Publications by authors named "Shuguang Jiang"

32 Publications

Effect of Ionic Liquid Surfactants on Coal Oxidation and Structure.

J Anal Methods Chem 2019 22;2019:1868265. Epub 2019 Apr 22.

School of Mines, China University of Mining & Technology, Xuzhou 221116, Jiangsu, China.

The effects of six ionic liquids with surfactant property (1-hydroxyethyl-3-methyl imidazolium bis(trifluoromethylsulfonyl)imide ([HOEtMIm][NTf]), 1-hydroxyethyl-3-methyl imidazolium tetrafluoroborate ([HOEtMIm][BF]), 1-dodecyl-3- methyl imidazolium bromide ([CMIm]Br), 1-tetradecyl-3- methyl imidazolium bromide ([CMIm]Br), trioctyl methyl ammonium chloride ([N])Cl, and tetraethyl ammonium chloride ([N]Cl)) on the oxidation characteristics and functional groups of coal were studied by means of critical micelle concentration, surface tension, thermogravimetric analysis, temperature-programmed oxidation, and Fourier transform infrared spectroscopy (FTIR) measurements. The lower critical micelle concentration for the ionic liquids except the [N]Cl suggests the favorable surface activity of these ionic liquids. The surface activities of [N]Cl, [CMIm]Br, [CMIm]Br, and [HOEtMIm][NTf] were high, while that of [N]Cl was relatively lower. The thermal stabilities of [HOEtMIm][NTf] and [HOEtMIm][BF] were high, while those of [N]Cl and [N]Cl were lower. The oxidation activities of ionic liquid-mixed coals were weakened to different degrees except [N]Cl-mixed coal, because of the poor thermal stability and decomposition of [N]Cl accelerating the coal oxidation. The other five ionic liquids were suitable for inhibiting coal oxidation, particularly the [HOEtMIm][BF] and [HOEtMIm][NTf] with higher inhibition rate, longer inhibition time, and also better thermal stabilities. The activation energy results further confirmed such inhibition effect. The functional group results showed that treatment of ionic liquids on coal can change the contents of hydrogen bonds, aliphatic groups, and aromatic groups in coal. It was inferred that the [HOEtMIm][BF], [HOEtMIm][NTf], and [CMIm]Br were more effectively to affect coal structure and decrease coal oxidation activity.
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http://dx.doi.org/10.1155/2019/1868265DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6501251PMC
April 2019

Analysis of the spatial distribution of collectors in dust scrubber based on image processing.

J Air Waste Manag Assoc 2019 06 11;69(6):764-777. Epub 2019 Apr 11.

d School of Safety Engineering , China University of Mining and Technology , Xuzhou , Jiangsu , People's Republic of China.

The spatial distribution of the collectors in dust scrubber is key in determining the effectiveness of the dust removal process. In the present study, a high-speed camera was used to capture images of the distribution of the collectors. Some of the image information was extracted by image processing, such as the gray mean (GM), the angular second moment (ASM), and the entropy (ENT) from the gray-level co-occurrence matrix of the image. Subsequently, the spatial distribution rules of the collectors were studied by analyzing the spatial proportion, dispersion area, and uniformity and intensiveness of the collectors. It is an intuitive approach and a novel analysis method for the operating state of dust scrubber. The results show that the spatial distribution of the collectors could be better reflected by image processing methods. The dispersion area of the collectors expanded with an increase in the airflow velocity. When the initial liquid level (ILL) was higher, the collectors expanded in an approximate circular shape, and when the ILL was lower the collectors expanded in an approximate sector shape. In general, the variation trend in the spatial proportion enhanced with an increase in ILL and airflow velocity, which is consistent with the uniformity of collectors. When the liquid level was 0-20 mm and the airflow velocity was greater than 6.5 m/sec, the spatial proportion and uniformity of the collectors reached the highest degree. However, the growth rate of the spatial proportion and uniformity of the collectors slowed down and even led to negative growth when the ILL was lower and the airflow velocity was higher. The intensiveness of the collectors was great when the ILL was higher, which was free from the apparent influence of the airflow velocity and the ILL. However, when the ILL was lower, the intensiveness of the collectors was poor, intensifying as the airflow velocity and ILL increased. When the liquid level was -5-10 mm and the airflow velocity was greater than 8 m/sec, the intensiveness of the collectors reached the highest degree, indicating that a liquid level greater than 0 mm and a higher airflow velocity improved the spatial distribution of the collectors. : This paper focuses on the spatial distribution of the collectors in dust scrubber. Some of the image information was extracted by image processing, such as the gray mean of the image, the angular second moment, and the entropy from the gray-level co-occurrence matrix of the image. The spatial distribution rules of the collectors were studied by analyzing the spatial proportion, the dispersion area, and the uniformity and intensiveness of the collectors.
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http://dx.doi.org/10.1080/10962247.2019.1586012DOI Listing
June 2019

Checkpoint Blockade Reverses Anergy in IL-13Rα2 Humanized scFv-Based CAR T Cells to Treat Murine and Canine Gliomas.

Mol Ther Oncolytics 2018 Dec 28;11:20-38. Epub 2018 Aug 28.

Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.

We generated two humanized interleukin-13 receptor α2 (IL-13Rα2) chimeric antigen receptors (CARs), Hu07BBz and Hu08BBz, that recognized human IL-13Rα2, but not IL-13Rα1. Hu08BBz also recognized canine IL-13Rα2. Both of these CAR T cell constructs demonstrated superior tumor inhibitory effects in a subcutaneous xenograft model of human glioma compared with a humanized EGFRvIII CAR T construct used in a recent phase 1 clinical trial (ClinicalTrials.gov: NCT02209376). The Hu08BBz demonstrated a 75% reduction in orthotopic tumor growth using low-dose CAR T cell infusion. Using combination therapy with immune checkpoint blockade, humanized IL-13Rα2 CAR T cells performed significantly better when combined with CTLA-4 blockade, and humanized EGFRvIII CAR T cells' efficacy was improved by PD-1 and TIM-3 blockade in the same mouse model, which was correlated with the levels of checkpoint molecule expression in co-cultures with the same tumor . Humanized IL-13Rα2 CAR T cells also demonstrated benefit from a self-secreted anti-CTLA-4 minibody in the same mouse model. In addition to a canine glioma cell line (J3T), canine osteosarcoma lung cancer and leukemia cell lines also express IL-13Rα2 and were recognized by Hu08BBz. Canine IL-13Rα2 CAR T cell was also generated and tested by co-culture with canine tumor cells and in an orthotopic model of canine glioma. Based on these results, we are designing a pre-clinical trial to evaluate the safety of canine IL-13Rα2 CAR T cells in dog with spontaneous IL-13Rα2-positive glioma, which will help to inform a human clinical trial design for glioblastoma using humanized scFv-based IL-13Rα2 targeting CAR T cells.
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http://dx.doi.org/10.1016/j.omto.2018.08.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6174845PMC
December 2018

Mesoscale behavior study of collector aggregations in a wet dust scrubber.

J Air Waste Manag Assoc 2018 Jan 9;68(1):73-91. Epub 2017 Nov 9.

b School of Chemical Engineering and Technology , China University of Mining and Technology , Xuzhou , People's Republic of China.

In order to address the bottleneck problem of low fine-particle removal efficiency of self-excited dust scrubbers, this paper is focused on the influence of the intermittent gas-liquid two-phase flow on the mesoscale behavior of collector aggregations. The latter is investigated by the application of high-speed dynamic image technology to the self-excited dust scrubber experimental setup. The real-time-scale monitoring of the dust removal process is provided to clarify its operating mechanism at the mesoscale level. The results obtained show that particulate capturing in self-excited dust scrubber is provided by liquid droplets, liquid films/curtains, bubbles, and their aggregations. Complex spatial and temporal structures are intrinsic to each kind of collector morphology, and these are considered as the major factors controlling the dust removal mechanism of self-excited dust scrubbers. For the specific parameters of gas-liquid two-phase flow under study, the evolution patterns of particular collectors reflect the intrinsic, intermittent, and complex characteristics of the temporal structure. The intermittent initiation of the collector and the air hole formation-collapse cyclic processes provide time and space for the fine dust to escape from being trapped by the collectors. The above mesoscale experimental data provide more insight into the factors reducing the dust removal efficiency of self-excited dust scrubbers.

Implications: This paper focuses on the reconsideration of the capturer aggregations of self-excited dust scrubbers from the mesoscale. Complex structures in time and space scales exist in each kind of capturer morphology. With changes of operating parameters, the morphology and spatial distributions of capturers diversely change. The change of the capturer over time presents remarkable, intermittent, and complex characteristics of the temporal structure.
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http://dx.doi.org/10.1080/10962247.2017.1370037DOI Listing
January 2018

Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation.

Protein Cell 2017 07 18;8(7):514-526. Epub 2017 May 18.

Center for Cellular Immunotherapies, University of Pennsylvania Cancer Center, Philadelphia, PA, 19104, USA.

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.
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http://dx.doi.org/10.1007/s13238-017-0422-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5498341PMC
July 2017

A versatile system for rapid multiplex genome-edited CAR T cell generation.

Oncotarget 2017 Mar;8(10):17002-17011

Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.

The therapeutic potential of CRISPR system has already been demonstrated in many instances and begun to overlap with the rapidly expanding field of cancer immunotherapy, especially on the production of genetically modified T cell receptor or chimeric antigen receptor (CAR) T cells. Efficient genomic disruption of multiple gene loci to generate universal donor cells, as well as potent effector T cells resistant to multiple inhibitory pathways such as PD-1 and CTLA4 is an attractive strategy for cell therapy. In this study, we accomplished rapid and efficient multiplex genomic editing, and re-directing T cells with antigen specific CAR via a one-shot CRISPR protocol by incorporation of multiple gRNAs in a CAR lentiviral vector. High efficient double knockout of endogenous TCR and HLA class I could be easily achieved to generate allogeneic universal CAR T cells. We also generated Fas-resistant universal CAR T cells by triple gene disruption. Simultaneous gene editing of four gene loci using the one-shot CRISPR protocol to generate allogeneic universal T cells deficient of both PD1 and CTLA-4 was also attempted.
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http://dx.doi.org/10.18632/oncotarget.15218DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5370017PMC
March 2017

Multiplex Genome Editing to Generate Universal CAR T Cells Resistant to PD1 Inhibition.

Clin Cancer Res 2017 May 4;23(9):2255-2266. Epub 2016 Nov 4.

Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania.

Using gene-disrupted allogeneic T cells as universal effector cells provides an alternative and potentially improves current chimeric antigen receptor (CAR) T-cell therapy against cancers and infectious diseases. The CRISPR/Cas9 system has recently emerged as a simple and efficient way for multiplex genome engineering. By combining lentiviral delivery of CAR and electro-transfer of Cas9 mRNA and gRNAs targeting endogenous TCR, β-2 microglobulin (B2M) and PD1 simultaneously, to generate gene-disrupted allogeneic CAR T cells deficient of TCR, HLA class I molecule and PD1. The CRISPR gene-edited CAR T cells showed potent antitumor activities, both and in animal models and were as potent as non-gene-edited CAR T cells. In addition, the TCR and HLA class I double deficient T cells had reduced alloreactivity and did not cause graft-versus-host disease. Finally, simultaneous triple genome editing by adding the disruption of PD1 led to enhanced antitumor activity of the gene-disrupted CAR T cells. Gene-disrupted allogeneic CAR and TCR T cells could provide an alternative as a universal donor to autologous T cells, which carry difficulties and high production costs. Gene-disrupted CAR and TCR T cells with disabled checkpoint molecules may be potent effector cells against cancers and infectious diseases. .
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http://dx.doi.org/10.1158/1078-0432.CCR-16-1300DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5413401PMC
May 2017

A Chimeric Switch-Receptor Targeting PD1 Augments the Efficacy of Second-Generation CAR T Cells in Advanced Solid Tumors.

Cancer Res 2016 Mar;76(6):1578-90

Division of Pulmonary, Allergy, and Critical Care, Department of Medicine, Perelman School of Medicine, The University of Pennsylvania, Philadelphia, Pennsylvania.

Chimeric antigen receptor (CAR)-modified adoptive T-cell therapy has been successfully applied to the treatment of hematologic malignancies, but faces many challenges in solid tumors. One major obstacle is the immune-suppressive effects induced in both naturally occurring and genetically modified tumor-infiltrating lymphocytes (TIL) by inhibitory receptors (IR), namely PD1. We hypothesized that interfering with PD1 signaling would augment CAR T-cell activity against solid tumors. To address this possibility, we introduced a genetically engineered switch receptor construct, comprising the truncated extracellular domain of PD1 and the transmembrane and cytoplasmic signaling domains of CD28, into CAR T cells. We tested the effect of this supplement, "PD1CD28," on human CAR T cells targeting aggressive models of human solid tumors expressing relevant tumor antigens. Treatment of mice bearing large, established solid tumors with PD1CD28 CAR T cells led to significant regression in tumor volume due to enhanced CAR TIL infiltrate, decreased susceptibility to tumor-induced hypofunction, and attenuation of IR expression compared with treatments with CAR T cells alone or PD1 antibodies. Taken together, our findings suggest that the application of PD1CD28 to boost CAR T-cell activity is efficacious against solid tumors via a variety of mechanisms, prompting clinical investigation of this potentially promising treatment modality.
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http://dx.doi.org/10.1158/0008-5472.CAN-15-2524DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4800826PMC
March 2016

A Combined Raman Spectroscopic and Thermogravimetric Analysis Study on Oxidation of Coal with Different Ranks.

J Anal Methods Chem 2015 23;2015:306874. Epub 2015 Nov 23.

School of Safety Engineering, China University of Mining and Technology, Xuzhou 221116, China.

Raman spectroscopy and nonisothermal thermogravimetric analysis (TGA) measurements have been reported for different rank coals (lignite, bituminous coal, and anthracite) and the relationship between the measurements was examined. It was found that the Raman spectra parameters can be used to characterize structure changes in the different rank coals, such as the band area ratios based on the curve-fitted results. Higher ranked coal was found to have higher values of I GR/I All and I (G + GR)/I All but lower values of I D/I (G+GR), I DL/I (G+GR), I (S + SL)/I (G+GR), and I (GL+GL')/I (G+GR). The oxidation properties of the coal samples were characterized by the reactivity indexes T ig, T 20%, and T max from TGA data which were found to correlate well with the band area ratios of I GR/I All, I (G + GR)/I All, and I (S + SL)/I (G+GR). Based on these correlations, the Raman band area ratios were found to correlate with the oxidation activity of coal providing additional structural information which can be used to understand the changes in the TGA measurements.
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http://dx.doi.org/10.1155/2015/306874DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4670864PMC
December 2015

Affinity-Tuned ErbB2 or EGFR Chimeric Antigen Receptor T Cells Exhibit an Increased Therapeutic Index against Tumors in Mice.

Cancer Res 2015 Sep;75(17):3596-607

Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania. Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.

Target-mediated toxicity is a major limitation in the development of chimeric antigen T-cell receptors (CAR) for adoptive cell therapy of solid tumors. In this study, we developed a strategy to adjust the affinities of the scFv component of CAR to discriminate tumors overexpressing the target from normal tissues that express it at physiologic levels. A CAR-expressing T-cell panel was generated with target antigen affinities varying over three orders of magnitude. High-affinity cells recognized target expressed at any level, including at levels in normal cells that were undetectable by flow cytometry. Affinity-tuned cells exhibited robust antitumor efficacy similar to high-affinity cells, but spared normal cells expressing physiologic target levels. The use of affinity-tuned scFvs offers a strategy to empower wider use of CAR T cells against validated targets widely overexpressed on solid tumors, including those considered undruggable by this approach.
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http://dx.doi.org/10.1158/0008-5472.CAN-15-0159DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4560113PMC
September 2015

Nature of tumor control by permanently and transiently modified GD2 chimeric antigen receptor T cells in xenograft models of neuroblastoma.

Cancer Immunol Res 2014 Nov 7;2(11):1059-70. Epub 2014 Aug 7.

Abramson Family Cancer Research Institute, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania.

Chimeric antigen receptor (CAR) therapy has begun to demonstrate success as a novel treatment modality for hematologic malignancies. The success observed thus far has been with T cells permanently engineered to express chimeric receptors. T cells engineered using RNA electroporation represent an alternative with the potential for similar efficacy and greater safety when initially targeting novel antigens. Neuroblastoma is a common pediatric solid tumor with the potential to be targeted using immunotherapy. We performed xenograft studies in NSG mice in which we assessed the efficacy of both permanently modified and transiently modified CAR T cells directed against the neuroblastoma antigen GD2 in both local and disseminated disease models. Disease response was monitored by tumor volume measurement and histologic examination, as well as in vivo bioluminescence. RNA-modified GD2 CAR T cells mediated rapid tumor destruction when delivered locally. A single infusion of lentivirally modified GD2 CAR T cells resulted in long-term control of disseminated disease. Multiple infusions of RNA GD2 CAR T cells slowed the progression of disseminated disease and improved survival, but did not result in long-term disease control. Histologic examination revealed that the transiently modified cells were unable to significantly penetrate the tumor environment when delivered systemically, despite multiple infusions of CAR T cells. Thus, we demonstrate that RNA-modified GD2 CAR T cells can mediate effective antitumor responses in vivo, and permanently modified cells are able to control disseminated neuroblastoma in xenograft mice. Lack of long-term disease control by RNA-engineered cells resulted from an inability to penetrate the tumor microenvironment.
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http://dx.doi.org/10.1158/2326-6066.CIR-14-0051DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584373PMC
November 2014

Relation of clinical culture method to T-cell memory status and efficacy in xenograft models of adoptive immunotherapy.

Cytotherapy 2014 May 16;16(5):619-30. Epub 2014 Jan 16.

Abramson Family Cancer Research Institute, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA.

Background Aims: Cytotoxic T lymphocytes modified with chimeric antigen receptors (CARs) for adoptive immunotherapy of hematologic malignancies are effective in pre-clinical models, and this efficacy has translated to success in several clinical trials. Many early trials were disappointing in large part because of the lack of proliferation and subsequent persistence of transferred cells. Recent investigations have pointed to the importance of delivering highly proliferative cells, whether of naive or early memory phenotypes.

Methods: We investigated the influence of two common cell culturing methods used in early trials and their relationship to T-cell phenotype and pre-clinical efficacy.

Results: We observed that stimulation with soluble anti-CD3 antibody OKT-3 and high-dose interleukin-2 produces more effector memory-type T cells with shorter average telomeres when compared with cells generated with the use of CD3/CD28 beads. When used in xenograft models of leukemia, bead-stimulated cells proliferated earlier and to a higher degree than those generated with the use of OKT-3/IL2 and resulted in better disease control despite no difference in distribution or migration throughout the mouse. Inclusion of the known successful clinical 4-1BB endodomain in the CAR could not rescue the function of OKT-3/IL-2-cultured cells. T cells isolated from animals that survived long-term (>120 days) retained a central memory-like phenotype and demonstrated a memory response to a large re-challenge of CD19-positive leukemia.

Conclusions: In summary, we confirm that cells with a younger phenotype or higher proliferative capacity perform better in pre-clinical models and that cell culturing influences cell phenotype seemingly independent of the 4-1BB endodomain in the CAR structure.
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http://dx.doi.org/10.1016/j.jcyt.2013.10.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3988256PMC
May 2014

The potent oncogene NPM-ALK mediates malignant transformation of normal human CD4(+) T lymphocytes.

Am J Pathol 2013 Dec;183(6):1971-80

With this study we have demonstrated that in vitro transduction of normal human CD4(+) T lymphocytes with NPM-ALK results in their malignant transformation. The transformed cells become immortalized and display morphology and immunophenotype characteristic of patient-derived anaplastic large-cell lymphomas. These unique features, which are strictly dependent on NPM-ALK activity and expression, include perpetual cell growth, proliferation, and survival; activation of the key signal transduction pathways STAT3 and mTORC1; and expression of CD30 (the hallmark of anaplastic large-cell lymphoma) and of immunosuppressive cytokine IL-10 and cell-surface protein PD-L1/CD274. Implantation of NPM-ALK-transformed CD4(+) T lymphocytes into immunodeficient mice resulted in formation of tumors indistinguishable from patients' anaplastic large-cell lymphomas. Our findings demonstrate that the key aspects of human carcinogenesis closely recapitulating the features of the native tumors can be faithfully reproduced in vitro when an appropriate oncogene is used to transform its natural target cells; this in turn points to the fundamental role in malignant cell transformation of potent oncogenes expressed in the relevant target cells. Such transformed cells should permit study of the early stages of carcinogenesis, and in particular the initial oncogene-host cell interactions. This experimental design could also be useful for studies of the effects of early therapeutic intervention and likely also the mechanisms of malignant progression.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745542PMC
http://dx.doi.org/10.1016/j.ajpath.2013.08.030DOI Listing
December 2013

The potent oncogene NPM-ALK mediates malignant transformation of normal human CD4(+) T lymphocytes.

Am J Pathol 2013 Dec;183(6):1971-80

With this study we have demonstrated that in vitro transduction of normal human CD4(+) T lymphocytes with NPM-ALK results in their malignant transformation. The transformed cells become immortalized and display morphology and immunophenotype characteristic of patient-derived anaplastic large-cell lymphomas. These unique features, which are strictly dependent on NPM-ALK activity and expression, include perpetual cell growth, proliferation, and survival; activation of the key signal transduction pathways STAT3 and mTORC1; and expression of CD30 (the hallmark of anaplastic large-cell lymphoma) and of immunosuppressive cytokine IL-10 and cell-surface protein PD-L1/CD274. Implantation of NPM-ALK-transformed CD4(+) T lymphocytes into immunodeficient mice resulted in formation of tumors indistinguishable from patients' anaplastic large-cell lymphomas. Our findings demonstrate that the key aspects of human carcinogenesis closely recapitulating the features of the native tumors can be faithfully reproduced in vitro when an appropriate oncogene is used to transform its natural target cells; this in turn points to the fundamental role in malignant cell transformation of potent oncogenes expressed in the relevant target cells. Such transformed cells should permit study of the early stages of carcinogenesis, and in particular the initial oncogene-host cell interactions. This experimental design could also be useful for studies of the effects of early therapeutic intervention and likely also the mechanisms of malignant progression.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745542PMC
http://dx.doi.org/10.1016/j.ajpath.2013.08.030DOI Listing
December 2013

Efficient clinical scale gene modification via zinc finger nuclease-targeted disruption of the HIV co-receptor CCR5.

Hum Gene Ther 2013 Mar 6;24(3):245-58. Epub 2013 Mar 6.

Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.

Since HIV requires CD4 and a co-receptor, most commonly C-C chemokine receptor 5 (CCR5), for cellular entry, targeting CCR5 expression is an attractive approach for therapy of HIV infection. Treatment of CD4(+) T cells with zinc-finger protein nucleases (ZFNs) specifically disrupting chemokine receptor CCR5 coding sequences induces resistance to HIV infection in vitro and in vivo. A chimeric Ad5/F35 adenoviral vector encoding CCR5-ZFNs permitted efficient delivery and transient expression following anti-CD3/anti-CD28 costimulation of T lymphocytes. We present data showing CD3/CD28 costimulation substantially improved transduction efficiency over reported methods for Ad5/F35 transduction of T lymphocytes. Modifications to the laboratory scale process, incorporating clinically compatible reagents and methods, resulted in a robust ex vivo manufacturing process capable of generating >10(10) CCR5 gene-edited CD4+ T cells from healthy and HIV+ donors. CD4+ T-cell phenotype, cytokine production, and repertoire were comparable between ZFN-modified and control cells. Following consultation with regulatory authorities, we conducted in vivo toxicity studies that showed no detectable ZFN-specific toxicity or T-cell transformation. Based on these findings, we initiated a clinical trial testing the safety and feasibility of CCR5 gene-edited CD4+ T-cell transfer in study subjects with HIV-1 infection.
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http://dx.doi.org/10.1089/hum.2012.172DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3609630PMC
March 2013

Enhanced function of redirected human T cells expressing linker for activation of T cells that is resistant to ubiquitylation.

Hum Gene Ther 2013 Jan 6;24(1):27-37. Epub 2012 Nov 6.

Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104-5156, USA.

It is likely that the enhancement of signaling after antigenic stimulation, particularly in the tumor microenvironment, would improve the function of adoptively transferred T cells. Linker for activation of T cells (LAT) plays a central role in T cell activation. We hypothesized that the ubiquitylation-resistant form of LAT in cells would enhance T cell signaling and thus augment antitumor activity. To test this, human CD4(+) or CD8(+) T cells were electroporated with small interfering RNA (siRNA) to repress endogenous LAT and ubiquitylation-resistant LAT 2KR or wild-type LAT mRNA was introduced for reexpression. Significantly enhanced phosphorylation of LAT and phospholipase C-γ (PLCγ) was observed, and augmented calcium signaling after T cell receptor (TCR) triggering was observed in LAT 2KR-expressing T cells. TCR-induced calcium signaling was abrogated in LAT knockdown cells, but the baseline was higher than that of control siRNA-electroporated cells, suggesting a fundamental requirement of LAT to maintain calcium homeostasis. Redirected LAT 2KR T cells expressing a chimeric antigen receptor or an MHC class I-restricted TCR showed augmented function as assessed by enhanced cytokine secretion and cytotoxicity. These results indicate that interruption of LAT ubiquitylation is a promising strategy to augment effector T cell function for adoptive cell therapy.
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http://dx.doi.org/10.1089/hum.2012.130DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3555093PMC
January 2013

Treatment of advanced leukemia in mice with mRNA engineered T cells.

Hum Gene Ther 2011 Dec 23;22(12):1575-86. Epub 2011 Sep 23.

Division of Oncology, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.

Cytotoxic T lymphocytes (CTLs) modified with chimeric antigen receptors (CARs) for adoptive immunotherapy of hematologic malignancies are effective in preclinical models and are being tested in several clinical trials. Although CTLs bearing stably expressed CARs generated by integrating viral vectors are efficacious and have potential long-term persistence, this mechanism of CAR expression can potentially result in significant toxicity. T cells were electroporated with an optimized in vitro transcribed RNA encoding a CAR against CD19. These RNA CAR CTLs were then tested in vitro and in vivo for efficacy. We found that T cells expressing an anti-CD19 CAR introduced by electroporation with optimized mRNA were potent and specific killers of CD19 target cells. CD19 RNA CAR T cells given to immunodeficient mice bearing xenografted leukemia rapidly migrated to sites of disease and retained significant target-specific lytic activity. Unexpectedly, a single injection of CD19 RNA CAR T cells reduced disease burden within 1 day after administration, resulting in a significant prolongation of survival in an aggressive leukemia xenograft model. The surface expression of the RNA CARs may be titrated, giving T cells with potentially tunable levels of effector functions such as cytokine release and cytotoxicity. RNA CARs are a genetic engineering approach that should not be subject to genotoxicity, and they provide a platform for rapidly optimizing CAR design before proceeding to more costly and laborious stable expression systems.
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http://dx.doi.org/10.1089/hum.2011.070DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3237694PMC
December 2011

[Effects of phytohormones on plant regeneration and production of flavonoids in transgenic Saussurea involucrata hairy roots].

Sheng Wu Gong Cheng Xue Bao 2011 Jan;27(1):69-75

Key Laboratory for Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.

We investigated the plant regeneration and production of flavonoids in three high-yield flavonoids transgenic Saussurea involucrata hairy roots C17, C27 and C46 by quantification of two phytohormones GA3 and IAA. The results showed that GA3 concentration at more than 1.0 mg/L could induce adventitious shoots in the hairy root lines. The highest shoot regeneration rate, about 82%, was obtained when the hairy roots C17 were cultured with 2.0 mg/L GA3. The results on HPLC and UV spectrophotometry showed that exogenous application of both GA3 and IAA increased the content of flavonoids in the hairy roots. The contents of flavonoids and apigenin in the hormone-treated hairy roots and regenerates were higher comparing with those in the untreated hairy roots and the regenerates. However, the content of flavonoids was not related to tissue weight, and was negatively related to the regeneration efficiency.
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January 2011

Endothelial cells mediate the regeneration of hematopoietic stem cells.

Stem Cell Res 2010 Jan 13;4(1):17-24. Epub 2009 Aug 13.

Division of Hematology and Medical Oncology, Hematologic Malignancies Program, Oregon Health & Science University, Portland, OR 97239, USA.

Recent studies suggest that endothelial cells are a critical component of the normal hematopoietic microenvironment. Therefore, we sought to determine whether primary endothelial cells have the capacity to repair damaged hematopoietic stem cells. Highly purified populations of primary CD31(+) microvascular endothelial cells isolated from the brain or lung did not express the pan hematopoietic marker CD45, most hematopoietic lineage markers, or the progenitor marker c-kit and did not give rise to hematopoietic cells in vitro or in vivo. Remarkably, the transplantation of small numbers of these microvascular endothelial cells consistently restored hematopoiesis following bone marrow lethal doses of irradiation. Analysis of the peripheral blood of rescued recipients demonstrated that both short-term and long-term multilineage hematopoietic reconstitution was exclusively of host origin. Secondary transplantation studies revealed that microvascular endothelial cell-mediated hematopoietic regeneration also occurs at the level of the hematopoietic stem cell. These findings suggest a potential therapeutic role for microvascular endothelial cells in the self-renewal and repair of adult hematopoietic stem cells.
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http://dx.doi.org/10.1016/j.scr.2009.08.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2938793PMC
January 2010

Hematopoietic stem cells contribute to lymphatic endothelium.

PLoS One 2008 26;3(11):e3812. Epub 2008 Nov 26.

Division of Hematology and Medical Oncology, Department of Medicine, Oregon Health & Science University, Portland, Oregon, United States of America.

Background: Although the lymphatic system arises as an extension of venous vessels in the embryo, little is known about the role of circulating progenitors in the maintenance or development of lymphatic endothelium. Here, we investigated whether hematopoietic stem cells (HSCs) have the potential to give rise to lymphatic endothelial cells (LEC).

Methodology/principal Findings: Following the transfer of marked HSCs into irradiated recipients, donor-derived LEC that co-express the lymphatic endothelial markers Lyve-1 and VEGFR-3 were identified in several tissues. HSC-derived LEC persisted for more than 12 months and contributed to approximately 3-4% of lymphatic vessels. Donor-derived LECs were not detected in mice transplanted with common myeloid progenitors and granulocyte/macrophage progenitors, suggesting that myeloid lineage commitment is not a requisite step in HSC contribution to lymphatic endothelium. Analysis of parabiotic mice revealed direct evidence for the existence of functional, circulating lymphatic progenitors in the absence of acute injury. Furthermore, the transplantation of HSCs into Apc(Min/+) mice resulted in the incorporation of donor-derived LEC into the lymphatic vessels of spontaneously arising intestinal tumors.

Conclusions/significance: Our results indicate that HSCs can contribute to normal and tumor associated lymphatic endothelium. These findings suggest that the modification of HSCs may be a novel approach for targeting tumor metastasis and attenuating diseases of the lymphatic system.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0003812PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2583952PMC
February 2009

Adoptive immunotherapy: good habits instilled at youth have long-term benefits.

Immunol Res 2008 ;42(1-3):182-96

Abramson Family Cancer Research Institute, University of Pennsylvania, BRB II/III, Room 554, 421 Curie Boulevard, Philadelphia, PA, 19104-6160, USA.

Many recent advances in basic cell biology and immunology are a harbinger of progress in adoptive cell therapy (ACT) including (1) the finding that host lymphodepletion enhances engraftment and efficacy, (2) the recognition that in vitro T cell functions may not correlate with in vivo efficacy, and (3) the development of advanced ex vivo culture methods to expand lymphocytes to therapeutically effective numbers. In this article, we focus on the development of artificial antigen presenting cells (aAPCs) in our laboratory and their applicability to augment ACT protocols. We also describe how aAPCs can be used to broaden ACT to treat patients with a wide variety of cancers, chronic infectious diseases, and autoimmune manifestations.
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http://dx.doi.org/10.1007/s12026-008-8070-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3809041PMC
March 2009

Distinct effects of IL-18 on the engraftment and function of human effector CD8 T cells and regulatory T cells.

PLoS One 2008 Sep 26;3(9):e3289. Epub 2008 Sep 26.

Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.

IL-18 has pleotropic effects on the activation of T cells during antigen presentation. We investigated the effects of human IL-18 on the engraftment and function of human T cell subsets in xenograft mouse models. IL-18 enhanced the engraftment of human CD8(+) effector T cells and promoted the development of xenogeneic graft versus host disease (GVHD). In marked contrast, IL-18 had reciprocal effects on the engraftment of CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) in the xenografted mice. Adoptive transfer experiments indicated that IL-18 prevented the suppressive effects of Tregs on the development of xenogeneic GVHD. The IL-18 results were robust as they were observed in two different mouse strains. In addition, the effects of IL-18 were systemic as IL-18 promoted engraftment and persistence of human effector T cells and decreased Tregs in peripheral blood, peritoneal cavity, spleen and liver. In vitro experiments indicated that the expression of the IL-18Ralpha was induced on both CD4 and CD8 effector T cells and Tregs, and that the duration of expression was less sustained on Tregs. These preclinical data suggest that human IL-18 may have use as an adjuvant for immune reconstitution after cytotoxic therapies, and to augment adoptive immunotherapy, donor leukocyte infusions, and vaccine strategies.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0003289PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2538560PMC
September 2008

Transvection mediated by the translocated cyclin D1 locus in mantle cell lymphoma.

J Exp Med 2008 Aug 14;205(8):1843-58. Epub 2008 Jul 14.

Center for Hematologic Malignancies, Oregon Cancer Institute, Portland, OR 97239, USA.

In mantle cell lymphoma (MCL) and some cases of multiple myeloma (MM), cyclin D1 expression is deregulated by chromosome translocations involving the immunoglobulin heavy chain (IgH) locus. To evaluate the mechanisms responsible, gene targeting was used to study long-distance gene regulation. Remarkably, these targeted cell lines lost the translocated chromosome (t(11;14)). In these MCL and MM cells, the nonrearranged cyclin D1 (CCND1) locus reverts from CpG hypomethylated to hypermethylated. Reintroduction of the translocated chromosome induced a loss of methylation at the unrearranged CCND1 locus, providing evidence of a transallelic regulatory effect. In these cell lines and primary MCL patient samples, the CCND1 loci are packaged in chromatin-containing CCCTC binding factor (CTCF) and nucleophosmin (NPM) at the nucleolus. We show that CTCF and NPM are bound at the IgH 3' regulatory elements only in the t(11;14) MCL cell lines. Furthermore, NPM short hairpin RNA produces a specific growth arrest in these cells. Our data demonstrate transvection in human cancer and suggest a functional role for CTCF and NPM.
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http://dx.doi.org/10.1084/jem.20072102DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2525596PMC
August 2008

Myeloid lineage progenitors give rise to vascular endothelium.

Proc Natl Acad Sci U S A 2006 Aug 18;103(35):13156-61. Epub 2006 Aug 18.

Oregon Stem Cell Center, Center for Hematologic Malignancies, Division of Hematology and Medical Oncology, Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, 97239, USA.

Despite an important role in vascular development and repair, the origin of endothelial progenitors remains unknown. Accumulating evidence indicates that cells derived from the hematopoietic system participate in angiogenesis. However, the identity and functional role of these cells remain controversial. Here we show that vascular endothelial cells can differentiate from common myeloid progenitors and granulocyte/macrophage progenitors. Endothelial cells derived from transplanted bone marrow-derived myeloid lineage progenitors expressed CD31, von Willebrand factor, and Tie2 but did not express the hematopoietic markers CD45 and F4/80 or the pericyte markers desmin and smooth muscle actin. Lineage tracing analysis in combination with a Tie2-driven Cre/lox reporter system revealed that, in contrast to bone marrow-derived hepatocytes, bone marrow-derived endothelial cells are not the products of cell fusion. The establishment of both hematopoietic and endothelial cell chimerism after parabiosis demonstrates that circulating cells can give rise to vascular endothelium in the absence of acute radiation injury. Our findings indicate that endothelial cells are an intrinsic component of myeloid lineage differentiation and underscore the close functional relationship between the hematopoietic and vascular systems.
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http://dx.doi.org/10.1073/pnas.0604203103DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1559769PMC
August 2006

In vivo genetic selection of renal proximal tubules.

Mol Ther 2006 Jan 10;13(1):49-58. Epub 2005 Oct 10.

Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR 97239, USA.

Repopulation by transplanted cells can result in effective therapy for several regenerative organs including blood, liver, and skin. In contrast, cell therapies for renal diseases are not currently available. Here we developed an animal model in which cells genetically resistant to a toxic intermediate of tyrosine metabolism, homogentisic acid (HGA), were able to repopulate the damaged proximal tubule epithelium of mice with fumarylacetoacetate hydrolase (Fah) deficiency. HGA resistance was achieved by two independent mechanisms. First, Fah+ transplanted bone marrow cells produced significant replacement of damaged proximal tubular epithelium (up to 50%). The majority of bone marrow-derived epithelial cells were generated by cell fusion, not transdifferentiation. In addition to regeneration by fusion-derived epithelial cells, proximal tubular repopulation was also observed by host epithelial cells, which had lost the homogentisic acid dioxygenase gene. These data demonstrate that extensive regeneration of the renal proximal tubule compartment can be achieved through genetic selection of functional cells.
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http://dx.doi.org/10.1016/j.ymthe.2005.09.004DOI Listing
January 2006

Donor marker infidelity in transgenic hematopoietic stem cells.

Stem Cells 2005 May;23(5):638-43

Center for Hematologic Malignancies, Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, OR, USA.

Transgenic marking approaches are increasingly used to evaluate the developmental potential of stem cells. However, cell fate mapping studies using different transgenic marking systems have produced conflicting results. These disparate findings may be due in part to the infidelity of donor marker gene expression. Analysis of hematopoietic stem cells (c-Kit+, Sca-1+, lineage marker- [KSL]) from a transgenic mouse (1Osb) engineered to ubiquitously express the enhanced green fluorescent protein (EGFP) reveals two distinct populations. Forty percent of KSL cells demonstrate intermediate levels of EGFP fluorescence and differentiate into subpopulations of B cells, T cells, and myeloid cells that do not express EGFP. By contrast, progeny of the remaining 60% of KSL cells are almost exclusively EGFP bright. Long-term multilineage hematopoietic reconstitution and serial transplantation experiments show that these differences in EGFP are a property of self-renewing stem cells. Furthermore, both the transgene integration site and the activation status of a cell are important determinants of EGFP expression. These results indicate that a combination of donor cell markers is required to reliably track the full differentiation potential of transgenic stem cells.
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http://dx.doi.org/10.1634/stemcells.2004-0325DOI Listing
May 2005

Transplanted human bone marrow contributes to vascular endothelium.

Proc Natl Acad Sci U S A 2004 Nov 17;101(48):16891-6. Epub 2004 Nov 17.

Center for Hematologic Malignancies, Division of Hematology and Medical Oncology, Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA.

Recent evidence indicates that bone marrow is a source of endothelial progenitor cells that are mobilized into the peripheral blood in response to cytokines or tissue injury. Previously, we showed that functional endothelial cells (ECs) can be clonally derived from phenotypically defined hematopoietic stem cells. To determine the EC potential of human bone marrow and peripheral blood stem cells, blood vessels in sex-mismatched transplant recipients were evaluated. EC outcomes were identified by using a combination of immunohistochemistry and XY interphase FISH. Donor-derived ECs were detected in the skin and gut of transplant recipients with a mean frequency of 2% and could readily be distinguished from CD45-expressing hematopoietic stem cells. None of the >4,000 ECs examined had more than two sex chromosomes, consistent with an absence of cell fusion. Y chromosome signals were not detected in sex-matched female recipients, excluding the vertical transmission of male cells. None of the recipients evaluated before hematopoietic engraftment demonstrated donor-derived ECs, indicating a close linkage between the recovery of hematopoiesis and EC outcomes. Transplantable bone marrow-derived endothelial progenitor cells may represent novel therapeutic targets for hematopoietic and vascular disease.
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http://dx.doi.org/10.1073/pnas.0404398101DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC534718PMC
November 2004

Transplanted adult hematopoietic stems cells differentiate into functional endothelial cells.

Blood 2004 Jan 4;103(1):13-9. Epub 2003 Sep 4.

Department of Medicine, Oregon Health and Science University, Portland, OR 97239, USA.

During early embryogenesis, blood vessels and hematopoietic cells arise from a common precursor cell, the hemangioblast. Recent studies have identified endothelial progenitor cells in the peripheral blood, and there is accumulating evidence that a subset of these cells is derived from precursors in the bone marrow. Here we show that adult bone marrow-derived, phenotypically defined hematopoietic stem cells (c-kit+, Sca-1+, lineage-) give rise to functional endothelial cells. With the exception of the brain, donor-derived cells are rapidly integrated into blood vessels. Durably engrafted endothelial cells express CD31, produce von Willebrand factor, and take up low-density lipoprotein. Analysis of DNA content indicates that donor-derived endothelial cells are not the products of cell fusion. Self-renewal of stem cells with hematopoietic and endothelial cell potential was revealed by serial transplantation studies. The clonal origin of both hematopoietic and endothelial cell outcomes was established by the transfer of a single cell. These results suggest that adult bone marrow-derived hematopoietic stem cells may serve as a reservoir for endothelial cell progenitors.
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http://dx.doi.org/10.1182/blood-2003-05-1684DOI Listing
January 2004

Cryptic MBP epitope 1-20 is inducing autoimmune anterior uveitis without EAE in Lewis rats.

Cell Immunol 2002 May-Jun;217(1-2):87-94

Neurological Sciences Institute, Oregon Health and Science University, 505 NW 185th Avenue, Beaverton, OR 97006, USA.

Lewis rats immunized with myelin basic protein (MBP) developed experimental autoimmune encephalomyelitis (EAE) and associated anterior uveitis (AU). Although several cryptic epitopes of MBP have strong encephalitogenic and uveitogenic properties, the peptide corresponding to the MBP residues 1-20 was uniquely capable of inducing AU without EAE. In this study, we showed that acetylation of the N-terminal amino acid did not produce encephalitogenicity, did not enhance uveitogenicity, and did not improve T cell proliferation in Lewis rats. The cytokine production profile induced by MBP(1-20) immunization was consistent with a Th1 response. In MBP-injected rats and in peptide-injected rats, the frequency of the IFN-gamma-secreting cells in MBP(69-89)-stimulated T cells was significantly higher than the frequency of IFN-gamma-secreting cells in MBP(1-20)-stimulated T cells. However, similar numbers of IFN-gamma-producing specific cells were found in the eyes of MBP(69-89) and MBP(1-20) immunized rats. In these rats, the iris-infiltrating cells consisted of a much higher percentage of CD4(+) T cells expressing L-selectin (CD62L) than did those cells found in the spinal cord. The results demonstrate that MBP(1-20) is immunogenic and uveitogenic, although it induced only weak proliferation and weak Th1 reaction. The fact that T cells with the same specificity have different effects on target organs suggested that, in the eye and spinal cord, a distinct mechanism might mediate the recruitment of cells to these organs.
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http://dx.doi.org/10.1016/s0008-8749(02)00514-2DOI Listing
December 2002

Crucial role of CCL3/MIP-1alpha in the recurrence of autoimmune anterior uveitis induced with myelin basic protein in Lewis rats.

J Autoimmun 2002 Jun;18(4):259-70

Neurological Sciences Institute, Oregon Health & Science University, Portland, Oregon 97006, USA.

In this study, we examined the role of CCL2/MCP-1, CCL3/MIP-1alpha, CCL4/MIP-1beta, and CCL5/RANTES during recurrent anterior uveitis (RAU). LEW rats injected with myelin basic protein (MBP) developed experimental autoimmune encephalomyelitis (EAE) and associated anterior uveitis (AU), which was mediated by CD4(+) T cells. After recovery, rats become resistant to EAE but developed RAU. Rats reinjected with MBP developed RAU without EAE. The chemokines tested were detected in the eye at RAU accelerated onset, increased as the disease progressed, and fell as clinical signs improved. At the same time, in the spinal cords of rats, these chemokines were still detected but at reduced levels. Administration of anti-MIP-1alpha neutralizing antibodies resulted in almost complete suppression of clinical RAU and significant reduction of inflammatory cell recruitment into the iris. Anti-MIP-1beta and anti-MCP-1 antibodies were effective in suppression of RAU but to lesser degree. Treatment with anti-RANTES antibodies was not effective in protecting against the recurrent development of the disease. In the eyes, the message for CCR1 and CCR5 was considerably elevated prior to the onset of AU and decreased after treatment with anti-chemokine antibodies. Our results suggest a crucial role of CCL3/MIP-1alpha in the development of RAU in Lewis rats. In addition, CCL2/MCP-1 and CCL4/MIP-1beta may also play a role in immunopathogenesis of RAU.
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http://dx.doi.org/10.1006/jaut.2002.0591DOI Listing
June 2002