Immune evasion of Neisseria gonorrhoeae 淋球菌的免疫逃避...

Immune evasion of Neisseria gonorrhoeae

淋球菌的免疫逃避（淋球菌利用（剥削）宿主的先天免疫）

Tie Chen, M.D. ProfessorTongji Hospital

Tongji Medical CollegeHuazhong University of Science and Technology

Visiting Associate ProfessorIndiana University School of Medicine

华中科技大学同济医学院附属同济医院陈铁

N. gonorrhoeae, (gonococci, GC, the cause of gonorrhea)

奈氏淋球菌

Gonococci Invade Host Cells 淋球菌侵入属主细胞

Some Facts about Neisseria gonorrhoeae (gonococci, GC)

1. Genital Infection: gonorrhea

2. Cervical- pelvicinflammatory disease (PID)

3. Pharyngeal, Rectal Infection, eye and systemic spread

4. Other noted names:Clap, Flowing seeds, Happydisease and Mother of STD

6. It is an ancient disease. 古老的疾病

5. 78 million cases each year.

7. No animal model 没有动物模型8. Does not generate protective immune response不产生保护性免疫9. Ability to facilitate HIV infection?能促进 HIV 的感染

GC Forms Opaque Colonies

Opa Protein Expression

Interaction of Neisseria gonorrhoeae with Host Cells

1. Initial adherence

2. Tight adherence

3. Invasion/uptake

4. Transcytosis

奈氏淋球菌与宿主细胞的相互作用

初期粘附

紧密粘附

侵袭 / 吞噬

胞吞转运

1. Neisseria gonorrhoeae inhibits host responses.证明 CEA(CD66, carcinoembryonic antigen)抗原是奈氏淋球菌的受体

R.J. Belland*, T. Chen, J. Swanson and S.H. Fischer. 1992. Mol. Microbiol. 6:1729-1737.

T. Chen, R. Belland, J. Wilson, and J. Swanson*. 1995. J. Exp. Med. 182:511-517.

T. Chen, J. Swanson, J. Wilson, and R. Belland*. 1995. Infect. Immun. 63:1790-1795.

T. Chen* and E. Gotschlich. 1996. Proc. Natl. Acad. Sci. USA. 93:14851-14856.

T. Chen*, F. Grunert, A. Medina-Marino and E. C. Gotschlich. 1997. J. Exp. Med. 185:1557-1564.

CEA (CEACAM, CD66, carcinoembryonic antigen) Ag Serve as Receptors for Opa+ GC

.

s

s

s

s

s

s

s

s

s

s

s

s

s

s

N

s

s

s

s

s

s

s

sN

s

s

s

s

s

sN

s

s

s

s

s

sN

s

sN

s

s

s

s

s

s

s

sN

BGPa

CGM6

CGM1a

NCA

CEA

PSG1

Plasmic membrane

Cytoplasmic domain

E x t r a c e l l u l a r D o m a i n

GPI linkageB1

A1

A1A1

A1

B1B2

B1

B1

A1

B2A2

A2

A2 B3

A3

CEA (CD66) Family Antigens

(CD66a)

(CD66b) (CD66c)

(CD66d)

(CD66e)

(CD66f)CEACAM1, (BGPa CD66a)

CEACAM3, (CGM1a,CD66d)

MembraneExtracellular

Cytoplasmic

CEACAM6CEACAM8

ITAMITIM ActivationInhibition

CEA(CD66) 抗原是奈氏淋球菌的受体

阳阴

PP

B Cellreceptor

PTKs

INTERNALIZATION

ITAM

ss

PP

CEACAM3 (CD66d)

ss

ss

ss

ss

PP

SHP-1SHIP

SHP-2

ITIM

CEACAM1 (CD66a)

Activation Versus Inhibition

PROLIFERATION

GC

ANTIBODY PRODUCTION

GC

PLC

Ca2+

Ca2+

P

FcRIIB

APOPTOSIS

Extracellular

Intracellular

?

BTK

T. Chen*, W. Zimmermann, J. Parker, I. Chen, A. Maeda and S. Bolland. 2001 J. Leuko. Biol. 70:335-340.T. Chen*, S. Bolland, I. Chen, J. Parker, M. Pantelic, F. Grunert, and W. Zimmermann. 2001. J. Biol. Chem. 276:17413-17419.

阳性和阴性的信号传递

抑制 抗体产生

Potential ITAM or ITIM Motif in the Cytoplasmic Domain Of BGPa and CGM1a

Immunoreceptor Tyrosine-based Activation ( or Inhibition) Motif

ITAM D/ExxxxxxxD/ExxYxxLxxxxxxxYxxL/I ITIM V/IxYxxL

BGPa DPPNKMNEVTYSTLNFEAQQPTQPTSASPSLTATEIIYSEV (CD66a) (may contain ITAM)

CGM1a PLPNPRTAASIYEELLKHDTNIYCRMDHKAVAS (CD66d) (may contain ITAM)

Extracellular Cytoplasmic

Y196F

Y207F

Y207FY196F

TM

CGM1a

CGM1a-Y207F

CGM1a-Y196F/Y207F

CGM1a-Y196F

Construction of CGM1a (CD66d)-Tyrosine Mutants

ITAM D/ExxxxxxxD/ExxYxxLxxxxxxxYxxL/I CGM1a PLPNPRTAASIYEELLKHDTNIYCRMDHKAVAS

| |Y207 Y196

CGM1a Contains a Functional ITAM

OpaI GC

1.9 100 200 300 400 500 604.2

25.4

40

50

60

70

80

90

100

109.4

S

[CA++]

DT40-CGM1a

CGM1a-Y196F

CGM1a-Y207F

DT40 B-Cell

CGM1a-Y196F/Y207F

Time, seconds

[Ca++]

PP

B Cellreceptor

PTKs

INTERNALIZATION

ITAM

ss

PP

CEACAM3 (CD66d)

ss

ss

ss

ss

PP

SHP-1SHIP

SHP-2

ITIM

CEACAM1 (CD66a)

Activation Versus Inhibition

PROLIFERATION

GC

ANTIBODY PRODUCTION

GC

PLC

Ca2+

Ca2+

P

FcRIIB

APOPTOSIS

Extracellular

Intracellular

?

BTK

T. Chen*, W. Zimmermann, J. Parker, I. Chen, A. Maeda and S. Bolland. 2001 J. Leuko. Biol. 70:335-340.T. Chen*, S. Bolland, I. Chen, J. Parker, M. Pantelic, F. Grunert, and W. Zimmermann. 2001. J. Biol. Chem. 276:17413-17419.

阳性和阴性的信号传递

抑制 抗体产生

Assay of Inhibition by ITIM

FcRIIB-BGPa

Primary antibody

Secondary antibody IgM

PP

B cellreceptor (BCR)

IT A M PP

IT IM

-BCR MAb (A)(stimulates ITAM)

B CellReceptor(BCR)

-Mouse Ab (I)(stimulates ITIM)

FcRIIB-BGPa

FcRIIB-BGP (WT)

FcRIIB-BGP (Y459F)

FcRIIB-BGP (Y486F)

FcRIIB-BGP (Y459-486F)

Extracellular Cytoplasmic

Y459F

Y486F

Y486FY459F

Flow CytometryTM

FcRIIB-BGPa Chimeric Molecules

FcRIIB BGPa

1.9 50 100 150 200 250 302.13.2

20

40

60

80

100

120

146.6

S

[CA++]

1.9 50 100 150 200 250 302.1

-7.1

50

100

150

191.6

S

[CA++]

1.9 50 100 150 200 250 302.1

0.0

20

40

60

80

100

120

136.4

S

[CA++]

1.9 50 100 150 200 250 304.0

0.4

20

40

60

80

100

120

140

160173.7

S

[CA++]

Y459 is Essential for Inhibiting Ca++ Influx

FcRIIB-BGPa FcRIIB-BGPa-Y459F

FcRIIB-BGPa-Y486F FcRIIB-BGPa-Y459/486F

A A

AA

I I

II

Hypothesis: The interactions of GC with CEACAM on

neutrophils （白血球） or B cells shape host responses.

.

0

200

400

600

800

1000

1200

0

20

40

60

80

100

ControlOpa- GCOpaI GC

No.

bac

teri

a/w

ell 1

0-2

DT40-ce

ll

Y207F

Y196F

CEACAM1

0

20

40

60

80

100

ControlOpa- GCOpaIGC

phagocytosis

cell death

CEACAM3 (CD66d) Controls Cell Death.

阳性信号传递导致细胞死亡

Hu

man

Ig

(µg/

ml)

stim

ulator

s + O

pa- G

C

stim

ulator

s + O

paI G

C0.00

0.10

0.20

0.30

0.40

0.50

0.60

no stim

ulator

s

stim

ulator

s

GC Inhibits Antibody Production by Primary Human B Cells

阴性的信号传递抑制 抗体产生

Neisseria gonorrhoeae inhibits host responses of CEACAM-expressing host cells through either activating or inhibitory signal transduction.

Summary:

奈氏淋球菌借助了人体的阴性 / 阳性的信号转导过程，抑制了人体的免疫反应