@article{adams_collins_williams_holmes_hess_atkins_scheidemantle_liu_lodge_johnson_et al._2024, title={Myeloid cell MHC I expression drives CD8+ T cell activation in nonalcoholic steatohepatitis}, volume={14}, ISSN={["1664-3224"]}, url={https://doi.org/10.3389/fimmu.2023.1302006}, DOI={10.3389/fimmu.2023.1302006}, abstractNote={Background & aimsActivated CD8+ T cells are elevated in Nonalcoholic steatohepatitis (NASH) and are important for driving fibrosis and inflammation. Despite this, mechanisms of CD8+ T cell activation in NASH are largely limited. Specific CD8+ T cell subsets may become activated through metabolic signals or cytokines. However, studies in NASH have not evaluated the impact of antigen presentation or the involvement of specific antigens. Therefore, we determined if activated CD8+ T cells are dependent on MHC class I expression in NASH to regulate fibrosis and inflammation.MethodsWe used H2Kb and H2Db deficient (MHC I KO), Kb transgenic mice, and myeloid cell Kb deficient mice (LysM Kb KO) to investigate how MHC class I impacts CD8+ T cell function and NASH. Flow cytometry, gene expression, and histology were used to examine hepatic inflammation and fibrosis. The hepatic class I immunopeptidome was evaluated by mass spectrometry.ResultsIn NASH, MHC class I isoform H2Kb was upregulated in myeloid cells. MHC I KO demonstrated protective effects against NASH-induced inflammation and fibrosis. Kb mice exhibited increased fibrosis in the absence of H2Db while LysM Kb KO mice showed protection against fibrosis but not inflammation. H2Kb restricted peptides identified a unique NASH peptide Ncf2 capable of CD8+ T cell activation in vitro. The Ncf2 peptide was not detected during fibrosis resolution.ConclusionThese results suggest that activated hepatic CD8+ T cells are dependent on myeloid cell MHC class I expression in diet induced NASH to promote inflammation and fibrosis. Additionally, our studies suggest a role of NADPH oxidase in the production of Ncf2 peptide generation.}, journal={FRONTIERS IN IMMUNOLOGY}, author={Adams, Victoria R. and Collins, Leonard B. and Williams, Taufika Islam and Holmes, Jennifer and Hess, Paul and Atkins, Hannah M. and Scheidemantle, Grace and Liu, Xiaojing and Lodge, Mareca and Johnson, Aaron J. and et al.}, editor={Williams, Taufika Islam and Collins, Leonard B. and Kennedy, ArionEditors}, year={2024}, month={Jan} } @article{feng_hess_tompkins_hildebrand_zhao_2023, title={A Kmer-based paired-end read de novo assembler and genotyper for canine MHC class I genotyping}, volume={26}, ISSN={["2589-0042"]}, DOI={10.1016/j.isci.2023.105996}, abstractNote={The major histocompatibility complex class I (MHC-I) genes are highly polymorphic. MHC-I genotyping is required for determining the peptide epitopes available to an individual's T-cell repertoire. Current genotyping software tools do not work for the dog, due to very limited known canine alleles. To address this, we developed a Kmer-based paired-end read (KPR) de novo assembler and genotyper, which assemble paired-end RNA-seq reads from MHC-I regions into contigs, and then genotype each contig and estimate its expression level. KPR tools outperform other popular software examined in typing new alleles. We used KPR tools to successfully genotype152 dogs from a published dataset. The study discovers 33 putative new alleles, finds dominant alleles in 4 dog breeds, and builds allele diversity and expression landscapes among the 152 dogs. Our software meets a significant need in biomedical research.}, number={2}, journal={ISCIENCE}, author={Feng, Yuan and Hess, Paul R. and Tompkins, Stephen M. and Hildebrand, William H. and Zhao, Shaying}, year={2023}, month={Feb} } @article{doka_suter_mastromauro_bennett_hess_2022, title={Doxorubicin for treatment of histiocytic sarcoma in dogs: 31 cases (2003-2017)}, volume={260}, ISSN={["1943-569X"]}, DOI={10.2460/javma.21.11.0498}, abstractNote={Abstract OBJECTIVE To evaluate the efficacy of doxorubicin for treatment of histiocytic sarcoma (HS) in dogs, whether administered as the sole treatment or as an adjunct to surgery or radiation therapy. ANIMALS 31 client-owned dogs with localized or disseminated HS examined between 2003 and 2017. PROCEDURES Medical records were reviewed retrospectively, and data were collected. The Kaplan-Meier method was used to estimate time-to-progression from the date of first doxorubicin administration and survival time from initial diagnosis. Factors that could be associated with poorer outcomes with doxorubicin treatment were analyzed with log-rank tests. RESULTS The objective response rate (ORR) was 26%. When stratified by disease status, dogs with localized and disseminated forms experienced 43% and 21% ORRs, respectively. Median time to progression after initiating doxorubicin treatment (n = 30 dogs) was 42 days. Median survival time from initial diagnosis to death (n = 29 dogs) was 169 days. Complete responses were obtained in only 2 dogs that had localized disease and received multimodality therapy. CLINICAL RELEVANCE Benefits of doxorubicin administration in canine HS are modest, with a limited ORR and delay in tumor progression, and are comparable to effects attained with other single-agent regimens. }, number={14}, journal={JAVMA-JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Doka, Rhiannon M. and Suter, Steven E. and Mastromauro, Michael L. and Bennett, Ashley L. and Hess, Paul R.}, year={2022}, month={Nov}, pages={1827–1833} } @article{nemec_holmes_hess_2021, title={Dog leukocyte antigen-88*034:01 presents nonamer peptides from canine distemper virus hemagglutinin, large polymerase, and matrix proteins}, volume={97}, ISSN={["2059-2310"]}, url={https://doi.org/10.1111/tan.14197}, DOI={10.1111/tan.14197}, abstractNote={Canine spontaneous cancers may offer greater fidelity than rodent models in advancing clinical immunotherapies. Boxers in particular are distinguished as study subjects by their popularity, and high incidence of human‐relevant cancers. Further, the MHC class I allele DLA‐88*034:01, with a known motif, dominates the breed, facilitating discovery of shared CTL responses against mutation‐origin neoepitopes by standard prediction methods. We experimentally confirmed the allomorph's binding motif by developing an MHC surface stabilization assay. The assay validated four DLA‐88*034:01‐presented peptides from canine distemper virus, ubiquitously administered in routine vaccines, for positive controls in future CTL studies. In turn, these viral peptides substantiated motif‐based prediction for DLA‐88*034:01. The study adds new tools for studying neoepitope‐specific CTL in Boxers to foster canine comparative oncology.}, number={5}, journal={HLA}, publisher={Wiley}, author={Nemec, Paige S. and Holmes, Jennifer C. and Hess, Paul R.}, year={2021}, month={May}, pages={428–434} } @article{holmes_scholl_dickey_hess_2021, title={High-resolution characterization of the structural features and genetic variation of six feline leukocyte antigen class I loci via single molecule, real-time (SMRT) sequencing}, volume={6}, ISSN={["1432-1211"]}, DOI={10.1007/s00251-021-01221-w}, abstractNote={Of the 12 full-length feline leukocyte antigen class I (FLAI) loci, 3 are presumed to be classical: FLAI-E, FLAI-H, and FLAI-K. As diversity is a class Ia hallmark, multi-allelism is an important surrogate supporting a classical designation, in the absence of direct demonstration of T-cell restriction. Conversely, limited polymorphism at an expressed locus suggests regulation of immune effectors with invariant receptors, and non-classical status. FLAI-A, FLAI-J, FLAI-L, and FLAI-O are putative class Ib genes in cats. For both classes, identifying prevalent variants across outbred populations can illuminate specific genotypes to be prioritized for immune studies, as shared alleles direct shared responses. Since variation is concentrated in exons 2 and 3, which encode the antigen-binding domains, partial-length cloning/sequencing can be used for allele discovery, but is laborious and occasionally ambiguous. Here we develop a targeted approach to FLAI genotyping, using the single-molecule real-time (SMRT) platform, which allows full-length (3.4-kb) reads without assembly. Consensus sequences matched full-length Sanger references. Thirty-one new class Ia genes were found in 17 cats. Alleles segregated strongly by loci, and the origins of formerly difficult-to-assign sequences were resolved. Although not targeted, FLAI-L and FLAI-J, and the pseudogene FLAI-F, were also returned. Eighteen class Ib alleles were identified. Diversity was restricted and outside hypervariable regions. Both class Ib genes were transcriptionally active. Novel alternative splicing of FLAI-L was observed. SMRT sequencing of FLAI amplicons is useful for full-length genotyping at feline class Ia loci. High-throughput sequencing could allow highly accurate allele surveys in large cat cohorts.}, journal={IMMUNOGENETICS}, author={Holmes, Jennifer C. and Scholl, Elizabeth H. and Dickey, Allison N. and Hess, Paul R.}, year={2021}, month={Jun} } @article{nemec_kapatos_holmes_stowe_hess_2019, title={Cancer‐testis antigens in canine histiocytic sarcoma and other malignancies}, volume={17}, ISSN={1476-5810 1476-5829}, url={http://dx.doi.org/10.1111/vco.12475}, DOI={10.1111/vco.12475}, abstractNote={Cancer‐testis antigens (CTAs) are a category of self proteins aberrantly expressed in diverse malignancies, mostly solid tumours, due to epigenetic de‐repression. Normally expressed only in fetal or gametogenic tissues, CTAs are tantalizing immunotherapy targets, since autoimmunity risks appear minimal. Few prevalent CTAs have been identified in human hematologic cancers, and just two in their veterinary counterparts. We sought to discover new CTAs in canine hematologic cancers such as histiocytic sarcoma (HS) and lymphoma to foster immunotherapy development. To accomplish this, the ligandome binding the dog leukocyte antigen (DLA)‐88*508:01 class I allele overexpressed in an HS line was searched by mass spectrometry to identify possible CTA‐derived peptides, which could serve as CD8+ T‐cell epitopes. Twenty‐two peptides mapped to 5 human CTAs and 12 additional proteins with CTA characteristics. Expression of five promising candidates was then evaluated in tumour and normal tissue by quantitative and end‐point RT‐PCR. The ortholog of an established CTA, IGF2BP3, had unexpectedly high expression in peripheral blood mononuclear cells (PBMCs). Four other testis‐enhanced proteins were also assessed. AKR1E2, SPECC1 and TPX2 were expressed variably in HS and T‐cell lymphoma biopsies, but also at high levels in critical tissues, including kidney, brain and marrow, diminishing their utility. A more tissue‐restricted candidate, NT5C1B, was detected in T‐cell lymphomas, but also at low levels in some normal dog tissues. These results illustrate the feasibility of discovering canine CTAs by a reverse approach, proceeding from identification of MHC class I‐presented peptides to a comparative RNA expression survey of tumours and normal tissues.}, number={3}, journal={Veterinary and Comparative Oncology}, publisher={Wiley}, author={Nemec, Paige S. and Kapatos, Alexander and Holmes, Jennifer C. and Stowe, Devorah M. and Hess, Paul R.}, year={2019}, month={Jun}, pages={317–328} } @article{hallman_hauck_williams_hess_suter_2019, title={Incidence and risk factors associated with development of clinical cardiotoxicity in dogs receiving doxorubicin}, volume={33}, ISSN={0891-6640 1939-1676}, url={http://dx.doi.org/10.1111/jvim.15414}, DOI={10.1111/jvim.15414}, abstractNote={BackgroundDoxorubicin (DOX) can cause cumulative cardiotoxicity in dogs, but the incidence of clinical cardiotoxicity in dogs receiving DOX has not been determined.Hypothesis/ObjectivesTo determine if the duration of DOX infusion influences the incidence of cardiotoxicity, to characterize the incidence of clinical cardiotoxicity in dogs during or after DOX chemotherapy, and to identify any risk factors associated with cardiotoxicity.AnimalsFour‐hundred ninety‐four dogs that received at least 1 dose of DOX for the treatment of cancer.MethodsRetrospective study of dogs that received DOX from 2006 to 2015.ResultsOf 494 dogs, 20 (4.0%) developed clinical cardiotoxicity. The duration of DOX infusion was not significantly associated with clinical cardiotoxicity, whereas a higher cumulative dose of DOX, higher body weight, decreases in fractional shortening after 5 doses of DOX, and development of ventricular premature contractions were significantly associated with clinical cardiotoxicity. High‐risk breeds for developing dilated cardiomyopathy had an incidence of 15.4%, whereas low‐risk breeds had an incidence of 3.0%.Conclusions and Clinical ImportanceAlthough the duration of DOX infusion did not influence the incidence of cardiotoxicity, premature contractions and decreases in fractional shortening should raise concern for the development of clinical cardiotoxicity. Overall, the incidence of clinical DOX‐induced cardiotoxicity is low, but Boxers and other breeds at high risk for dilated cardiomyopathy may be at an increased risk.}, number={2}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Hallman, Briana E. and Hauck, Marlene L. and Williams, Laurel E. and Hess, Paul R. and Suter, Steven E.}, year={2019}, month={Jan}, pages={783–791} } @article{ross_nemec_kapatos_miller_holmes_suter_buntzman_soderblom_collins_hess_et al._2018, title={The canine MHC class Ia allele DLA-88*508:01 presents diverse self- and canine distemper virus-origin peptides of varying length that have a conserved binding motif}, volume={197}, ISSN={0165-2427}, url={http://dx.doi.org/10.1016/j.vetimm.2018.01.005}, DOI={10.1016/j.vetimm.2018.01.005}, abstractNote={Ideally, CD8+ T-cell responses against virally infected or malignant cells are defined at the level of the specific peptide and restricting MHC class I element, a determination not yet made in the dog. To advance the discovery of canine CTL epitopes, we sought to determine whether a putative classical MHC class Ia gene, Dog Leukocyte Antigen (DLA)-88, presents peptides from a viral pathogen, canine distemper virus (CDV). To investigate this possibility, DLA-88*508:01, an allele prevalent in Golden Retrievers, was expressed as a FLAG-tagged construct in canine histiocytic cells to allow affinity purification of peptide-DLA-88 complexes and subsequent elution of bound peptides. Pattern analysis of self peptide sequences, which were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS), permitted binding preferences to be inferred. DLA-88*508:01 binds peptides that are 9-to-12 amino acids in length, with a modest preference for 9- and 11-mers. Hydrophobic residues are favored at positions 2 and 3, as are K, R or F residues at the C-terminus. Testing motif-matched and -unmatched synthetic peptides via peptide-MHC surface stabilization assay using a DLA-88*508:01-transfected, TAP-deficient RMA-S line supported these conclusions. With CDV infection, 22 viral peptides ranging from 9-to-12 residues in length were identified in DLA-88*508:01 eluates by LC-MS/MS. Combined motif analysis and surface stabilization assay data suggested that 11 of these 22 peptides, derived from CDV hemagglutinin, large polymerase, matrix, nucleocapsid, and V proteins, were processed and presented, and thus, potential targets of anti-viral CTL in DLA-88*508:01-bearing dogs. The presentation of diverse self and viral peptides indicates that DLA-88 is a classical MHC class Ia gene.}, journal={Veterinary Immunology and Immunopathology}, publisher={Elsevier BV}, author={Ross, Peter and Nemec, Paige S. and Kapatos, Alexander and Miller, Keith R. and Holmes, Jennifer C. and Suter, Steven E. and Buntzman, Adam S. and Soderblom, Erik J. and Collins, Edward J. and Hess, Paul and et al.}, year={2018}, month={Mar}, pages={76–86} } @article{nemec_kapatos_holmes_hess_2018, title={The prevalent Boxer MHC class Ia allotype dog leukocyte antigen (DLA)-88*034:01 preferentially binds nonamer peptides with a defined motif}, volume={92}, ISSN={2059-2302}, url={http://dx.doi.org/10.1111/tan.13398}, DOI={10.1111/tan.13398}, abstractNote={Development of effective immunotherapy for chemoresistant malignancies can be advanced by studies in spontaneous cancer models, such as the dog. A crucial first step, T‐cell epitope discovery, can be assisted by determination of binding motifs of common dog leukocyte antigen (DLA) class Ia allotypes. Boxers are popular, inbred dogs with increased risks of relevant target cancers and restricted MHC diversity. We sought to identify the motif of DLA‐88*034:01, a breed‐dominant allotype, to assist peptide prediction from tumor antigens. Mass spectrometry of eluted peptides showed a preference for nonamers with conserved amino acid preferences: basic at position (P)1; hydrophobic at P2; acidic at P4; histidine at P6; and phenylalanine at P9. This data should expedite finding epitopes restricted by this DLA‐88 allotype.}, number={6}, journal={HLA}, publisher={Wiley}, author={Nemec, Paige S. and Kapatos, Alexander and Holmes, Jennifer C. and Hess, Paul R.}, year={2018}, month={Nov}, pages={403–407} } @article{dorman_foster_fernhoff_hess_2017, title={Canine scent detection of canine cancer: A feasibility study}, volume={8}, journal={Veterinary Medicine-Research and Reports}, author={Dorman, D. C. and Foster, M. L. and Fernhoff, K. E. and Hess, P. R.}, year={2017}, pages={1–7} } @article{dorman_foster_fernhoff_hess_2017, title={Canine scent detection of canine cancer: a feasibility study}, volume={Volume 8}, ISSN={2230-2034}, url={http://dx.doi.org/10.2147/vmrr.s148594}, DOI={10.2147/VMRR.S148594}, abstractNote={The scent detection prowess of dogs has prompted interest in their ability to detect cancer. The purpose of this study was to determine whether dogs could use olfactory cues to discriminate urine samples collected from dogs that did or did not have urinary tract transitional cell carcinoma (TCC), at a rate greater than chance. Dogs with previous scent training (n=4) were initially trained to distinguish between a single control and a single TCC-positive urine sample. All dogs acquired this task (mean =15±7.9 sessions; 20 trials/session). The next training phase used four additional control urine samples (n=5) while maintaining the one original TCC-positive urine sample. All dogs quickly acquired this task (mean =5.3±1.5 sessions). The last training phase used multiple control (n=4) and TCC-positive (n=6) urine samples to pro-mote categorical training by the dogs. Only one dog was able to correctly distinguish multiple combinations of TCC-positive and control urine samples suggesting that it mastered categorical learning. The final study phase evaluated whether this dog would generalize this behavior to novel urine samples. However, during double-blind tests using two novel TCC-positive and six novel TCC-negative urine samples, this dog did not indicate canine TCC-positive cancer samples more frequently than expected by chance. Our study illustrates the need to consider canine olfactory memory and the use of double-blind methods to avoid erroneous conclusions regarding the ability of dogs to alert on specimens from canine cancer patients. Our results also suggest that sample storage, confounding odors, and other factors need to be considered in the design of future studies that evaluate the detection of canine cancers by scent detection dogs.}, journal={Veterinary Medicine: Research and Reports}, publisher={Dove Medical Press Ltd.}, author={Dorman, David and Foster, Melanie and Fernhoff, Katherine and Hess, Paul}, year={2017}, month={Oct}, pages={69–76} } @article{mastromauro_suter_hauck_hess_2017, title={Oral melphalan for the treatment of relapsed canine lymphoma}, volume={16}, ISSN={1476-5810}, url={http://dx.doi.org/10.1111/vco.12356}, DOI={10.1111/vco.12356}, abstractNote={Oral melphalan has been included in multi‐agent rescue protocols for canine lymphoma but its activity as a single‐agent for this purpose has not been established. Inexpensive cost, ease of administration and tolerability make oral melphalan an attractive candidate for single‐agent rescue therapy of canine lymphoma. Retrospective evaluation of 19 cases of relapsed canine lymphoma treated with oral melphalan was performed. Melphalan was primarily administered (n = 16) via a high dose protocol (HDM) with a median dosage of 19.4 mg m−2. Fifteen dogs (78.9%) were treated concurrently with corticosteroids. Response evaluation was possible for all dogs with a calculated overall clinical benefit (partial response [PR] + stable disease [SD]) of 31.6% (PR 3/19; SD 3/19). Times to progression following melphalan (TTP‐M) were 14, 24 and 34 days for responders and 20, 28 and 103 days for dogs experiencing SD. Twelve of 17 dogs evaluable for toxicity experienced an adverse event (AE) with only 3 dogs experiencing a grade III or higher AE. Haematologic toxicity was common (11/17) while gastrointestinal toxicity was rare (1/17). Although treatment resulted in limited clinical benefit and non‐durable responses, oral melphalan was well‐tolerated and may be a reasonable rescue option in cases where minimal effective agents remain.}, number={1}, journal={Veterinary and Comparative Oncology}, publisher={Wiley}, author={Mastromauro, M. L. and Suter, S. E. and Hauck, M. L. and Hess, P. R.}, year={2017}, month={Sep}, pages={E123–E129} } @article{saporin-conjugated tetramers identify efficacious anti-hiv cd8+ t-cell specificities._2017, url={http://europepmc.org/articles/PMC5636067}, DOI={10.1371/journal.pone.0184496}, abstractNote={Antigen-specific T-cells are highly variable, spanning potent antiviral efficacy and damaging auto-reactivity. In virus infections, identifying the most efficacious responses is critical to vaccine design. However, current methods depend on indirect measures or on ex vivo expanded CTL clones. We here describe a novel application of cytotoxic saporin-conjugated tetramers to kill antigen-specific T-cells without significant off-target effects. The relative efficacy of distinct antiviral CD8+ T-cell specificity can be directly assessed via antigen-specific CD8+ T-cell depletion. The utility of these reagents is demonstrated here in identifying the CD8+ T-cell specificity most effective in preventing HIV progression in HIV-infected HLA-B*27-positive immune controllers.}, journal={PloS one}, year={2017}, month={Oct} } @article{leitman_palmer_buus_chen_riddell_sims_klenerman_saez-cirion_walker_hess_et al._2017, title={Saporin-conjugated tetramers identify efficacious anti-HIV CD8+T-cell specificities}, volume={12}, number={10}, journal={PLoS One}, author={Leitman, E. M. and Palmer, C. D. and Buus, S. and Chen, F. and Riddell, L. and Sims, S. and Klenerman, P. and Saez-Cirion, A. and Walker, B. D. and Hess, P. R. and et al.}, year={2017} } @article{bennett_williams_ferguson_hauck_suter_lanier_hess_2016, title={Canine acute leukaemia: 50 cases (1989-2014)}, volume={15}, ISSN={1476-5810}, url={http://dx.doi.org/10.1111/vco.12251}, DOI={10.1111/vco.12251}, abstractNote={AbstractAcute leukaemia (AL) is a bone marrow malignancy of hematopoietic progenitors that historically is poorly responsive to treatment. With the widespread adoption of dose‐intense chemotherapy, more human patients attain long‐term survivals, but whether comparable progress has been made in canine AL is unknown. To investigate this question, medical records from three academic veterinary hospitals were reviewed. Fifty dogs met the criteria for AL, having excess circulating or marrow blasts, a major cytopenia(s), and no substantial lymphadenopathy. Thirty‐six dogs received cytotoxic chemotherapy; 23 achieved a complete or partial response for a median of 56 days (range, 9–218). With failure or relapse, 14 dogs were rescued. Median survival with treatment was poor at 55 days (range, 1–300). Untreated (n = 6) and palliatively‐treated (n = 8) dogs lived a median of 7.5 days. Most dogs developed chemoresistance within weeks of initiating treatment, and consequently, survival times for AL remain disappointingly short.}, number={3}, journal={Veterinary and Comparative Oncology}, publisher={Wiley}, author={Bennett, A. L. and Williams, L. E. and Ferguson, M. W. and Hauck, M. L. and Suter, S. E. and Lanier, C. B. and Hess, P. R.}, year={2016}, month={Jul}, pages={1101–1114} } @article{gojanovich_ross_holmer_holmes_hess_2013, title={Characterization and allelic variation of the transporters associated with antigen processing (TAP) genes in the domestic dog (Canis lupus familiaris)}, volume={41}, ISSN={0145-305X}, url={http://dx.doi.org/10.1016/j.dci.2013.07.011}, DOI={10.1016/j.dci.2013.07.011}, abstractNote={The function of the transporters associated with antigen processing (TAP) complex is to shuttle antigenic peptides from the cytosol to the endoplasmic reticulum to load MHC class I molecules for CD8+ T-cell immunosurveillance. Here we report the promoter and coding regions of the canine TAP1 and TAP2 genes, which encode the homologous subunits forming the TAP heterodimer. By sampling genetically divergent breeds, polymorphisms in both genes were identified, although there were few amino acid differences between alleles. Splice variants were also found. When aligned to TAP genes of other species, functional regions appeared conserved, and upon phylogenetic analysis, canine sequences segregated appropriately with their orthologs. Transfer of the canine TAP2 gene into a murine TAP2-defective cell line rescued surface MHC class I expression, confirming exporter function. This data should prove useful in investigating the association of specific TAP defects or alleles with immunity to intracellular pathogens and cancer in dogs.}, number={4}, journal={Developmental & Comparative Immunology}, publisher={Elsevier BV}, author={Gojanovich, Gregory S. and Ross, Peter and Holmer, Savannah G. and Holmes, Jennifer C. and Hess, Paul R.}, year={2013}, month={Dec}, pages={578–586} } @article{cora_neel_grindem_kissling_hess_2013, title={Comparison of automated versus manual neutrophil counts for the detection of cellular abnormalities in dogs receiving chemotherapy: 50 cases (May to June 2008)}, volume={242}, ISSN={["0003-1488"]}, url={https://doi.org/10.2460/javma.242.11.1539}, DOI={10.2460/javma.242.11.1539}, abstractNote={Abstract Objective—To determine the frequency of clinically relevant abnormalities missed by failure to perform a blood smear evaluation in a specific subset of dogs receiving chemotherapy and to compare automated and manual neutrophil counts in the same population Design—Retrospective case series Animals—50 dogs receiving chemotherapy with a total nucleated cell count > 4,000 nucleated cells/μL. Procedures—50 blood smears were evaluated for abnormalities that have strong potential to change the medical plan for a patient: presence of blast cells, band neutrophils, nucleated RBCs, toxic change, hemoparasites, schistocytes, and spherocytes. Automated and manual neutrophil counts were compared. Results—Blood smears from 10 (20%) patients had ≥ 1 abnormalities. Blast cells were identified on 4 (8%) blood smears, increased nucleated RBCs were identified on 5 (10%), and very mild toxic change was identified on 2 (4%). Correlation coefficient of the neutrophil counts was 0.96. Analysis revealed a slight bias between the automated and manual neutrophil counts (mean ± SD difference, −0.43 × 103/μL ± 1.10 × 103/μL) Conclusions and Clinical Relevance—In this series of patients, neutrophil count correlation was very good. Clinically relevant abnormalities were found on 20% of the blood smears. An automated CBC appears to be accurate for neutrophil counts, but a microscopic examination of the corresponding blood smear is still recommended; further studies are needed to determine whether the detection or frequency of these abnormalities would differ dependent on chemotherapy protocol, neoplastic disease, and decision thresholds used by the oncologist in the ordering of a CBC without a blood smear evaluation.}, number={11}, journal={JAVMA-JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Cora, Michelle C. and Neel, Jennifer A. and Grindem, Carol B. and Kissling, Grace E. and Hess, Paul R.}, year={2013}, month={Jun}, pages={1539–1543} } @article{hess_young_miller_vincent_buntzman_collins_frelinger_hess_2013, title={Deletion of naïve T cells recognizing the minor histocompatibility antigen HY with toxin-coupled peptide-MHC class I tetramers inhibits cognate CTL responses and alters immunodominance}, volume={29}, ISSN={0966-3274}, url={http://dx.doi.org/10.1016/j.trim.2013.10.005}, DOI={10.1016/j.trim.2013.10.005}, abstractNote={Alloreactive T-cell responses directed against minor histocompatibility (H) antigens, which arise from diverse genetic disparities between donor and recipient outside the MHC, are an important cause of rejection of MHC-matched grafts. Because clinically significant responses appear to be directed at only a few antigens, the selective deletion of naïve T cells recognizing donor-specific, immunodominant minor H antigens in recipients before transplantation may be a useful tolerogenic strategy. We have previously demonstrated that peptide-MHC class I tetramers coupled to a toxin can efficiently eliminate specific TCR-transgenic T cells in vivo. Here, using the minor histocompatibility antigen HY as a model, we investigated whether toxic tetramers could inhibit the subsequent priming of the two H2-Db-restricted, immunodominant T-cell responses by deleting precursor CTL. Immunization of female mice with male bone marrow elicited robust CTL activity against the Uty and Smcy epitopes, with Uty constituting the major response. As hypothesized, toxic tetramer administration prior to immunization increased survival of cognate peptide-pulsed cells in an in vivo CTL assay, and reduced the frequency of corresponding T cells. However, tetramer-mediated decreases in either T-cell population magnified CTL responses against the non-targeted epitope, suggesting that Db-Uty+ and Db-Smcy+ T cells compete for a limited common resource during priming. Toxic tetramers conceivably could be used in combination to dissect manipulate CD8+ T-cell immunodominance hierarchies, and to prevent the induction of donor-specific, minor H antigen CTL responses in allotransplantation.}, number={1-4}, journal={Transplant Immunology}, publisher={Elsevier BV}, author={Hess, Sabrina M. and Young, Ellen F. and Miller, Keith R. and Vincent, Benjamin G. and Buntzman, Adam S. and Collins, Edward J. and Frelinger, Jeffrey A. and Hess, Paul R.}, year={2013}, month={Dec}, pages={138–145} } @article{herzog_fu_wilson_hess_sen_mcdaniel_pan_sheng_yago_silasi-mansat_et al._2013, title={Podoplanin maintains high endothelial venule integrity by interacting with platelet CLEC-2}, volume={502}, ISSN={0028-0836 1476-4687}, url={http://dx.doi.org/10.1038/NATURE12501}, DOI={10.1038/NATURE12501}, abstractNote={A transmembrane O-glycoprotein podoplanin (PDPN) expressed on fibroblastic reticular cells is the activating ligand for platelet receptor CLEC-2; this interaction leads to perivenular release of sphingosine-1-phosphate and expression of VE-cadherin on high endothelial venules, a key process for the maintenance of vascular integrity in lymph nodes. The lymph nodes, essential sites for immune responses, are supplied with circulating lymphocytes that enter via specialized post-capillary blood vessels known as high endothelial venules (HEVs). How the integrity of this vascular barrier is maintained in the face of the increase in lymphocyte homing during immune responses is not known. Lijun Xia and colleagues now show that the mechanism involves cross-talk between HEVs, platelets and the fibroblastic reticular cells (FRCs) that surround the HEVs. The transmembrane protein podoplanin, expressed on FRCs, interacts with the platelet activation receptor CLEC-2, prompting the release of sphingosine-1-phosphate that in turn preserves the adhesion molecule VE-cadherin on the HEVs. This finding may suggest new approaches to the regulation of lymph node homeostasis in conditions of increased HEV proliferation, such as chronic inflammation. Circulating lymphocytes continuously enter lymph nodes for immune surveillance through specialized blood vessels named high endothelial venules1,2,3,4,5, a process that increases markedly during immune responses. How high endothelial venules (HEVs) permit lymphocyte transmigration while maintaining vascular integrity is unknown. Here we report a role for the transmembrane O-glycoprotein podoplanin (PDPN, also known as gp38 and T1α)6,7,8 in maintaining HEV barrier function. Mice with postnatal deletion of Pdpn lost HEV integrity and exhibited spontaneous bleeding in mucosal lymph nodes, and bleeding in the draining peripheral lymph nodes after immunization. Blocking lymphocyte homing rescued bleeding, indicating that PDPN is required to protect the barrier function of HEVs during lymphocyte trafficking. Further analyses demonstrated that PDPN expressed on fibroblastic reticular cells7, which surround HEVs, functions as an activating ligand for platelet C-type lectin-like receptor 2 (CLEC-2, also known as CLEC1B)9,10. Mice lacking fibroblastic reticular cell PDPN or platelet CLEC-2 exhibited significantly reduced levels of VE-cadherin (also known as CDH5), which is essential for overall vascular integrity11,12, on HEVs. Infusion of wild-type platelets restored HEV integrity in Clec-2-deficient mice. Activation of CLEC-2 induced release of sphingosine-1-phosphate13,14 from platelets, which promoted expression of VE-cadherin on HEVs ex vivo. Furthermore, draining peripheral lymph nodes of immunized mice lacking sphingosine-1-phosphate had impaired HEV integrity similar to Pdpn- and Clec-2-deficient mice. These data demonstrate that local sphingosine-1-phosphate release after PDPN–CLEC-2-mediated platelet activation is critical for HEV integrity during immune responses.}, number={7469}, journal={Nature}, publisher={Springer Science and Business Media LLC}, author={Herzog, Brett H. and Fu, Jianxin and Wilson, Stephen J. and Hess, Paul R. and Sen, Aslihan and McDaniel, J. Michael and Pan, Yanfang and Sheng, Minjia and Yago, Tadayuki and Silasi-Mansat, Robert and et al.}, year={2013}, month={Sep}, pages={105–109} } @article{holmes_holmer_ross_buntzman_frelinger_hess_2013, title={Polymorphisms and tissue expression of the feline leukocyte antigen class I loci FLAI-E, FLAI-H, and FLAI-K}, volume={65}, ISSN={0093-7711 1432-1211}, url={http://dx.doi.org/10.1007/s00251-013-0711-z}, DOI={10.1007/s00251-013-0711-z}, abstractNote={Cytotoxic CD8+ T-cell immunosurveillance for intracellular pathogens, such as viruses, is controlled by classical major histocompatibility complex (MHC) class Ia molecules, and ideally, these antiviral T-cell populations are defined by the specific peptide and restricting MHC allele. Surprisingly, despite the utility of the cat in modeling human viral immunity, little is known about the feline leukocyte antigen class I complex (FLAI). Only a few coding sequences with uncertain locus origin and expression patterns have been reported. Of 19 class I genes, three loci--FLAI-E, FLAI-H, and FLAI-K--are predicted to encode classical molecules, and our objective was to evaluate their status by analyzing polymorphisms and tissue expression. Using locus-specific, PCR-based genotyping, we amplified 33 FLAI-E, FLAI-H, and FLAI-K alleles from 12 cats of various breeds, identifying, for the first time, alleles across three distinct loci in a feline species. Alleles shared the expected polymorphic and invariant sites in the α1/α2 domains, and full-length cDNA clones possessed all characteristic class Ia exons. Alleles could be assigned to a specific locus with reasonable confidence, although there was evidence of potentially confounding interlocus recombination between FLAI-E and FLAI-K. Only FLAI-E, FLAI-H, and FLAI-K origin alleles were amplified from cDNAs of multiple tissue types. We also defined hypervariable regions across these genes, which permitted the assignment of names to both novel and established alleles. As predicted, FLAI-E, FLAI-H, and FLAI-K fulfill the major criteria of class Ia genes. These data represent a necessary prerequisite for studying epitope-specific antiviral CD8+ T-cell responses in cats.}, number={9}, journal={Immunogenetics}, publisher={Springer Science and Business Media LLC}, author={Holmes, Jennifer C. and Holmer, Savannah G. and Ross, Peter and Buntzman, Adam S. and Frelinger, Jeffrey A. and Hess, Paul R.}, year={2013}, month={Jun}, pages={675–689} } @article{ross_holmes_gojanovich_hess_2012, title={A cell-based MHC stabilization assay for the detection of peptide binding to the canine classical class I molecule, DLA-88}, volume={150}, ISSN={["0165-2427"]}, url={http://europepmc.org/articles/PMC3494747}, DOI={10.1016/j.vetimm.2012.08.012}, abstractNote={Identifying immunodominant CTL epitopes is essential for studying CD8+ T-cell responses in populations, but remains difficult, as peptides within antigens typically are too numerous for all to be synthesized and screened. Instead, to facilitate discovery, in silico scanning of proteins for sequences that match the motif, or binding preferences, of the restricting MHC class I allele – the largest determinant of immunodominance – can be used to predict likely candidates. The high false positive rate with this analysis ideally requires binding confirmation, which is obtained routinely by an assay using cell lines such as RMA-S that have defective transporter associated with antigen processing (TAP) machinery, and consequently, few surface class I molecules. The stabilization and resultant increased life-span of peptide–MHC complexes on the cell surface by the addition of true binders validates their identity. To determine whether a similar assay could be developed for dogs, we transfected a prevalent class I allele, DLA-88*50801, into RMA-S. In the BARC3 clone, the recombinant heavy chain was associated with murine β2-microglobulin, and importantly, could differentiate motif-matched and -mismatched peptides by surface MHC stabilization. This work demonstrates the potential to use RMA-S cells transfected with canine alleles as a tool for CTL epitope discovery in this species.}, number={3-4}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Ross, Peter and Holmes, Jennifer C. and Gojanovich, Gregory S. and Hess, Paul R.}, year={2012}, month={Dec}, pages={206–212} } @article{ross_buntzman_vincent_grover_gojanovich_collins_frelinger_hess_2012, title={Allelic diversity at the DLA-88 locus in Golden Retriever and Boxer breeds is limited}, volume={80}, ISSN={["1399-0039"]}, url={http://europepmc.org/articles/PMC3407292}, DOI={10.1111/j.1399-0039.2012.01889.x}, abstractNote={In the dog, previous analyses of major histocompatibility complex class I genes suggest a single polymorphic locus, dog leukocyte antigen (DLA)‐88. While 51 alleles have been reported, estimates of prevalence have not been made. We hypothesized that, within a breed, DLA‐88 diversity would be restricted, and one or more dominant alleles could be identified. Accordingly, we determined allele usage in 47 Golden Retrievers and 39 Boxers. In each population, 10 alleles were found; 4 were shared. Seven novel alleles were identified. DLA‐88*05101 and *50801 predominated in Golden Retrievers, while most Boxers carried *03401. In these breeds, DLA‐88 polymorphisms are limited and largely non‐overlapping. The finding of highly prevalent alleles fulfills an important prerequisite for studying canine CD8+ T‐cell responses.}, number={2}, journal={TISSUE ANTIGENS}, author={Ross, P. and Buntzman, A. S. and Vincent, B. G. and Grover, E. N. and Gojanovich, G. S. and Collins, E. J. and Frelinger, J. A. and Hess, P. R.}, year={2012}, month={Aug}, pages={175–183} } @article{campbell_hess_williams_2012, title={Chronic lymphocytic leukaemia in the cat: 18 cases (2000-2010)}, volume={11}, ISSN={1476-5810}, url={http://dx.doi.org/10.1111/j.1476-5829.2011.00315.x}, DOI={10.1111/j.1476-5829.2011.00315.x}, abstractNote={AbstractThere is little information regarding the presentation, biologic behaviour, treatment and prognosis in cats with chronic lymphocytic leukaemia (CLL), and further investigation is needed to characterize this disease in cats. The goal of this study was to describe the clinical presentation, response to treatment and prognosis of feline CLL. A multi‐institutional retrospective study of 18 cats diagnosed with CLL between 2000 and 2010 was performed. CLL was defined as the presence of a mature lymphocytosis (>9000 lymphocytes µL−1) and confirmation of an immunophenotypically monomorphic or clonal lymphoid population. Each patient was required to also have at least one of the two following criteria: (1) concurrent cytopenia of at least one cell line and/or (2) >15% mature lymphocytes in the bone marrow. Data on signalment, history, clinical signs, clinicopathologic features and response to treatment were reviewed. Median age of the cats at initial presentation was 12.5 years (range: 5–20 years). The most common presenting complaint was chronic weight loss, which was present in 8/18 (44%) cats. Sixteen of 18 (89%) cats were treated with chlorambucil and prednisolone; four of these cats also received vincristine. Two (11%) cats were treated with multi‐agent injectable chemotherapy (L‐CHOP, l‐asparaginase, cyclophosphamide, doxorubicin, vincristine, prednisolone). Eighty‐eight percent of cats evaluable for response achieved a complete (nine cats) or partial (six cats) remission. Median overall remission was 15.7 months (range: 1.3–22.8 months). The median overall survival in the 17 cats with follow‐up data was 14.4 months (range: 0.9–25.3 months). Results of this study suggest that CLL affects older‐aged cats and responds favourably to treatment with oral chlorambucil and prednisolone.}, number={4}, journal={Veterinary and Comparative Oncology}, publisher={Wiley}, author={Campbell, M. W. and Hess, P. R. and Williams, L. E.}, year={2012}, month={Feb}, pages={256–264} } @article{kidd_ross_buntzman_hess_2012, title={Development of an ELISA to detect circulating anti-asparaginase antibodies in dogs with lymphoid neoplasia treated with Escherichia colil-asparaginase}, volume={13}, ISSN={1476-5810}, url={http://dx.doi.org/10.1111/vco.12014}, DOI={10.1111/vco.12014}, abstractNote={AbstractResistance to Escherichia coli l‐asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an enzyme‐linked immunosorbent assay (ELISA) to measure plasma anti‐asparaginase immunoglobulin G responses. Using samples from dogs that had received multiple doses, specific reactivity against l‐asparaginase was demonstrated, while naïve patients' samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1 600 000 in positive control dogs. Intra‐ and inter‐assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA‐positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. l‐asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.}, number={2}, journal={Veterinary and Comparative Oncology}, publisher={Wiley}, author={Kidd, J. A. and Ross, P. and Buntzman, A. S. and Hess, P. R.}, year={2012}, month={Dec}, pages={77–88} } @article{gojanovich_hess_2012, title={Making the Most of Major Histocompatibility Complex Molecule Multimers: Applications in Type 1 Diabetes}, volume={2012}, ISSN={1740-2522 1740-2530}, url={http://dx.doi.org/10.1155/2012/380289}, DOI={10.1155/2012/380289}, abstractNote={Classical major histocompatibility complex (MHC) class I and II molecules present peptides to cognate T-cell receptors on the surface of T lymphocytes. The specificity with which T cells recognize peptide-MHC (pMHC) complexes has allowed for the utilization of recombinant, multimeric pMHC ligands for the study of minute antigen-specific T-cell populations. In type 1 diabetes (T1D), CD8+ cytotoxic T lymphocytes, in conjunction with CD4+ T helper cells, destroy the insulin-producingβcells within the pancreatic islets of Langerhans. Due to the importance of T cells in the progression of T1D, the ability to monitor and therapeutically target diabetogenic clonotypes of T cells provides a critical tool that could result in the amelioration of the disease. By administering pMHC multimers coupled to fluorophores, nanoparticles, or toxic moieties, researchers have demonstrated the ability to enumerate, track, and delete diabetogenic T-cell clonotypes that are, at least in part, responsible for insulitis; some studies even delay or prevent diabetes onset in the murine model of T1D. This paper will provide a brief overview of pMHC multimer usage in defining the role T-cell subsets play in T1D etiology and the therapeutic potential of pMHC for antigen-specific identification and modulation of diabetogenic T cells.}, journal={Clinical and Developmental Immunology}, publisher={Hindawi Limited}, author={Gojanovich, Greg S. and Hess, Paul R.}, year={2012}, pages={1–9} } @misc{gojanovich_hess_2012, title={Making the most of major histocompatibility complex molecule multimers: Applications in type 1 diabetes}, journal={Clinical & Developmental Immunology}, author={Gojanovich, G. S. and Hess, P. R.}, year={2012} } @article{takaesu_inagaki_takubo_mishina_hess_dean_yoshimura_matsumoto_suda_ninomiya-tsuji_et al._2012, title={TAK1 (MAP3K7) Signaling Regulates Hematopoietic Stem Cells through TNF-Dependent and -Independent Mechanisms}, volume={7}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0051073}, DOI={10.1371/journal.pone.0051073}, abstractNote={A cytokine/stress signaling kinase Tak1 (Map3k7) deficiency is known to impair hematopoietic progenitor cells. However, the role of TAK1 signaling in the stem cell function of the hematopoietic system is not yet well defined. Here we characterized hematopoietic stem cells (HSCs) harboring deletion of Tak1 and its activators, Tak1 binding proteins 1 and 2 (Tab1 and Tab2) using a competitive transplantation assay in a mouse model. Tak1 single or Tab1/Tab2 double deletions completely eliminated the reconstitution activity of HSCs, whereas Tab1 or Tab2 single deletion did not cause any abnormality. Tak1 single or Tab1/Tab2 double deficient lineage-negative, Sca-1+, c-Kit+ (LSK) cells did not proliferate and underwent cell death. We found that Tnfr1 deficiency restored the reconstitution activity of Tak1 deficient bone marrow cells for 6–18 weeks. However, the reconstitution activity of Tak1- and Tnfr1-double deficient bone marrow cells declined over the long term, and the number of phenotypically identified long-term hematopoietic stem cells were diminished. Our results indicate that TAB1- or TAB2-dependent activation of TAK1 is required for maintenance of the hematopoietic system through two mechanisms: one is prevention of TNF-dependent cell death and the other is TNF-independent maintenance of long-term HSC.}, number={11}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Takaesu, Giichi and Inagaki, Maiko and Takubo, Keiyo and Mishina, Yuji and Hess, Paul R. and Dean, Gregg A. and Yoshimura, Akihiko and Matsumoto, Kunihiro and Suda, Toshio and Ninomiya-Tsuji, Jun and et al.}, editor={Tjwa, MarcEditor}, year={2012}, month={Nov}, pages={e51073} } @article{the use of peptide-major-histocompatibility-complex multimers in type 1 diabetes mellitus._2012, url={http://europepmc.org/articles/PMC3440061}, DOI={10.1177/193229681200600305}, abstractNote={Major histocompatibility complex (MHC) class I and MHC class II molecules present short peptides that are derived from endogenous and exogenous proteins, respectively, to cognate T-cell receptors (TCRs) on the surface of T cells. The exquisite specificity with which T cells recognize particular peptide-major-histocompatibility-complex (pMHC) combinations has permitted development of soluble pMHC multimers that bind exclusively to selected T-cell populations. Because the pathogenesis of type 1 diabetes mellitus (T1DM) is driven largely by islet-reactive T-cell activity that causes β-cell death, these reagents are useful tools for studying and, potentially, for treating this disease. When coupled to fluorophores or paramagnetic nanoparticles, pMHC multimers have been used to visualize the expansion and islet invasion of T-cell effectors during diabetogenesis. Administration of pMHC multimers to mice has been shown to modulate T-cell responses by signaling through the TCR or by delivering a toxic moiety that deletes the targeted T cell. In the nonobese diabetic mouse model of T1DM, a pMHC-I tetramer coupled to a potent ribosome-inactivating toxin caused long-term elimination of a specific diabetogenic cluster of differentiation 8+ T-cell population from the pancreatic islets and delayed the onset of diabetes. This review will provide an overview of the development and use of pMHC multimers, particularly in T1DM, and describe the therapeutic promise these reagents have as an antigen-specific means of ameliorating deleterious T-cell responses in this autoimmune disease.}, journal={Journal of diabetes science and technology}, year={2012}, month={May} } @article{grosenbaugh_leard_bergman_klein_meleo_susaneck_hess_jankowski_jones_leibman_et al._2011, title={Safety and efficacy of a xenogeneic DNA vaccine encoding for human tyrosinase as adjunctive treatment for oral malignant melanoma in dogs following surgical excision of the primary tumor}, volume={72}, url={https://doi.org/10.2460/ajvr.72.12.1631}, DOI={10.2460/ajvr.72.12.1631}, abstractNote={Abstract Objective—To evaluate the safety and efficacy of a vaccine containing plasmid DNA with an insert encoding human tyrosinase (ie, huTyr vaccine) as adjunctive treatment for oral malignant melanoma (MM) in dogs. Animals—111 dogs (58 prospectively enrolled in a multicenter clinical trial and 53 historical controls) with stage II or III oral MM (modified World Health Organization staging scale, I to IV) in which locoregional disease control was achieved. Procedures—58 dogs received an initial series of 4 injections of huTyr vaccine (102 μg of DNA/injection) administered transdermally by use of a needle-free IM vaccination device. Dogs were monitored for adverse reactions. Surviving dogs received booster injections at 6-month intervals thereafter. Survival time for vaccinates was compared with that of historical control dogs via Kaplan-Meier survival analysis for the outcome of death. Results—Kaplan-Meier analysis of survival time until death attributable to MM was determined to be significantly improved for dogs that received the huTyr vaccine, compared with that of historical controls. However, median survival time could not be determined for vaccinates because < 50% died of MM before the end of the observation period. No systemic reactions requiring veterinary intervention were associated with vaccination. Local reactions were primarily limited to acute wheal or hematoma formation, mild signs of pain at the injection site, and postvaccination bruising. Conclusions and Clinical Relevance—Results support the safety and efficacy of the huTyr DNA vaccine in dogs as adjunctive treatment for oral MM. Impact for Human Medicine—Response to DNA vaccination in dogs with oral MM may be useful in development of plasmid DNA vaccination protocols for human patients with similar disease.}, number={12}, journal={American Journal of Veterinary Research}, author={Grosenbaugh, D.A. and Leard, T. and Bergman, P.J. and Klein, M.K. and Meleo, K. and Susaneck, S. and Hess, P.R. and Jankowski, M.K. and Jones, P.D. and Leibman, N.F. and et al.}, year={2011}, month={Dec}, pages={1631–1638} } @article{mccleary-wheeler_williams_hess_suter_2010, title={Evaluation of an in vitro telomeric repeat amplification protocol assay to detect telomerase activity in canine urine}, volume={71}, ISSN={0002-9645}, url={http://dx.doi.org/10.2460/ajvr.71.12.1468}, DOI={10.2460/ajvr.71.12.1468}, abstractNote={Abstract Objective—To evaluate the usefulness of a PCR-based telomeric repeat amplification protocol (TRAP) assay for detecting telomerase activity in cells from a canine transitional cell carcinoma (TCC) cell line and, ultimately, in the urine of dogs with TCC. Animals—11 dogs with histologic or cytologic evidence of TCC, 10 dogs with benign lower urinary tract disease, and 9 healthy dogs. Procedures—Telomerase activity was initially evaluated in cells from canine TCC (K9TCC) and telomerase-negative (WI-38) cell lines. Following assay optimization, telomerase stability was evaluated at various storage durations and temperatures. Urine samples were then obtained prospectively from study dogs. Results—Telomerase activity was detected in the K9TCC cell line. The TRAP assay detected telomerase activity in as few as 10 K9TCC cells alone and as low as 2% of a total cell population in K9TCC and WI-38 mixing experiments. A loss of telomerase activity was detected with increasing urine storage durations at various temperatures. Telomerase activity was clearly detected in samples collected from 10 of 11 dogs with TCC, 2 of 10 dogs with benign lower urinary tract disease, and none of the 9 healthy dogs. Conclusions and Clinical Relevance—The TRAP-based assay detected telomerase activity in the canine TCC cell line and revealed that the telomerase ribonucleoprotein complex was inherently unstable at various storage durations and conditions. Telomerase activity was also detectable in urine samples obtained from dogs with TCC, which suggested the TRAP assay may be useful in diagnosing TCC in dogs.}, number={12}, journal={American Journal of Veterinary Research}, publisher={American Veterinary Medical Association (AVMA)}, author={McCleary-Wheeler, Angela L. and Williams, Laurel E. and Hess, Paul R. and Suter, Steven E.}, year={2010}, month={Dec}, pages={1468–1474} } @article{palgrave_hunter_clarke_hess_2010, title={Pathology in Practice}, volume={237}, ISSN={["0003-1488"]}, DOI={10.2460/javma.237.8.911}, abstractNote={An 11-year-old 20-kg (44-lb) spayed female Basset Hound was submitted for necropsy following euthanasia because of a chronic, progressive disease that had been managed clinically for approximately 3 years.The dog' s antemortem clinical signs included polyuria, polydipsia, signs of musculoskeletal pain, and recurrent urinary tract infections. Clinical and Gross FindingsAntemortem serum biochemical analyses indicated that the dog had hypercalcemia (16.5 mg/dL; reference range, 8.9 to 11.4 mg/dL), hyperproteinemia (9.2 g/dL; reference range, 5.0 to 7.4 g/dL), hyperglobulinemia (6.1 g/dL; reference range, 1.6 to 3.6 g/dL), and a low albumin-to-globulin concentration ratio (0.4; reference range, 0.8 to 2.0).Hematologic findings included evidence of anemia (erythrocyte count, 3.32 X 10 6 erythrocytes/µL [reference range, 4.8 X 10 6 erythrocytes/µL to 9.3 X 10 6 erythrocytes/µL]; Hct, 23.1% [reference range, 36% to 60%]), panleukopenia (2.}, number={8}, journal={JAVMA-JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Palgrave, Christopher J. and Hunter, Stuart A. and Clarke, Dawn M. and Hess, Paul R.}, year={2010}, month={Oct}, pages={911–913} } @article{vincent_young_buntzman_stevens_kepler_tisch_frelinger_hess_2010, title={Toxin-Coupled MHC Class I Tetramers Can Specifically Ablate Autoreactive CD8+ T Cells and Delay Diabetes in Nonobese Diabetic Mice}, volume={184}, ISSN={0022-1767 1550-6606}, url={http://dx.doi.org/10.4049/jimmunol.0903931}, DOI={10.4049/jimmunol.0903931}, abstractNote={Abstract There is compelling evidence that self-reactive CD8+ T cells are a major factor in development and progression of type 1 diabetes in animals and humans. Hence, great effort has been expended to define the specificity of autoimmune CD8+ T cells and to alter their responses. Much work has focused on tolerization of T cells using proteins or peptides. A weakness in this approach is that residual autoreactive T cells may be activated and exacerbate disease. In this report, we use a novel approach, toxin-coupled MHC class I tetramers. Used for some time to identify Ag-specific cells, in this study, we use that same property to delete the Ag-specific cells. We show that saporin-coupled tetramers can delete islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)-reactive T cells in vitro and in vivo. Sequence analysis of TCRβ-chains of IGRP+ cells reveals the repertoire complexity in the islets is markedly decreased as NOD mice age and significantly altered in toxic tetramer-treated NOD mice. Further tetramer+ T cells in the islets are almost completely deleted, and, surprisingly, loss of tetramer+ T cells in the islets is long lasting. Finally, we show deletion at 8 wk of age of IGRP+ CD8+ T cells, but not dystophia myotonica kinase- or insulin B-reactive cells, significantly delays diabetes in NOD mice.}, number={8}, journal={The Journal of Immunology}, publisher={The American Association of Immunologists}, author={Vincent, Benjamin G. and Young, Ellen F. and Buntzman, Adam S. and Stevens, Rosemary and Kepler, Thomas B. and Tisch, Roland M. and Frelinger, Jeffrey A. and Hess, Paul R.}, year={2010}, month={Mar}, pages={4196–4204} } @article{young_hess_arnold_tisch_frelinger_2009, title={Islet lymphocyte subsets in male and female NOD mice are qualitatively similar but quantitatively distinct}, volume={42}, ISSN={["0891-6934"]}, DOI={10.3109/08916930903213993}, abstractNote={Islet-infiltrating lymphocytes of individual male and female non-obese diabetic (NOD) mice were examined with the purpose of determining the differences that lead to a predominance of diabetes in female versus males NOD mice. When normalized for the amount of islet lymphocytes recovered, the infiltrating lymphocytes of female NOD mice were indistinguishable from those of male NOD mice. The only observed difference was that islet inflammation progressed at an increased rate in female compared to male NOD mice. There was no difference in the composition of islet infiltrates in male and female NOD mice. Unexpectedly, the ratio of CD4+:CD8+ T cells was tightly controlled in the islets throughout diabetogenesis. The frequency of IL-4+ CD4+ T cells started high but quickly fell to 3% of the population that was maintained with increasing inflammation. A significant portion of the CD8+ T cells were islet-specific glucose-6-phosphatase catalytic subunit-related protein specific in both male and female NOD mice and this population was antigen experienced and increased at high levels of islet inflammation. Surprisingly, a large pool of antigen inexperienced naïve T cells was detected in the islets. We conclude the underlying immunological processes in both male and female NOD mice are similar while the rates differ and the presence of naïve T cell in the islets may contribute to epitope spreading.}, number={8}, journal={AUTOIMMUNITY}, author={Young, Ellen F. and Hess, Paul R. and Arnold, Larry W. and Tisch, Roland and Frelinger, Jeffrey A.}, year={2009}, pages={678–691} } @article{hess_barnes_woolard_johnson_cullen_collins_frelinger_2007, title={Selective deletion of antigen-specific CD8(+) T cells by MHC class I tetramers, coupled to the type I ribosome-inactivating protein saporin}, volume={109}, ISSN={["0006-4971"]}, DOI={10.1182/blood-2006-06-028001}, abstractNote={AbstractCD8+ cytotoxic T lymphocytes (CTLs) are important effector cells responsible for tissue destruction in several autoimmune and allograft-related diseases. To discover if pathogenic T cells could be selectively deleted, we investigated the ability of a toxin coupled to major histocompatibility complex (MHC) class I tetramers to kill antigen-specific CD8+ T cells. H2-Db tetramers were assembled using streptavidin conjugated to the ribosome-inactivating protein (RIP) saporin (SAP). These tetramers inhibited ribosome activity in vitro, retained the T-cell receptor (TCR)–binding specificity of their nontoxic counterparts, and were internalized by 100% of target cells, leading to cell death in 72 hours. Cytotoxicity was dependent on the tetramer dose and avidity for the T cell. A single injection of the SAP-coupled tetramer eliminated more than 75% of cognate, but not control, T cells. This work demonstrates the therapeutic potential of cytotoxic tetramers to selectively eradicate pathogenic clonotypes while leaving overall T-cell immunity intact.}, number={8}, journal={BLOOD}, author={Hess, Paul R. and Barnes, Carie and Woolard, Matthew D. and Johnson, Michael D. L. and Cullen, John M. and Collins, Edward J. and Frelinger, Jeffrey A.}, year={2007}, month={Apr}, pages={3300–3307} } @article{hess_english_hegarty_brown_breitschwerdt_2006, title={Experimental Ehrlichia canis infection in the dog does not cause immunosuppression}, volume={109}, ISSN={["1873-2534"]}, DOI={10.1016/j.vetimm.2005.07.027}, abstractNote={A carrier state develops in some Ehrlichia canis-infected dogs due to ineffective host defenses. The subsequent development of immune-mediated diseases or opportunistic infections in chronic ehrlichiosis suggests dysregulation of immunity; however, the immunobiology of this infection has not been well characterized. In this study, eight dogs were infected with E. canis, and changes in seroreactivity, serum immunoglobulin (Ig) concentrations, peripheral blood T cell subsets, lymphocyte blastogenesis (LBT), and lymphokine-activated killer (LAK) activity were evaluated over 4 months. Infection, which was documented by seroconversion, polymerase chain reaction, and blood culture, caused self-limiting fever and thrombocytopenia. Infected dogs developed an anti-E. canis antibody response but were not immune to re-infection. Serum IgM, IgG, and IgA concentrations were unaffected by E. canis. The percentage of circulating CD4(+) T cells was similar in uninfected and infected dogs at all points. Infected dogs developed a CD8(+) lymphocytosis 6 weeks after inoculation that subsequently subsided, despite organism persistence. Functional defects of cell-mediated immunity, measured as suppression of LAK activity or mitogen-driven LBT, were not observed. These results suggest that immune responses are not grossly impaired in young dogs during the first several months following experimental E. canis infection.}, number={1-2}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Hess, PR and English, RV and Hegarty, BC and Brown, GD and Breitschwerdt, EB}, year={2006}, month={Jan}, pages={117–125} } @article{thrall_larue_yu_samulski_sanders_case_rosner_azuma_poulson_pruitt_et al._2005, title={Thermal dose is related to duration of local control in canine sarcomas treated with thermoradiotherapy}, volume={11}, number={14}, journal={Clinical Cancer Research}, author={Thrall, D. E. and Larue, S. M. and Yu, D. H. and Samulski, T. and Sanders, L. and Case, B. and Rosner, G. and Azuma, C. and Poulson, J. and Pruitt, A. F. and et al.}, year={2005}, pages={5206–5214} } @article{hess_boczkowski_nair_snyder_gilboa_2006, title={Vaccination with mRNAs encoding tumor-associated antigens and granulocyte-macrophage colony-stimulating factor efficiently primes CTL responses, but is insufficient to overcome tolerance to a model tumor/self antigen}, volume={55}, ISSN={["1432-0851"]}, DOI={10.1007/s00262-005-0064-z}, abstractNote={Immunization of mice with dendritic cells transfected ex vivo with tumor-associated antigen (TAA)-encoding mRNA primes cytotoxic T lymphocytes (CTL) that mediate tumor rejection. Here we investigated whether direct injection of TAA mRNA, encapsulated in cationic liposomes, could function similarly as cancer immunotherapy. Intradermal and intravenous injection of ovalbumin (OVA) mRNA generated specific CTL activity and inhibited the growth of OVA-expressing tumors. Vaccination studies with DNA have demonstrated that co-administration of antigen (Ag)- and cytokine-encoding plasmids potentiate the T cell response; in analogous fashion, the inclusion of granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA enhanced OVA-specific cytotoxicity. The ability of this GM-CSF-augmented mRNA vaccine to treat an established spontaneous tumor was evaluated in the Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) mouse, using the SV40 large T Ag (TAg) as a model tumor/self Ag. Repeated vaccination elicited vigorous TAg-specific CTL activity in nontransgenic mice, but tumor-bearing TRAMP mice remained tolerant. Adoptive transfer of naïve splenocytes into TRAMP mice prior to the first vaccination restored TAg reactivity, and slowed tumor progression. The data from this study suggests that vaccination with TAA mRNA is a simple and effective means of priming antitumor CTL, and that immunogenicity of the vaccine can be augmented by co-delivery of GM-CSF mRNA. Nonetheless, limitations of such vaccines in overcoming tolerance to tumor/self Ag may mandate prior or simultaneous reconstitution of the autoreactive T cell repertoire for this form of immunization to be effective.}, number={6}, journal={CANCER IMMUNOLOGY IMMUNOTHERAPY}, author={Hess, PR and Boczkowski, D and Nair, SK and Snyder, D and Gilboa, E}, year={2006}, month={Jun}, pages={672–683} } @inbook{hess_bunch_2000, title={Diagnostic approach to hepatobiliary disease}, volume={13}, booktitle={Kirk's current meterinary therapy XIII, small animal practice}, publisher={Philadelphia: WB Saunders Co.}, author={Hess, P. R. and Bunch, S. E.}, year={2000}, pages={659–664} } @article{hess_hawkins_drost_1999, title={What is your diagnosis? Nodular lung disease}, volume={214}, number={2}, journal={Journal of the American Veterinary Medical Association}, author={Hess, P. R. and Hawkins, E. C. and Drost, W. T.}, year={1999}, pages={193–194} } @article{hess_sellon_1997, title={Steroid-responsive, cervical, pyogranulomatous pachymeningitis in a dog}, volume={33}, ISSN={["1547-3317"]}, DOI={10.5326/15473317-33-5-461}, abstractNote={Syndromes of steroid-responsive meningitis have been described in the dog and typically are characterized by neutrophilic pleocytosis and an elevated protein concentration of the cerebrospinal fluid. In a minority of cases, histopathology has demonstrated suppurative leptomeningeal (i.e., arachnoid and pia) inflammation. A case of compressive, cervical, pyogranulomatous inflammation of undetermined cause affecting the dura mater (i.e., pachymeningitis), accompanied by fever and hyperpathia, is presented. The pachymeningitis ultimately regressed with long-term immunosuppressive therapy. This case shares features with hypertrophic spinal pachymeningitis of humans, an uncommon, frequently idiopathic, chronic inflammatory disorder causing dural hypertrophy, radiculopathy, and spinal cord compression.}, number={5}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Hess, PR and Sellon, RK}, year={1997}, pages={461–468} } @article{hess_bunch_1995, title={MANAGEMENT OF PORTAL-HYPERTENSION AND ITS CONSEQUENCES}, volume={25}, ISSN={["0195-5616"]}, DOI={10.1016/S0195-5616(95)50037-3}, abstractNote={Increased pressure in the protal venous system results from impedance to blood flow at any point along it's course from the splanchnic circulation through the liver to the right heart. Typical manifestations of sustained increases in portal venous pressure commonly may include accumulation of abdominal fluid and development of acquired portosystemic shunts. Pathophysiology of altered portal vascular dynamics, diagnostic approach for animals suspected of having an intra-abdominal source of portal hypertension and treatment options are discussed.}, number={2}, journal={VETERINARY CLINICS OF NORTH AMERICA-SMALL ANIMAL PRACTICE}, author={HESS, PR and BUNCH, SE}, year={1995}, month={Mar}, pages={461–483} } @article{rhoades_chiaia_hess_miller_1988, title={Effects of neonatal transection upon substance P- and leucine enkephalin-like immunoreactivities in trigeminal subnucleus caudalis of the rat}, volume={8}, DOI={10.1523/jneurosci.08-07-02234.1988}, abstractNote={The distributions of substance P-like immunoreactivity (SPLI) and leucine-enkephalin-like immunoreactivity (LENKLI) in subnucleus caudalis of normal adult rats were compared with those observed in the adult rats that sustained transection of the infraorbital (IO) nerve either on the day of birth or in adulthood. All immunocytochemical experiments in the neonatally nerve damaged rats were carried out at least 60 d after the nerve transection. In the animals that sustained nerve transections as adults, brains were processed for immunohistochemistry between 7 and 60 d after the lesions. In the rats that sustained IO nerve transections as adults, there was a transient reduction in the density of the SPLI in layers I and II of ipsilateral subnucleus caudalis. It was most apparent about 2 weeks after the nerve transection and returned to near normal values by 60 d after the lesion. In the rats that sustained IO nerve transections on the day of birth, there was no reduction in the density of SPLI in caudalis, and the band of staining on the deafferented side of the brain stem was actually 40% wider than that on the intact side. Neither neonatal nor adult IO nerve transection had appreciable effects upon the distribution of LENKLI in the rat's trigeminal brain-stem complex. In another series of experiments, rats that sustained neonatal transection of the IO nerve had this same nerve recut in adulthood. Twelve days after the second lesion, the brains of these animals were processed for SPLI. There was a marked reduction in the density of the staining for this peptide on the deafferented side. This last result is consistent with the interpretation that the increased distribution of SPLI in the neonatally nerve damaged rats is due, at least partially, to reorganization of primary afferents.}, number={7}, journal={Journal of Neuroscience}, author={Rhoades, R. W. and Chiaia, N. L. and Hess, Paul and Miller, M. W.}, year={1988}, pages={2234–2247} } @article{mooney_nikoletseas_hess_allen_lewin_rhoades_1988, title={The projection from the superficial to the deep layers of the superior colliculus: an intracellular horseradish peroxidase injection study in the hamster}, volume={8}, DOI={10.1523/jneurosci.08-04-01384.1988}, abstractNote={Intracellular recording and horseradish peroxidase (HRP) injection techniques were employed to examine the projections of superficial layer [stratum griseum superficiale (SGS) and stratum opticum (SO)] superior collicular (SC) neurons in the hamster that sent axon collaterals into the deep laminae (those ventral to the SO) of this structure. Sixty-nine neurons were studied, selected from a sample of over 185 HRP-filled superficial layer cells on the basis of having heavily stained axons. Of the 69 cells included in the study, 43.4% (n = 30) sent at least one axon collateral to the deep laminae. Not all cell types in the superficial layers contributed equally to this interlaminar projection: 78.6% (n = 11) of the recovered wide-field vertical cells, 55.0% (n = 11) of the narrow-field vertical cells, 16.7% (n = 2) of the stellate cells, 40.0% (n = 2) of the marginal cells, 18.2% (n = 2) of the horizontal cells, and 28.6% (n = 2) of neurons we could not classify on the basis of their somadendritic morphology projected to the deep layers. Within a given cell class, there were no significant morphological or physiological differences between the neurons that possessed deep axon collaterals and those that did not. The deep axon collaterals of most of the interlaminar projection neurons were restricted to the stratum griseum intermediate (SGI). In this layer, the largest segment of the axon arbor was located lateral to a projection line that was orthogonal to the SC surface and that passed through the soma of the cell in question. These results, along with those of a previous study (Mooney et al., 1984), which demonstrated that the dendrites of deep layer cells may extend through the SO and into the SGS, indicate that there is an extensive anatomical substrate by which sensory information may be communicated from superficial to deep layer SC neurons.}, number={4}, journal={Journal of Neuroscience}, author={Mooney, R. D. and Nikoletseas, M. M. and Hess, Paul and Allen, Z. and Lewin, A. C. and Rhoades, R. W.}, year={1988}, pages={1384–1399} } @inbook{rhoades_mooney_chiaia_szczepanik_hess_renehan_klein_nagele_jacquin_1987, title={Development and lesion induced reorganization of the peripheral projections of the trigeminal ganglion in rat}, ISBN={0845127322}, booktitle={Effects of injury on trigeminal and spinal somatosensory systems}, publisher={New York: Alan R. Liss, Inc.}, author={Rhoades, R. W. and Mooney, R. D. and Chiaia, N. L. and Szczepanik, A. M. and Hess, P. R. and Renehan, W. E. and Klein, B. G. and Nagele, R. G. and Jacquin, M. F.}, editor={L. M. Pubols and Sessle, B. J.Editors}, year={1987}, pages={49–60} } @article{chiaia_hess_hosoi_rhoades_1987, title={MORPHOLOGICAL-CHARACTERISTICS OF LOW-THRESHOLD PRIMARY AFFERENTS IN THE TRIGEMINAL SUBNUCLEI INTERPOLARIS AND CAUDALIS (THE MEDULLARY DORSAL HORN) OF THE GOLDEN-HAMSTER}, volume={264}, ISSN={["0021-9967"]}, DOI={10.1002/cne.902640407}, abstractNote={AbstractIntra‐axonal recording and horseradish peroxidase (HRP) injection techniques were employed to define the response characteristics of low‐threshold, rapidly conducting trigeminal primary afferents and the morphological features of their axon arbors in subnucleus interpolaris and subnucleus caudalis (or the medullary dorsal horn; these last two terms are used synonomously throughout the paper). A total of 61 such afferents were characterized and recovered. Of these, ten gave rapidly adapting (RA) and 17 slowly adapting (SA type I) responses to vibrissa deflection. Twenty were sensitive to guard hair deflection and 14 were responsive to indentation of the hairy skin.The vibrissa‐sensitive primary afferents were all quite similar morphologically. Primary collaterals proceeded directly, in a radial fashion, to their zone of termination and gave rise to dense and compact arbors. These tended to be larger in the medullary dorsal horn (MDH) than in interpolaris and they also gave rise to more boutons in the former nucleus. Guard hair afferents generally had smaller arbors and gave rise to fewer boutons than vibrissa‐sensitive axons. Like vibrissa afferents, their arbor were generally circumscribed in both interpolaris and MDH, but they were larger in the latter nucleus. Skin‐sensitive afferents had arbors that tended to be somewhat larger than those of vibrissa‐ or guard‐hair‐related fibers. Unlike the other fiber types, the arbors of skin‐sensitive afferents were on average larger in interpolaris than MDH. Quantitative analysis of the morphological data from well‐filled examples from each of these four functional types verified our qualitative impressions regarding differences between interpolaris and MDH collaterals of a given fiber type. Statistical comparison of data from different functional classes indicated trends that supported our qualitative impressions, but none of these was statistically significant.The topography of the trigeminal primary afferent input to interpolaris was organized such that the head was inverted and fibers with caudal receptive fields terminated in the lateral portion of the nucleus. This was true for all of the functional afferent types that we examined. Vibrissa‐related fibers differed from nonvibrissa afferents in that they tended to avoid the most rostral portion of interpolaris. In the MDH, the primary afferent representation of the head was also inverted, but fibers with caudal facial receptive fields tended to terminate medially rather than laterally. The primary afferent input to the MDH also provided some support for the “onion‐leaf” somatotopic organization that has been suggested for this nucleus (e.g., Kunc, '70). Fibers with rostral receptive fields tended to terminate in the rostral portion of the MDH while primary afferents with caudal fields gave off most of their collaterals in the caudal MDH and the rostral portion of the first cervical segment. There were, however, a number of exceptions to this generalization.}, number={4}, journal={JOURNAL OF COMPARATIVE NEUROLOGY}, author={CHIAIA, NL and HESS, PR and HOSOI, M and RHOADES, RW}, year={1987}, month={Oct}, pages={527–546} } @article{chiaia_hess_rhoades_1987, title={PREVENTING REGENERATION OF INFRAORBITAL AXONS DOES NOT ALTER THE GANGLIONIC OR TRANSGANGLIONIC CONSEQUENCES OF NEONATAL TRANSECTION OF THIS TRIGEMINAL BRANCH}, volume={36}, ISSN={["0165-3806"]}, DOI={10.1016/0165-3806(87)90066-6}, abstractNote={Retrograde and transganglionic tracing with a combination of horseradish peroxidase (HRP) and wheatgerm agglutinin (WGA) — conjugated HRP (WGA-HRP) was employed to determine whether transection of the infraorbital (IO) nerve on the day of birth and prevention of regeneration by retransecting it at weekly intervals until the time of a terminal anatomical experiment had effects upon ganglion cell survival and innervation of the brainstem by this trigeminal (V) branch that differed from those which followed a single transection of the same nerve on the day of birth without any attempt to prevent peripheral regeneration of the cut axons. Counts of labelled ganglion cells and examination of the brainstem labelling produced by application of HRP and WGA-HRP to the IO nerve proximal to the point of transection(s) at 6 weeks of age demonstrated no differential effects of preventing regeneration of the cut nerve. In animals subjected to a single transection of the nerve (n = 9), we counted an average of 5001.2 (S.D. = 1286.9) labelled ganglion cells and these had an average diameter of 22.7 μm (S.D. = 6.3). In the rats (n = 9) that sustained multiple nerve cuts, the average number of labelled ganglion cells was 4447.8 (S.D. = 1060.9). The mean diameter for these primary afferent neurons was 21.5 μm (S.D. = 6.6). Neither of these values were significantly different from those from the rats subjected to a single nerve cut. The cell counts from both of these groups were significantly lower than those obtained after application of HRP and WGA-HRP to the IO nerve in normal rats (n = 3, ¯X = 12,553.3, S.D. = 1454.8), but the average cell diameter in the normals (X = 23.2, S.D. = 6.6) was not significantly greater than that in the nerve-damaged animals. The pattern of brainstem labelling observed in the rats subjected to multiple nerve cuts was the same as that in the rats which sustained a single transection of the IO nerve on the day of birth16. Very little terminal labelling was observed in nucleus principalis, subnucleus oralis, subnucleus interpolaris or the magnocellular portion of caudalis. There was, however, very heavy labelling in laminae I and II of the latter nucleus.}, number={1}, journal={DEVELOPMENTAL BRAIN RESEARCH}, author={CHIAIA, NL and HESS, PR and RHOADES, RW}, year={1987}, month={Nov}, pages={75–88} }