I- Niirn. Sci. Cul~n. Sli  LaliLa 1951 5 (1) : 

Sali~ary Xn3mnun~glob~iins annd L,actofeeurisr is  Dersttal Caries 

S- DISSANAYAKE, L- P. SAMARAN A YAKE 

Faculty of k / L r ! i c i n d ,  7 :n i~-?r j l !~  o i  ['rrri&:,ni7c, P r ~ g r ! i r ~ i : ; : ,  S7i li n2 .. 

)'.N r> 

li. M. BENNETT 
Il~liartli ,s:lt  D; Mei!ic:;le. ! 'ni<:e:-sity O{ Chiingc, C,!l icClgo,  i!/ili,,is, i:. S. ,-1 

(Date of ::c~.ipt ; 07 bioi~ll  1080) 

(Date o! ,::cceptm:i- : .<.j J n n r : a i )  i gBr )  

Abstract: :'Ile I c r ~ c ! ~  of seru::-: JpG, [PA, saiiv;,~'; IgA :.n-! salivary lactcifcrrin wcrr: 
dcl.ernlin& high unii. io\v cnrics r u b j r c t s .  The !c.se!s of i n ~ r i ~ u r r ~ ~ ~ l ~ ~ ~ - u ! i r r  i~rtd 
!:!ctt;fcrrilr m2l-c fL:und ti) be e;ex~ntci: in l i i ~ h  car-irs ~ubjccrs .  

Nrsmcrous studfes have shown ~ a r i a t i o n s  i c ;  serum a d  salivary immunoglobulins 

i n  dental caries, 4 . 7 3 " s 1 5 7  some reports suggesting a prorectivc function for salivary 

imrr~irr-\oglobulins. "4.5.9-i:i. 

Lactrjlcrrin, t he  iron a n d  cospcr bindjng protein in  ~ e c r e t i o n s , ~  is present in 

saliva and has bttm described as a bacteriostatic: agent.' As bacteria play asigr-ai- 

iicant r o l e  in  th:: csrablishmrnt :!nd progress o!: dental  c;~ries, ir is imprp~tn r~ t  re, 
knnu, wherket- Iactoferrin levels show a n y  relztiort to the scveritv of ciiseasc. 

in this cornmunicariun, we summarise o u r  I?relimi;~ary dara o n  serum irnxnu- 

r~oglob~i l ins ,  salivary immunoglobulins and lai toferrin in denra! caries in man. 

2. Materials and Methods 

2.1, Subjects  

The series consizrei! of 122 patients selected from those a l terding an POtlt 

Paeietlcs' clinic, at the Genera! iiospiral, Ka.ndy and 28 caries .free Cirst year 

lnedica1 stuciellts f rom Peradeni)-a. The  medical s t r td i r~ts  werc freshmen an6 
were studied within one nlorarh of rheir arrival in rhe campus. 'Their family 
income was used ro assess t l>e socio-t'conomic backgrourlds arid 28  students of 

sacio-economic s ta tus  ce>mpara'o!e to that of the test group were selected. All 
subjects were oxamined by the  same clcnta! surgeon anci those patients wit11 
evidence uf othcr oral diseases, ejpecialiy gingivitis, wcrr excluded. 'The dcgrec 

wS denta l  caries was measured hi: [-he  umber of decayed, missing a n d  filjed teeth 

[DMF Index). 'The subjects were classified accorcling to ~11c DMF lndcx, 



2 .2, Collection of saliva a d  sexam, 

About 5 to 10 1x1 of saliva v,:ere collected und:r ra i -a f f i i~  sri;:-.ulatioc f rcm i . a c l ~  

subjcct. The time of: saliva col!ecricn mas bctxvecn 10 ::In and 12 nocm. The 
saliva samples wtre cIarifieci by centri~ugaticn (2C.0 g) and scored n t  -2,CI"C. Blocd 
%as coilected into Hacrnatocrit tubes by  :rhefir?ger prick rneti~od, serum s~,p:~racetj 

~ ~ n d  stored at  -20" C. 

l.Jnconccntrateci saliva was used in al l  dctcn-:lin:itions SigA Ic~:cIs k.1 salivz 

vzcrc dctcrmined by Single Radial immu;lodiiiusicn (SRIII)) in I.lij:," itga;.o~t. 

using a rabbit ar:tiser u ~ n  (sFecilic) i.0 Luman S IgA ( ?Jar-dic l i~ i in~~i io lo~: ic :~I  

Laborntorics; clae Net-!ler!a~~ds). Thc 1 1  Igi\  stacdar-ii  u.:?s 1:rr.par-rd in .ii>is 

laboratory. Ser"jm TgG asid laA icvcis xv(:ic d~tci-rni~zc~i  by SZlC ill 256 Ag:~;ost: 

usi!~::, cliiss specific zritisera (Nordic I r n r ~ ~ i a o l o ~ i c d  2,:tb:)r2to:-icr;j u : i t?~  rcirrcr~ce 

to t b e ' ~ i - 1 0  Stn~zdard Se:um 6'119'3. 

IiS IgA (as determined by gel filtration on. :i cnlibrilticl Scr>l-~:ic!cs C;-ZClj 
c o l u ~ ~ n )  was isijlatcd f rom hurK~a i l  colostrum. ( ; r ; i ~ ! ~ ; ~ - - g ; l ~ L u l i i ~ : ;  in Piui\l;i~l 
coli>strurn were isolatcd by r?peated precipitation with 3.3% a : i imu!~i~im r:riiph:iic 
in  phcspl~atc buffer, pH 7.4. 11s I g A  was  olotxir~ec! b y  ;rcl filtration in t11c c:!ii 
'u~-a-i-eJ Sc.ilihai'ie,; G--2CO column 2nd f i ? i ~ l : ~ r  pu!- i ( icdhj~ afliriity i i l roi l inr~; : , , r ;?~I~~ 
c?si CNi3r--Sepiiarosl- 4C i i~sol~~bi l izcd i-ahbit- anti.- l ~ ~ i r i - i o ~  SI!;A ( f r o ~ i ~  Nordii: 
imn\.cinslosical L;i%orz:ories, tlir: N c ~ h e i l ~ n d : , ) .  '1 l ~ c  ! IS i::,4 thus  pi-i.puretl 
showed one precipitation line o n  irnmr:noeTec.trc~pl~oi'i'sis a~~ai i ls t  r:il->bir anti- 
h u i l l n ~ l  Fab and rabbit a n t i - l ~ u ~ n ~ ~ n  S igA  (Nordic Immuno!u;;ii:li L:!l:i~ra~orics]. 
C;;lculaiion of standard c o n c c n ~ r a i i o ~ ; ~  sins baticci oil thc nssuir:cd cx~ii!c;:ion 
cotlfficienr for SIgA of F: :,h':m 1 oi, =-: 13. i i . ' V 7 h i s  p;cpa;-:~:ion \xT,ias 3lt.o c31i 

bratcd against the WHO Standard Srrui:~ 6'ii97 ~ ~ i t h  a corrci-tioil factor  applied 
for  differences in rates of diffusion." 

Dztcrxnir>ation cf salivary lnctcierritl i cvc ls  was p f r f~ rmcd  by su!id ?3Jih)- 

~ ! I ~ ~ w I I I ) ; ~ s s ~ ~  exactly 2s desci.ibic! previoujly.~ 

The number of subiects in fach caries grc,up, .ihc nlran S. D, of their :12;ci 

UMF Index, Ieve!s of seru1a.l IgG, iF;A, salivsry ipA,  a n d  ':acca~c.;:in arc shou~n j11 

'Table !. In Tsb!e 2 are slliocvrx :he statiztic.1 correlations (by linear rcgrc;sii-.(-I 

anzlysis) bctwecn salivary !gA, lactof'errin and t h e  DMF I:ldex, 



\)rjhen the  U k F  Ii-\des was greater than 15, the mean serum IgG i r v e \ s  w c ~ c  
sigr:ificantly grca:er th;:n t h a t  of the control  group w i th  a DMF :Indcx=:O. T!?e 
salivnl-y lgA ai>d 1nc:ofzrrin !eire!s sllov~ed a gradunl increase wirh the iccreasc 
ur' DMF Index. In high czries si lhjccts {DMF l n i e x  - 11-15 a n d  15) the 
mcnr! Icvel:; of ~a! i -L '~r~ 19.4 ail(! lac:olcrriil cvevc sigi?ificantly higher than t l ; ~  
corrcspor~ding i-aluss {or the  concro! group with DMF %ndcx - 0. 
-7 ~ . ~ ~ b ! c  3 - 'f!,e T'iv/l~zil, -:- 3. D. of &e, D?../lF' Xndex Scrum Serum IgA, 

- 
.7 

hn! i .~n r \ i  !gh ari:! Sal ivar .~  L3.cioTei-rin. 

. ~ ~ .-- -~ -- - - -  - - ,  . .- 

'" : d.iffrrc:lie il.<:rn DMF 1nd . e~  :- O , :isailiir:nt at 20% level, 
:; .i : dIi;crci:cc irt : i~i  OMF Incicu - I? , big;lifLcanr :xt OO.Cf.% ?etel. 

Su!~jcct C r u u p  Salivary 1gA and !..>ctofe;riti acd. l.zctufcrl.in ar?d 
9 M F  h a e x  DMF Index I g h  

c o r r s ' n r i o ~ ~  currelacion corrclar ion . < l>b!F l!xcl?x = J-3 , ; .--0.163 , NS ,: 0-063 N S  ,, 0.062 NS 

DMF Inii.::< - 6 -10 ,, 2-348 ,? - 0.01 , , 0.036 NS , , 0.035 NS 

NS : i!oe significant. 



Subjects with a DMF !ndex greatcr than 15  exhibited a significantly eEevnted 
lrvel of serum IgG: thz group crith a DMF Indes = I1 :o 15 (but not s?~orre 15) 
exhibited a significant elevation of se rum I g h .  These findings a re  in general. 
agreement. with orher reports."' I n  corrtrast, salivary IgA. a n d  i;rctofcrrir~ 
demonstrated. a g r a d u ~ l  increase :vitll the increase of the DMF Index. Thestan-- 
dard deviarigns were so high, ho~vevrr ,  that only in thc grous with DMF Index 
greater t h a ~ i  15 (grearer than 10 for :actoferrin) was the difference fro-ns) tkc 
conerc>l group silt'ficient to be statistically significanr. To our kncwlcdge, this 
is the tirst report on elcvated levels of lacrocerrin in ifenti1 caries. 

The corrcentratioil of salivary proteins arc kno-r~n t o  be a lfectccl by the  rate 
of: saliva ~ec re t i on .~ .  '' Hos~cver,  it is not known to what estcnt t.hes.2 cxpcxi- 
mentally determined differences -refiest true diiierences irr [he glandular- secrr- 
t ion of salivary prcttins. In  the  present investigation, M ~ C  \\rere nor zble to 
il-reasure t h e  rates of saliva secretiorl and r:hereiore, it was not pcrssihJr to correct 
for differences in rates of flow. 

The ahsolute levels of salivary L ~ K  observed in the present series are Pzighcr 
th.an the values reported b y  other:. 4.6,10,12-" S S U C ~   lariati ions   IT^ to  be expected 
for  reasons of cthn.ic, social and econonlic diffcrcnccs, Howc\;er, tcchz-rica! 
exrors dare to differel~ccs in Standard SIgA preparations canriot of cor1i:se be 
exclt~dccl. TJnfor?:unately an international standard for SIgA is not avsi!ablc 
and therefore accurtitc standardizations coultl not hr madc. 

The p:.ltr.reri of iiari.;tion of salivary 134.  it^^ rciotio~> ro thr DMl; 1od.e:"; pro-- 
bab'lv reflects the stImujation of salivary ZgA production by antigerrs af cariogenic 
organisms. A deficiency of secre:or\: immunogIobulii-rs in saliva p~eripitatin,a 
dental. caries seems vniikely. Tile significance of tine observed slcvatiasn ni  sali- 
vary lactuferrin in high caries subjects is n.ot cicar, Further studics TO clncidate 
the ~.cXk: of lactofcrriar in dental caries a.rrd also the  antibody spec,ificity of salFbrary 
IgA. arc. irii progress. 

References 
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1.95. 
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MAhSOh, P. C. B HEREMANS, 1. F. 11956) 1';otlilcs Biol. FIuir!s Pm;. Co l lug  B: ugcr. IJ : 
115. 
MICHALEK, S. M .>  >.,icGFlLE. J.  R.: MESTECKY, J., AP.PJ31-D, K. R. t? EOZZO, L. (1976) 
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