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Neonatal Hypersusceptibility To Endotoxin Correlates With Increased Tumor Necrosis Factor Production In Mice, Vitaliano Cusumano, Giuseppe Mancuso, Francesco Genovese, Maria Cuzzola, Maria Carbone, James A. Cook, Joel B. Cochran, Giuseppe Teti

MUSC Faculty Journal Articles

Septic shock is a major cause of mortality in neonates. The hypothesis was tested that neonatal, age is associated with altered sensitivity to shock-inducing bacterial products or proinflammatory, cytokines (or both). Mice of different ages were inoculated with various doses of lipopolysaccharide, (LPS), superantigenic staphylococcal enterotoxin B (SEB), or recombinant tumor necrosis factor-α, (rTNF-α), alone or in combination with the sensitizing agent D-galactosamine. Neonatal mice were, markedly more susceptible to LPS-induced lethality but more resistant to SEB than were adults (P, < .05). Mice of different ages did not differ, however, in their sensitivity to lethal activities of rTNFα. Neonatal susceptibility to LPS and SEB correlated directly with plasma TNF-α but not IFN-γ, levels, which was confirmed by TNF-α and IFN-γ blockade experiments. These data document, marked age-related differences in the pathophysiology of septic shock and suggest that IFN-γ is, not an obligatory mediator of either LPS- or SEB-induced lethality in neonates.

Chronicity In Strongyloides Stercoralis Infections: Dichotomy Of The Protective Immune Response To Infective And Autoinfective Larvae In A Mouse Model., R. A. Brigandi, H. L. Rotman, T. J. Nolan, G. A. Schad, D. Abraham

Department of Microbiology and Immunology Faculty Papers

Strongyloidiasis is an intestinal disease that can last for decades due to the occurrence of autoinfective larvae (L3a) in an infected person, which contribute to the maintenance of the population of adult worms in the intestine. The goal of the present study was to determine if L3a are susceptible to the protective immunity that targets the infective stage of the worm, the third-stage larvae (L3). Mice immunized and challenged with Strongyloides stercoralis L3 kill more than 90% of challenge larvae contained within diffusion chambers. The L3 do not remain antigenically static in mice, however, but undergo some degree of antigenic …

Local Delivery Of Interleukin 4 By Retrovirus-Transduced T Lymphocytes Ameliorates Experimental Autoimmune Encephalomyelitis., Michael K. Shaw, James B. Lorens, Archana Dhawan, Richard Dalcanto, Harley Y. Tse, Alyssa B. Tran, Colleen Bonpane, Shanti L. Eswaran, Stefan Brocke, Nora Sarvetnick, Lawrence Steinman, Garry P. Nolan, C. Garrison Fathman

Journal Articles: Regenerative Medicine

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory autoimmune disease of the central nervous system which serves as a model for the human disease multiple sclerosis. We demonstrate here that encephalitogenic T cells, transduced with a retroviral gene, construct to express interleukin 4, and can delay the onset and reduce the severity of EAE when adoptively transferred to myelin basic protein-immunized mice. Thus, T lymphocytes transduced with retroviral vectors can deliver "regulatory cytokines" in a site-specific manner and may represent a viable therapeutic strategy for the treatment of autoimmune disease.

Transgenic Mice Which Overexpress Nerve Growth Factor, Kathryn M. Albers, Brian M. Davis

Neuroscience Faculty Patents

Transgenic mice that express increased levels of nerve growth factor (NGF) in the epidermis and other stratified, keratinized epithelium. The nerve growth factor expressing transgenic mice are useful in the study of neurodegenerative disorders of the brain such as Parkinson's syndrome and Alzheimer's disease and for testing for drug candidates for the treatment of these diseases.

Loss Of Dcc Expression And Glioma Progression., M Reyes-Mugica, K Rieger-Christ, H Ohgaki, B C Ekstrand, M Helie, G Kleinman, Alan M. Yahanda Md, Facs, E R Fearon, P Kleihues, M A Reale

PCI Publications and Projects

The deleted in colorectal cancer (DCC) gene, a candidate tumor suppressor gene on chromosome 18q21, encodes a neural cell adhesion molecule family protein that is most highly expressed in the nervous system. To address the hypothesis that DCC may play a role in glioma development and/or progression, we examined DCC expression by immunohistochemistry in 57 resected human astrocytic tumors. Overall, low-grade astrocytomas were predominantly DCC positive (15 of 16, or 94%), whereas high-grade tumors significantly less often expressed the DCC protein (27 of 41, or 66%; P = 0.03). We were able to directly assess the relationship between DCC expression …

Ifn-Gamma, Igif, And Iddm., Nora Sarvetnick

Journal Articles: Regenerative Medicine

No abstract provided.

Divergent Selection For Heat Loss In Mice: Ii. Correlated Responses In Feed Intake, Body Mass, Body Composition, And Number Born Through Fifteen Generations, Merlyn K. Nielsen, B. A. Freking, L. D. Jones, S. M. Nelson, T. L. Vorderstrasse, B. A. Hussey

Department of Animal Science: Faculty Publications

Divergent selection for heat loss (kcal•kg^-.75•d^-1) , measured in 9- to 11-wk-old male mice, was conducted for 15 generations. Selection for high (MH) and low (ML) heat loss and unselected control (MC) occurred in each of three replicates for a total of nine unique lines. Feed intake in males was measured during Generations 9 through 15. Body mass at commencement of mating in females and at time of measurement of heat loss in males was recorded. Body fat percentage at 12 wk for animals of Generations 6, 10, and 14 was predicted as a function of …

A Simulation Model Including Ovulation Rate, Potential Embryonic Viability, And Uterine Capacity To Explain Litter Size In Mice: Ii. Responses To Alternative Criteria Of Selection, E. L. De A. Ribeiro, Merlyn K. Nielsen, Kreg A. Leymaster, Gary Bennett

Department of Animal Science: Faculty Publications

Direct selection for litter size was compared with selection for ovulation rate, ova success, or uterine capacity and for indexes of ovulation rate with ova success or uterine capacity. Selection was simulated for 10 generations in a mouse population based on a model integrating ovulation rate, potential embryonic viability, and uterine capacity. Two indexes including ovulation rate (OR) and ova success (OS) were I = .291 x OR + 2.19 x OS and I = .165 x OR + .736 x OS. Heritabilities for ovulation rate and ova success, assumed in the simulation and to derive the indexes, were .25 …

Divergent Selection For Heat Loss In Mice: I. Selection Applied And Direct Response Through Fifteen Generations, Merlyn K. Nielsen, L. D. Jones, B. A. Freking, J. A. Deshazer

Department of Animal Science: Faculty Publications

Divergent selection for heat production/ loss (kcal•kg^-.75•d^-1) , measured in 9- to 11-wk-old male mice, was conducted for 15 generations. Heat loss was measured for 15 h on individual animals placed overnight in direct, gradient-layer calorimeters. Selection for high (MH) and low (ML) heat loss and unselected control (MC) occurred in each of three replicates for a total of nine unique lines. Repeatability of the heat loss measurement was .45 and the CV was 10.5%. Cumulative realized selection differentials, averaged for the three replicates, were 145.1 and -105.0 (kcal•kg^-.75•d^-1) and ranged from …

A Simulation Model Including Ovulation Rate, Potential Embryonic Viability, And Uterine Capacity To Explain Litter Size In Mice: I. Model Development And Implementation, E. L. De A. Ribeiro, Merlyn K. Nielsen, Gary Bennett, Kreg A. Leymaster

Department of Animal Science: Faculty Publications

Litter size in mice was studied using a model including ovulation rate, potential embryonic viability, and uterine capacity. Simulated results were compared with experimental results from a selection experiment with mice. The four criteria of selection were selection on number born (LS), selection on an index of ovulation rate and ova success (IX), selection on number born to unilaterally ovariectomized females (UT), and unselected control (LC). Comparisons were made to statistics of the base generation and to responses after 13 generations of selection. Phenotypic and genetic statistics for uterine capacity were generated so that simulations produced the experimental means, standard …

Cellular Basis Of Decreased Immune Responses To Pneumococcal Vaccines In Aged Mice, Manju Garg, Wei Luo, Alan M. Kaplan, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Previously, model systems were developed in our laboratory to study murine immune responses to the 23-valent pneumococcal polysaccharide vaccine Pnu-Imune, both in vivo and in vitro (M. Garg and B. Subbarao, Infect. Immun. 60:2329-2336, 1992; M. Garg, A. M. Kaplan, and S. Bondada, J. Immunol. 152: 1589-1596, 1994). Using these systems, we found that aged mice did not respond to the vaccine in vivo or in vitro. Cell separation studies showed that the unresponsiveness of the aged spleen cells to the vaccine was not due to an intrinsic B-cell defect or to T-cell-mediated immunosuppression but resulted from an accessory cell …

Mechanisms Of Cytokine-Mediated Localized Immunoprotection., Nora Sarvetnick

Journal Articles: Regenerative Medicine

No abstract provided.

Pancreatic Expression Of Interleukin-4 Abrogates Insulitis And Autoimmune Diabetes In Nonobese Diabetic (Nod) Mice., Regula Mueller, Troy Krahl, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Diabetes in nonobese diabetic (NOD) mice is a T cell-dependent autoimmune disease. The destructive activities of autoreactive T cells have been shown to be tightly regulated by effector molecules. In particular, T helper (Th) 1 cytokines have been linked to diabetes pathogenesis, whereas Th2 cytokines and the cells that release them have been postulated to be protective from disease. To test this hypothesis, we generated transgenic NOD mice that express interleukin (IL) 4 in their pancreatic beta cells under the control of the human insulin promoter. We found that transgenic NOD-IL-4 mice, both females and males, were completely protected from …

Cloning Of The Mammalian Type Ii Iodothyronine Deiodinase. A Selenoprotein Differentially Expressed And Regulated In Human And Rat Brain And Other Tissues., Walburga Croteau, Jennifer C. Davey, Valerie Anne Galton, Donald L. St Germain

Dartmouth Scholarship

The deiodination of thyroid hormones in extrathyroidal tissues plays an important role in modulating thyroid hormone action. The type II deiodinase (DII) converts thyroxine to the active hormone 3,5,3'-triiodothyronine, and in the rat is expressed in the brain, pituitary gland, and brown adipose tissue (BAT). Complementary DNAs (cDNAs) for the types I and III deiodinases (DI and DIII, respectively) have been isolated and shown to code for selenoproteins. However, information concerning the structure of the mammalian DII remains limited, and the pattern of its expression in human tissues is undefined. We report herein the identification and characterization of rat and …

Il-10 Is Necessary And Sufficient For Autoimmune Diabetes In Conjunction With Nod Mhc Homozygosity., Myung-Shik Lee, Regula Mueller, Linda S. Wicker, Laurence B. Peterson, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Contrary to expectations based on in vitro experiments, we previously found that pancreatic IL-10 did not inhibit autoimmune diabetes but accelerated it in an MHC-dependent manner. Therefore, the ability of IL-10 to overcome the absence of all non-MHC diabetes susceptibility (Idd) alleles was studied in transgenic mice expressing pancreatic IL-10 backcrossed to B10.H2g7 congenic mice, which have no Idd alleles other than NOD MHC (H2g7). IL-10 transgenic backcross 1 (BC1) mice with H2g7/g7 haplotype developed clear-cut insulitis and diabetes, but neither transgenic mice with the H2g/b haplotype nor nontransgenic BC1 mice did so. Further implicating IL-10 in autoimmune diabetes, anti-IL-10 …

Disruption Of The Cbfa2 Gene Causes Necrosis And Hemorrhaging In The Central Nervous System And Blocks Definitive Hematopoiesis., Qing Wang, Terryl Stacy, Michael M Binder, Miguel Marin-Padilla, Arlene H. Sharpe, Nancy A. Speck

Dartmouth Scholarship

The CBFA2 (AML1) gene encodes a DNA-binding subunit of the heterodimeric core-binding factor. The CBFA2 gene is disrupted by the (8;21), (3;21), and (12;21) chromosomal translocations associated with leukemias and myelodysplasias in humans. Mice lacking a CBF alpha 2 protein capable of binding DNA die between embryonic days 11.5 and 12.5 due to hemorrhaging in the central nervous system (CNS), at the nerve/CNS interfaces of cranial and spinal nerves, and in somitic/intersomitic regions along the presumptive spinal cord. Hemorrhaging is preceded by symmetric, bilateral necrosis in these regions. Definitive erythropoiesis and myelopoiesis do not occur in Cbfa2-deficient embryos, and disruption …

A Tandem Duplication Within The Fibrillin 1 Gene Is Associated With The Mouse Tight Skin Mutation., Linda D. Siracusa, Rodney Mcgrath, Qing Ma, John J. Moskow, Jayanthi Manne, Paul J. Christner, Arthur M. Buchberg, Sergio A. Jimenez

Department of Medicine Faculty Papers

Mice carrying the Tight skin (Tsk) mutation have thickened skin and visceral fibrosis resulting from an accumulation of extracellular matrix molecules. These and other connective tissue abnormalities have made Tskl + mice models for scleroderma, hereditary emphysema, and myocardial hypertrophy. Previously we localized Tsk to mouse chromosome 2 in a region syntenic with human chromosome 15. The microfibrillar glycoprotein gene, fibrillin 1 (FBN1), on human chromosome 15q, provided a candidate for the Tsk mutation. We now demonstrate that the Tsk chromosome harbors a 30- to 40-kb genomic duplication within the Fbn1 gene that results in a larger than normal in-frame …

Cd40-Cd40 Ligand Interactions In Experimental Allergic Encephalomyelitis And Multiple Sclerosis., Koen Gerritse, Jon D. Laman, Randolph J. Noelle, Alejandro Aruffo

Dartmouth Scholarship

We investigated the role of CD40-CD40 ligand (CD40L) interactions in multiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE). Activated helper T cells expressing CD40L (gp39) surface protein were found in MS patient brain sections, but not in brain tissue sections of normal controls or patients with other neurological disease. CD40L-positive cells were co-localized with CD40-bearing cells in active lesions (perivascular infiltrates). Most of these CD40-bearing cells proved to be of the monocytic lineage (macrophages or microglial cells), and relatively few were B cells. To functionally evaluate CD40-CD40L interactions, EAE was elicited in mice by means of proteolipid-peptide immunization. Treatment with …

Biochemical And Mutational Analysis Of The Histidine Residues Of Staphylococcal Enterotoxin A., M Hoffman, M Tremaine, J Mansfield, M Betley

Manuscripts, Articles, Book Chapters and Other Papers

The goal of this study was to examine the role of histidine residues in the biological activities of staphylococcal enterotoxin A (SEA). Carboxymethylated SEA was unable to stimulate murine T-cell proliferation but was resistant to monkey stomach lavage fluid degradation, suggesting that native conformation was intact. Site-directed mutagenesis of the histidine residues of SEA was subsequently performed. SEA-H44A (SEA with histidine 44 replaced with alanine), SEA-H44D, SEA-H50A, SEA-H50D, SEA-H114A, SEA-H114D, SEA-H187A, and SEA-H187D retained superantigen and emetic activities, whereas SEA-H225A and SEA-H225D were defective in the ability to stimulate T-cell proliferation. These mutants were unable to compete with SEA for …

Analysis Of The Effects Of Overexpression Of Metallothionein-I In Transgenic Mice On The Reproductive Toxicology Of Cadmium., T Dalton, Kai Fu, G C Enders, R D Palmiter, G K Andrews

Journal Articles: Pathology and Microbiology

Exposure to low levels of cadmium reduces fertility. In male mice spermatogenesis is highly sensitive to cadmium, whereas in females the peri-implantation period of pregnancy is sensitive. To examine the potential roles of the cadmium-binding protein, metallothionein (MT), in the reproductive toxicology of cadmium, we examined a transgenic mouse strain that overexpresses metallothionein-I (MT-I). These mice had dramatically increased steady-state levels of MT-I mRNA and MT in the testes and in the female reproductive tract during the peri-implantation period of pregnancy, and this overexpression occurred in a cell-specific and temporally regulated manner similar to that of the endogenous MT-I gene. …

Embryonal Survival To 6 Days In Mice Selected On Different Criteria For Litter Size, E. L. De A. Ribeiro, M. A. J. Van Engelen, Merlyn K. Nielsen

Department of Animal Science: Faculty Publications

Embryonal survival was compared in mice resulting from four criteria of selection: LS = selection on number born; IX = selection on an index of ovulation rate and ova success; UT = selection on number born to unilaterally ovariectomized females; and LC = unselected control. Selection occurred for 21 generations with three replicates of the four criteria; thereafter, relaxed selection was practiced. The evaluation was performed using mice of two replicates at Generation 35 and one replicate at Generation 36. Data on a total of 289 female mice were recorded. Females, at an average age of 9 wk, were mated …

Estimates Of Heritabilities And Genetic And Environmental Correlations For Left- And Right-Side Uterine Capacity And Ovulation Rate In Mice, Merlyn K. Nielsen, Y. L. Kochera Kirby, A. C. Clutter

Department of Animal Science: Faculty Publications

Heritabilities for and genetic and environmental correlations between uterine capacity, ovulation rate, and body mass (BM) were estimated in mice. Uterine capacity was defined as the number of fetuses (LUC or RUC for left or right side) in one uterine horn for unilaterally ovariectomized females. Ovulation rate (corpora lutea, LCL or RCL for left or right ovary) was measured on the remaining single ovary in these same females. Data on 1,931 mice from four selection populations were used. Left ovulation rate and LUC were measured on 958 animals, and RCL and RUC of another 972 animals were recorded. Genetic and …

Inactivation Of Raf-1 By A Protein-Tyrosine Phosphatase Stimulated By Gtp And Reconstituted By G Subunits, Paul Dent, Dean B. Reardon, Steven L. Wood, Margaret A. Lindorfer, Stephen G. Graber, James C. Garrison, David L. Brautigan, Thomas W. Sturgill

Faculty & Staff Scholarship

No abstract provided.

A Requirement For Bone Morphogenetic Protein-7 During Development Of The Mammalian Kidney And Eye., Andrew T. Dudley, K. M. Lyons, E. J. Robertson

Journal Articles: Genetics, Cell Biology & Anatomy

BMP-7/OP-1, a member of the transforming growth factor-beta (TGF-beta) family of secreted growth factors, is expressed during mouse embryogenesis in a pattern suggesting potential roles in a variety of inductive tissue interactions. The present study demonstrates that mice lacking BMP-7 display severe defects confined to the developing kidney and eye. Surprisingly, the early inductive tissue interactions responsible for establishing both organs appear largely unaffected. However, the absence of BMP-7 disrupts the subsequent cellular interactions required for their continued growth and development. Consequently, homozygous mutant animals exhibit renal dysplasia and anophthalmia at birth. Overall, these findings identify BMP-7 as an essential …

Stimulation Of Alpha 1(I) Procollagen Gene Expression In Nih-3t3 Cells By The Human T Cell Leukemia Virus Type 1 (Htlv-1) Tax Gene., E. Munoz, D. Suri, S. Amini, K. Khalili, Sergio A. Jimenez

Selected Works of Sergio Jiménez, MD, MACR

The mechanisms that regulate the expression of genes encoding extracellular matrix proteins in fibroblasts and other mesenchymal cells have remained elusive. Studies from several laboratories have indicated that Tax, a trans-regulatory protein from the human T cell leukemia virus type I not only augments viral gene expression but also triggers the expression of various cellular genes. Here, we examined the hypothesis that the expression of collagen genes may also be modulated by Tax. NIH-3T3 cells were simultaneously transfected with a Tax expressor plasmid and a chimeric construct containing regulatory sequences (-804 to +42 bp) of the alpha 1(I) procollagen gene …

Survival Of Mouse Pancreatic Islet Allografts In Recipients Treated With Allogeneic Small Lymphocytes And Antibody To Cd40 Ligand., David C. Parker, Dale L. Greiner, Nancy E. Phillips, Michael C. Appel, Alan W. Steele, Fiona H. Durie, Randolph J. Noelle, John P. Mordes, Aldo A. Rossini

Dartmouth Scholarship

Combined treatment with allogeneic small lymphocytes or T-depleted small lymphocytes plus a blocking antibody to CD40 ligand (CD40L) permitted indefinite pancreatic islet allograft survival in 37 of 40 recipients that differed from islet donors at major and minor histocompatibility loci. The effect of the allogeneic small lymphocytes was donor antigen-specific. Neither treatment alone was as effective as combined treatment, although anti-CD40L by itself allowed indefinite islet allograft survival in 40% of recipients. Our interpretation is that small lymphocytes expressing donor antigens in the absence of appropriate costimulatory signals are tolerogenic for alloreactive host cells. Anti-CD40L antibody may prevent host T …

Expression Of Genes Encoding Antioxidant Enzymes In Preimplantation Mouse And Cow Embryos And Primary Bovine Oviduct Cultures Employed For Embryo Coculture., M B Harvey, M Y Arcellana-Panlilio, X Zhang, G A Schultz, A J Watson

Obstetrics & Gynaecology Publications

Preimplantation embryos from a variety of mammalian species contrast markedly in their response to culture in vitro. Murine preimplantation embryos display a wider tolerance than other mammalian species to culture environments, and this has contributed to the development of several effective defined culture media. Embryo coculture on somatic cells remains the most effective method of supporting reasonable rates of bovine preimplantation development in vitro. The patterns of gene expression for several antioxidant enzymes during preimplantation murine and bovine development were examined by use of the reverse transcription-polymerase chain reaction technique to determine whether the differential developmental capacity of mammalian preimplantation …

The Effect Of Ionizing Irradiation On Type I Collagen Of The Tail In Growing Mice: A Histology And Electron Microscopy Study, M. Q. Yang, E. Kjellén, C. H. Håkansson, M. Palmegren

Scanning Microscopy

In order to examine the effect of radiation on growing tissue, especially the fibroblasts and their end-product, the collagen fibres, tails from 24 mice were irradiated at an age of 8 days with 20 Gy and 30 Gy (>°Co). Tails from 18 animals served as controls. Six mice from each group were sacrificed on day 8, 20 and 30. Transmission electron microscopy was used to examine the fibroblasts and the collagen fibrils. Non-irradiated fibroblasts had a nucleus rich in chromatin and an abundant endoplasmic reticulum with cistemae and condensing vacuoles.

On day 20, approximately 50 % , and on …

Astrocytes Derived From P53-Deficient Mice Provide A Multistep In Vitro Model For Development Of Malignant Gliomas., Alan M. Yahanda Md, Facs, J M Bruner, L A Donehower, R S Morrison

PCI Publications and Projects

Loss or mutation of p53 is thought to be an early event in the malignant transformation of many human astrocytic tumors. To better understand the role of p53 in their growth and transformation, we developed a model employing cultured neonatal astrocytes derived from mice deficient in one (p53 +/-) or both (p53 -/-) p53 alleles, comparing them with wild-type (p53 +/+) cells. Studies of in vitro and in vivo growth and transformation were performed, and flow cytometry and karyotyping were used to correlate changes in growth with genomic instability. Early-passage (EP) p53 -/- astrocytes achieved higher saturation densities and had …

Increased Alpha 1(I) Procollagen Gene Expression In Tight Skin (Tsk) Mice Myocardial Fibroblasts Is Due To A Reduced Interaction Of A Negative Regulatory Sequence With Ap-1 Transcription Factor., Neena Philips, Reza I. Bashey, Sergio A. Jimenez

Department of Medicine Faculty Papers

The TSK mouse, a model of fibrosis, displays exaggerated connective tissue accumulation in skin and visceral organs including the heart. To study the mechanisms of myocardial fibrosis in TSK mice, we established several strains of TSK mice myocardial fibroblasts in culture and examined the regulation of collagen gene expression in these cells. These strains displayed increased collagen gene expression in comparison with myocardial fibroblasts established from normal mice. On an average, the TSK myocardial fibroblast cultures showed a 4-fold increase in collagen synthesis and 4.4- and 3.6-fold increases, respectively, in alpha 1(I) and alpha 1(III) collagen mRNA steady state levels. …