Immunology in the Animal: Microbial Interaction, Stress and

Nutrition

Chris ChaseDept. Vet. & Biomed Sci

South Dakota State UniversityBrookings, SD

Credits� Kuby Immunology

� Immunobiology, 9th edition

� Bruns et al, 2010 PLoS Pathog

� David Topham, University of Rochester

� Mike Kogut, ARS

� Lance Baumgard, Iowa State

Topics� Three Levels of Immune Protection

� Mucosa-Barrier- New Frontier, Good

� Innate Immunity-Good, Bad, Ugly

� Acquired Immunity-Good, Bad

“I Know It When I See It”

Supreme Court Justice Stewart Potterto describe his threshold test for

Obscenity in Jacobellis v. Ohio (1964)

Gut Health Symposium Dec. 3-5 , 2012

What is gut health ?

The ”Gut”� the largest interface between the external environment & the internal

host milieu

� constitutes the major barrier through which molecules can either be absorbed or secreted

� largest residence of immune cells in body; acts as a physical & immune barrier to pathogens

� natural habitat for a large & dynamic community of microbes that participate & regulate gut & systemic functions

1. Effective digestion/food absorption

2. Stable microbial population

3. Effective immune status

4. Effective gut barrier

5. Effective neuroendocrine system

Components of a Healthy GI System

Kogut Gut Health Symposium Dec. 3-5 , 2012

Neuro-Immune Interaction

Margolis KG, Gershon MD, Bogunovic M. Cellular Organization of Neuroimmune Interactions in the Gastrointestinal Tract. Trends Immunol 2016;37:487–501.

MAMPs microbial-associated molecule patterns

Think of the body as a hollow plastic tube…

The food is digested within the hole in the tube, but it never actually enters into the solid plastic material.

Tube inner surface ~Digestive System~

Plastic interior ~Body~

Tube outer surface ~Skin~

Immune responses

Invasion& infection

Barriers

Innate immunity

Adaptive immunity

+

+

Inflammation

From David J. Topham, Introduction to Viral Immunology:Part I

1st Line of DefenseBarriers

mucous, tears, gastric pH,saliva, skin

Cellular and humoral defensesinterferon, cytokines (pro-inflammatory and

T stimulatory), complement proteins, phagocytosis, NK cells

2nd Line of Defense

Cellular and humoral defensesAntibodies, cytokines, T helper cells,

cytotoxic T cells

3rd Line of Defense

Stressors at the Farm

� Co-mingling

� Injury

� Water - palability and supply

� Feed - time to first

� Pen density

� Pen total number

� Heat Stress

� Handling and movements

� Fear and Flight

� New �add-ons�

� Weather extremes

� Dust

� COMPETITION

� Vaccination

Adapted from Reiche et al. 2004. The Lancet Oncology 5:617-625.

Stress

Hypothalamus

CNS

Pituitary

Inflammation/Innate Immunity

AdaptiveImmunity

Hormones and cytokinesNeuropeptides and neurotransmitters

AutonomicNerves

EndocrineSystem

Immune System

InfectionsTrauma Fear & Flight

Mucosal Immune Responses

2nd Line of DefenseHumoral and Cellular Defenses

Cellular, cytokine and protein defensesInterferons, defensins, chemokines, cytokines

(pro-inflammatory and T stimulatory), complement proteins, TLRs, phagocytosis, NK

cells

Invasion& infection

Barriers

Innate immunity

Adaptive immunity

+

+

Inflammation3rd Line of Defense

Humoral and Cellular Defenses Cellular and humoral defenses

Antibodies, cytokines, chemokines,T helper cells, cytotoxic T cells

•1st Line of Defense•Barriers

Skin & Mucous membranes and secretions

Barrier, rapidly regenerating surfaces, peristaltic movement, lysozyme, sebaceous/mucous

secretions, stomach acid, commensal organisms

Epithelial Cells and First Line of Defense

Decrease water intake- dehydration barrierê

Epithelium and Kill Zone

Epithelial Cells� Location

� GI tract� Respiratory tract� Reproductive tract� Skin

Epithelium and Immune Function

Microbiota (Microflora)� The collection of organisms found in and on our body- very location

specific and individual specific (genetic component)

� Gastrointestinal microbiota is the most diverse and has the largest interaction both with mucosa and ingesta- “superorganism”

Commensal Organisms� 1014 organisms-eukarya, archae and bacteria

� 1012 commensal bacteria/ml- 10X higher than the combined somatic and stem cells in man

� Commensals are essential for immune development

� Composition of commensal bacteria influenced by the host immune status

Microbiota and Immune Development

Global Effect of Microbiota

Reynolds, L. A., & Finlay, B. B. (2013).. Expert Review of Clinical Immunology, 9(11), 1019–1030.

Commensal Bacteria

Front. Vet. Sci., 23 September 2015 | https://doi.org/10.3389/fvets.2015.00036The gut microbiome and its potential role in the development and function of newborn calf gastrointestinal tract

Commensals Mucosal Barrier and Immunity

Obata, Y., Furusawa, Y., & Hase, K. (2015). Epigenetic modifications of the immune system in health and disease. Immunology and Cell Biology, 93(3), 226–232. http://doi.org/10.1038/icb.2014.114

Adapted from Cell 156, Issues 1–2, 2014, Pages 7

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Microbes and Regulating Innate Immunity

Anti-inflammatory response is keyfor healthy mucosa

Commensals Mucosal Barrier and Immunity

Timsit E, Holman DB, Hallewell J, et al. The nasopharyngeal microbiota in feedlot cattle and its role in respiratory health. Animal Frontiers 2016;6:44–50.

Stress and Dysbiosis

Commensals and their Induced Immune Changes

Khosravi and Mazmanian, Current Opinion in Microbiology 2013, 16:221–227

Microflora, Diet and Stress� Nutrition isn’t just feeding the “animal” it is also feeding the

“microbes”

� Production of vitamins

� More efficient bacteria- increase opportunity for obesity

• Innate Immunity - 2nd Line of Defense

• the troops are called to battle…– injury & infection– macrophages are on patrol– cytokine chemicals attract other �troops�– Absolutely essential for vaccine responses

Inflammatory Response

Neuro-Immune Interaction

Obata Y, Pachnis V. The Effect of Microbiota and the Immune System on the Development and Organization of the Enteric Nervous System. Gastroenterology 2016;151:836–844.

Epithelial Cells- Immunomodulation� Pro-inflammatory cytokines-

� Epithelial Cell Enemy# 1� Tumor necrosis factor-alpha

Leaky Gut� Leaky gut explains the negative consequences of heat stress and off-

feed events (all farm animals)

Baumgard L, International Symposium on Dairy Cattle Nutrition, Wageningen NL October 26, 2017

Leaky Gut Mucosa

Inflammatory Response

Systemic response

First Immune Organ

Neutrophils-Blowing Up-Collateral Damage

Bruns S, Kniemeyer O, Hasenberg M, et al. . PLoS Pathog 2010;6:e1000873.

Tissue Damage- Overactive Immune System

Overactive Inflammatory Response

Heat Stress and Gut Health� Diversion of blood flow to skin and extremities

� Coordinated vasoconstriction in intestinal tissues� Reduced nutrient and oxygen delivery to enterocytes� Hypoxia increases reactive oxygen species (ROS)

� Reduced nutrient uptake increases intestinal osmolarity

Baumgard L, International Symposium on Dairy Cattle Nutrition, Wageningen NL October 26, 2017

Cytokine Storm

Tisoncik J R et al. Microbiol. Mol. Biol. Rev. 2012;76:16-32

Cytokine Storm� High Temps- 104˚-106˚F

� Respiratory Disease-� Acute Lung Injury� Acute Respiratory Disease Syndrome

� Is BRSV???

� Is Vaccination or Aspirin the Answer?

Acute Respiratory Disease Syndrome

Tisoncik J R et al. Microbiol. Mol. Biol. Rev. 2012;76:16-32

Acute

Chronic

Brain-Gut-Microbiota axisInflammation and Obesity- Overconditioned Weaned

Calves, Fat Cattle, Transition Cow

Cluny, N.L., et al. Cannabinoid signaling regulates inflammation and energy balance: The importance of the brain–gut axis. Brain Behav. Immun. (2012), doi:10.1016/j.bbi.2012.01.004

DietChange Microbiome

CB-cannabinoid receptor

Endocannabinoid system

Documented Causes of Increased Intestinal Permeability (“leaky gut”)Large Intestine

Acidosis

Distant Inflammation

Weaning

Small Intestine Bacteria OvergrowthLarge Intestine PTN Fermentation

Feed Restriction

Transition Period Heat Stress

Leaky Gut

Psychological Stress

Baumgard L, International Symposium on Dairy Cattle Nutrition, Wageningen NL October 26, 2017

Inflammation and Production� Proinflammatory Cytokines

� Tumor Necrosis Factor-! (TNF-!)� Interleukin 1-" (IL-")� Interleukin 6 (IL-6)

� Proinflammatory Cytokines- Turn on Acute Phase Proteins

Inflammation and Effect on Production Parameters

� Increase Sickness Behavior-listlessness

� Decrease feed intake- Inappetence-

� Increase body temperature sweats

� Decreased feed conversion

� Decrease gain

� Decrease milk production

� Increased Mastitis

� Increased Metritis

� Increased BRD

Consequences of Leaky Gut and LPS

Johnson JS, Abuajamieh M, Fernandez MVS, et al. Thermal stress alters postabsorptive metabolism during pre-and postnatal development. In: Sejian V, ed. Climate Change Impact on Livestock: Adaptation and Mitigation. New Delhi: Springer India; 2015:61–79.

Overactive Inflammatory Response

Tissue Damage

Proinflammatory Summary

� Innate Pro-inflammatory response� Cell Recruitment� Neutrophil Migration� Acute Inflammation� Humoral Immunity IgG� Cell mediate immunity-Memory

Pro-Inflammatory

Disease Immunity

Anti-inflammatory Summary

� Innate Anti-inflammatory response� Maintains gut homeostasis� Humoral immunity-IgA� Cell Mediated Regulatory T cells- Helps to Control

Acute Inflammation-

Anti-Inflammatory

Disease Function-IgA

Innate Immunity Interventions

� NSAIDS� Aspirin-� Meloxicam

� Cytokine� Imrestor (pegbovigrastim injection)- Granulocyte Colony Stimulating Factor

(PEG bG-CSF)- Elanco- Off the Market

� Immunomodulators� Immunoboost- MCWF Amplimune Bioniche- Nova Vive� Zelnate- Liposome-DNA- CpG-Bayer-

� Gut Health- Nutriceuticals� Prebiotics� Probiotics

Mucosal Vaccine Responses

2nd Line of DefenseHumoral and Cellular Defenses

Cellular, cytokine and protein defensesInterferons, defensins, chemokines, cytokines

(pro-inflammatory and T stimulatory), complement proteins, TLRs, phagocytosis, NK

cells

Invasion& infection

Barriers

Innate immunity

Adaptive immunity

+

+

Inflammation3rd Line of Defense

Humoral and Cellular Defenses Cellular and humoral defenses

Antibodies, cytokines, chemokines,T helper cells, cytotoxic T cells

•1st Line of Defense•Barriers

Skin & Mucous membranes and secretions

Barrier, rapidly regenerating surfaces, peristaltic movement, lysozyme, sebaceous/mucous

secretions, stomach acid, commensal organisms

Where Does The Intranasal Vaccine Response Occur?

Brandtzaeg P. Potential of nasopharynx-associated lymphoid tissue for vaccine responses in the airways. Am J Respir Crit Care Med 2011;183:1595–1604.

T cells B cells

From: Kuby Immunology 4/E by Goldsby, Kindt & Osborne, 1992, 1994, 1997, 2000 by W. H. Freeman and Company. Used with permission

2

1

Danger and Activation of the Acquired Immune System

3rd Line of Defense “mucosal immune system�

Nasal Cavity

Lymph Node

BloodVessel

Lymphatic Duct

Spleen

LactatingMammary Glands

Reproductive Tract

Middle Ear

RespiratoryTract

Gastrointestinal Tract

TonsilsSalivaryGlands

LacrimalGlands

Take Home: A Healthy Gut is a Necessity- What About

Nutriceuticals?

� Essential Oils, Organic Acids, Bioactives (Plasma Proteins, colostrum), Bacterial cultures, yeast, cell wall products, metabolites

� Problem: how do we measure it

� Do we need them all the time? Depends- times of stress

Summary

� Immune System is the Body’s Defense System- 3 lines of defense

� Mucosa-Barrier- New Frontier, � Good- Antimicrobial proteins, IgA, cytokines� Bad-inflammatory cells- leaky gut

� Innate Immunity-� Good-Primary “reactive” defense, necessary for acquired� Bad-leaky gut� Ugly-collateral damage

� Acquired Immunity-� Good- specific immunity, duration� Bad-allergy, chronic inflammation

Harvey Dunn (1884-1952) Prairie is My Garden, South Dakota Art Museum

Cattle First, Then PARTY