LN patients with the nephrotic syndrome as compared with those of normal controls. In the present study, IL-10 inhibited the ConA-stimulated VPF secretion of PBMC in LN. A significant inhibitory effect of IL-10 was observed in cultures of PBMC from LN and IgA nephropathy patients as well as from normal persons. When both ConA and anti-IL-10 antibody were added together to the PBMC, a further increase in ConA-stimulated secretion of VPF occurred. These data suggest that IL-10 is a powerful inhibitor of the secretion of VPF in LN patients.

Lnterleukin-13 Inhibits Cytokine Secretion by Blood Monocytes from Patients with IgA Nephropathy

In this study we have examined the effects of recombinant human interleukin (IL)-13 on peripheral blood monocytes (PBM) from patients with IgA nephropathy (IgAN). Significantly increased spontaneous and lipopolysaccharide (LPS)-stimulated secretion of tumor necrosis factor-alpha (TNF) and IL-8 was determined in PBM cultures of IgAN patients compared to those of normal controls. In the present study, IL-13 inhibited the spontaneous as well as the LPS-stimulated cytokine secretion of PBM in IgAN. Significant inhibitory effect of IL-13 was observed in cultures of PBM from IgAN patients as well as from normal persons. When both LPS and anti-IL-13 antibody were added together to the PBM, a further increase of LPS-enhanced secretion of cytokines occurred. Taken together, these results indicate that IL-13 down-regulates the secretion of TNF and IL-8 from IgAN PBM.

IL-4 and TNF-α-MEDIATED PROLIFERATION OF THE HUMAN MEGAKARYOCYTIC LINE M-O7E IS REGULATED BY INDUCED AUTOCRINE PRODUCTION OF GM-CSF

In this study, the authors examined the effects of recombinant human interleukin 4 (rhIL-4) and recombinant human tumour necrosis factor alpha (rhTNF-α) alone or in combination on proliferation of the human cytokine dependent myeloid cell line, M-O7e. While rhIL-4 or rhTNF-α alone induced only a weak proliferative response, a synergistic proliferative signal was clearly evident on stimulation of cells with a combination of both cytokines. The stimulatory effect of rhTNF-α is mediated predominantly by the 55-kDa TNF receptor because the agonistic monoclonal antibody Htr-9 and the Trp 32Thr 86TNF-α mutant protein specific for this receptor type produced similar results to rhTNF-α. In contrast, the Asn 143Arg 145TNF-α mutant protein specific for the 75-kDa TNF receptor produced only minimal proliferation of M-O7e cells. Using RT-PCR, we found that rhTNF-α rapidly and strongly induced granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA production, while rhIL-4 was a slow and less efficient inducer of GM-CSF mRNA. However, there was little evidence of the TNF-α/IL-4 combination acting synergistically on GM-CSF mRNA production as the levels of GM-CSF mRNA increased only marginally compared with IL-4 or TNF-α alone. Thus, the observed synergistic effect of TNF-α/IL-4 costimulation of M-O7e cells seems to be mediated via induction of GM-CSF secretion rather than an enhanced production of GM-CSF mRNA. Higher levels of GM-CSF were detectable in supernatants of cells treated with both rhIL-4 and rhTNF-α than in cells stimulated with either cytokine alone. Furthermore, addition of a neutralising antibody against GM-CSF abrogated the observed synergistic effect of rhIL?4 and rhTNF-α treatment, indicating that the rhIL-4/THF-α combination acts to significantly increase GM-CSF release which then acts in an autocrine manner to enhance the proliferation of M-O7e cells.

Differential effects of recombinant human interleukin-13 on the in vitro growth of human haemopoietic progenitor cells.

Effects of recombinant human interleukin (IL)-13 on in vitro haemopoiesis from non-adherent mononuclear cells (NAMC) or highly enriched CD34+ cells of human cord blood (CB) were studied. IL-13 significantly increased megakaryocyte (MK) colony formation from either NAMC or CD34+ cells cultured in a plasma clot system supplemented with aplastic anaemia serum (AAS) and phytohaemagglutinin-stimulated human peripheral blood leucocyte-conditioned medium (PHA-LCM) in a dose-dependent manner. Experiments using a modified plasma clot culture, in which normal AB serum and various cytokines were added to replace AAS and PHA-LCM, demonstrated an increased MK colony number in the presence of IL-13, especially in combination with IL-3. However, IL-13 had no stimulatory effect, but rather a slight inhibitory effect in some cases on granulocyte-macrophage (GM) colony formation in both plasma clot cultures. Furthermore, the growth of GM progenitor cells in a methylcellulose culture system in the presence of IL-3, GM-CSF, Epo, G-CSF or in combination was significantly inhibited by the addition of IL-13. On the other hand, high concentrations (100 ng/ml) of IL-13 were needed to cause a slight inhibition on the growth of BFU-E-derived colonies under the same methylcellulose culture. These results indicate that IL-13, alone and synergistically with the effect of IL-3, promotes MK colony formation, but it inhibits the growth of GM and erythroid progenitor cells in vitro.

The effects of recombinant human interleukin (IL)-1 alpha, IL-1 beta or IL-6 on hypothalamo-pituitary-adrenal axis activation.

We have investigated the effects of recombinant human interleukin (IL)-1 alpha, IL-1 beta and IL-6 on the activation of the hypothalamo-pituitary-adrenal axis. We have determined the effects of a single i.p. injection of cytokine on circulating ACTH and corticosterone levels, corticotrophin-releasing factor (CRF) mRNA in the parvocellular cells of the paraventricular nucleus and pro-opiomelanocortin (POMC) mRNA in the anterior pituitary at both 4 h and 24 h after injection. IL-1 alpha had no effect on any of the parameters measured at either time-point. In contrast, IL-1 beta increased CRF mRNA in the parvocellular paraventricular nucleus and POMC mRNA in the anterior pituitary 4 h after injection. Plasma ACTH and corticosterone were increased at 4 h and circulating ACTH was still increased at 24 h after treatment with IL-1 beta. IL-6 had no effect on message levels but did increase circulating ACTH and corticosterone levels both 4 h and 24 h after injection. The mechanism responsible for the increase in circulating ACTH after IL-6 injection is unclear but would appear to be different from that which is activated by IL-1 beta which also results in increased CRF and POMC gene expression.

Cytokine Regulation of Proliferation and ICAM-1 Expression of Human Dermal Microvascular Endothelial Cells In Vitro

The effects of recombinant human interleukin 1 alpha (IL-1 alpha), interleukin 1 beta (IL-1 beta), interleukin 6 (IL-6), granulocyte/macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor-alpha (TNF) on the cell proliferation and the expression of intercellular adhesion molecule-1 (ICAM-1) were assessed in cultured human dermal microvascular endothelial cells (HDMEC). IL-1 alpha and IL-1 beta stimulated the proliferation of HDMEC in a dose-dependent manner, whereas in control experiments using human umbilical vein endothelial cells (HUVEC), IL-1 alpha and IL-1 beta did not stimulate HUVEC growth. Also GM-CSF stimulated the proliferation of HDMEC, whereas IL-6 did not affect endothelial cell growth in vitro. Treatment with IL-1 alpha, IL-1 beta, and TNF markedly increased the expression of ICAM-1 on HDMEC in a time- and dose-dependent manner, in contrast to IL-6 and GM-CSF. By pre-embedding immunoelectron microscopy, membrane-bound expression of ICAM-1 was visualized with pronounced labeling in areas of microvillous cell protrusions. The TNF-induced expression of ICAM-1 on HDMEC was blocked by co-incubation with a neutralizing antibody against TNF, but not with neutralizing antibodies against IL-1 alpha, IL-1 beta, or IL-6. In addition, co-incubation of HDMEC with TNF and the retinoid compound acitretin, dexamethasone, or indomethacin did not abrogate the TNF-induced ICAM-1 expression. These results disclose IL-1 as a major, multifunctional endothelial cell-targeted cytokine and further confirm the concept that pro-inflammatory cytokines exert differential regulatory effects on dermal microvascular endothelial cell proliferation and expression of cell-adhesion molecules.

Interleukin 3 Stimulation Enhances Tyrosine Phosphorylation and Proliferative Activity of Human Fetal Thymocytes and Leukemic T-cell Precursors at Multiple Developmental Stages of T-Cell Ontogeny

The purposes of this study were to elucidate the biologic effects of recombinant human interleukin 3 (rhIL-3) on leukemia T-cell precursors and fetal thymocytes corresponding to discrete and sequential developmental stages of human T-cell ontogeny, and to examine the biochemical nature of the IL-3 receptor linked transmembrane signal in human T-cell precursor populations. The specific binding of biosynthetically labeled 35S-rhIL-3 to leukemic T-cell precursors from T-lineage ALL patients was initially investigated. In 5 of 9 cases, the binding of 35S-rhIL-3 was significantly blocked by excess cold rhIL-3, and the percentage of inhibitable binding ranged from 40% to 64% (mean ± SE = 53 ± 4%). In these cases, 5–13 femtomols (mean + SE = 7.0 ± 1.5 fms) of 35S-rhIL-3/107 cells were specifically bound. rhIL-3 stimulated in a dose dependent fashion the in vitro clonal proliferation of leukemic T-cell precursors with composite immunophenotypes corresponding to the developmental stages of prothymocytes/double negative immature thymocytes (3 of 5 cases) and corticothymocytes/double positive CD4+CD8+ immature thymocytes (4 of 4 cases). By contrast, leukemic T-cell precursors from 2 T-lineage ALL cases with a composite immunophenotype of medullary thymocytes/single positive CD4+CD8−/CD4−CD8+ mature thymocytes did not show an enhanced proliferative activity after stimulation with increasing concentrations of rhIL-3. All of the 5 cases with significant 35S-rhIL-3 binding and none of the 4 cases with no 35S-rhIL-3 binding showed a proliferative response to rhIL-3. Thus, there was a high correlation between 35S-rhIL-3 binding and proliferative response of leukemic T-cell precursors in colony assays, indicating that functional IL-3 receptors were detected in ligand binding assays. rhIL-3 also stimulated the proliferation of immature double positive CD4+CD8+ thymocytes from 9 of 10 fetal thymuses without inducing differentiation and with no selective advantage for the development of CD4+CD8− or CD4− or CD4−CD8+ single positive thymocytes. The observed differences in IL-3 responsiveness among T-cell precursor populations at distinct developmental stages indicates that IL-3 receptors may be expressed only during the early stages of human T-cell ontogeny preceding the negative or positive selection events within the thymic microenvironment. Stimulation of fetal thymocytes with rhIL-3 resulted in enhanced tyrosine phosphorylation of 8 distinct cellular substrates with molecular weights of 44 kDa, 55 kDa, 60 kDa, 69 kDa, 98 kDa, 123 kDa, 150 kDa and 190 kDa, but it did not result in stimulation of phosphoinositide turnover and increased Ins-1,4,5-P3 production. The induction of tyrosine phosphorylation by rhIL-3 was augmented by further crosslinking surface bound IL-3 molecules with an anti-IL-3 antibody and it was abrogated by the tyrosine kinase inhibitor genistein. The ligation of the CD45 protein tyrosine phosphatase markedly suppressed the tyrosine phosphorylation of specific substrates induced by IL-3 stimulation. Thus, the mitogenic transmembrane signal triggered by the engagement of the IL-3 R on human T-cell precursors is linked to a functional protein tyrosine kinase (PTK)/protein tyrosine phosphatase (PTP) regulatory pathway. Taken together, these results indicate that IL-3 may have an important growth regulatory role in early stages of human T-cell ontogeny.

Effects of human recombinant interleukin 2 on in vitro tumor cytotoxicity in dogs

SUMMARY In these studies, the effects of recombinant human interleukin 2 (rHuil-2) were examined on in vitro tumor cytotoxicity by canine blood lymphocytes obtained from peripheral vessels through use of a chromium release microcytotoxicity assay. Cytotoxic activity by lymphokine-activated killer cells was significantly increased, compared with that by untreated lymphocytes in a dose-dependent manner. The maximal effect was attained with 300,000 IU of rHuil-2/ml. Lymphokine-activated killing also was dependent on the duration of incubation with rHuil-2. After 1 day of rHuil-2 incubation, cytotoxicity was significantly increased, compared with that of untreated lymphocytes. Of the 3 times examined, cytotoxicity peaked after 3 days of rHuil-2 incubation. High levels of cytotoxic activity were still present at 7 days of incubation. Numbers of granular lymphocytes increased over the times examined. These results demonstrate functional and morphologic changes in canine peripheral blood lymphocytes obtained from peripheral vessels after incubation with rHuil-2 in a dose- and time-dependent manner.