scholarly journals Bobel-24 Activity against Cryptosporidium parvum in Cell Culture and in a SCID Mouse Model

2007 ◽  
Vol 52 (3) ◽  
pp. 1150-1152 ◽  
Author(s):  
Cristina Rueda ◽  
Soledad Fenoy ◽  
Fernando Simón ◽  
Carmen del Aguila
Keyword(s):  
Cell Culture ◽  
Mouse Model ◽  
Cryptosporidium Parvum ◽  
Scid Mouse ◽  
In Vitro Growth ◽  
Scid Mouse Model ◽  
First Time

ABSTRACT The anticryptosporidial activity of Bobel-24 (2,4,6-triiodophenol) was studied for the first time, resulting in a reduction of the in vitro growth of Cryptosporidium of up to 99.6%. In a SCID mouse model of chronic cryptosporidiosis, significant differences (P < 0.05) in oocyst shedding were observed in animals treated with 125 mg/kg/day. These results merit further investigation of Bobel-24 as a chemotherapeutic option for cryptosporidiosis.

Role of Il-2, Il-7, and IL-15 in T Cell Mediated Graft-vs-Leukemia Against Human Acute Lymphoblastic Leukemia in a NOD/scid Chimeric Mouse Model.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5256-5256
Author(s):  
Doug Cipkala ◽  
Kelly McQuown ◽  
Lindsay Hendey ◽  
Michael Boyer
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
T Cells ◽  
Mouse Model ◽  
Scid Mouse ◽  
In Vitro Cytotoxicity ◽  
Scid Mouse Model

Abstract The use of cytotoxic T-lymphocytes (CTL) has been attempted experimentally with various tumors to achieve disease control. Factors that may influence GVT include CTL cytotoxicity, ability to home to disease sites, and survival of T cells in the host. The objective of our study is to evaluate the GVL effects of human alloreactive CTL against ALL in a chimeric NOD/scid mouse model. CTL were generated from random blood donor PBMCs stimulated with the 697 human ALL cell line and supplemented with IL-2, -7, or -15. CTL were analyzed for in vitro cytotoxicity against 697 cells, phenotype, and in vitro migration on day 14. NOD/scid mice were injected with 107 697 ALL cells followed by 5x106 CTL. Mice were sacrificed seven days following CTL injection and residual leukemia was measured in the bone marrow and spleen via flow cytometry. The ratios of CD8/CD4 positive T cells at the time of injection were 46/21% for IL-2, 52/31% for IL-7, and 45/14% for IL-15 cultured CTL (n=13). Control mice not receiving CTL had a baseline leukemia burden of 2.01% and 0.15% in the bone marrow and spleen, respectively (n=15). Mice treated with IL-15 cultured CTL had a reduction in tumor burden to 0.2% (n=13, p=0.01) and 0.05% (n=13, p=0.01) in bone marrow and spleen, respectively. Those treated with IL-2 or IL-7 cultured CTL showed no significant difference in leukemia burden in either the bone marrow (IL-2 1.28%, Il-7 5.97%) or spleen (IL-2 0.4%, IL-7 0.33%). No residual CTL could be identified in the bone marrow or spleen at the time of sacrifice in any CTL group. CTL grown in each cytokine resulted in similar in vitro cytotoxicity at an effector:target ratio of 10:1 (IL-2 41.3%, IL-7 37.7%, IL-15 45.3%, n=12–15, p&gt;0.05 for all groups) and had statistically similar intracellular perforin and granzyme-B expression. In vitro CTL migration to a human mesenchymal stem cell line was greatest with IL-15 CTL (30.5%, n=4), followed by IL-7 CTL (18.9%, n=4), and least in IL-2 CTL (17.9%, n=4), though the differences were not significant. In vitro CTL migration was analyzed to an SDF-1α gradient as CXCR4/SDF-1α interactions are necessary for hematopoietic progenitor cell homing to the bone marrow. IL-15 cultured CTL showed the highest migration (48.8%, n=8) as compared to IL-2 (21.7%, n=6, p=0.048) or IL-7 CTL (35.9%, n=8, p&gt;0.05). However, surface expression of CXCR4 measured by flow cytometry was significantly higher in IL-7 CTL (89.4%, n=9) compared to IL-2 CTL (52.2%, n=9, p&lt;0.001) and IL-15 CTL (65.4%, n=10, p=0.002). Experiments are currently underway to further evaluate the role of CXCR4/SDF-1α in GVL. Preliminary in vivo experiments do not suggest any significant differences in CTL engraftment when evaluated at 24 hours post injection. Expression of the anti-apoptotic bcl-2 protein was greatest on IL-7 (MFI=5295, n=13) and IL-15 (MFI=4865, n=14) when compared to IL-2 CTL (MFI=3530, n=13, p=0.02 vs. IL-7, p=0.05 vs. IL-15), suggesting an increased in vivo survival ability. We hypothesize that IL-15 cultured CTL have greater GVL effects due to either higher in vivo survival, greater bone marrow homing efficiency, or both. Future experiments are planned to evaluate in vivo administration of IL-2 to enhance CTL survival in the host. In conclusion, IL-15 cultured CTL had significantly greater in vivo GVL effects compared to IL-2 and IL-7 CTL in the NOD/scid mouse model. This model can be utilized to evaluate the mechanism of T cell mediated GVL against ALL and potentially other human malignancies.

Therapeutic Efficacy of Natural Compounds From Viscum Album l. In Acute Lymphoblastic Leukemia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3261-3261
Author(s):  
Catharina I. Delebinski ◽  
Sebastian Jaeger ◽  
Kristin Kemnitz-Hassanin ◽  
Arend von Stackelberg ◽  
Günter Henze ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Scid Mouse ◽  
Lymphoblastic Leukemia ◽  
Viscum Album ◽  
Mitochondrial Potential ◽  
Scid Mouse Model ◽  
Mistletoe Extracts ◽  
First Time

Abstract Abstract 3261 Viscum album L. (mistletoe) is one of the most widely used complementary cancer therapies. Due to their low solubility, aqueous extracts contain hardly any triterpenes which are known to possess anti-tumoral properties. Using cyclodextrins it was possible to solubilisate mistletoe triterpenes (mainly oleanolic acid (OA)) and achieve a plant extract with high levels of OA and mistletoe lectins (ML). In the present study, we determined for the first time the effect of clearly defined mistletoe extracts against human acute lymphoblastic leukemia (ALL) in vitro and in vivo. These mistletoe extracts contain either lectins (aqueous extract, viscum) or cyclodextrin solubilised triterpenes (STE) such as oleanolic - and betulinic acid and combinations thereof (viscumTT). We used the C.B-17/SCID mouse model and tested efficacy and mechanisms of the treatment with these preparations in vitro and in vivo. The human leukemia cell line NALM-6 was incubated with increasing concentrations of mistletoe preparations (10-60 μg/ml OA; 0.8–8 ng/ml ML) and tested for their cytotoxicity in vitro. Apoptosis was determined using mitochondrial potential, DNA fragmentation and Annexin/PI assays. In vivo efficacy was determined in the C.B-17/SCID mouse model. For this purpose, 1×106 NALM-6 cells were injected IV into groups of C.B-17/SCID mice (n=8) and STE extracts were administered three times per week for 14 days by intraperitoneal (IP) injection. Viscum album L. extracts inhibited cell proliferation and show cytotoxic properties in vitro. The highest level of apoptosis with a decrease of the mitochondrial potential was observed with STE preparation at a concentration of 50 μg/ml OA and for lectin-treated cells for 4.7 ng/ml (IC50). To exclude an unwanted cell death via necrosis, LDH release was measured after 4h of incubation with different doses and extracts of Viscum album L without significant LDH release. Based on these data, we investigated the effect of Viscum album L. extracts in vivo. For this purpose 40 mg/kg/day oleanolic acid (STE), 3 μg/kg/day lectin (viscum) or a combination thereof (viscumTT) were administered IP. In line with the in vitro results, mice treated with viscumTT showed a significant longer survival. Mice receiving PBS had a mean survival time of 38 days whereas mice treated with viscumTT had a mean survival of 50,5 days (p=0,005). In summary, we demonstrate for the first time that either solubilised triterpenes or lectins and combinations thereof induce dose- and time-dependent apoptosis in the ALL cell line NALM-6. Based on the in vivo data we believe that triterpene containing Viscum album L. extracts may possess an impressive therapeutic potential. Thus, our investigations provide an important base line for the design of further experimental studies and clinical trials to investigate the effects of individual components and potential synergisms in ALL. Disclosures: No relevant conflicts of interest to declare.

Growth Potential of Human Leukemia Blast Cells in SCID Mice and Association with Prognosis of Human Acute Leukemias.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1900-1900
Author(s):  
Ying Yan ◽  
Peter Steinherz ◽  
Xiuqin Guan ◽  
Ann Jakubowski ◽  
Joesph P. McGuirk ◽  
...  
Keyword(s):  
Mouse Model ◽  
Scid Mouse ◽  
Scid Mice ◽  
Growth Potential ◽  
Leukemia Cells ◽  
Human Leukemia ◽  
Acute Leukemias ◽  
Blast Cells ◽  
Scid Mouse Model

Abstract We have described a severe combined immunodeficiency (SCID) mouse model that permits the subcutaneous growth of primary human acute leukemia blast cells into a measurable subcutaneous nodule which may be followed by the development of disseminated disease. Utilizing the SCID mouse model, we examined the ability of patient-derived leukemic blasts to generate leukemic growth in the animals after subcutaneous inoculation without conditioning treatment. Leukemia blasts derived from 133 patients with acute leukemias, (67 acute lymphoblastic leukemia (ALL) and 66 acute myeloid leukemia (AML)), were inoculated into SCID mice. The leukemias displayed three distinct growth patterns: "aggressive", "indolent", or "no tumor growth". Out of 133 patients, leukemia samples 46 (34.5%) displayed an aggressive growth pattern, 14 (10.5%) indolent growth and 73 (55.0%) did not grow in SCID mice. The growth probability of leukemias from relapsed and/or refractory disease was 3 fold higher than that from patients with newly diagnosed disease. Serial observations found that leukemic blasts from the same individual, which did not initiate tumor growth at initial presentation and/or at early relapse, may engraft and grow in the later stages of disease, suggesting that the ability of leukemia cells for engraftment and proliferation was gradually acquired following the process of leukemia progression. Nine autonomous growing leukemia cell lines were established in vitro. These displayed an aggressive proliferation pattern, suggesting a possible correlation between the capacity of human leukemia cells for autonomous proliferation in vitro and an aggressive growth potential in SCID mice. It was demonstrated that patients whose leukemic blasts displayed an aggressive growth and dissemination pattern in SClD mice had a poor clinical outcome in patients with ALL as well as AML. Patients whose leukemic blasts grew indolently or whose leukemia cells failed to induce growth had a significantly longer DFS and more favorable clinical course.

scholarly journals P38 mitogen activated protein kinase inhibitor improves platelet in vitro parameters and in vivo survival in a SCID mouse model of transfusion for platelets stored at cold or temperature cycled conditions for 14 days

PLoS ONE ◽  
2021 ◽  
Vol 16 (5) ◽  
pp. e0250120
Author(s):  
Andrey Skripchenko ◽  
Monique P. Gelderman ◽  
Jaroslav G. Vostal
Keyword(s):  
Mouse Model ◽  
Cold Storage ◽  
Scid Mouse ◽  
Room Temperature ◽  
Human Platelet ◽  
Human Platelets ◽  
Bacterial Proliferation ◽  
Scid Mouse Model

Platelets for transfusion are stored at room temperature (20–24°C) up to 7 days but decline in biochemical and morphological parameters during storage and can support bacterial proliferation. This decline is reduced with p38MAPK inhibitor, VX-702. Storage of platelets in the cold (4–6°C) can reduce bacterial proliferation but platelets get activated and have reduced circulation when transfused. Thermocycling (cold storage with brief periodic warm ups) reduces some of the effects of cold storage. We evaluated in vitro properties and in vivo circulation in SCID mouse model of human platelet transfusion of platelets stored in cold or thermocycled for 14 days with and without VX-702. Apheresis platelet units (N = 15) were each aliquoted into five storage bags and stored under different conditions: room temperature; cold temperature; thermocycled temperature; cold temperature with VX-702; thermocycled temperature with VX-702. Platelet in vitro parameters were evaluated at 1, 7 and 14 days. On day 14, platelets were infused into SCID mice to assess their retention in circulation by flow cytometry. VX-702 reduced negative platelet parameters associated with cold and thermocycled storage such as an increase in expression of activation markers CD62, CD63 and of phosphatidylserine (marker of apoptosis measured by Annexin binding) and lowered the rise in lactate (marker of increase in anaerobic metabolism). However, VX-702 did not inhibit agonist-induced platelet aggregation indicating that it does not interfere with platelet hemostatic function. In vivo, VX-702 improved initial recovery and area under the curve in circulation of human platelets infused into a mouse model that has been previously validated against a human platelet infusion clinical trial. In conclusion, inhibition of p38MAPK during 14-days platelet storage in cold or thermocycling conditions improved in vitro platelet parameters and platelet circulation in the mouse model indicating that VX-702 may improve cell physiology and clinical performance of human platelets stored in cold conditions.

Combination Therapy of Humanized Chimeric-Anti-Human VLA4 Monoclonal Antibodies and Anti-Cancer Drugs for AML.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4518-4518
Author(s):  
Takuya Matsunaga ◽  
Kazunori Kato ◽  
Maki Tanaka ◽  
Yukari Masuta ◽  
Kageaki Kuribayashi ◽  
...  
Keyword(s):  
Mouse Model ◽  
Scid Mouse ◽  
Variable Region ◽  
Human Leukemia ◽  
Region Gene ◽  
Anti Cancer ◽  
Variable Region Gene ◽  
Scid Mouse Model

Abstract Bone marrow (BM) minimal residual disease causes relapse after chemotherapy in AML. We have previously reported that VLA4-positive leukemic cells acquired resistance to drug-induced apoptosis through the PI-3K/AKT/Bcl-2 signaling pathway, which is activated by the interaction of VLA4 and fibronectin on BM stromal cells. This resistance was negated by mouse-anti-human VLA4Ab (mouse VLA4Ab). In human leukemia SCID mouse model, we demonstrated a 100% survival rate with combination of mouse VLA4 Ab and AraC, while with AraC alone, only slight prolongation of survival was attained. In clinical study, overall survival at 5 years was 90% for 10 VLA4− patients and 25% for 15 VLA4+ patients (Matsunaga T et al, Nature Med 2003, 9, 1158–1165). In the present study, to perform the translational research, we first examined the myelosuppressive effect of the combination of rat-anti-mouse VLA4 Ab and AraC in C57/BL6 mice, and found that CBC data were almost the same as those of the mice treated with AraC alone. We next produced humanized chimeric-anti-human VLA4 Ab (chimeric-VLA4Ab), and examined its efficacy in combination with anti-cancer drugs in vitro and in vivo (human leukemia SCID mouse model). Chimeric-VLA4Abs were produced as follows: (i) total RNA of mouse VLA4 Abs were extracted from two hybridomas (SG/17 and SG/73), (ii) cDNA were synthesized by reverse transcriptase, (iii) variable region gene of mouse VLA4 Abs were amplified by 5′RACE method, (iv) TA cloning of amplified gene was performed, (v) sequence of mouse VLA4Abs gene was determined, (v) cloned variable region gene of mouse VLA4 Abs and constant region gene of human IgG1 were inserted into expression vector, and the expression vector was transfected into 293T cells, (vi) supernatant of the 293T cells was collected and purified to obtain chimeric-VLA4 Abs. The effects of chimeric-VLA4 Abs thus obtained in in vitro and in vivo (human leukemia SCID mouse model) were comparable to those of mouse VLA4 Abs. To perform the clinical study, we are presently producing the GMP-graded chimeric-VLA4 Abs.

Significant in Vivo Efficacy of the SINE KPT-330 in Mouse Models of CLL.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2452-2452
Author(s):  
Rosa Lapalombella ◽  
Virginia Goettl ◽  
Katie Williams ◽  
Larissa Tangeman ◽  
Shruti Jha ◽  
...  
Keyword(s):  
Mouse Model ◽  
Stromal Cells ◽  
Nuclear Export ◽  
Scid Mouse ◽  
Phase 1 ◽  
Vehicle Control ◽  
Leukemia Cells ◽  
Scid Mouse Model

Abstract Abstract 2452 CRM1 (chromosomal regional maintenance 1) or XPO1 (exportin 1) is the major protein that mediates nuclear export. XPO1 has been shown to be over-expressed in cancer cells leading to ineffective cytoplasmic localization of multiple tumor suppressor genes such us p53, FOX03a, and Iκ-β. Chronic Lymphocytic Leukemia (CLL) is characterized by disrupted apoptosis caused both by microenviromental stimuli and aberrant activation of survival pathways including PI3K/Akt, NF-kB, and p53. We previously showed that first generations Selective Inhibitor of Nuclear Export (SINE), KPT-185 and KPT-251, specifically bind to and block XPO1 function resulting in re-activation of tumor suppressor pathways and apoptosis in CLL. Furthermore, we showed that KPT-251 at 75mg/Kg, 3 times per week (QODX3) slowed disease progression, and improved overall survival in the Eμ-TCL1-SCID mouse model of CLL (median OS = 130 and 72 days for KPT-330 and vehicle respectively; p=0.01) with minimal weight loss or other toxicities. The goal of this study was to determine the efficacy of KPT-330, a new generation SINE currently in Phase 1 human studies, with improved pharmacokinetic and oral bioavailability, in the Eμ-TCL1-SCID mouse model of CLL. We first compared the in-vitro activity of KPT-330 to KPT-251. KPT-330 induced improved levels of cytotoxicity on CLL cells compared to KPT-251 with retained selective cytotoxicity against tumor cells when compared to peripheral blood mononuclear cells and isolated B, and T cells. Similar to KPT-251, KPT-330 showed enhanced cytotoxic potency on CLL cells under coculture conditions with stromal cells compared to CLL alone (p<0.05). Interestingly KPT-330 prevented CPG induced proliferation of CLL cells (p<0.005) and migration of CLL cells driven by CXCL12 and CXCL13, homing factors for hematopoietic cells to the marrow that are constitutively secreted by bone marrow stromal cells. KPT-330 was also more effective than KPT-251 in mediating cytotoxicity of murine TCL1+ cells in-vitro. We therefore tested the ability of KPT-330 to slow disease progression in Eμ-TCL1-SCID mouse model of CLL in which CD19+ TCL1 leukemia cells were engrafted into SCID mice via tail vein injection. Treatments began 2 weeks post engraftment. 9 or 10 mice in each treatment group were treated with: a) vehicle control twice weekly (BIW), b) KPT-330 5 mg/kg BIW, c) KPT-330 15 mg/kg BIW d) vehicle control QODX3, e) KPT-330 1 mg/kg QODX3 for 9 weeks followed by 15 mg/kg QODX3, f) KPT-330 3 mg/kg QODX3, g) KPT-330 10 mg/kg QODX3 for up to a maximum of 36 weeks. In surviving mice, treatment was stopped after 36 weeks and animals were followed up for 30 additional days. Overall survival (OS) was the primary endpoint. Within the BIW groups, only KPT-330 at 15 mg/kg (median OS = undefined p=0.0002) showed a significant improvement over vehicle (median OS = 98 days) with 100% of the mice still alive at the end of the study (week 40). Within the QODX3 groups, KPT-330 10 and 15 mg/kg QODX3 (median OS = undefined, 213 days respectively) both showed a significant improvement (p<0.0001) over vehicle (median OS = 98.5). No improvement in survival compared to vehicle treated animals was induced by KPT-330 3 mg/kg. KPT-330 was well tolerated in mice, weight loss suggestive of drug toxicity was seen at the beginning of the study only in groups treated with KPT-330 at doses of >10 mg/kg QODX3 but was fully reversed by the end of the study and did not appear to adversely affect the animals. Moreover, KPT-330 at doses of >10 mg/kg BIW and QODX3 significantly prevented an increase in circulating CLL cells compared to vehicle. To further validate KPT-330 in mice with leukemic phase (ie, very high tumor burdens), 20 additional mice were engrafted with CD19+ TCL1 leukemia cells and treatment with vehicle or 15 mg/kg KPT-330 (QoDx3/wk) was initiated 70 days post engraftment. Mice treated with KPT-330 had a significant survival advantage (p=0.0008, median OS = undefined; 99 days, for KPT-330 and vehicle respectively). KPT-330 was strikingly active in prolonging survival of TCL1-SCID mice. Using CLL as a disease model we validate the clinical effectiveness of targeting XPO1 with use of SINE, with favorable therapeutic index in-vivo. These data strongly support the ongoing Phase 1 clinical studies of KPT-330 in patients with advanced hematologic malignancies including CLL. Disclosures: Sandanayaka: Karyopharm Therapeutics: Employment. Shechter:Karyopharm Therapeutics: Employment. McCauley:Karyopharm Therapeutics Inc: Employment. Shacham:Karyopharm Therapeutics: Employment. Kauffman:Karyopharm Therapeutics Inc: Employment.

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