Adaptive Immune

Response in Periodontitis

A/Prof Glen Scholz

glenms@unimelb.edu.au

17 August 2018
 Outline of Today’s Lecture

1. Brief re-cap of adaptive immunity

2. T-lymphocyte subsets in periodontitis

• Cytotoxic T-lymphocytes (CTLs)

• Helper T-lymphocytes (Th cells)

• Regulatory T-lymphocytes (Treg cells)

3. B-lymphocytes and antibodies in periodontitis

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 Adaptive Immunity

• Provides specific protection against pathogens (antigen-specific

receptors; TCR and BCR)

• Provides immunological memory

• Consists of:
▪ T-lymphocytes (aka "T cells")
❖ Provide cell-mediated immunity (cytokines, other effector
proteins)
▪ B-lymphocytes (aka "B cells")
❖ Provide humoral immunity (antibodies)

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 T-lymphocytes

• CD8+ Cytotoxic T-lymphocytes (CTLs):

▪ Kill infected cells; also kill tumour cells

• CD4+ Helper T-lymphocytes (Th cells):

▪ Provide additional "signals" (e.g. IFNg and IL-4) to:
➢ Enhance killing of intracellular pathogens by macrophages
➢ Enhance antibody production by B-lymphocytes

• CD4+ Regulatory T-lymphocytes (Treg cells):

▪ Suppress the activity of other lymphocytes

See slides 34-42 of Adaptive Immunity lecture for a refresher 4

 B-lymphocytes

• Antigen-activated B-lymphocytes proliferate and differentiate into

antibody secreting plasma cells, and memory cells

• Antibody effector functions:

▪ Neutralisation: Prevent interaction of pathogens and products
(e.g. toxins) with host cells

▪ Opsonisation: Enhance phagocytosis and killing/destruction

of pathogens and products (by neutrophils and macrophages)

▪ Complement activation: Activate classical pathway

See slides 19-27 of Adaptive Immunity lecture for a refresher 5

 Disease Progression in Periodontitis
Healthy gingiva Severe gingivitis Chronic periodontitis

Ohlrich EJ et al, Australian Dental Journal (2009) 54 Suppl 1:S2-10

Innate Immunity Adaptive Immunity

Kept under control? Or does it become dysregulated?

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 Disease Progression in Periodontitis
Healthy gingiva Severe gingivitis Chronic periodontitis

Ohlrich EJ et al, Australian Dental Journal (2009) 54 Suppl 1:S2-10

• During the early phase (gingivitis), the inflammatory infiltrate consists

mostly of T-lymphocytes (also innate immune cells – mainly neutrophils)

• In non-susceptible individuals, T-lymphocyte mediated immunity

(together with innate immunity) prevents periodontal "infection"

• Stable lesion with limited tissue damage, restoration of tissue health

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 Disease Progression in Periodontitis
Healthy gingiva Severe gingivitis Chronic periodontitis

Ohlrich EJ et al, Australian Dental Journal (2009) 54 Suppl 1:S2-10

• In susceptible individuals, the bacteria cannot be kept under control by

T-lymphocyte mediated immunity (and innate immune cells)

• Development of B-lymphocyte mediated immunity NB: B-lymphocytes

are the most abundant immune cell in chronic periodontitis lesions.

• Tissue damage progressively worsens (e.g. connective tissue breakdown

and alveolar bone loss, resulting in tooth loss)
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 Disease Progression in Periodontitis
Healthy gingiva Severe gingivitis Chronic periodontitis

Ohlrich EJ et al, Australian Dental Journal (2009) 54 Suppl 1:S2-10

• Shift from a predominantly "T-lymphocyte lesion" to a predominantly

"B-lymphocyte lesion" as the patient progresses from gingivitis to
periodontitis T-cell response - gingivitis
B-cel response - periodontitis

• What about the innate immune response? Macrophages become more

predominant

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T-lymphocyte subsets
in Periodontitis

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 CD8+ Cytotoxic T cells in Periodontitis
• No convincing evidence that CD8+ T cells directly contribute to
tissue destruction during disease progression
• Antibody-mediated depletion of CD8+ T cells in mice prior to
infection with a periodontal pathogen did not reduce alveolar
bone loss
• b2-microglobulin-deficient mice (lack MHC class I) developed
normal alveolar bone loss following infection

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 CD8+ Cytotoxic T cells in Periodontitis
• No convincing evidence that CD8+ T cells directly contribute to
tissue destruction during disease progression
• Antibody-mediated depletion of CD8+ T cells in mice prior to
infection with a periodontal pathogen did not reduce alveolar
bone loss
• b2-microglobulin-deficient mice (lack MHC class I) developed
normal alveolar bone loss following infection
• Most periodontal pathogens are extracellular; therefore,
pathogen-derived peptides are presented via MHC class II to CD4+
T cells

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 CD8+ Cytotoxic T cells in Periodontitis
• No convincing evidence that CD8+ T cells directly contribute to
tissue destruction during disease progression
• Antibody-mediated depletion of CD8+ T cells in mice prior to
infection with a periodontal pathogen did not reduce alveolar
bone loss
• b2-microglobulin-deficient mice (lack MHC class I) developed
normal alveolar bone loss following infection
• Most periodontal pathogens are extracellular; therefore,
pathogen-derived peptides are presented via MHC class II to CD4+
T cells
• And, antibody-mediated depletion of CD4+ T cells in mice reduces
suggests CD4 t-cells play a role in bone loss
alveolar bone loss 13
 CD4+ T-lymphocyte subsets

Intracellular bacteria
Cell-mediated immunity

Helminth (parasitic
worm) infections
Allergy type diseases

Zhou & Restifo, Nature Reviews Immunology (2010) 10, 248-256 14

 Th1 cells

• Development driven by IL-12

• Secrete IFNg (signature "Th1 cytokine")

• Activate bactericidal functions of macrophages, and promotes

maturation of dendritic cells

• Suppress B cells and antibody-producing plasma cells

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 Th1 cells in Periodontitis

• DCs in the gingival epithelium process foreign peptide antigens

and present them to CD4+ T cells

• In the presence of antigen and IL-12, CD4+ T cells proliferate and

differentiate into effector Th1 cells

• Antigen-activated Th1 cells secrete IFNg, which enhances the

innate immune response, and reinforces the Th1 response

• Inhibition of IFNg in vivo (in mice) results in increased P. gingivalis

survival and virulence

• IL-12 levels often inversely correlate with severity of human

periodontitis
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 Th1 cells in Periodontitis
important mediator of disease
• However, Porphyromonas gingivalis can impair/inhibit:
▪ IL-12 expression (Th1)
▪ IFNg expression (Th1)
▪ Th1 chemokine expression (e.g. CXCL10)
IL-12
• IL-12 consists of two subunits: p40 + p35

• TLR2 activates p40 but not p35 expression

• TLR4 induces p40 & p35 expression

• Impaired Th1 cell-mediated response?

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 Th2 cells

• Development driven by IL-4

• Secrete IL-4, IL-5, IL-10 (signature "Th2 cytokines")

• Regulate humoral (antibody-mediated) immunity by providing

cytokines required for B cell proliferation and activation

• Th2 cytokines (e.g. IL-4) reinforce the Th2 response

• Involved in allergic reactions

• Suppress Th1 cell-mediated immunity

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 Th2 cells in Periodontitis

• Majority of studies indicate that Th2 cells are more abundant than
Th1 cells in periodontitis lesions

• However, no consistent evidence of distinct T-helper cell

populations causing pathology in periodontitis:
▪ Some studies found "Th1 cytokines" (e.g. IFNγ) predominated
over "Th2 cytokines" (e.g. IL-4)

▪ Other studies showed the presence of both Th1 and Th2

cytokines

• Dynamic interaction between Th1 and Th2 cell subsets that results
in fluctuations in disease activity?
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 Th1 and Th2 cells in Periodontitis

Therefore, maintenance of an appropriate

balance between Th1 and Th2 responses is
required for periodontal health?

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 Th1 and Th2 cells in Periodontitis

Therefore, maintenance of an appropriate

balance between Th1 and Th2 responses is
required for periodontal health?

Unfortunately, it’s not that simple.....

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 CD4+ T cell subsets

Intracellular bacteria
Cell-mediated immunity

Helminth (parasitic
worm) infections
Allergy type diseases

Down-regulation of the immune

response in autoimmunity &
infection

Extracellular bacteria, fungi

Cell-mediated autoimmunity

Zhou & Restifo, Nature Reviews Immunology (2010) 10, 248-256 22

 Th17 cells

• Development driven by TGFb, IL-6, IL-1β, IL-23

• Secrete IL-17, IL-6, IL-22 (signature "Th17 cytokines")

• Amplify inflammatory responses

• Important for immunity against extracellular bacteria

• Prominent in many chronic inflammatory diseases

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 Th17 cells in Periodontitis

• Th17 cells reported to be the dominant T helper cell subset in

human periodontitis lesions

• IL-17 stimulates expression of MMPs and inflammatory cytokines

by gingival fibroblasts

• Also stimulates RANKL expression (e.g. by osteoblasts)

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Th17 cells in Periodontitis

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Hajishengallis & Korostoff, Periodontology 2000 (2017) 75, 116-151
 Th17 cells in Periodontitis

• Th17 cells reported to be the dominant T helper cell subset in

human periodontitis lesions

• IL-17 stimulates expression of MMPs and inflammatory cytokines

by gingival fibroblasts

• Also stimulates RANKL expression (e.g. by osteoblasts)

• Th17 cells also provide "help" to B cells (for antibody responses)

• IL-17 expression is up-regulated in human periodontitis lesions

• IL-17 levels correlate with periodontitis severity in humans

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 Treg cells

• Development driven by TGFb (also important for Th17 cells)

• Secrete TGFb and IL-10 (signature "Treg cytokines")

• Regulate other T cell subsets, to maintain tolerance to self-antigens

• Dampen immune responses

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 Treg cells in Periodontitis

• Treg cell numbers are increased in periodontitis (together with

increased numbers of B cells)

• Different Treg subsets (e.g. "natural" and "inducible")

• TGFb and IL-10 expression are elevated in periodontal lesions

• But are the Treg cells doing their job effectively?

• Could Treg cells convert into Th17 cells?

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CD4+ T cell subsets in Periodontitis

Hajishengallis & Korostoff, Periodontology 2000 (2017) 75, 116-151

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B-lymphocytes and Antibodies
in Periodontitis

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 B-lymphocytes in Periodontitis
Healthy gingiva Severe gingivitis Chronic periodontitis

Ohlrich EJ et al, Australian Dental Journal (2009) 54 Suppl 1:S2-10

• B cells (and plasma cells) most abundant immune cell in periodontitis

lesions associated with bone loss (~60% of total leukocytes)

• Production of an antibody response is thought to be beneficial in

preventing periodontal infection

• But remember, B cells also produce RANKL and IL-1b, which promote
osteoclastogenesis (stimulate alveolar bone resorption) 31
 Antibodies in Periodontitis

• IgA and IgG antibodies can be produced locally in the periodontium

• Antibodies to all suspected periodontal pathogens have been

detected in human serum and gingival crevicular fluid

• Antibody titres tend to increase following therapy, considered a

favourable response

• But antibody titres vary greatly between patients

• Different people can produce different isotype antibodies

(remember, the isotype determines the effector function of the
antibody)

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 Antibodies in Periodontitis

• Quality of the antibody (humoral) response may affect the

progression of periodontal infection

• Are the antibodies produced relevant or non-specific?

• For example, are they:

▪ Protective – i.e. provide periodontal immunity

▪ Non-protective – Remember, immune complexes are potent

activators of inflammation (promote connective tissue
destruction and bone loss?)

▪ What happens if the antibodies are not protective?

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 Antibodies in Periodontitis

• Antibody titres to P. gingivalis increased in periodontitis patients

• Serum antibody titres to P. gingivalis reduced following successful

treatment of periodontitis patients

• IgG avidities to P. gingivalis increase following treatment

• Several studies reported a predominance of IgG2 antibodies

• IgG2 antibodies:
▪ Primary antibody subclass produced in response to bacterial
carbohydrates and LPS

▪ Weak complement activation and opsonisation (see slide 26 of

Adaptive Immunity lecture)
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 Antibodies in Periodontitis

• Different patterns of immunoreactivity towards periodontal

pathogens have been reported

• Antibody response to P. gingivalis fails to eliminate the pathogen

• Non-protective, low avidity anti-P. gingivalis antibodies that may

not be capable of effectively mediating a variety of immune
responses (complement activation, opsonisation/phagocytosis)

35
 Antibodies in Periodontitis

• Different patterns of immunoreactivity towards periodontal

pathogens have been reported

• Antibody response to P. gingivalis fails to eliminate the pathogen

• Non-protective, low avidity anti-P. gingivalis antibodies that may

not be capable of effectively mediating a variety of immune
responses (complement activation, opsonisation/phagocytosis)

❖ Are B-lymphocyte/antibody responses protective against

periodontitis? (Could we vaccinate people?)
❖ Or do they cause periodontitis?
❖ Or could they be both protective and causative?
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 Summary

• Shift from a predominantly "T cell lesion" to a "B cell lesion" in the
progression from gingivitis to periodontitis

• Probably a shift from cell-mediated immunity (Th1) to humoral

immunity (Th2)

• IgG2 antibodies tend to predominate in periodontitis

▪ But likely to be ineffective in clearing the infection

• Want an IgG1 response (complement activation, neutralisation,

opsonisation)

• *Much of the (functional) data come from animal studies

• *Many conflicting human studies

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Interaction between Innate and Adaptive Immunity
in Periodontitis

IL-8

IL-8 Dysregulated
immune
response

Hajishengallis G, Trends in Immunology (2014) 35:3-11

 Immunology Lecture Series

Lecture 4: Introduction to Immunology (3:00 pm, 16 July 2018) ✓

Lecture 5: Innate Immunity (8:00 am, 18 July 2018) ✓

Lecture 7: Adaptive Immunity (3:00 pm, 23 July 2018) ✓

Lecture 8: Periodontal Inflammation (10:00 am, 25 July 2018) ✓

Lecture 12: Innate Immune Response in Periodontitis (9:00 am, 15 Aug 2018) ✓

Lecture 13: Adaptive Immune Response in Periodontitis (12:00 pm, 17 Aug 2018) ✓

Lecture 4: Immune Subversion by Bacterial Pathogens in Periodontitis (9:00 am, 10 Sep 2018)

Lecture 5: Vaccination in Disease Control (9:00 am, 19 Sep 2018)

Lecture 11: Revision (9:00 am, 2 Oct 2018)