USDA.gov National Agricultural Library
Animal Welfare Information Center
HomeAbout AWICPublicationsWorkshopsServicesNews and EventsHelpContact Us
Search AWIC
Search all of the United States Department of Agriculture
Advanced search
Browse by Subject
Research Animals
Farm Animals
Zoo, Circus and Marine Animals
Companion Animals
Government and Professional Resources
Alternatives
Literature Searching and Databases
Pain and Distress
Humane Endpoints and Euthanasia
 
You are here: Home / Publications / Bibliographies and Resource Guides / Canine Models in Biomedical Research, 1990-2009  / Immune System  Printer Friendly Page
Publications
 
Canine Models in Biomedical Research,  1990-2009
<< Table of Contents << Previous |  Next >>

 

Immune System

Barrett, E.G., K. Rudolph, S.Z. Wang, B.A. Muggenburg, M. Nysus, K. Clarke, and C. Mccall (2002). Epicutaneous sensitization with house dust mites leads to pulmonary immune responses in dogs. Journal of Allergy and Clinical Immunology 109(1 Supplement): S35. ISSN: 0091-6749.
NAL Call Number: 448.8 J8236
Descriptors: biochemistry and molecular biophysics, immune system, chemical coordination and homeostasis, respiratory system, respiration, epicutaneous sensitization, pulmonary immunity, meeting abstract.

Bauer, T.R.J., K.E. Creevy, Y.C. Gu, L.M. Tuschong, R.E. Donahue, M.E. Metzger, L.J. Embree, T. Burkholder, J.D. Bacher, C. Romines, M.L.I. Thomas, L. Colenda, and D.D. Hickstein (2004). Very low levels of donor cd18+ neutrophils following allogeneic hematopoietic stem cell transplantation reverse the disease phenotype in canine leukocyte adhesion deficiency. Blood 103(9): 3582-3589. ISSN: 0006-4971.
NAL Call Number: RB145.A1B57
Descriptors: large animal model, clinical immunology, human medicine, medical sciences, hematology, surgery, bacterial infection, infectious disease, leukocyte adhesion deficiency, blood and lymphatic disease, immune system disease, allogeneic hematopoietic stem cell transplantation, clinical techniques, gene therapy, genetic techniques, laboratory techniques, nonmyeloablative hematopoietic stem cell transplantation, disease phenotype.

Bauer, T.R.J., Y.C. Gu, K.E. Creevy, L.M. Tuschong, L. Embree, S.M. Holland, R.A. Sokolic, and D.D. Hickstein (2004). Leukocyte adhesion deficiency in children and irish setter dogs. Pediatric Research 55(3): 363-367. ISSN: 0031-3998.
NAL Call Number: RJ1.P4
Abstract: Children with the genetic immunodeficiency disease leukocyte adhesion deficiency, or LAD, develop life-threatening bacterial infections as a result of the inability of their leukocytes to adhere to the vessel wall and migrate to the sites of infection. Recently, the canine counterpart to LAD, known as canine leukocyte adhesion deficiency, or CLAD, has been described in Irish setter dogs. This review describes how the clinical phenotype of dogs with CLAD closely parallels that of children with the severe deficiency phenotype of LAD, thus enabling the CLAD dog to provide a disease-specific, large-animal model for testing novel hematopoietic stem cell and gene therapy strategies before their translation to children with LAD.
Descriptors: immune system, leukocyte adhesion deficiency syndrome, genetic disease, gene therapy, clinical techniques, genetic techniques, phenotype, animal model, genetic immunodeficiency disease leukocyte hematopoietic stem cell therapy.

Bethge, W.A., D.S. Wilbur, R. Storb, D.K. Hamlin, E.B. Santos, M.W. Brechbiel, D.R. Fisher, and B.M. Sandmaier (2003). Selective t-cell ablation with bismuth-213-labeled anti-tcralphabeta as nonmyeloablative conditioning for allogeneic canine marrow transplantation. Blood 101(12): 5068-5075. ISSN: 0006-4971.
NAL Call Number: RB145.A1B57
Abstract: Two major immunologic barriers, the host-versus-graft (HVG) and graft-versus-host (GVH) reactions, have to be overcome for successful allogeneic hematopoietic cell transplantation. T cells were shown to be primarily involved in these barriers in the major histocompatibility complex identical setting. We hypothesized that selective ablation of T cells using radioimmunotherapy together with postgrafting immunosuppression would suffice to ensure stable allogeneic engraftment. We had described a canine model of nonmyeloablative marrow transplantation in which host immune reactions were impaired by a single dose of 200 cGy total body irradiation (TBI), and both GVH and residual HVG reactions were controlled by postgraftifig immunosuppression with mycophenolate mofetil (MMF) and cyclosporine (CSP). Here, we substituted the alpha-emitter bismuth-213 (213Bi) linked to a monoclonal antibody (mAb) against T-cell receptor (TCR) alphabeta, using the metal-binding chelate diethylenetriaminepentaacetic acid (DTPA) derivative cyclohexyl-(CHX)-A", for 200 cGy TBI. Biodistribution studies using a gamma-emitting indium-111-labeled anti-TCRalphabeta mAb showed uptake primarily in blood, marrow, lymph nodes, spleen, and liver. Four dogs were treated with 0.13 to 0.46 mg/kg TCRalphabeta mAb labeled with 3.7 to 5.6 mCi/kg (137-207 MBq/kg) 213Bi. The treatment was administered in 6 injections on days -3 and -2 followed by transplantation of dog leukocyte antigen-identical marrow on day 0 and postgrafting immunosuppression with MMF/CSP. The therapy was well tolerated except for elevations of transaminases that were transient in all but one of the dogs. No other organ toxicities or signs of graft-versus-host disease were noted. The dogs had prompt allogeneic hematopoietic engraftment and achieved stable mixed donor-host hematopoietic chimerism with donor contributions ranging from 5% to 55% after more than 30 weeks of follow up.
Descriptors: immune system, chemical coordination and homeostasis, methods and techniques, graft vs host disease, immune system disease, allogeneic marrow transplantation, clinical techniques, therapeutic and prophylactic techniques, radioimmunotherapy, immunologic techniques, laboratory techniques, selective t cell ablation.

Brady, C.A. and C.M. Otto (2001). Systemic inflammatory response syndrome, sepsis, and multiple organ dysfunction. Veterinary Clinics of North America. Small Animal Practice 31(6): 1147-1162. ISSN: 0195-5616.
NAL Call Number: SF601.V523
Abstract: Companion animals with sepsis and multiple organ dysfunction can be the most challenging of all patients to treat. Current research in humans and laboratory models offers some exciting insights into the pathophysiology behind some of our most frustrating clinical challenges. This article applies several current concepts to a clinical case of pancreatitis and secondary sepsis to illustrate some of the cardiovascular, immune, and coagulation abnormalities commonly seen.
Descriptors: sepsis, multiple organ dysfunction, treatment, common problems, pathophysiology.

Caminis, A., C. Diez Fernandez, and P. Prieto (1999). Cell-surface expression of heat shock proteins in dog neutrophils after oxidative stress. Toxicology In Vitro 13(3): 437-443. ISSN: 0887-2333.
NAL Call Number: RA1190.T692
Abstract: The effect of oxidative stress induced by different concentrations of hydrogen peroxide on dog neutrophils was studied. This effect was measured using dichlorofluorescein-diacetate (DCFH-DA) and by the cell surface membrane expression of heat shock protein (HSP) 27 kDa, HSP 72 kDa and HSP 90 kDa families. Hydrogen peroxide induced a concentration-dependent increase in DCFH oxidation (from 10-6 M to 10-4 M), and an increase in the cell surface expression of HSPs families. At a concentration of 10-4 M, the percentage of positive cells that showed an oxidation of DCFH was 94.7% +- 5.2 (n = 3). Only vitamin E (but not vitamin C) at a concentration of 0.5 mM was able to inhibit the intracellular oxidative stress induced by hydrogen peroxide. The percentage of positive cells that express these proteins after the treatment with hydrogen peroxide (10-4 M) was: 74% +- 3.5 for HSP 27, 72% +- 2.6 for HSP 72 and 73% +- 1.2 for HSP 90 (n = 3). This cell surface expression was not abolishedby either vitamin C or vitamin E. Localization of HSPs in plasma membrane is of immunological interest because they have been implicated in autoimmune diseases.
Descriptors: biochemistry and molecular biophysics, immune system, chemical coordination and homeostasis, metabolism, oxidative stress.

Casal, M., S. Ryan, J. Rhodes, and J. Scheidt (2003). Immunological aspects of x-linked ectodermal dysplasia: a dog model for the human disease. American Journal of Human Genetics 73(5): 454. ISSN: 0002-9297.
NAL Call Number: QH431.A1A54
Descriptors: molecular genetics, biochemistry and molecular biophysics, X linked ectodermal dysplasia, genetic disease, immune system disease, integumentary system disease, respiratory system disease, genetics, immunology, pathology, transmission, pulmonary disorders, respiratory system disease, respiratory tract infection, infectious disease, cytokine production, immune system defects, immunological aspects, systemic, localized humoral, cellular responses.

Chaprazov, T. (2006). Changes in some clinical and laboratory indices in dogs with experimental Staphylococcus aureus sepsis. Bulgarian Journal of Veterinary Medicine 9(1): 51-60. ISSN: 1311-1477.
Abstract: The study was conducted in 6 clinically healthy dogs that were experimentally infected via intravenous injection of 5 mL broth culture of a field Staphylococcus aureus strain (1.2x109 cells/mL). The dynamics of rectal temperature, heart and respiratory rates and haematological parameters (erythrocyte counts, haematocrit, haemoglobin, erythrocyte sedimentation rate, total and differential leukocyte counts, total protein, fibrinogen, bilirubin, urea, creatinine, pyruvate and lactate) were observed for 28-day period. It was observed that after 2 h of post-infection an increase in body temperature, tachycardia and tachypnea occurred. Followed by a marked leukopenia with an increased leukocyte counts at the 24th h with a left shift were noticed. Statistically significant increased in the values of blood biochemical parameters were observed for fibrinogen and bilirubin, while the concentrations of pyruvate decreased. The levels of total protein, creatinine, urea and lactate remained stable without significant deviations during the entire period of the survey..
Descriptors: bilirubin, blood chemistry, body temperature, clinical aspects, creatinine, disease markers, erythrocytes, experimental infections, fibrinogen, hematocrit, hematology, hemoglobin, heart rate, lactic acid, leukocytes, leukopenia, pathogenesis, pyruvic acid, respiration, septicaemia, urea, dogs, Staphylococcus aureus.

Christopher, S.A., B. Chung, J. Kennedy, S. Krakowka, P.F. Moore, K.I. Weinberg, and P.J. Felsburg (2004). Enhanced t cell regeneration following allogeneic bone marrow transplantation (bmt) in xscid dogs co-transplanted with il-7 transduced mesenchymal stem cells (msc). FASEB Journal 18(4-5): Abst. 560.4. ISSN: 0892-6638.
Online: http://www.fasebj.org/
NAL Call Number: QH301.F3
Descriptors: immune system, chemical coordination and homeostasis, methods and techniques, lymphopenia, blood and lymphatic disease, allogenic bone marrow transplantation, clinical techniques, therapeutic and prophylactic techniques, gene therapy, genetic techniques, transfection, laboratory techniques, t cell regeneration, enhancement, thymopoiesis.

Creevy, K.E., T.R.J. Bauer, L.M. Tuschong, L.J. Embree, L. Colenda, K. Cogan, M.F. Starost, M.E. Haskins, and D.D. Hickstein (2003). Canine leukocyte adhesion deficiency colony for investigation of novel hematopoietic therapies. Veterinary Immunology and Immunopathology 94(1/2): 11-22. ISSN: 0165-2427.
NAL Call Number: SF757.2.V38
Abstract: The genetic immunodeficiency disease canine leukocyte adhesion deficiency (CLAD) was originally described in juvenile Irish Setters with severe, recurrent bacterial infections. CLAD was subsequently shown to result from a mutation in the leukocyte integrin CD18 subunit which prevents leukocyte surface expression of the CD11/CD18 complex. We describe the development of a mixed-breed CLAD colony with clinical features that closely parallel those described in Irish Setters. We demonstrate that the early identification of CLAD heterozygotes and CLAD-affected dogs by a combination of flow cytometry and DNA sequencing allows the CLAD-affected animals to receive life-saving antibiotic therapy. The distinct clinical phenotype in CLAD, the ability to detect CD18 on the leukocyte surface by flow cytometry, and the history of the canine model in marrow transplantation, enable CLAD to serve as an attractive large-animal model for the investigation of novel haematopoietic stem cell and gene therapy strategies.
Descriptors: diagnosis, DNA sequencing, flow cytometry, genetic disorders, heterozygotes, leukocyte disorders, leukocytes.

Felsburg, P.J., B.J. Hartnett, T.A. Gouthro, and P.S. Henthorn (2003). Thymopoiesis and t cell development in common gamma chain-deficient dogs. Immunologic Research 27(2-3): 235-245. ISSN: 0257-277X.
NAL Call Number: QR180.S88
Abstract: Our laboratory has identified an X-linked severe combined immunodeficiency (XSCID) in dogs that is the result of mutations in the common gamma chain (gammac) subunit of the interleukin-2 (IL-2), IL-4, IL-7, IL-9, IL-15, and IL-21 receptors. Canine XSCID, unlike genetically engineered gammac-deficient mice, has a clinical and immunologic phenotype virtually identical to human XSCID, suggesting species-specific differences exist in the role of the gammac and its associated cytokines in mice in comparison to their role in humans and dogs. This review compares and contrasts thymopoiesis and postnatal T cell development in gammac-deficient (XSCID) dogs raised in a conventional environment, with gammac-deficient dogs raised in a gnotobiotic environment. Therapy to accelerate T cell regeneration following hematopoietic stem cell transplantation or gene therapy is also discussed.
Descriptors: cell biology, genetics, immune system, chemical coordination and homeostasis, methods and techniques, X linked severe combined immunodeficiency, genetic disease, immune system disease, gene therapy, clinical techniques, genetic techniques, laboratory techniques, therapeutic and prophylactic techniques, genetic engineering, hematopoietic stem cell transplantation, thymopoiesis.

Georges, G.E., R. Storb, J.M. Zaucha, A.G. Taranova, T. Gooley, and R.A. Nash (2003). Il-2 does not enhance the conversion to complete donor chimerism following nonmyeloablative hematopoietic cell transplantation in dogs. Bone Marrow Transplantation 31(11): 1027-1031. ISSN: 0268-3369.
Abstract: A dog model of stable mixed hematopoietic chimerism was established in which leukocyte-antigen-identical littermates receive nonmyeloablative total body irradiation before hematopoietic cell transplantation and postgrafting immunosuppression with mycophenolate mofetil and cyclosporine. Unmodified donor lymphocyte infusion (DLI) into stable mixed chimeras failed to increase donor chimerism, while DLI from donors sensitized to recipient minor-histocompatibility antigens promptly converted all recipients to complete donor chimerism. This established a model for studying approaches to enhance the graft-versus-host (GVH)-effect, a potential surrogate for graft-versus-leukemia activity. We asked if interleukin-2 (IL-2) given after unmodified DLI could result in reliable conversion to complete donor chimerism. IL-2, 4X105 IU/kg/day, was administered to six mixed chimeric dogs for 14 days. Four dogs received unmodified DLI with IL-2. At 20-40 weeks after DLI, all dogs remained mixed chimeras. For the two recipients of IL-2 only, mixed chimerism also remained unchanged. These results show that IL-2 given with DLI after nonmyeloablative transplantation in dogs is not effective in reliably converting mixed to complete donor chimerism.
Descriptors: immune system, chemical coordination and homeostasis, pharmacology, nonmyeloablative hematopoietic cell transplantation, clinical techniques, therapeutic and prophylactic techniques, total body irradiation, laboratory techniques, unmodified donor lymphocyte infusion, complete donor chimerism, graft vs host effect, immunosuppression, mixed donor chimerism.

Hammerberg, B. and A.K. Smith (2003). Membrane-bound and soluble stem cell factor expression by skin cells during atopic dermatitis in dogs. Journal of Allergy and Clinical Immunology 111(2 Abstract Supplement): S342-S343. ISSN: 0091-6749.
NAL Call Number: 448.8 J8236
Descriptors: cell biology, immune system, chemical coordination and homeostasis, integumentary system, chemical coordination and homeostasis, immunohistochemistry, immunologic techniques, laboratory techniques, reverse transcriptase polymerase chain reaction, genetic techniques.

Higgins, M.A., B.R. Berridge, B.J. Mills, A.E. Schultze, H. Gao, G.H. Searfoss, T.K. Baker, and T.P. Ryan (2003). Gene expression analysis of the acute phase response using a canine microarray. Toxicological Sciences 74(2): 470-484. ISSN: 1096-6080.
NAL Call Number: RA1190.F8
Abstract: The safety of pharmaceuticals is typically assessed in the dog and rat prior to investigation in humans. As a result, a greater understanding of adverse effects in these preclinical testing species would improve safety assessment. Despite this need, there is a lack of tools to examine mechanisms and identify biomarkers in the dog. To address this issue, we developed an Affymetrix-based oligonucleotide microarray capable of monitoring the expression of thousands of canine genes in parallel. The custom canine array contains 22,774 probe sets, consisting of 13,729 canine and 9045 human-derived probe sets. To improve cross-species hybridization with human-derived probes, the detection region was moved from the variable 3' UTR to the more homologous coding region. Testing of this strategy was accomplished by comparing hybridization of naive dog liver RNA to the canine array (coding region design) and human U133A array (standard 3' design). Although raw signal intensity was greater with canine-specific probe sets, human-derived probes detected the expression of additional liver transcripts. To assess the ability of this tool to detect differential gene expression, the acute phase response was examined in beagle dogs given lipopolysaccharide (LPS). Hepatic gene expression 4 and 24 h post-LPS administration was compared to gene expression profiles of vehicle-treated dogs (n = 3/group). Array data was consistent with an acute inflammatory response, with transcripts for multiple cytokines and acute phase proteins markedly induced 4 h after LPS challenge. Robust changes in the expression of transcripts involved with glucose homeostasis, biotransformation, and extracellular matrix remodeling were observed 24 h post-dose. In addition, the canine array identified several potential biomarkers of hepatic inflammation. Strong correlations were found between gene expression data and alterations in clinical chemistry parameters such as serum amyloid A (SAA), albumin, and alkaline phosphatase (ALP). In summary, this new genomic tool successfully detected basal canine gene expression and identified novel aspects of the acute phase response in dog that shed new light on mechanisms underlying inflammatory processes.
Descriptors: immune system, chemical coordination and homeostasis, molecular genetics, biochemistry and molecular biophysics, toxicology, affymetrix based oligonucleotide microarray, genetic techniques, laboratory techniques, canine microarray, gene expression analysis, acute phase response, biotransformation, canine genomics, extracellular matrix remodeling, glucose homeostasis, hepatic gene expression, inflammation, toxicogenomics.

Hogan, W.J., M.L. Sorror, R. Diaconescu, E. Zellmer, M.T. Little, and R. Storb (2003). Marrow engraftment after nonmyeloablative conditioning followed by postgrafting immunosuppression with mycophenolate mofetil, sirolimus and cyclosporine in a canine model. Blood 102(11): 396b. ISSN: 0006-4971.
NAL Call Number: RB145.A1B57
Abstract: Previous studies have demonstrated that postgrafting immunosuppression with cyclosporine (CSP) for 5 weeks combined with mycophenolate mofetil (MMF) for 4 weeks will allow durable donor engraftment after 200 cGy TBI but not after 100 cGy TBI. Similarly, a combination of CSP for 5 weeks combined with sirolimus (SRL) for 4 weeks can secure engraftment after 200 cGy. We sought to determine whether the combination of all three agents to the post-grafting immunosuppression regimen would enhance durable engraftment after non-myeloablative conditioning in a canine model at 100 cGy TBI. Four dogs received 100 cGy TBI followed by a dog leukocyte antigen (DLA) identical marrow graft and post-grafting immunosuppression with MMF (10mg/kg sc bid, day 0 to 27), SRL (0.05 mg/d sc, day 0 to 27) and CSP (10 mg/kg po bid, day-1 to +35). All dogs showed initial engraftment with a median of 5% donor representation in both the granulocyte and lymphocyte fractions at the completion of immunosuppression. Three dogs went on to reject their grafts at a median of 10 (range 9-12) weeks and all three had autologous reconsitiution without severe or prolonged cytopenias. The fourth dog remains engrafted with 6% donor representation in both granulocyte and mononuclear lineages at 5 weeks. The regimen was well tolerated, all dogs remain alive and well, none experienced significant hepatic or renal dysfunction throughout the peritransplant period despite high CSP and SRL levels consistent with a pharmacokinetic interaction between SRL and CSP. Dose reductions were performed as necessary to maintain therapeutic levels of both CSP and SRL. Gastrointestinal toxicity predominated with anorexia, diarrhea and weight loss which was transient. Mild myelosuppression was observed with median (range) absolute neutrophil count (ANC) nadirs of 3.3 (2.1-4.1)X109/L and median (range) platelet nadirs of 34 (9-72)X109/L respectively. We conclude that the addition of MMF to SRL/CSP is associated with acceptable toxicity but rates of durable engraftment are not increased beyond that achieved by SRL/CSP alone. It is likely that 200 cGy TBI represents a threshold for conditioning below which engraftment cannot be secured by intensification of post-grafting immunosuppression alone. Alternative strategies to further reduce the intensity of conditioning may include the use of peripheral blood derived stem cell products or promotion of tolerance to donor cells prior to HCT by co-stimulatory blockade or antimetabolite blockade after pretransplant exposure to donor cells.
Descriptors: blood and lymphatics, transport and circulation, immune system, chemical coordination and homeostasis, pharmacology, absolute neutrophil count, histology and cytology techniques, laboratory techniques, nonmyeloablative conditioning, clinical techniques, therapeutic and prophylactic techniques, marrow engraftment.

Horton, P.J., W.J. Hawthorne, S. Walters, T. Patel, G.J. Stewart, R.D. Allen, and J.R. Chapman (2008). Tolerance induction in a large-animal model using total lymphoid irradiation and intrathymic bone marrow. Transplantation 86(12): 1830-6.
NAL Call Number: QP89.T73
Abstract: BACKGROUND: Immunological unresponsiveness of T cells to alloantigen can be induced by intrathymic injection of donor-specific antigen in small-animal models. Intrathymic tolerance to vascularized grafts in large animals has not previously been reported. METHODS: Thirty-two dogs were allocated into dog leukocyte antigen DP locus allele (class II)-matched donor-recipient pairs. Female recipients were paired with male donors. Tissue typing was based on restriction fragment length polymorphism. Recipients were given 18 Gy total lymphoid irradiation in 16 fractions (1.125 Gy each) over 4 weeks. Thoracotomy after the 6th fraction permitted perithymic (n=4) or intrathymic (n=4) injection of donor bone marrow (BM) or intrathymic injection of saline (n=5). Another group received intravenous peripheral BM infusion (n=3). Fifty days postthoracotomy recipients underwent bilateral nephrectomy and donor-specific kidney transplantation. Acute rejection, suspected when serum creatinine was more than 600 mumol/L or urea was more than 40 mmol/L, was confirmed histologically. Full-thickness skin grafts followed more than 100 days posttransplantation. Tissue samples were taken for Y-chromosome polymerase chain reaction. RESULTS: One intrathymic (25%) and three perithymic (75%) BM recipients developed tolerance to renal allografts. Three intrathymic BM recipients rejected after 27, 32, and 54 days and one perithymic BM recipient rejected after 42 days. All recipients given peripheral BM or saline had rejected by 29 and 38 days, respectively. All recipients surviving more than 100 days posttransplantation, accepted donor specific and rejected dog leukocyte antigen-DP locus allele (class II) identical third-party skin grafts. Polymerase chain reaction detected intrathymic but not hematopoietic chimerism in sex-mismatched pairs. CONCLUSIONS: Fractionated total lymphoid irradiation and perithymic or intrathymic donor-specific BM induced tolerance to renal and skin allografts without inducing hematopoietic chimerism.
Descriptors: histocompatibility testing methods, immune tolerance, isoantigens pharmacology, kidney transplantation immunology, lymphatic system radiation effects, t lymphocytes immunology, bone marrow cells pathology, bone marrow cells physiology, bone marrow transplantation, computer simulation, dogs, histocompatibility antigens class ii immunology, lymphatic system immunology, models, animal, nephrectomy, polymorphism, restriction fragment length, skin transplantation immunology.

Huang, J.C., W.T. Lu, B.R. Hsu, C.H. Kuo, S.H. Fu, H.M. Chen, N.K. Yao, and J.H. Juang (2003). Canine islet isolation, cryopreservation, and transplantation to nude mice. Chang Gung Medical Journal 26(10): 722-8.
Abstract: BACKGROUND: Successful human islet transplantation has led to insulin independence in type 1 diabetes. Dogs constitute an animal model for preclinical studies. We present our recent experience in canine islet isolation, cryopreservation and transplantation. METHODS: Twenty-seven pancreases from mongrel dogs, weighing 9-31 kg, were removed. Each pancreas was digested with collagenase, and then purified by density gradients. Islet number and purity were counted, and the viability of isolated islets was assessed in vitro by static incubation, perifusion study and in vivo transplantation into nondiabetic or diabetic nude mice. Additionally. freshly isolated islets were cryopreserved for 1 week, and then studied in vitro. RESULTS: The islet yield and purity were 121,000 +/- 135,000 IEQ per pancreas and 81.4 +/- 1.2%, respectively. The stimulation index (insulin release in 300 mg/dl glucose/insulin release in 100 mg/dl glucose) of the isolated islets was 6.6 +/- 1.9 (N = 7), and first and second phases of insulin secretion were demonstrated during perifusion study. After 1-week cryopreservation, the islet number decreased from 1,000 to 540 (N = 1) and insulin content decreased from 50.95 to 39.23 microg/150 islets (N = 1). These islets maintained their insulin response to high glucose. Four weeks after transplantation, the grafts showed abundant beta-cells and significant insulin content. Normoglycemia was achieved in 14 of 23 diabetic recipients after transplantation with 2,000 freshly isolated islets. CONCLUSION: Canine islets isolated at our laboratory were viable and maintained their physiological function both in vitro and in vivo.
Descriptors: animal model, preclinical studies, canine islet isolation, cryopreservation, transplantation, human islet transplantation, diabetes.

Kemming, G., J.B. Messick, W. Mueller, G. Enders, F. Meisner, S. Muenzing, H. Kisch Wedel, A. Schropp, C. Wojtczyk, K. Packert, K. Messmer, and E. Thein (2004). Can we continue research in splenectomized dogs? Mycoplasma haemocanis: old problem - new insight. European Surgical Research 36(4): 198-205. ISSN: 0014-312X.
Descriptors: cardiovascular system, transport and circulation, infection, Mycoplasma haemocanis infection, bacterial disease, diagnosis, therapy, blood smear, diagnostic techniques, laboratory techniques, specific polymerase chain reaction, genetic techniques.

Kosaka, T., Y. Kaneko, Y. Nakada, M. Matsuura, and S. Tanaka (1996). Effect of chitosan implantation on activation of canine macrophages and polymorphonuclear cells after surgical stress. Journal of Veterinary Medical Science 58(10): 963-967. ISSN: 0916-7250.
NAL Call Number: SF604.J342
Abstract: The effect of cotton type chitosan implantation under the skin on the immunological response mediated by macrophages and whole blood was evaluated chemiluminescence (CL) in dogs. The number of white blood cells was significantly decrease until 120 hr after operation in the control group (p lt 0.05), while chitosan implantation increased the number of white blood cells, particularly neutrophils, from 24 to 96 hr after implantation (p lt 0.05). The CL response of whole blood in the control group seemed to be reduced at 48 and 96 after operation (p lt 0.05), but in the chitosan groups it maintained higher activity until 120 hr after implantation (p lt 0.05). The macrophage activity measured by CL assay in the control group was markedly decreased form 24 to 120 hr after operation, and that for the 5 mg/kg chitosan group was also decreased at 24 and 48 hr after implantation (p lt 0.05), although high activity was observed from 72 to 120 hr after implantation (p lt 0.05). Neither 5 mg/kg nor 10 mg/kg chitosan groups showed and reduction in CL response of macrophages after operation, and the 20 mg/kg chitosan group retained high CL response of macrophages until 120 hr after operation (p lt 0.05). These findings suggest that chitosan may be and efficacious and useful immunopotentiator for preventation of immunosuppression after surgery.
Descriptors: chemiluminescence, implantation, macrophages, stress, chitosan, nonspecific immunostimulation, surgery, blood and lymphatics, transport and circulation, cell biology, immune system, chemical coordination and homeostasis, physiology , veterinary medicine, medical sciences, analytical method, blood and lymphatics, implantation, immunosuppressant method, macrophage , polymorphonuclear cell, surgical stress.

Kuhr, C.S., M. Yunusov, G. Sale, C. Loretz, and R. Storb (2007). Long-term tolerance to kidney allografts in a preclinical canine model. Transplantation 84(4): 545-7. ISSN: 0041-1337.
NAL Call Number: QP89.T73
Abstract: Durable immune tolerance supporting vascularized allotransplantation offers the possibility of extending graft survival and avoiding harmful complications of chronic immunosuppression. Immune tolerance to renal allografts was induced in a preclinical canine model through engraftment of donor hematopoietic cells using a combination of low-dose total body irradiation and a short course of immunosuppression. Subsequently, donor renal allografts were transplanted accompanied by bilateral native nephrectomies. With 5-year follow up, we found normal renal function in all recipients and no histological evidence of acute or chronic rejection. This tolerance does not extend universally to donor skin grafts, however, with two of four animals rejecting delayed donor skin grafts. Hematopoietic chimerism produces durable and robust immune tolerance to kidney allografts, although incomplete tolerance to donor skin grafting.
Descriptors: graft survival immunology, kidney transplantation immunology, transplantation tolerance immunology, dogs, immune tolerance immunology, immunosuppression, kidney pathology, kidney physiology, kidney transplantation pathology, kidney transplantation physiology, models, animal, skin transplantation immunology, time factors.

Lesnikova, M., M.C. Jensen, A. Bukovsky, A. Nikitine, B. Thomasson, J. Morris, R.F. Storb, H.P. Kiem, R.A. Nash, and G.E. Georges (2003). Resistance to mycophenolate mofetil (mmf) after lentiviral transduction of canine t cells with a mutant impdh vector. Blood 102(11): 499B. ISSN: 0006-4971.
NAL Call Number: RB145.A1B57
Abstract: Several clinical studies of adoptive immunotherapy with genetically modified (GM) donor T cells infused after allogeneic hematopoietic cell transplantation (HCT) have shown limited in vivo function of the ex vivo expanded T cells. Multiple factors including duration of ex vivo cell culture, immunogenicity of the transgene, lack of a selective advantage for the GM T cells and insufficient preclinical data in large animal models may have impaired successful clinical translation of this approach. To help address these problems, we constructed a self-inactivating, replication incompetent lentiviral vector expressing a (Thr-333fwdarwIle, Ser-351fwdarwTyr) trans-dominant mutant inosine monophosphate dehydrogenase II (IMPDH*), which renders transduced T cells resistant to MMF with intact de novo guanosine synthesis. MMF administration after infusion of IMPDH* transduced T cells would inhibit host immune responses to the GM T cells and provide a proliferative advantage for transduced T cells. Lentiviral vector transduction can reduce the time in culture since T cell proliferation is not required. In preparation for in vivo studies in the dog model of allogeneic HCT, we first confirmed function of the IMPDH* vector in immortalized T cell lines and then optimized conditions for lentiviral transduciton of fresh, primary dog T cells to achieve 20-30% transduction efficiency. Replication incompetent, VSV-G pseudotyped IMPDH* lentiviral vector, pRRLsin.cPPT.MSCV.IMPDH*.Wpre, was prepared from transiently transfected 293T cells. Immortalized dog T lymphocyte CLGL-90 cells were transduced with IMPDH* lentiviral vector and selected in 1 muM MMF for 14 days. Transduced cells were cloned by limiting dilution and assayed for MMF resistance. All IMPDH* CLGL-90 clones were uniformly resistant to MMF 0.1 to 10 muM, compared to control CLGL cells. Quantitative TaqMan PCR assay with lentiviral-specific primers showed transduced CLGL clones had up to 3 lentiviral vector insertion copies per clone. Next, fresh CD3+ T cells were isolated from dog peripheral blood with immunomagnetic selection (Miltenyi) or plastic adherence to >90% purity. Cells were cultured with IL-2 (10 IU/mL) and IL-7 (20 ng/mL) for 72 hours. T cells were transduced once at an MOI of 20 with the spinoculation method. Transduced cells were stimulated after 24 hours with MHC-mismatched dendritic cells (DC) (10:1 responder: stimulator ratio) and selected in 0.5 muM MMF 2 days later. After 2nd stimulation with DCs, cells were assessed for proliferation with 3H-thymidine uptake. In the presence of MMF, IMPDH* transduced cells had 3-fold increased proliferation over nontransduced control T cells. In summary, lentiviral vector expressing trans-dominant mutant IMPDH* rendered transduced dog T cells resistant to MMF. These results establish the IMPDH* vector as a tool to confer in vivo resistance to MMF. We are planning adoptive immunotherapy studies with infusion of IMPDH* transduced donor T cells into recipient dogs treated with MMF after allogeneic HCT to determine the feasibility of in vivo selection of GM T cells.
Descriptors: blood and lymphatics, transport and circulation, immune system, chemical coordination and homeostasis, molecular genetics, biochemistry and molecular biophysics, pharmacology, adoptive immunotherapy, clinical techniques, immunologic techniques, laboratory techniques, therapeutic and prophylactic techniques, allogeneic hematopoietic cell transplantation, clinical techniques, lentivirus transduction, drug resistance mechanism, host immune response, transduction efficiency.

Lester, T., M. Passage, R. Yang, C. Tanaka, V. Anand, J. Lemontt, and E. Kakkis (2003). Reduction of the immune response to therapeutic enzyme replacement therapy in a canine model of a lysosomal storage disorder. Journal of Inherited Metabolic Disease 26(Supplement 2): 143. ISSN: 0141-8955.
NAL Call Number: RC627.8.J68
Descriptors: enzymology, biochemistry and molecular biophysics, immune system, chemical coordination and homeostasis, metabolism, pharmacology, canine mucopolysaccharidosis I, lysosomal storage disorder, connective tissue disease, genetic disease, metabolic disease, therapy, enzyme replacement therapy, clinical techniques, therapeutic and prophylactic techniques, drug regimen.

Linfert, D., T. Chowdhry, and H. Rabb (2009). Lymphocytes and ischemia-reperfusion injury. Transplantation Reviews 23(1): 1-10.
NAL Call Number: QP89.T7
Abstract: Ischemia reperfusion injury (IRI) is a common and important clinical problem in many different organ systems, including kidney, brain, heart, liver, lung, and intestine. IRI occurs during all deceased donor organ transplants. IRI is a highly complex cascade of events that includes interactions between vascular endothelium, interstitial compartments, circulating cells, and numerous biochemical entities. It is well established that the innate immune system, such as complement, neutrophils, cytokines, chemokines, and macrophages participate in IRI. Recent data demonstrates an important role for lymphocytes, particularly T cells but also B cells in IRI. Lymphocytes not only participate in augmenting injury responses after IRI, but could also be playing a protective role depending on the cell type and stage of injury. Furthermore, lymphocytes appear to be participating in the healing response from IRI. These new data open the possibility for lymphocyte targeted therapeutics to improve the short and long term outcomes from IRI.
Descriptors: b lymphocytes immunology, organ transplantation adverse effects, reperfusion injury immunology, t lymphocytes immunology, cd4 positive t lymphocytes immunology, cadaver, dogs, immunosuppressive agents therapeutic use, mice, mice, scid immunology, models, animal, reperfusion injury physiopathology, reperfusion injury prevention and control, retrospective studies, tissue donors.

Maeda, S., K. Ohmori, N. Yasuda, K. Kurata, M. Sakaguchi, K. Masuda, K. Ohno, and H. Tsujimoto (2004). Increase of CC chemokine receptor 4-positive cells in the peripheral CD4 cells in dogs with atopic dermatitis or experimentally sensitized to Japanese cedar pollen. Clinical and Experimental Allergy 34(9): 1467-1473. ISSN: 0954-7894.
Abstract: BACKGROUND: Since dogs frequently develop allergic diseases, similar to those in humans, dogs represent a possible animal model for allergy in humans. In human atopic dermatitis (AD), CC chemokine receptor 4 (CCR4) has been shown to play an important role in the development of allergic inflammation of AD; however, the association between allergic reaction and CCR4 is not well understood in dogs. OBJECTIVE: To examine CCR4 expression in peripheral blood CD4+ cells in dogs that had AD and were experimentally sensitized with Japanese cedar pollen. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from 17 dogs with AD. The proportion of CCR4+ cells in peripheral blood CD4+ cells (CCR4/CD4) was evaluated by flow cytometry and compared with that in 10 healthy dogs. Similarly, in dogs that were experimentally sensitized to Japanese cedar pollen antigen, the proportion of CCR4/CD4 was examined pre- and post-sensitization. RESULTS: The proportion of CCR4/CD4 in dogs with AD was 40.3+/-3.3%, which was significantly higher than that in normal dogs (23.6+/-4.3%) (P<0.01). In the experimentally sensitized dogs, the proportion of CCR4/CD4 was 25.4+/-2.6% at pre-sensitization and it was significantly increased (29.8+/-2.9%) at post-sensitization (P<0.01). CONCLUSION: The proportion of CCR4+ cells in peripheral blood CD4+ cells was measured in dogs with allergic conditions. The present findings indicate that CCR4+ cells may be involved in the pathogenesis of allergy in dogs as in humans.
Descriptors: animal model, allergies, atopic dermatitis, cedar pollen, chemokine receptor 4 (CCR4).

Marsella, R. and C.F. Nicklin (2001). Sulphido-leukotriene production from peripheral leukocytes and skin in clinically normal dogs and house dust mite positive atopic dogs. Veterinary Dermatology 12(1): 3-12. ISSN: 0959-4493.
NAL Call Number: SF901.V47
Descriptors: atopy, leukotrienes, leukocytes, skin, skin diseases, allergies, allergic reactions, house dust mites, dermatophagoides, atopy, dermatitis, leukocytes, skin, leukotrienes, pruritus, IgE, blood serum, allergens.
Language of Text: Summaries in German, Spanish, French.

Marsella, R., C.F. Nicklin, S. Saglio, and J. Lopez (2004). Investigation on the effects of topical therapy with 0.1% tacrolimus ointment (Protopic) on intradermal skin test reactivity in atopic dogs. Veterinary Dermatology. 15(4): 218-224. ISSN: 0959-4493.
NAL Call Number: SF901.V47
Descriptors: atopy, dermatitis, dermatological agents, drug therapy, ointments, pharmacology, skin diseases, skin tests, topical application.
Language of Text: Summaries in German; Spanish; French.

Petersen, S.M., M. Gendelman, K.M. Murphy, M. Torbenson, R.J. Jones, J.E. Althaus, G. Stetten, C. Bird, and K.J. Blakemore (2007). Use of T-cell antibodies for donor dosaging in a canine model of in utero hematopoietic stem cell transplantation. Fetal Diagnosis and Therapy 22(3): 175-9. ISSN: 1015-3837.
Abstract: AIM: Microchimerism following canine in utero hematopoietic stem cell transplantation (IUHSCT) development of T-cell dosing regimens. OBJECTIVE: To investigate the use of anti-T-cell antibodies for cell dosing of the donor graft in a canine model of IUHSCT. STUDY DESIGN: Canine IUHSCT was performed by ultrasound-guided intraperitoneal injection in days 35-38 of fetal canines with CD34(+) cells at doses of 4.5 x 10(8) to 1.3 x 10(9) cells/kg and T cells (CD3(+) CD5(+)) at doses of 8 x 10(6) to 8.8 x 10(8) cells/kg. Postnatal studies included tissue histology and polymerase chain reaction-based chimerism analysis. RESULTS: Term survival was 86-100%. Microchimerism (0-2%) was detected in five of eight recipients in multiple tissues. Histopathology revealed no evidence of graft-versus-host disease (GVHD). CONCLUSION: Canine IUHSCT is a useful model to investigate the role of donor T cells in engraftment and GVHD. IUHSCT at early gestational ages with high doses of donor T cells in the graft yields microchimerism in multiple tissues without GVHD. (c) 2007 S. Karger AG, Basel.
Descriptors: fetal therapies methods, fetal tissue transplantation methods, hematopoietic stem cell transplantation methods, animals, newborn, antilymphocyte serum administration and dosage, base sequence, dna primers genetics, dogs, fetal tissue transplantation immunology, graft survival immunology, models, animal, t lymphocytes immunology, tissue donors, transplantation chimera genetics, transplantation chimera immunology.

Pumarola, M., P.F. Moore, and G.D. Shelton (2004). Canine inflammatory myopathy: analysis of cellular infiltrates. Muscle and Nerve 29(6): 782-789. ISSN: 0148-639X.
Abstract: Inflammatory myopathies (IMs) occur relatively frequently in dogs, and, with the exception of masticatory muscle myositis (MMM), have not been characterized. This study analyzed the distribution and types of cellular infiltrates in 21 cases of generalized IM, 3 cases of focal IM (MINIM), and 1 case with features of both generalized and focal IM, using a panel of monoclonal antibodies to cell surface markers. In generalized IM, mononuclear cells showed an endomysial and perimysial distribution with invasion of non-necrotic fibers similar to human IM. T lymphocytes with T-cell receptor (TCR)alphabeta predominated. Distinct differences were seen in MMM including prominent B-cell infiltration, dendritic cells and macrophages in greater numbers than T cells, and numerous T cells with TCRgammadelta. Thus, generalized IM and MMM appear to be distinct diseases with different mechanisms. Canine generalized IM may be an important animal model for human IM.
Descriptors: immune system, chemical coordination and homeostasis, infection, muscular system, movement and support, veterinary medicine, medical sciences, dermatomyositis, connective tissue disease, integumentary system disease, muscle disease, polymyositis, connective tissue disease, muscle disease.

Ramiro, M.J., J.J. Zarate, T. Hanke, D. Rodriguez, J.R. Rodriguez, M. Esteban, J. Lucientes, J.A. Castillo, and V. Larraga (2003). Protection in dogs against visceral leishmaniasis caused by leishmania infantum is achieved by immunization with a heterologous prime-boost regime using dna and vaccinia recombinant vectors expressing lack. Vaccine 21(19-20): 2474-2484. ISSN: 0264-410X.
NAL Call Number: QR189.V32
Abstract: A heterologous prime-boost vaccination regime with DNA and recombinant vaccinia virus (rVV) vectors expressing relevant antigens has been shown to enhance specific cellular immune responses and to elicit protection against a variety of pathogens in animal models. In this paper, we describe the effectiveness of the prime-boost strategy by immunizing dogs with a plasmid carrying the gene for the LACK antigen from Leishmania infantum (DNA-LACK) followed by a booster with a rVV containing the same gene (rVV-LACK). Thereafter, animals were challenged with L. infantum to induce visceral leishmaniasis (VL). In the vaccinated dogs as compared with the controls, the outcome of the infection after challenge with a high inoculum (108) of L. infantum stationary promastigotes was assessed by tissue parasite load, specific anti-Leishmania antibody production, cytokine level and development of clinical signs of leishmaniasis. We observed a 60% protection against infection in dogs immunized by DNA-LACK prime/rVV/-LACK boost while two doses of DNA-LACK did not elicit protection against the disease. The interleukin 4 (IL-4), interferon gamma (IFNgamma) and IL-12 (p40 subunit) cytokine mRNA expression profiles in PBMC as well as lymphocyte proliferative response and the IgG2/IgG1 ratios specific for LACK suggest that in vaccinated animals there is triggering of cellular immune responses. This type of DNA/rVV prime/boost immunization approach may have utility against visceral leishmaniasis-in dogs.
Descriptors: immune system, methods and techniques, parasitology, visceral leishmaniasis, parasitic disease, heterologous prime boost regime immunization, clinical techniques, therapeutic and prophylactic techniques.

Shengwu, Z., L. Qingfeng, J. Hao, J. Banich, F. Kaiding, C. Benson, W. Huiyong, Z. Danning, G. Bing, L. Qinxiu, T. Lujia, Z. Tao, L. Yuping, and Z. Tisheng (2007). Developing a canine model of composite facial/scalp allograft transplantation. Annals of Plastic Surgery 59(2): 185-194. ISSN: 0148-7043.
Abstract: The study developed a model of composite facial and scalp allograft transplantation in canines. Dog cadavers were used for anatomy study. Three types of autotransplantations and 2 types of allotransplantations were performed. Cyclosporin A and methylprednisolone or prednisone were given for immunosuppression. Two long-term-surviving dogs with autologous facial transplantation developed leakage of salivary secretions. In the allotransplantation group (n = 5), 1 dog presented rejection at 28 postoperative days but was successfully treated and survived long term (>402 days); 1 dog died of pulmonary infection at 29 postoperative days; 3 dogs survived (>252, >222, and >201 days). Serial electromyelogram studies revealed progressive improvement of the function of the orbicularis oculi muscle. The study indicated that the unilateral superior half of the composite facial and scalp, including one third of the inferior tarsal plate and palpebral conjunctiva (type IV flap) allograft transplantation model, was an ideal model for the study of facial allotransplantation.
Descriptors: face surgery, models, animal, scalp transplantation, tissue transplantation methods, dogs.

Suter, S.E., T.A. Gouthro, P.A. Mcsweeney, R.A. Nash, M.E. Haskins, P.J. Felsburg, and P.S. Henthorn (2004). Isolation and characterization of pediatric canine bone marrow cd34+ cells. Veterinary Immunology and Immunopathology 101(1-2): 31-47. ISSN: 0165-2427.
NAL Call Number: SF757.2.V38
Descriptors: animal models, bone marrow, characterization, genetic disorders, hematopoiesis, isolation, transplantation, blood and lymphatics, transport and circulation, immune system, chemical coordination and homeostasis, methods and techniques, veterinary medicine, medical sciences, bone marrow transplantation, clinical techniques, therapeutic and prophylactic techniques, gene therapy, clinical techniques, genetic techniques, laboratory techniques, therapeutic and prophylactic techniques, gene transfer, liquid suspension cultures, culturing techniques, methylcellulose assay, bioassay techniques.

Tang, L., T. Morales, K.L. Boroughs, K.C. Lo Keiser, K. Sellins, K. Stedman, C. Mccall, and M.J. Mcdermott (2003). Expression and characterization of recombinant canine il-13 receptor alpha2 protein and its biological activity in vitro. Molecular Immunology 39(12): 719-727. ISSN: 0161-5890.
NAL Call Number: 448.3 Im62
Abstract: Our previous study showed that recombinant canine IL-13 (rcaIL-13) stimulated production of allergen-specific IgE in vitro by peripheral blood mononuclear cells (PBMC) from flea allergen-sensitized dogs. This has also been demonstrated using human IL-13 (huIL-13) and PBMC isolated from human allergy patients. The stimulatory activity of rcaIL-13 was specifically inhibited by a fusion protein of the extracellular domain of canine IL-13Ralpha2 and the Fc fragment of canine IgG heavy chain (rcaIL-13Ralpha2-Fc). In this communication, we report the construction and expression of a non-fused recombinant extracellular domain of canine IL-13Ralpha2 (rcaIL-13Ralpha2) in an E. coli expression system. The E. coli expressed rcaIL-13Ralpha2 was isolated in inclusion bodies, then solubilized in buffer containing denaturants and reducing agents. After refolding and purification, the biological activity of rcaIL-13Ralpha2 was found in the monomer fraction resulting from gel filtration and ion exchange chromatographies. Biological activity of purified rcaIL-13Ralpha2 was demonstrated by the specific inhibition of rcaIL-13 activity in a TF-1 cell proliferation assay. Additionally, rcaIL-13Ralpha2 was found to be active in neutralizing rcaIL-13 induced upregulation of IgE mRNA levels in PBMCs of 'high IgE' dogs, which have been bred to exhibit a predisposition for high IgE production and are used as a model for allergic asthma. The data confirm our previous report that the regulatory effects of IL-13 on IgE production in canine PBMCs are similar to those reported in humans. Thus, allergic dogs, such as the 'high IgE' producing dogs, may be excellent models for research on IgE-mediated diseases in humans.
Descriptors: cell biology, immune system, chemical coordination and homeostasis, molecular genetics, biochemistry and molecular biophysics, allergic asthma, cell proliferation assay, histology and cytology techniques, laboratory techniques, gel filtration, genetic techniques, ion exchange chromatography, chromatographic techniques.

Ting, S.S., S. Christopher, U. Choi, J. Deleon, G. Linton, N. Whiting Theobald, K. Yamashita, M.L. Harry, and P. Felsburg (2003). In vivo gene transfer by intravenous injection of RD114-pseudotyped mfgs-gammac-ires-gfphmgmt achieves myeloid and lymphoid marking in XSCID dogs. Blood 102(11): 249A-250A. ISSN: 0006-4971.
NAL Call Number: RB145.A1B57
Abstract: Dogs are an informative large animal model for hematopoietic stem cell therapies (transplantation or gene therapy). Relevent to the current study are dogs with mutant common cytokine receptor gamma chain (gammac) that have clinical phenotype similar to human X-linked severe combined immnunodeficiency (XSCID). Restoration of immune function in children with XSCID was achieved by ex vivo transduction of marrow stem cells with retrovirus vector encoding normal gammac. However, lymphoid malignancy occurred in 2 of 11 children at two to three years following gene therapy. This appeared to be associated with insertional mutagenesis activation of LMO2. The contributory role of unregulated expression of gammac transgene remains unclear. Of note is that the leukemias occurred in two patients who were youngest infants at time of gene therapy. Malignancies have not been seen with gene therapy correction of murine XSCID, but it is possible that leukemogenesis requires a longer period than the lifetime of mice. Gene therapy of XSCID dogs may provide an opportunity for prolonged followup required to see leukemogenesis. Efficient ex vivo retroviral gene transfer to hematopoietic stem cells using murine oncoretrovirus vectors requires growth factor mediated cell division, which induces cell differentiation and loss of homing and self-renewal capabilities. Recently, sustained marking of leukocytes has been reported following intravenous administration of ultrafiltration concentrated amphotropic envelope pseudotyped murine retrovirus vector for gene therapy of canine mucosaccharidosis VII (O'Malley et al., Molecular Therapy 7:S82, 2003). In the current study we used intravenous administration of the RD114 pseudotyped murine retrovirus vector MFGS-gammac-IRES-eGFPhMGMT* to three day old XSCID dogs to transduce hematopoietic stem cells in vivo, achieving marking of blood cells. This may prove to be a useful large animal model to observe and study long term effects that might arise. The vector encodes canine gc but also expresses downstream of an IRES element a reporter GFP fused to human P140K mutant methyl guanine methyl transfersase (MGMT*). This fusion protein confers both green fluorescence and benzyl guanine resistance. RD114 MFGS-gammac-IRES-GFPhMGMT* derived from FLYRD18 was concentrated 10 fold by tangential flow ultrafiltration/diafiltration, and further concentrated by ultracentrifugation. Three newborn XSCID dogs received 1.2X107 infectious units of resuspended virus intravenously twice a day for 2-3 days. GFP expression was first evident in small numbers of myeloid and lymphoid cells by initial flow cytometry assessment of peripheral blood at 10 days following the last injection. At 4 weeks lymphocyte counts in all three dogs were above 1500/mul. 7-21% of lymphocytes and 0.08-0.1% of myeloid cells were GFP positive. Preliminary studies using LAM-PCR demonstrated multiple insertion events in blood cells. Similar analysis in specific cellular subtypes is in progress. We demonstrate that direct intravenous administration of concentrated RD114 pseudotyped MFGS-gammac-IRES-GFPhMGMT* oncoretroviral vector achieves marking of both lymphoid and myeloid compartments with enhanced marking of the lymphoid compartment relative to the myeloid compartment. This is associated with steadily improving lymphocyte counts and polyclonal insertion pattern. We will monitor integrant pattern and clinical profile of these animals.
Descriptors: blood and lymphatics, transport and circulation, immune system, chemical coordination and homeostasis, molecular genetics, biochemistry and molecular biophysics, lymphoid malignancy, blood and lymphatic disease, immune system disease, neoplastic disease, genetics, gene therapy, clinical techniques, genetic techniques, laboratory techniques, therapeutic and prophylactic techniques, gene transfer, ultrafiltration, laboratory techniques, immune function, leukemogenesis.

Trepanier, L.A. (2004). Idiosyncratic toxicity associated with potentiated sulfonamides in the dog. Journal of Veterinary Pharmacology and Therapeutics 27(3): 129-138. ISSN: 0140-7783.
NAL Call Number: SF915.J63
Abstract: Idiosyncratic toxicity to potentiated sulfonamides occurs in both humans and dogs, with considerable clinical similarities. The syndrome in dogs can consist of fever, arthropathy, blood dyscrasias (neutropenia, thrombocytopenia, or hemolytic anemia), hepatopathy consisting of cholestasis or necrosis, skin eruptions, uveitis, or keratoconjunctivitis sicca. Other manifestations seen less commonly include protein-losing nephropathy, meningitis, pancreatitis, pneumonitis, or facial nerve palsy. The pathogenesis of these reactions is not completely understood, but may be due to a T-cell-mediated response to proteins haptenated by oxidative sulfonamide metabolites. Our laboratory is working on tests to characterize dogs with possible idiosyncratic sulfonamide reactions, to include ELISA for anti-drug antibodies, immunoblotting for antibodies directed against liver proteins, flow cytometry for drug-dependent anti-platelet antibodies, and in vitro cytotoxicity assays. The management of idiosyncratic sulfonamide toxicity involves client education to identify clinical signs early and allow rapid drug discontinuation, supportive care to include possibly ascorbate and glutathione precursors, and avoidance of subsequent re-exposure. It is important to realize that only antimicrobial sulfonamides, such as sulfamethoxazole, sulfadiazine, and sulfadimethoxine, share this clinical syndrome. There is no evidence for cross-reactivity with drugs that have different underlying structures but share a sulfonamide moiety, such as acetazolamide, furosemide, glipizide, or hydrochlorothiazide.
Descriptors: adverse effects, antibodies, clinical aspects, dosage effects, drug allergies, drug toxicity, pathogenesis, reviews, sulfadiazine, sulfadimethoxine, sulfamethoxazole, sulfonamides, T lymphocytes, treatment, pharmacology, elisa, immunologic techniques, laboratory techniques, cytotoxicity assay, bioassay techniques, flow cytometry.

Weck, A.L.d., P. Mayer, B. Stumper, B. Schiessl, and L. Pickart (1997). Dog allergy, a model for allergy genetics. International Archives of Allergy and Immunology 113(1/3): 55-57. ISSN: 1018-2438.
NAL Call Number: RC583.I57
Abstract: Experimental sensitization in dogs has revealed that the capacity to produce high levels of IgE against a variety of allergens (high IgE responders), an essential characteristic of the atopic state, is a genetic trait inherited in a dominant manner. In high IgE responder dogs spontaneous development of IgE to inhaled allergens, such as house dust mites [usually Dermatophagoides spp.], on the other hand, represents an apparent phenotype very similar to that observed in human atopic families. The full potential of the high IgE response gene appears to be fulfilled only under some conditions such as early and repeated exposition to allergens. It is, therefore, quite possible that the true phenotype of human atopy would also be inherited in a dominant fashion, but not constantly expressed. This would explain why the increase in the prevalence of allergic diseases started long before the environmental factors currently accused could have been at play. This hypothesis, which can be verified experimentally, has important implications for the future of allergy.
Descriptors: IgE, genetics, allergic reactions, house dust mites, disease models, laboratory animals, arthropod allergies, dogs, Dermatophagoides, Arachnida, Acari.

Weyer, J., C.E. Rupprecht, J. Mans, G.J. Viljoen, and L.H. Nel (2007). Generation and evaluation of a recombinant modified vaccinia virus Ankara vaccine for rabies. Vaccine 25(21): 4213-4222. ISSN: 0264-410X.
NAL Call Number: QR189.V32
Abstract: Modified vaccinia virus Ankara (MVA) has become a vaccine vector of choice for recombinant vaccine development. A MVA-based rabies vaccine would be advantageous for use as a vaccine for dogs (and wildlife), particularly if it proves innocuous and efficacious by the oral route. Here, the generation and immunological testing of a recombinant MVA expressing a rabies virus glycoprotein gene is described. In a murine model, higher dosages of recombinant MVA were needed to induce equivocal immune responses as with Vaccinia Copenhagen or Vaccinia Western Reserve recombinants, when administered by a parenteral route. The MVA recombinant was not immunogenic or efficacious when administered per os in naive mice. The ability of the recombinant MVA to induce anamnestic responses in dogs and raccoons was also investigated. Recombinant MVA boosted humoral immune responses in these animals when administered peripherally, but not when administered orally.
Descriptors: antigens, viral immunology, glycoproteins immunology, rabies prevention and control, rabies vaccines immunology, vaccines, synthetic immunology, vaccinia virus immunology, viral envelope proteins immunology, antibodies, viral, antigens, viral genetics, cell line, chickens, cricetinae, dogs, glycoproteins genetics, immunologic memory, mesocricetus, mice, mice, inbred balb c, models, animal, neutralization tests, rabies vaccines genetics, rabies virus genetics, rabies virus immunology, raccoons, vaccines, synthetic genetics, vaccinia virus genetics, viral envelope proteins genetics.

Yeh, H.I., Y.J. Lai, Y.N. Lee, Y.J. Chen, Y.C. Chen, C.C. Chen, S.A. Chen, C.I. Lin, and C.H. Tsai (2003). Differential expression of connexin43 gap junctions in cardiomyocytes isolated from canine thoracic veins. Journal of Histochemistry and Cytochemistry 51(2): 259-266. ISSN: 0022-1554.
NAL Call Number: 381 J8222
Abstract: We investigated the phenotypic features of cardiomyocytes, including the gap junctions, in the myocardial sleeve of thoracic veins. Single cardiomyocytes, isolated from the canine pulmonary veins (PV) and superior vena cava (SVC) using digestive enzymes, were examined by immunoconfocal microscopy using antisera against connexin43 (Cx43), Cx40, and other cell markers. The results showed that isolated cardiomyocytes displayed rod shapes of various sizes, ranging from <50 mum to >200 mum in length, and all the cells expressed alpha-actinin and vinculin. Gap junctions made of various amounts of Cx43 and Cx40 were found at the cell borders. These two connexins were extensively co-localized. Comparison between the thoracic veins showed that cells of the SVC contained more Cx43 gap junctions (total Cx43 gap junctions area per cell surface area, 4.0+-0.2% vs 1.5+-0.2%; p<0.01). In addition, for single-nucleus cells, those from the PV were longer (103.7+-3.6 vs 85.0+-3.1 mum; p<0.01) but narrower (14.4+-0.5 vs 16.9mu0.9 mum; p<0.01). In conclusion, canine thoracic veins contain cardiomyocytes with differences in shape and gap junctions, suggesting that the electrical conduction properties may be different between the thoracic veins.
Descriptors: cardiovascular system, transport and circulation, cell biology, immunoconfocal microscopy, imaging and microscopy techniques, immunologic techniques, laboratory techniques.

Yuasa, K., M. Yoshimura, N. Urasawa, S. Ohshima, J.M. Howell, A. Nakamura, T. Hijikata, Y. Miyagoe Suzuki, and S. Takeda (2007). Injection of a recombinant AAV serotype 2 into canine skeletal muscles evokes strong immune responses against transgene products. Gene Therapy 14(17): 1249-1260. ISSN: 0969-7128.
Abstract: Using murine models, we have previously demonstrated that recombinant adeno-associated virus (rAAV)-mediated microdystrophin gene transfer is a promising approach to treatment of Duchenne muscular dystrophy (DMD). To examine further therapeutic effects and the safety issue of rAAV-mediated microdystrophin gene transfer using larger animal models, such as dystrophic dog models, we first investigated transduction efficiency of rAAV in wild-type canine muscle cells, and found that rAAV2 encoding beta-galactosidase effectively transduces canine primary myotubes in vitro. Subsequent rAAV2 transfer into skeletal muscles of normal dogs, however, resulted in low and transient expression of beta-galactosidase together with intense cellular infiltrations in vivo, where cellular and humoral immune responses were remarkably activated. In contrast, rAAV2 expressing no transgene elicited no cellular infiltrations. Co-administration of immunosuppressants, cyclosporine and mycophenolate mofetil could partially improve rAAV2 transduction. Collectively, these results suggest that immune responses against the transgene product caused cellular infiltration and eliminated transduced myofibers in dogs. Furthermore, in vitro interferon-gamma release assay showed that canine splenocytes respond to immunogens or mitogens more susceptibly than murine ones. Our results emphasize the importance to scrutinize the immune responses to AAV vectors in larger animal models before applying rAAV-mediated gene therapy to DMD patients.
Descriptors: dependovirus genetics, gene therapy adverse effects, genetic vectors adverse effects, muscle, skeletal immunology, muscular dystrophy, animal therapy, muscular dystrophy, duchenne therapy, base sequence, calmodulin genetics, cyclosporine administration and dosage, dogs, dystrophin genetics, dystrophin metabolism, gene therapy methods, genetic engineering, genetic vectors genetics, immunosuppressive agents administration and dosage, injections, intramuscular, interferon gamma immunology, lymphocyte activation, mice, mice, inbred c57bl, models, animal, molecular sequence data, muscle fibers, skeletal immunology, muscle fibers, skeletal virology, muscular dystrophy, animal immunology, muscular dystrophy, duchenne immunology, parvoviridae infections immunology, rna, messenger analysis, reverse transcriptase polymerase chain reaction, species specificity, t lymphocytes, cytotoxic immunology, transduction, genetic methods, transgenes, beta galactosidase genetics.

Back to Top  
<< Table of Contents << Previous |  Next >>
Last Modified: Jan 23, 2014  
 
AWIC Home | NAL Home | USDA | AgNIC | ARS | Web Policies and Important Links | RSS Feeds | Site Map
FOIA | Accessibility Statement | Privacy Policy | Non-Discrimination Statement | Information Quality | USA.gov | White House