Detection of amyloid beta protein precursor immunoreactivity in normal and Alzheimer's disease cerebrospinal fluid

338

NEUROBIOLOGY OF AGING, VOLUME 1 t. 19c)4~ ABSTRACTS OF SECOND INTERNATIONAL CONFERENCE ON ALZHEIMER'S DISEASE BIOLOGICAL MARKERS/SYSTEMIC MARKERS

Chong1, Henry J. HaiglerI and William O. Whetsell, Jr.2 1Abbott Laboratories, Abbott Par~ IL. 60064, USA, zVanderbih University, Nashville, ~ 37232, USA The distribution of an Alzheimer's disease associated protein (ADAP) was studied in post-mortem brain tissue from a group of five Alzheimer's disease (AD) and a group of five Non-Alzheimer's disease (NAD). Eighteen different brain regions from each of these ten brains were examined. An ALZ50 based enzyme immunoassay was used to measure ADAP level. ADAP consists of at least three proteins including A68. The NAD group showed essentially no detectable ADAP in the 18 regions. The AD group (diagnosis established by clinical history and post-mortem neuropathological findings) had substantial levels of ADAP in frontal, parietal and temporal cortical, and subcortical regions with lower levels of activity found in the occipital cortex. The lowest ADAP levels found in the AD group were in the medulla and in the cerebellum. The levels of ADAP in 15 of the 18 areas examined in the AD group were significantly higher than those of the same areas for the NAD group (t-test, p<0.05). The ADAP levels in the areas globus pallidus, medulla and cerebellum were the same for both groups. The ADAP levels in four regions of AD brains were compared to the neuritic plaque (NP) counts in the corresponding brain regions of the matching formaldehyde fixed hemispheres. Although the NP count in each case met the criteria for a diagnosis of AD, the numbers of NP's in these four regions examined (i.e. frontal cortex, parietal cortex, hippocampus and subiculum) did not quantitatively correlate with the ADAP levels.

Ghanbari. M. I. N. D. Diagnostics, D-9MA, AP20, Abbott Laboratories, Abbott Park, Illinois, USA. 60064. The amyloid A4 (or beta protein), a 4.2 kD polypeptide, is a major component of amyloid deposits in the brains of patients with Alzheimer's Disease (AD). The selfaggregating amyloid A4 protein of AD is encoded as part of three larger proteins by the amyloid A4 precursor gene. The corresponding of amyloid beta protein precursor (ABPP) as a diagnostic marker for AD an antiserum against a synthetic peptide (175-186), predicted from cDNA sequence for AgPP, was used. The immunoreactivity of ABPP in normal and AD cerebrospinal fluid (CSF) was measured by Western blot and detected with radiolabeled protein A. A total of fifty,seven CSF samples (AD=27 and norfi~al=30) were analyzed for ABPP immunoreactivity. A polyclonal antibody detected two major protein bands with apparent molecular weights of 96kD and 84kD both in normal and AD CSF. The difference between normal and AD CSF was not significant. These results indicate that immunoreactivity of AgPP is present both in normal and AD CSF, and that the difference is too small to be used as a diagnostic marker,

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351 ~l zHmMER AMYLOID B A4.PROTE~ AWI'IBODIES lH H U M A N S E R A A N D CSF *U. MOnningl, U.Schmiter-Gasser2, C Hflbich l, D. Bunket, R. prior1, C LMasters 3 and K. Beyrcuther t 1Centel"foxMolecularBiololff,Uai~mtityd Hl~ddb~rg(ZMBH), lm NcmenheimcxFeld D-6900Heid¢l~,l, F.R.O 2Central lmtitutefor Miatal Health,J 5, ~ Mlllnheim,F.R.G. 3Department of Plithok~, Univeriityof Melbourne, parkvitl¢,Vict(~a 3052,Atltralia.

Since Alzheimer's disease is associated with a massive deposition of serf-aggregating BA4-protein we investigated the occurence of antibody against 8A4-protein in order to determine whether immunological mechanisms are involved in plaque formation. We identified a speeitie set of immunogiobulins in body fluids of controlpatients and Alzheimer-patients which showed a high affinity to synthetic and native amyioid BA4-proteins. The titer of these naturally oeeuring antibodies was increased in CSF of two thirds of the Alzheimer patients included in our study. In contrast in human sera the immunoreaetivity against amyloid BA4-proteins did not correlate with the clinical features of Alzheimer's di~ase, The broad oeeurence of these antibodies suggests that amyloid 8A4-protein reactive anuqx)dies represent a specific and perhaps obligatory component of the natural antibody repertoire. The elevate~ titer of these a n n ~ d i e s in CSF is poss~ly related to the amyloid BA4 depositions in the brain of Alzheimer patients.

352

DETECTION OF AMYLOID BETA PROTEIN PRECURSOR IMMUNOREACTIVlTY IN NORMAL AND ALZHEIMER'S DISEASE CEREBROSPINAL FLUID. Jonathan K. Chong*, Barney E. Miller, and Hossein A.

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353 I M M U N O R E A C T I V I T Y TO B E T A - A M Y L O [ D PROTEIN [N '['ttE SKINS OF AD AND DS PEOPLE. ¢-B. B l o n d a I , E. B e n e d i k z , G. Y. Wen, H. M. W i s n i e w s k i , K. S. Kim, G. G u d m u n d s s o n , P . M e h t a . D e p t . oi Anatomy, U n i v e r s i t y ~,f £celand, V/Suourgotu, 101 R e y k j a v i k , I c e l a n d (HB,EB)., NYS i n s t i t u t e for B a s i c R e s e a r c h , gtat:e~ Island, N Y , I 0 3 1 4 USA (GYW, HMW,KSK, PM)., BepL. of N e u r o l o g y , U n i v e r s i t y of Iceland, R e y k i a v i k , [(eland

(GG).

Recently i t was r e p o r t e d t h a t B - a m ) l o i d p r o L e i n was p r e s e n t in t h e a u t e p s i e d s k i n , ~ n t e s t i n e and c o l o n of A l z h e i m e r ' s d i s e a s e (AD) p a k J e n [ s (]oachi[;] et al, Nature 341:226-230, 19891. We s t u d i e d s k i n b i o p s i e s of 9 AD 63-77 y e a r s old and ~ c o n L r o l s 8-1q y e a r s o l d w i t h 4G8 ( m o n o c l o n a l ankLhody r e a c t i v e to amino a c i d r e s i d u e # 1 7 - 2 4 of s y n t h e t i c B - a m y l o i d p e p t i d e ) . When Elite ABC (Vector I,ab) was used as the b r i d g i n g a n t i b o d y , the e p i d e r m i s ol 6/9 AD, 1/13 c o n t r o l is p o s i t i v e l y r e a c t i v e to 4G8. We c u r r e n t l y i n v e s t i g a t e ii cases of Down S y n d r o m e (DS) age 50-77 years using A B C o m p l e x HRP from D a k o p a t t s as the bridge a n t i b o d y . T h e r e is no immunoreact ivttv in any c o m p o n e n t of the skins of these II De patients, except

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c y t o p l a s m of sweat gland cells as seen in AD skin b i o p s i e s . We also studied the epi(lermis of hamster skin w h i c h r e v e a l e d p o s i t i v e i m m u n o r e a c t i v i t y to I~-51 ( p o l y c l o n a l a n t i b o d y to amino acid residne #I-4(I o[! BAP), hut n e g a t i v e to 4G8, R-57 ( p o l y c l o n a l a n t i b o d y to C - t e r m i n a l of B - a m y l o i d p r e c u s o r protein) and a n t i - M E - 7 ( p o l y c l o n a l a n t i b o d y to s e r a p i e protein, PrP). More skin b i o p s i e s from old age control and AD patients are c o l l e c t e d to c o n f i r m our o b s e r v a t i o n .

354 Are Alpha-l-antichymotrypsin inhibitor peripheral markers

and Inter-alpha-trypsin of A l z h e i m e r ' s d i s e a s e ?

*A. Furby, D. Leys, A. Delacourte, L. Bu6e, G. Soetart, H.Petit.

Departments of neurology and neurosciences. 1NSERM & ADERMA. CHRU de Lille. H~pital B. 59037 Lille, FRANCE.