Health status. Microbial impact on research. Mouse hepatitis in the nude mouse. Microbiology Impact of infections on research
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1 Health status Micriobial status Microbial impact on research Specific infections Normal flora Axel Kornerup Hansen, Professor, DVM, Dr.Med.Vet. Department of Veterinary Disease Biology Section of Biomedicine Lab Animal Science and Welfare Mouse hepatitis in the nude mouse Nude mouse Immunedeficient No thymus No T-cells Mouse hepatitis Caused by a coronavirus Symptoms Wasting Cachexia Diarrhea Jaundice Microbiology Impact of infections on research DIRECT Clinical/pathological changes Immunemodulation Physiological modulation Oncological modulation Fertilitity modulation Microbiological competition INDIRECT Contamination of biological products Activation of direct effects Death Disease Misinterpretation 1
2 Tyzzer s disease Tyzzer s Disease in Mice Agent Clostridium piliforme Clinical signs Peracute deaths Diarrhea Weakness Pathology Multiple focal liver necrosis Enteritis Type 1 diabetes i BB rats infected with Mycoplasma pulmonis L. VOOT, S. SADEWASSER, and I. KLOETING. THE DEVELOPMENT OF BB-RAT DIABETES IS DELAYED OR PREVENTED BY INFECTIONS OR APPLICATIONS OF IMMUNOGENS. Zeitschrift fur Versuchstierkunde 31 (4):197, Physiological modulation Tyzzer s disease and pharmacokinetics 2
3 Examples Microbial modulation of oncogenesis Examples of published impact of rodent viruses on oncology H1 parvovirus Generally anti-cancerogenic Lactic dehydrogenase virus Enhanced growth of some tumortypes Prevents oncogenic effect of Mouse mammary tumor virus Mouse hepatitis virus Raised susceptibility to chemical carcinogenesis Reovirus type 3 Suppressed urethan pulmonary carcinogenesis Sendai virus Raised susceptibility to chemical carcinogenesis Reduce incidence/delay pulmonary adenomas Examples Microbial modulation of oncogenesis Helicobacter hepaticus Bacteria Citrobacter rodentium Raised susceptibility to chemical carcinogenesis Salmonella spp. Suppressed growth of transplantable tumors Helicobacter spp. Causes hepatic cancer Infectious impact on rodent reproduction Uterine infections Placental transfer Reduced fertility Neonatal mortality Mouse hepatitis virus Lymphocytic choriomeningitis virus Parvoviruses Ectromelia Rotaviruses Mycoplasma pulmonis Clostridium piliforme Salmonella Pasteurella pneumotropica
4 Microbiological competition Bia, F. J., Thornton, G. F., Main, A. J., Fong, C. K. Y., Hsiung, G. D., Western equine encephalitis mimic-king herpes simplex encephalitis, J.Amer.Med.Ass. 244, 367, A patient is sufferring from encephalitis A biopsy is taken from the brain of the patient The biopsy is inoculated into mice All mice dies Reinoculation from the dead mice All new mice dies Mouse hepatitis virus is recovered from the dead mice The patient is treated with arabinoside A against human hepatitis The patient recovers Serology is made on serum from the patient The patient has no antibodies against Mouse hepatitis virus The patient has antibodies against Western Equine Encephalitis The patient has not been suffering from a mouse disease The patient has been suffering from a horse disease 1 mouse different germs The composition of the mouse gut microbiota individual organisms 23 Hansen et al, 2011 (in preparation) 4
5 Organ related Type 1 diabetes Arthritis IBD Multiple sclerosis Stress Visceral pain Inflammatory pain Atherosclerosis Atopic dermatitis Contact hypersensitivity Systemic Allergy Obesity Lupus erythematosus Type 2 diabetes Lauritzen et al Scandinavian Journal of Laboratory Animal Science 37 (1):43-54, Relation between intestinal flora and inflammatory disease in animal models Impact of Organ germfree state specificity on prevalence Type 1 Diabetes β-cells None Animal models NOD mice BB rats Impact of the enteric microflora + Inflammatory bowel disease Bowel IL10 KO mice HLA-B27 TG rats +++ Experimental allergic encephalomyelitis Multiple sclerosis CNS -? (EAE) Rheumatoid arthritis Systemic Collagenor adjuvant induced arthritis +++ Type 2 Diabetes Systemic - Lep ob mouse? Obesity Systemic - Lep ob mouse ++ Atherosclerosis Systemic - ApoE KO Mice? Allergy/Asthma 27 Systemic - Anaphylactic shock Percutaneous anaphylaxis (PCA) +++ Hypothetic immunology of hygiene hypothesis Birth T reg
6 Hypothetic immunology of hygiene hypothesis T cells weak signals Type 1 diabetes Th1 Inflammatory IFN-γ IL-2 TNF-α α Th2 Protective T effector cells IL-10 IL-4 IL-5 IL-13 Allergy Birth Th1/Th T cells intermediate signals Th1/Th2 Balance FoxP3 T reg cells Microbe with MAMP Enterocytes Th1 IL-12 Dendritic cell Precursor T-cell Th2 33 Toll like receptors (TLR) and Microbial Associated Molecular Patterns (MAMPs) TLR s and MAMP s Recognize common microbial molecules distinguishable from host molecules microbial associated molecular patterns (MAMPs) different types (TLR1, TLR2,..) Present on various cell types Peptidoglycan Gram + Lipoarabinomannan Mycobacteria PSA Bacillus fragilis Doublestranded RNA Lipopolysaccharides Gram Lipoteichoic acid Gram + Flagellin TLR5 TLR3 TLR
7 Non-obese diabetic (NOD) mouse Autoimmune pancreatic insulitis > four weeks of age Type 1 diabetes Females > 3 months Makino et al, 1980 Non-obese diabetic mice (T1D) Experimental study design Non-obese diabetic mice (T1D) Experimental study design Groups Vancomycin Birth Vancomycin Adult treatment Neonatal treatment Age (weeks) Diabetes onset betes incidence (%) Cumulative diab Age (days) Control Adult treatment Neonatal treatment Hansen et al, 2011 (Manuscript) Hansen et al, 2011 (Manuscript) Non-obese diabetic mice (T1D) Experimental study design Early life priming of the regulatory immunity LPS Regulatory and innate immunity Hansen et al, 2011 (Manuscript) 7
8 Gut microbiota in relation to obesity and type 2 diabetes Pure hypothesis Leptin deficient obese mice Lep ob Obesity Type 2 diabetes Lipids LPS Bacteria Enterocytes TNF-α Jackson Laboratories, Influence of antibiotic manipulation on glucose intolerance in C57BL/6 mice Ampicillin-treated DIO mice Week (amp +) -) Sørensen et al, Manuscript 47 Bech-Nielsen et al,
9 Inbred animals Various genetically related conditions 9
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