GLOBULIN MIN DCH CU TRC V CHC NNG

TS. BS. Phan Ngc Tin

MC TIU Trnh by c v cc thuc tnh chung ca tt c cc globulin min dch M t c cu trc c bn ca globulin min dch Trnh by c mi lin quan n cu trc globulin min dch c chc nng Xc nh c vng siu bin ca globulin min dch v vng khun Xc nh c cc lp, loi globulin min dch M t c cc cu trc v tnh cht ca cc lp globulin min dch

T KHA

Globulin min dch (Immunoglobulin) Ha tr (Valence) Chui nng (Heavy chain) Chui nh (Light chain) Vng thay i (Variable region) Vng hng nh (Constant region) Vng ni (Hinge region) Domen (Domain) Vng siu bin (Hypervariable region) Vng khun (Framework region) Nhm v di nhm (Groups & subgroups) Fab & Fc, F(ab')2 Type v di type (Type & subtype) Lp v di lp (Class & subclass) Opsonin Chui J (J chain) Mnh tit (Secretory component)

I. NH NGHA
Globulin min dch (Ig): L phn t glycoprotein do tng bo sn xut chng li khng nguyn ( (ch hc nng kh khng th). ) in di huyt thanh

Hnh 1

II. CHC NNG CA GLOBULIN MIN DCH

A. Kt hp vi khng nguyn - Ig kt hp vi 1 hay nhiu loi khng nguyn. Mi Ig ch kt hp vi 1 quyt nh khng nguyn c hiu. u - Kt hp vi khng nguyn bo v c th l chc nng chnh ca khng th. - Ha tr ca khng th chnh l s lng quyt nh khng nguyn m khng th c th kt hp c. Ho tr ca khng th t nht l 2.

B. Chc nng tc ng: Hiu qu s sinh hc khng g c to ra a t s gn kt gi g a khng g nguyn guy v khng th m c to ra t tc ng th pht ca khng th. Khng phi tt c cc Ig u iu ho chc nng ny. Chc nng tc ng ny bao gm: 1. C nh b th: lm cho t bo b ly gii v phng thch c phn t c hot tnh sinh hc 2. Kt hp vi cc loi t bo: thc bo, bch cu, t bo mast, BC a baz ( c cc th th i vi Ig trn b mt) - Vic kt hp ny c th hot ha t bo thc hin chc nng ca chng. - Mt s Ig c kh nng gn kt i vi cc th th trn l nui ca mng nhau,v nhau v dn n mt vi Ig c kh nng xuyn qua mng nhau nhau. Kt qu l khng th p ng min dch c truyn sang thai nhi v tr s sinh

III.Cu trc c bn ca Ig: (hnh 2) Mc d cc Ig khc nhau th c cu trc khc nhau nhng u ging nhau v mt c bn. A. Chui nng, g, chui nh: Tt c cc Ig c cu trc chui gm 4 n v: gm 2 chui nng(5070kD) v 2 chui nh(23kD) B. Cu ni Disulfide: 1. Cu ni Disulfide gia: 2 chui nh v 2 chui nng c gn vi nhau bng cu ni Disulfide v lin kt cng ho tr . S lng cu ni di lfid th disulfide thay i t trong cc phn h t globulin l b li mi in dch h kh khc nhau. h 2. Cu ni Disulfide bn trong chui: Trong mi chui polypeptide cng c lin kt disulfide.

C. Vng bin i v vng hng nh: C hai chui nng g v nh u c th c chia thnh hai khu vc trn c s bin i trong cc chui axit amino. 1. Chui nh - VL (110 axit amin) v CL (110 axit amin) 2. Chui nng - VH (110 axit amin) v CH (330-440 axit amin) D. Vng bn l y l vng m nhnh h h ca phn h t kh khng th c dng mt Y, Y c gi l vng bn l bi v c tnh linh hot trong phn t ti im ny.

E. Vng (Domains) Hnh nh ba chiu ca Ig cho thy Ig khng c thng, ng cu trc c xp vo khu vc hnh cu, trong c mt mi lin kt disulfua trong chui. Nhng vng ny c gi l domain 1. Tn domain Chui nh - VL v CL 2. Tn domain Chui nng - VH, CH1 - CH3 (CH4) F. Oligosaccharides Carbohydrates c gn vo domain CH2 trong hu ht cc globulin min dch. Tuy nhin, trong mt s trng hp carbohydrates c th c gn ti cc v tr khc.

H 2 Cu trc c bn ca immunoglobulin

IV. Cu trc vng bin i A. Vng gs siu u b bin ( (HVR) ) hay ay vng g quyt nh b tc (CDR) (C ) - So snh cc trnh t acid amin trong cc vng bin i ca cc Ig cho thy hu ht cc bin i nm trong ba khu vc c gi l cc cc vng siu bin hay vng gn quyt nh b tc nh minh ha trong hnh 3. - Khng th c c hiu khc nhau (khc nhau ti v tr kt hp ) th c cc CDR khc nhau, cn cc khng th c tnh c hiu th c CDR ging nhau. h - CDR c chui nng ln chui nh B. Vng khung: L vng nm gia cc vng quyt nh b tc (hnh 3) 3).

H 3 Cu trc vng g bin i: Vng g siu bin i, Vng g khung g

V. Cc mnh: mi quan h cu trc/chc nng Mnh Ig c sn xut t vic phn gii protein A. Fab: phn hu vi papain lm ph v cc phn t globulin min dch trong vng bn l trc cu ni H-H disulfide lin chui. iu ny dn n s h h th hnh thnh h ca hai h i mnh h gi ing ht nhau h c ch ha cc chu h i nh h v VH v CH1 ca chui nng. B Fc: phn hu vi papain cng sn xut mt on c cha phn cn li ca B. hai chui nng, mi chui c cha mt CH2 v CH3. on ny c gi l Fc v c th kt tinh. C. F(ab')2: Ig nu c x l bng pepsin s phn ct chui nng sau cu ni disulfide thnh 2 mnh. Mt mnh c cha 2 v tr gn kt khng nguyn gi l F(ab ( ') ) 2 ( hoa tr hai). ) Mnh Fc cn li ca p phn t c p pepsin p phn gii thnh cc peptide.

H 4 Immunoglobulin: Cu trc v chc nng

H2

H5: Khng th x l bng men papain

H5: Khng th x l bng men pepsin

VI. CC LP, DI LP, TYPE V DI TYPE GLOBULIN MIN DCH NGI A. Phn lp Ig: Ig I c chia hi lm l 5 lp da trn t s khc kh bit trnh t h t axit it amin i t trong vng hng nh ca chui nng. Tt c cc globulin min dch trong mt lp nht nh s c nhng vng hng nh tng t nhau trong chui nng. ng Nhng khc bit ny c th c pht hin vi cc nghin cu huyt thanh hc: 1. IgG chui nng Gamma 1 2. IgM - chui nng Mu 3. IgA - chui nng Alpha 4. IgD g - chui nng g Delta 5. IgE - chui nng Epsilon

B. Cc di lp Ig: Cc lp ca Ig c chia thnh cc di lp da trn s khc bit nh trong trnh t axit amin trong vng hng nh trong chui nng. Tt c cc Ig trong mt di lp ny s c nhiu s ging nhau trong trnh t chui acid amin. Nhng khc bit ny cng c pht hin bng huyt thanh hc. 1 Cc 1. C d i lp ca IgG I G a) IgG1 - chui nng Gamma 1 b) IgG2 - chui nng Gamma 2 c) IgG3 - chui nng Gamma 3 d) IgG4 - chui nng Gamma 4 2 Cc di lp ca IgA 2. a) IgA1 - Alpha 1 chui nng b) IgA2 - Alpha 2 chui nng

C. Cc loi chui nh Globulin min dch cng c th c phn loi theo cc loi chui nh. Loi chui nh c da trn s khc bit trong trnh t axit amin trong vng hng nh. Nhng khc bit ny c pht hin bng huyt thanh hc. C hai loi chui nh 1. chui nh Kappa 2. chui nh Lambda E. Danh php: Cc globulin min dch c t tn da trn lp hoc phn lp ca chui nng v loi hoc di loi ca chui nh. F. Tnh a dng Globulin min dch l mt phn t a dng v chng bao gm cc lp v cc phn lp khc nhau, mi trong s li c cc loi chui nh Ngoi ra, ra cc phn t globulin min dch khc nhau c th c tnh cht gn kt khng nguyn khc nhau c quy nh bi cc vng VH v VL khc nhau.

VII. CU TRC V MT S C TNH CA LP IG CC PHN LP A. IgG gG 1. Cu trc: Tt c IgG u c cu trc n phn (Ig 7S). Cc phn lp khc nhau v s lng cc lin kt disulfide v chiu di ca vng bn l. 2. Tnh cht L I Ig a nng nh ht bi v n c kh nng thc hin tt c cc ch hc nng ca cc phn t globulin min dch. a) IgG l Ig chnh trong huyt thanh chim 75% b) IgG l Ig ch yu ngoi mch mu c) C th truyn qua nhau thai - IgG l lp duy nht ca Ig i qua nhau thai: nh th th trn t bo nhau thai dnh cho vng Fc ca IgG. (IgG2 khng qua c). c)

H6: IgG

d) Hot ha b th - Khng phi tt c cc phn lp u hot ha b th nh nhau; IgG4 khng hot ha b th. e) Kt hp vi cc t bo - Cc i thc bo, bch cu n nhn, PMN v mt s t bo lympho c cc th th Fc. (IgG2 v IgG4 khng lin kt vi th th Fc). Kt qu ca vic lin kt vi cc th th Fc trn PMN, bch cu n nhn v i thc bo l cc t bo c th tip nhn cc khng nguyn tt hn. Cc khng th gip t bo thc bo thc bo khng nguyn tt hn. Thut ng Opsonin c s dng m t tnh tng cng thc bo. IgG l mt opsonin tt. S kt hp ca IgG vi th th Fc trn cc loi t bo khc gip kch hot cc chc nng khc nhau

H7: Opsonisation

B. IgM 1.Cu trc - IgM I M thng th th ng l mt pentamer t (19S), (19S) nh hng n cng c th tn ti dng monomer. Trong dng pentamer tt c cc chui nng ging nhau v tt c cc chui nh ging nhau. Nh vy, ha tr l thuyt l 10. 10 - IgM c thm mt min trn chui Mu (CH4) v n c mt cu ha tr S-S c gi l chui J. 2. Tnh cht a) ) IgM g l Ig g c s l ng g nhiu th ba trong g huy yt thanh b) IgM l Ig u tin c to thnh bo thai v Ig u tin c sn xut khi c kch thch bi khng nguyn. c) IgM hot ho b th mnh. V vy, cc khng th IgM l rt hiu qu trong vic ly gii ca vi sinh vt

H8: IgM

d) IgM cng l mt cht kt dnh tt. V vy, cc khng th IgM rt tt trong vic kt t vi sinh s vt c t th lo oi b. e) IgM lin kt vi mt s t bo thng qua th th Fc. f) Ig trn b mt t bo B IgM trn b mt l mt monomer khng c chui J nhng n c thm 20 acid amin ti on cui C gn n vo mng t bo (H 9). I M b mt t bo IgM b c ch hc nng nh h mt th th cho h khng kh nguyn t trn t bo B. IgM b mt c kt hp khng ha tr vi hai loi protein trong mng ca t bo B c gi l Ig-alpha v Ig-beta. Nhng protein b sung hot ng nh phn t truyn tn hiu khi t bo cht ca cc phn t Ig qu ngn truyn mt tn hiu. Trc khi mt tn hiu c th c truyn i bi Ig-alpha v Ig-beta i hi cc Ig b mt phi lin kt vi khng nguyn. Trong trng hp khng nguyn khng ph thuc tuyn c, c ch cn s gn kt gia khng nguyn v Ig b mt l kch hot cc t bo B sn xut khng th. Tuy nhin, i vi cc khng nguyn ph thuc tuyn c, cn g p ca cc t bo T h tr trc khi cc t bo B c kch hot. c s tr gip

H9: IgM b mt t bo

H10: IgM b mt t bo v cc phn t truyn tn hiu

C. IgA 1. Cu trc - Trong huyt thanh IgA l mt monomer nhng trong cc dch tit Ig l mt dimer (Hnh 11) nh s kt ni ca mt chui J. - Ngoi ra c mt protein lin kt c gi l mnh tit hoc mnh T; sIgA l globulin l b li mi in dch h 11s. 11 IgA I A c to thnh th h trong t cc tng bo, b nh hng mnh tit c to ra trong cc t bo biu m v c gn vo cc IgA khi n i vo cc dch tit (hnh 12). Cc mnh tit gip IgA c vn chuyn qua nim mc v cng bo v IgA trong cc dch tit. t 2. Tnh cht: a) IgA l Ig ph bin th 2 trong huyt thanh. thanh b) IgA l c nhiu nht trong cc dch tit nh nc mt, nc bt, sa non, cht nhy. IgA c vai tr quan trng trong min dch ti ch (nim mc) ) c) Thng thng IgA khng hot ha b th, tr khi b kt tp. d) IgA c th lin kt vi t bo PMN v t bo lympho.

H11: IgA

D. IgD 1. Cu trc I D tn ti nh IgD h l mt monomer. 2. Tnh cht a) IgD c hm lng rt thp trong huyt thanh, thanh vai tr ca n cha c chc chn b) IgD ch yu c tm thy trn b mt t bo B c chc nng nh l mt th th cho khng nguyn nguyn. IgD trn b mt ca cc t bo B c thm axit amin ti on cui C gip bm mng t bo. N cng lin kt vi chui Ig-alpha v Ig-beta. c) ) IgD g khng g hot ha b th.

H12: IgD

E. IgE 1. Cu t trc c IgE dng monomer, v c thm mt vng trong vng hng nh. 2. Tnh cht a) IgE nng thp nht trong huyt thanh v n lin kt rt cht ch vi cc th th Fc trn cc bch cu a baz v cc t bo mast, ngay c trc khi tng tc vi khng nguyn b)Tham gia trong phn ng d ng IgE gn kt vi cc cht gy d ng v lm phng thch cc trung gian dc l khc nhau gy nn cc triu chng d ng trn lm sng sng. c) IgE cng ng vai tr quan trng trong cc bnh k sinh trng. IgE huyt thanh tng cao trong cc bnh k sinh trng trng, v vy o nng IgE huyt thanh hu ch trong chn on nhim k sinh trng. Bch cu i toan c th th Fc ca IgE v s gn kt gia bch cu i toan vi IgE gip tiu dit k g sinh trng. d) IgE khng hot ha b th.

H13: IgE

Isotypes Allotypes and Idiotypes Isotypes,

I.

ISOTYPES

A. nh ngha: l quyt nh khng nguyn c trng cho cc lp v di lp ca chui nng v cc loi v di loi ca chui nh. B. V tr: isotype chui nng c tm thy trn phn Fc ca vng hng nh, i t isotype chu h i nh h c tm t thy trong t vng hng nh. h (Hnh (H h 14) C. Hin din: isotype c tm thy trong tt c nhng c th bnh thng ca loi Tin t Iso loi. Iso- c ngha l ging nhau trong tt c cc thnh vin ca loi. Mt s c th suy gim min dch c th thiu mt hoc nhiu isotypes nhng c th bnh thng th c tt c cc isotypes. D. Vai tr: c dng trong nh lng cc lp v di lp Ig cc bnh khc nhau, trong cc c tnh ca bnh bch cu t bo B v trong chn on cc bnh suy y gi g m min dch.

Hnh 14: Isotype

II. ALLOTYPES A. nh ngh g a - Allotype l quyt nh khng nguyn c trng theo cc dng allen ca Ig. - Allotype i din cho s khc bit nh trong trnh t axit amin ca chui nng hoc chui nh ca cc c th khc nhau. Ngay c mt s khc bit mt amino acid c th lm pht sinh mt yu t quyt nh allotype, nhiu trng hp thay i vi amino acid. - S khc bit Allotype c pht hin bng cch s dng khng th chng li yu t quyt nh allotypic. Nhng khng th ny c th c to ra bng cch tim Ig cng loi c allotype khc lm KN gy min dch ch. B. V tr ngi i, allotype nm vng hng nh ca chui nng v chui nh (Hnh 15).

C. Occurrence - Allotypes c tm thy trong cc c th ca mt loi, nhng khng phi l tt c - Tin t Allo- c ngha l khc nhau trong nhng c th ca mt loi. D. Allotypes Ig ngi - Danh D h php h - allotypes ll t c t tn t da vo chu h i nng hay h chu h i nh h m n nm. - Mt allotype chui nng Gamma1 l: G1m. - Mt allotype trn mt chui nh Kappa l: Km (1). (1) (Bng 1).

Hnh 15: Allotype

Bng 1: Allotypes ngi

Chuoi IgG1

Domain CH1 CH1 CH1

Allotype G1m(f) = (3) G1m(z) = (17) G1m(a) = (1)

Amino Acid Arg Lys Arg, Asp, Glu, Leu

V tr 214

355-358

light g chain CL CL

Km(1) ( ) Km(3)

Val, Leu Ala, Val

153, 191 153,191

III. IDIOTYPES (Id) A. nh ngh g a - Yu t quyt nh khng nguyn duy nht biu hin trn cc phn t khng th c th hoc trn cc phn t c tnh c hiu ging nhau. - c hiu ging nhau c ngha l tt c cc phn t khng th c cng mt vng siu bin ng nht ging nhau - Yu t quyt nh khng nguyn c to ra bi vng kt hp (vi khng nguyn) ca mt khng th c gi l idiotypes v cc khng th to ra do d idiotype idi t c gi l kh khng th anti-Id. ti Id Idiotypes l yu t quyt nh khng nguyn to ra bi cc vng siu bin ca mt khng th v khng th chng idiotypic s trc tip chng li nhng vng siu bin . B. V tr - Idiotypes nm trn mnh Fab ca cc phn t Ig Ig. C th, chng nm ti hoc gn vng siu bin ca chui nng v chui nh. Trong nhiu trng hp cc quyt nh khng nguyn (idiotype) c th bao gm mt vi phn g siu bin. Idiotypes yp thng g l yu t quy q yt nh to ra bi cn li gn vng HVR ca c hai chui nng v chui nh HVR mc d i khi ch chui nng hoc chui nh n c th hin idiotype.

C. Vai tr 1. Vng V - Idiotype l mt du hiu hu ch xc nh vng bin i. 2. iu ha p ng min dch - c bng chng cho thy p ng min dch c th c ieu ha bi khng th anti-Id chng li Id ca bn thn. 3. Vc xin - Trong mt s trng hp khng th anti-Id kch thch t bo B to khng th v do chng c th c s dng nh mt loi vc xin. Cch tip cn ny ang c c gng chng nga chng li cc tc nhn gy bnh rt nguy him m khng th c s dng mt cch an ton nh vcxin 4. iu tr cc khi u t bo B Khng th Anti-Id chng li mt idiotype trn cc t bo B c tnh c th c s dng tiu dit cc t bo. bo C ch l do b th hoc v cc phn t gy c c gn vo khng th.

Hnh 16: Idiotype

IMMUNOGLOBULINS- ANTIGEN-ANTIBODY REACTIONS PHN NG KHNG NGUYN-KHNG TH

I.

BN CHT CA PHN NG KHNG NGUYN KHNG TH

A. Khi nim kha v cha kha: Vng kt hp ca mt khng th nm phn Fab ca phn t v c to thnh t cc bin i v vng siu bin ca chui nng v chui nh. Nghin cu v s tng tc gia khng nguyn v khng th cho thy rng cc quyt nh khng nguyn nm trong cu trc dng hm ch c hnh thnh bi cc vng kt hp ca khng th (hnh 16). Nh vy, khi nim v phn ng Ag-Ab c ngha l s n khp ca mt cha kha (Ag) vi mt kha (Ab). B Cu ni kh B. khng h ha t tr: Lin kt gi Ag trong vng kt hp khng th l tt c cc lin kt khng ha tr trong t nhin, bao gm y , - Lin kt hydro, - Lin kt tnh in - Lc Van der Waals - Lin kt k nc Cc lin kt ny m bo rng Ag s gn cht ch vo Ab. Ab C. Thun nghch g Ag-Ab g xy ra nh cc lin kt khng g ha tr, nh chng g cng g c tnh Phn ng thun nghch.

Hnh 16

II. AFFINITY AND AVIDITY A. Affinity y - i lc khng th l lc phn ng gia mt quyt nh khng nguyn v mt vng kt hp trn khng th - i lc l hng s cn bng m t phn ng Ag Ag-Ab. Ab Hu ht cc khng th c i lc cao vi khng nguyn. B. Avidity y - Avidity l tng ca cc lc s lin kt ni mt khng nguyn vi nhiu quyt nh khng nguyn v khng th a ha tr. - i lc cp n lc s lin kt s gia mt quyt nh khng nguyn vi vng kt ni ca mt khng th, trong khi Avidity cp n lc tng th gn kt gia khng nguyn v khng th a ha tr. - Avidity b nh hng bi c ha tr ca khng th v ha tr ca khng nguyn. Avidity l nhiu hn tng ca cc i lc

Figure 2

Hnh 17: Affinity

III. C HiU V PHN NG CHO A c hiu A. - L mt vng gn vi KN ca khng th ch phn ng vi mt quyt nh khng nguyn. - Thng thng c s c hiu cao trong phn ng KN KN-KT KT - Khng th c th phn bit s khc bit v: 1) Cu trc c bn ca mt khng nguyn 2) Cc hnh thc ng phn ca mt khng nguyn 3) Cu trc cp hai v cp ba ca mt khng nguyn. B. Phn ng cho: Kh nng ca mt vng kt hp khng th phn ng vi nhiu hn mt quyt nh khng nguyn

Figure 4

Hnh 18: Avidity v phn ng cho

Ti liu Roitt, Brostoff, Male. Immunology 6th Edition, Mosby, 2002

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