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Article ; Online: Molecular diagnosis using RNAscope

Shiomitsu, Keijiro / Bechtel, Sandra M / Thompson, Patrick M / Frasca, Salvatore

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire

2020 Volume 84, Issue 4, Page(s) 319–323

Abstract: Immunohistochemistry has been used extensively to evaluate protein expression in clinical and research settings. However, immunohistochemistry is not always successful in veterinary medicine due to the lack of reliable antibody options, poor tissue ... ...

Abstract	Immunohistochemistry has been used extensively to evaluate protein expression in clinical and research settings. However, immunohistochemistry is not always successful in veterinary medicine due to the lack of reliable antibody options, poor tissue preservation, labor-intensive staining, and antigen-retrieval optimization processes. RNAscope
MeSH term(s)	Animals ; Dog Diseases/diagnosis ; Dogs ; Immunohistochemistry/methods ; Immunohistochemistry/veterinary ; In Situ Hybridization/methods ; In Situ Hybridization/veterinary ; Neoplasms/diagnosis ; Neoplasms/veterinary ; RNA, Messenger
Chemical Substances	RNA, Messenger
Language	English
Publishing date	2020-09-02
Publishing country	Canada
Document type	Journal Article
ZDB-ID	632814-3
ISSN	1928-9022 ; 0830-9000
ISSN (online)	1928-9022
ISSN	0830-9000
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Article ; Online: Dogs with osteosarcoma have altered pro- and anti-inflammatory cytokine profiles.

Axiak-Bechtel, Sandra M / Mathew, Leanne M / Amorim, Juliana R / DeClue, Amy E

Veterinary medicine and science

2019 Volume 5, Issue 4, Page(s) 485–493

Abstract: Background: Current advances in immunotherapy are an exciting area of study in canine osteosarcoma (OSA). The objective of this study was to determine the immune response in dogs with osteosarcoma by measuring stimulated leukocyte production of tumor ... ...

Abstract	Background: Current advances in immunotherapy are an exciting area of study in canine osteosarcoma (OSA). The objective of this study was to determine the immune response in dogs with osteosarcoma by measuring stimulated leukocyte production of tumor necrosis factor (TNF), interleukin (IL)-6, IL-10 and TNF and IL-6 to IL-10 ratios. Methods: Whole blood was collected from dogs with osteosarcoma receiving non-steroidal anti-inflammatory drugs (NSAIDs, n = 11), dogs with osteosarcoma not receiving NSAIDs (n = 14) and healthy dogs (n = 5). Results: No difference in TNF production was found among healthy and OSA dogs regardless of NSAID administration following stimulation with lipopolysaccharide (LPS) (p = .410), lipoteichoic acid (LTA) (p = .693) or PBS (p = .120). Leukocyte IL-6 production was greater in all dogs with OSA after stimulation with LPS (p = .015), LTA (p = .014) and PBS (p = .034) with no difference between OSA dogs receiving NSAIDs and those not. No differences in IL-10 were found among healthy controls and dogs with OSA regardless of NSAID use. There was no difference among groups for LPS-stimulated TNF to IL-10 ratios (p = .407). For LTA-stimulated leukocytes, the TNF to IL-10 ratio was lower in dogs with OSA than in healthy dogs (p = .031) with no difference between OSA NSAID dogs compared to OSA non-NSAID dogs (p = .059). No differences were found in LPS (p = .310)- or LTA (p = .265)-stimulated leukocyte IL-6 to IL-10 production ratios among groups. Conclusions: Dogs with osteosarcoma have an altered pro- and anti-inflammatory immunologic profile compared to healthy dogs regardless of NSAID use. Further study is indicated to determine the potential prognostic and therapeutic implications of these findings.
MeSH term(s)	Animals ; Anti-Inflammatory Agents, Non-Steroidal/administration & dosage ; Dog Diseases/physiopathology ; Dogs ; Female ; Immunity, Innate/drug effects ; Interleukin-10 ; Interleukin-6/metabolism ; Leukocytes/metabolism ; Male ; Osteosarcoma/physiopathology ; Osteosarcoma/veterinary ; Pathogen-Associated Molecular Pattern Molecules/administration & dosage ; Tumor Necrosis Factor-alpha/metabolism
Chemical Substances	Anti-Inflammatory Agents, Non-Steroidal ; Interleukin-6 ; Pathogen-Associated Molecular Pattern Molecules ; Tumor Necrosis Factor-alpha ; Interleukin-10 (130068-27-8)
Language	English
Publishing date	2019-08-02
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2819409-3
ISSN	2053-1095 ; 2053-1095
ISSN (online)	2053-1095
ISSN	2053-1095
DOI	10.1002/vms3.191
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Article ; Online: Pharmacokinetics and safety of TCMCB07, a melanocortin-4 antagonist peptide in dogs.

Axiak-Bechtel, Sandra M / Leach, Stacey B / Scholten, David G / Newton-Northup, Jessica R / Johnson, Brendan J / Durham, H E / Gruber, Kenneth A / Callahan, Michael F

Pharmacology research & perspectives

2021 Volume 9, Issue 3, Page(s) e00777

Abstract: The melanocortin-4 receptor (MC4R) antagonistic peptide TCMCB07 was developed for the treatment of cachexia. The objectives of this study were to examine pharmacokinetics and safety of TCMCB07 administered subcutaneously to healthy dogs. Dogs were ... ...

Abstract	The melanocortin-4 receptor (MC4R) antagonistic peptide TCMCB07 was developed for the treatment of cachexia. The objectives of this study were to examine pharmacokinetics and safety of TCMCB07 administered subcutaneously to healthy dogs. Dogs were treated with high- (2.25 mg kg
MeSH term(s)	Animals ; Arrhythmias, Cardiac/chemically induced ; Arterial Pressure/drug effects ; Body Weight/drug effects ; Dogs ; Female ; Histamine/blood ; Injections, Subcutaneous ; Male ; Peptides, Cyclic/adverse effects ; Peptides, Cyclic/pharmacokinetics ; Phosphorus/blood ; Receptor, Melanocortin, Type 4/antagonists & inhibitors
Chemical Substances	Peptides, Cyclic ; Receptor, Melanocortin, Type 4 ; Phosphorus (27YLU75U4W) ; Histamine (820484N8I3)
Language	English
Publishing date	2021-05-20
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2740389-0
ISSN	2052-1707 ; 2052-1707
ISSN (online)	2052-1707
ISSN	2052-1707
DOI	10.1002/prp2.777
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Article ; Online: Safety of TCMCB07, a melanocortin-4 antagonist peptide, in dogs with naturally occurring cachexia.

Journal of veterinary internal medicine

2023 Volume 37, Issue 6, Page(s) 2344–2355

Abstract: Background: The melanocortin 4 antagonist TCMCB07 is safe and effective in reversing cachexia caused by sepsis or cancer in rodents. The safety and pharmacokinetics of TCMCB07 are demonstrated in healthy beagle dogs.: Hypothesis/objectives: The ... ...

Abstract	Background: The melanocortin 4 antagonist TCMCB07 is safe and effective in reversing cachexia caused by sepsis or cancer in rodents. The safety and pharmacokinetics of TCMCB07 are demonstrated in healthy beagle dogs. Hypothesis/objectives: The objectives of this study were to investigate the safety, peak plasma concentrations, and potential for efficacy of TCMCB07 in pet dogs with naturally occurring cachexia over a 4-week time period. Animals: Fourteen dogs with cachexia of any underlying cause, except cancer of the oral cavity or gastrointestinal tract, were eligible for enrollment with informed client consent. Methods: This study was a prospective, 1-armed open-label trial. Physical examination, complete blood count, chemistry panel, and owner-assessed quality of life surveys were checked at weeks 1, 2, and 4. Due to potential for bradycardia and hypotension, Holter monitoring and blood pressure evaluations were scheduled at pre-enrollment and week 4. Results: Fourteen dogs completed the trial. Significant changes detected included increased mean body weight (18.6-19.5 kg, P < .02), increased body condition score (median Tufts 5-point thin dog scale score P < .004 and WSAVA muscle condition score P < .02) and increased mean blood urea nitrogen (21.79-30.43 mg dL Conclusions and clinical importance: TCMCB07 was safe and has potential efficacy in pet dogs with cachexia.
MeSH term(s)	Humans ; Animals ; Dogs ; Cachexia/drug therapy ; Cachexia/veterinary ; Prospective Studies ; Quality of Life ; Melanocortins ; Peptides ; Neoplasms/veterinary ; Dog Diseases/drug therapy
Chemical Substances	Melanocortins ; Peptides
Language	English
Publishing date	2023-10-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	92798-3
ISSN	1939-1676 ; 0891-6640
ISSN (online)	1939-1676
ISSN	0891-6640
DOI	10.1111/jvim.16915
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Article: Autologous cancer cell vaccination, adoptive T‐cell transfer, and interleukin‐2 administration results in long‐term survival for companion dogs with osteosarcoma

Flesner, Brian K / Wood, Gary W / Gayheart‐Walsten, Pamela / Sonderegger, F. Lynn / Henry, Carolyn J / Tate, Deborah J / Bechtel, Sandra M / Donnelly, Lindsay L / Johnson, Gayle C / Kim, Dae Young / Wahaus, Tammie A / Bryan, Jeffrey N / Reyes, Noe

Journal of veterinary internal medicine. 2020 Sept., v. 34, no. 5

2020

Abstract: BACKGROUND: Osteosarcoma (OSA) in dogs is an aggressive bone tumor with frequent chemotherapy failure and translational relevance for human health. HYPOTHESIS/OBJECTIVES: We hypothesized that dogs with OSA could be treated safely by ex vivo activated T‐ ... ...

Abstract	BACKGROUND: Osteosarcoma (OSA) in dogs is an aggressive bone tumor with frequent chemotherapy failure and translational relevance for human health. HYPOTHESIS/OBJECTIVES: We hypothesized that dogs with OSA could be treated safely by ex vivo activated T‐cells that were generated by autologous cancer vaccination and supported by interleukin‐2 (IL‐2) treatment with survival more than twice that reported for amputation alone. ANIMALS: Osteosarcoma‐bearing dogs (n = 14) were enrolled in a single‐arm prospective trial after complete staging before amputation. Four healthy dogs also were treated in a safety study. METHODS: Autologous cancer cell vaccinations were administered intradermally and dogs underwent leukapheresis. Mononuclear cell products were stimulated ex vivo with a T‐cell‐activating agent. Activated product was transfused and 5 SC IL‐2 injections were administered q48h. Dogs were monitored for metastasis by thoracic radiography every 3 months. RESULTS: Autologous cancer cell vaccine and activated cellular therapy (ACT) products were successfully generated. Toxicity was minimal after premedicants were instituted before ACT. With premedication, all toxicities were grade I/II. Median disease‐free interval for all dogs was 213 days. One dog developed cutaneous metastasis but then experienced spontaneous complete remission. Median survival time for all dogs was 415 days. Five dogs survived >730 days. CONCLUSIONS AND CLINICAL IMPORTANCE: This immunotherapy protocol without cytotoxic chemotherapy is safe and tolerable. Compared to historical amputation reports, survival was notably prolonged in this group of patients. Additional prospective studies are warranted to elucidate active immunologic mechanisms and further improve disease response and survival.
Keywords	T-lymphocytes ; amputation ; cytotoxicity ; dogs ; drug therapy ; human health ; interleukin-2 ; metastasis ; neoplasm cells ; osteosarcoma ; radiography ; remission ; vaccination ; vaccines ; veterinary medicine
Language	English
Dates of publication	2020-09
Size	p. 2056-2067.
Publishing place	John Wiley & Sons, Inc.
Document type	Article
Note	NAL-AP-2-clean ; JOURNAL ARTICLE
ZDB-ID	92798-3
ISSN	1939-1676 ; 0891-6640
ISSN (online)	1939-1676
ISSN	0891-6640
DOI	10.1111/jvim.15852
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Identification of immunologic and clinical characteristics that predict inflammatory response to C. Novyi-NT bacteriolytic immunotherapy.

DeClue, Amy E / Axiak-Bechtel, Sandra M / Zhang, Yan / Saha, Saurabh / Zhang, Linping / Tung, David D / Bryan, Jeffrey N

BMC veterinary research

2018 Volume 14, Issue 1, Page(s) 119

Abstract: Background: Clostridium novyi-NT (CNV-NT), has shown promise as a bacterolytic therapy for solid tumors in mouse models and in dogs with naturally developing neoplasia. Factors that impact the immunologic response to therapy are largely unknown. The ... ...

Abstract	Background: Clostridium novyi-NT (CNV-NT), has shown promise as a bacterolytic therapy for solid tumors in mouse models and in dogs with naturally developing neoplasia. Factors that impact the immunologic response to therapy are largely unknown. The goal of this pilot study was to determine if plasma immune biomarkers, immune cell function, peripheral blood cytological composition and tumor characteristics including evaluation of a PET imaging surrogate of tumor tissue hypoxia could predict which dogs with naturally developing naïve neoplasia would develop an inflammatory response to CNV-NT. Results: Dogs that developed an inflammatory response to CNV-NT had a higher heart rate, larger gross tumor volume, greater tumor [ Conclusions: Development of inflammation in response to CNV-NT is best predicted by pretreatment unstimulated leukocyte IL-10 production, heart rate, and gross tumor volume.
MeSH term(s)	Animals ; Clostridium/immunology ; Clostridium Infections/immunology ; Clostridium Infections/therapy ; Clostridium Infections/veterinary ; Dog Diseases/immunology ; Dog Diseases/therapy ; Dogs ; Female ; Immunotherapy/methods ; Immunotherapy/veterinary ; Inflammation/immunology ; Inflammation/microbiology ; Inflammation/veterinary ; Male
Language	English
Publishing date	2018-04-02
Publishing country	England
Document type	Journal Article
ISSN	1746-6148
ISSN (online)	1746-6148
DOI	10.1186/s12917-018-1424-1
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Article ; Online: Immune response to C. novyi-NT immunotherapy.

DeClue, Amy E / Axiak-Bechtel, Sandra M / Zhang, Yan / Saha, Saurabh / Zhang, Linping / Tung, David / Bryan, Jeffrey N

Veterinary research

2018 Volume 49, Issue 1, Page(s) 38

Abstract: Clostridium novyi-NT (CVN-NT) spores germinate in hypoxic regions of tumors and have successfully cured induced neoplasia in mouse models and resulted in objective tumor responses in naturally developing neoplasia in the dog. The objective of this pilot, ...

Abstract	Clostridium novyi-NT (CVN-NT) spores germinate in hypoxic regions of tumors and have successfully cured induced neoplasia in mouse models and resulted in objective tumor responses in naturally developing neoplasia in the dog. The objective of this pilot, descriptive, prospective, clinical investigation, was to evaluate and describe the immune response to CNV-NT spores to better understand which immune pathways might play a role in the response to this bacteriolytic immunotherapy. Intratumoral injection of CNV-NT spores result in increased phagocytosis and NK cell-like function after treatment. Intravenous injection of CNV-NT spores resulted in increased LPS-induced TNF-α production, LTA-induced IL-10 production and NK cell-like function post-treatment. Increased NK cell-like function was sustained to 28 (intratumoral) or 56 (intravenous) days post-treatment, and increased phagocytic function was sustained to 28 days post-treatment suggesting that CNV-NT spores induce longer-term immune cell function changes. Future investigations evaluating long-term immune system changes and associations between immune function and tumor remission rates should include evaluation of these pathways.
MeSH term(s)	Animals ; Biomarkers/analysis ; Clostridium/immunology ; Dogs ; Female ; Immunity, Innate ; Immunotherapy/veterinary ; Injections, Intravenous/veterinary ; Male ; Neoplasms/etiology ; Neoplasms/therapy ; Pilot Projects ; Prospective Studies ; Spores, Bacterial/immunology
Chemical Substances	Biomarkers
Language	English
Publishing date	2018-04-24
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1146298-x
ISSN	1297-9716 ; 0928-4249
ISSN (online)	1297-9716
ISSN	0928-4249
DOI	10.1186/s13567-018-0531-0
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Article ; Online: Resveratrol administration increases phagocytosis, decreases oxidative burst, and promotes pro-inflammatory cytokine production in healthy dogs.

Mathew, Leanne M / Woode, Rowena A / Axiak-Bechtel, Sandra M / Amorim, Juliana R / DeClue, Amy E

Veterinary immunology and immunopathology

2018 Volume 203, Page(s) 21–29

Abstract: Resveratrol is a polyphenol that is safe to administer to dogs and has immunomodulating properties. Canine in vitro work indicated that resveratrol spared polymorphonuclear cell (PMN) phagocytosis but reduced the robustness of PMN oxidative burst and ... ...

Abstract	Resveratrol is a polyphenol that is safe to administer to dogs and has immunomodulating properties. Canine in vitro work indicated that resveratrol spared polymorphonuclear cell (PMN) phagocytosis but reduced the robustness of PMN oxidative burst and resulted in a pro-inflammatory leukocyte cytokine profile. The objective of this study was to determine the short-term effect of resveratrol on the healthy canine innate immune system in vivo. The hypothesis was that resveratrol would spare phagocytosis, depress the vigor of PMN oxidative burst, and result in a proinflammatory stimulated leukocyte cytokine profile in vivo. In an open-label study, whole blood was collected from 12 healthy, adult client-owned dogs on day 0 and 3. Six dogs received resveratrol, 200 mg kg
MeSH term(s)	Animals ; Cytokines/metabolism ; Dogs ; Female ; Immunologic Factors/pharmacology ; Interleukin-10/blood ; Interleukin-6/blood ; Leukocytes/drug effects ; Leukocytes/metabolism ; Male ; Phagocytosis/drug effects ; Respiratory Burst/drug effects ; Stilbenes/pharmacology ; Tumor Necrosis Factor-alpha/blood
Chemical Substances	Cytokines ; Immunologic Factors ; Interleukin-6 ; Stilbenes ; Tumor Necrosis Factor-alpha ; Interleukin-10 (130068-27-8) ; resveratrol (Q369O8926L)
Language	English
Publishing date	2018-07-30
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	754160-0
ISSN	1873-2534 ; 0165-2427
ISSN (online)	1873-2534
ISSN	0165-2427
DOI	10.1016/j.vetimm.2018.07.013
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Article: Immune response to C. novyi-NT immunotherapy

DeClue, Amy E / Axiak-Bechtel, Sandra M / Zhang, Yan / Saha, Saurabh / Zhang, Linping / Tung, David / Bryan, Jeffrey N

Veterinary research. 2018 Dec., v. 49, no. 1

2018

Abstract	Clostridium novyi-NT (CVN-NT) spores germinate in hypoxic regions of tumors and have successfully cured induced neoplasia in mouse models and resulted in objective tumor responses in naturally developing neoplasia in the dog. The objective of this pilot, descriptive, prospective, clinical investigation, was to evaluate and describe the immune response to CNV-NT spores to better understand which immune pathways might play a role in the response to this bacteriolytic immunotherapy. Intratumoral injection of CNV-NT spores result in increased phagocytosis and NK cell-like function after treatment. Intravenous injection of CNV-NT spores resulted in increased LPS-induced TNF-α production, LTA-induced IL-10 production and NK cell-like function post-treatment. Increased NK cell-like function was sustained to 28 (intratumoral) or 56 (intravenous) days post-treatment, and increased phagocytic function was sustained to 28 days post-treatment suggesting that CNV-NT spores induce longer-term immune cell function changes. Future investigations evaluating long-term immune system changes and associations between immune function and tumor remission rates should include evaluation of these pathways.
Keywords	Clostridium ; animal models ; dogs ; immune response ; immune system ; immunotherapy ; interleukin-10 ; intravenous injection ; neoplasms ; phagocytosis ; remission ; spores ; tumor necrosis factor-alpha ; veterinary medicine
Language	English
Dates of publication	2018-12
Size	p. 38.
Publishing place	BioMed Central
Document type	Article
ZDB-ID	1146298-x
ISSN	1297-9716 ; 0928-4249
ISSN (online)	1297-9716
ISSN	0928-4249
DOI	10.1186/s13567-018-0531-0
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Article ; Online: Autologous cancer cell vaccination, adoptive T-cell transfer, and interleukin-2 administration results in long-term survival for companion dogs with osteosarcoma.

Flesner, Brian K / Wood, Gary W / Gayheart-Walsten, Pamela / Sonderegger, F Lynn / Henry, Carolyn J / Tate, Deborah J / Bechtel, Sandra M / Donnelly, Lindsay L / Johnson, Gayle C / Kim, Dae Young / Wahaus, Tammie A / Bryan, Jeffrey N / Reyes, Noe

Journal of veterinary internal medicine

2020 Volume 34, Issue 5, Page(s) 2056–2067

Abstract: Background: Osteosarcoma (OSA) in dogs is an aggressive bone tumor with frequent chemotherapy failure and translational relevance for human health.: Hypothesis/objectives: We hypothesized that dogs with OSA could be treated safely by ex vivo ... ...

Abstract	Background: Osteosarcoma (OSA) in dogs is an aggressive bone tumor with frequent chemotherapy failure and translational relevance for human health. Hypothesis/objectives: We hypothesized that dogs with OSA could be treated safely by ex vivo activated T-cells that were generated by autologous cancer vaccination and supported by interleukin-2 (IL-2) treatment with survival more than twice that reported for amputation alone. Animals: Osteosarcoma-bearing dogs (n = 14) were enrolled in a single-arm prospective trial after complete staging before amputation. Four healthy dogs also were treated in a safety study. Methods: Autologous cancer cell vaccinations were administered intradermally and dogs underwent leukapheresis. Mononuclear cell products were stimulated ex vivo with a T-cell-activating agent. Activated product was transfused and 5 SC IL-2 injections were administered q48h. Dogs were monitored for metastasis by thoracic radiography every 3 months. Results: Autologous cancer cell vaccine and activated cellular therapy (ACT) products were successfully generated. Toxicity was minimal after premedicants were instituted before ACT. With premedication, all toxicities were grade I/II. Median disease-free interval for all dogs was 213 days. One dog developed cutaneous metastasis but then experienced spontaneous complete remission. Median survival time for all dogs was 415 days. Five dogs survived >730 days. Conclusions and clinical importance: This immunotherapy protocol without cytotoxic chemotherapy is safe and tolerable. Compared to historical amputation reports, survival was notably prolonged in this group of patients. Additional prospective studies are warranted to elucidate active immunologic mechanisms and further improve disease response and survival.
MeSH term(s)	Animals ; Bone Neoplasms/drug therapy ; Bone Neoplasms/veterinary ; Dog Diseases/drug therapy ; Dogs ; Interleukin-2/therapeutic use ; Osteosarcoma/drug therapy ; Osteosarcoma/veterinary ; Pets ; Prospective Studies ; T-Lymphocytes ; Treatment Outcome ; Vaccination/veterinary
Chemical Substances	Interleukin-2
Keywords	covid19
Language	English
Publishing date	2020-07-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	92798-3
ISSN	1939-1676 ; 0891-6640
ISSN (online)	1939-1676
ISSN	0891-6640
DOI	10.1111/jvim.15852
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