The Immunological Identification of Brain Proteins on Cellulose Nitrate in Human Demyelinating Disease

Abstract

An immunological technique has been employed to identify proteins, separated in polyacrylamide gels, which show changes in brain samples from cases of multiple sclerosis and subacute sclerosing panencephalitis. Sodium dodecylsulphate‐treated proteins in particulate and soluble fractions were separated in polyacrylamide slab gels, transferred electrophoretically onto cellulose nitrate sheets, incubated with specific antisera and visualized by an immunoperoxidase method. Protein bands showing changes were identified using antisera raised against the myelin basic and Wolfgram proteins, the neurofilament triplet proteins, tubulin and glial fibrillary acidic protein. In addition to the loss of myelin proteins, decreases in the neurofilament proteins and in tubulin were seen in both multiple sclerosis and subacute sclerosing panencephalitis samples. The distribution of glial fibrillary acidic protein polypeptides in the particulate and soluble fractions of plaque samples appeared to vary according to the degree of fibrosis. Changes in the levels of the myelin‐associated glycoprotein, the lower molecular weight component of the Wolfgram protein, albumin and immunoglobulin G, none of which were visualized by protein staining, were also seen. This immunological technique has allowed a closer examination of changes occurring in brain protein spectra in multiple sclerosis and subacute sclerosing panencephalitis.

Myelin in multiple sclerosis is developmentally immature.

“…Our studies have shown that the NH2-terminus is heterogeneously acylated with alkyl carboxylates varying in length from [2][3][4][5][6][7][8][9][10] detected at this time. From the age of 4 yr, the adult proportion of C-8 was obtained i.e., -20%.…”

“…Two (6). At least three variants have been reported in the human brain, the 21.5-, 18.5-, and 17.3-kD sizes (7)(8)(9)(10)(11). These variants are believed to arise by alternate splicing of a single message.…”

Cognitive Dysfunction and Histological Findings in Adult Rats One Year After Whole Brain Irradiation.

“…40) The immunohistological characteristics of multiple sclerosis, one of the most well-known demyelinating diseases, have frequently been described. 15,30) Degeneration of the corpus callosum was found in a patient with multiple sclerosis. 12) Furthermore, MR imaging of patients of Alzheimer's disease, which is typically characterized by dementia, showed white matter change in about 60% of the brain tissue.…”

Immunocytochemical Characterization of Primary Glial Cell Cultures From Normal Adult Human Brain

“…Monoclonal antibodies A2B5 and anti-human fibronectin F9 (Sera-Lab), human macrophage marker antibody EBM/II (Franklin et al, 1986) (Dakopatts), and normal mouse and rabbit serum (Nordic) were also used. The myelinoligodendrocyte and GFAP antibodies have been characterized on immunoblots and human brain tissue sections, and we have examined the specificity of the glutamine synthetase and EBM/11 antibodies on sections (Newcombe et al, 1982(Newcombe et al, , 1988. Anti-mouse or anti-rabbit immunoglobulin G (IgG) or F(ab), fragments conjugated to fluorescein or rhodamine isothiocyanate (FITC) (Serotec, Dakopatts and Sigma) were used at a 1 : 20 dilution.…”

“…The following mouse monoclonal antibodies were gifts: anti-neurofilament antibody RT97 (from Dr B. Anderton), anti-bovine myelin basic protein (MBP) (Groome, Harland & Dawkes, 1985) and anti-rat galactocerebroside (Ranscht et al, 1982). The following rabbit antisera were also gifts: anti-human Wolfgram protein from Dr E. Bock (Newcombe, Glynn & Cuzner, 1982), anti-human carbonic anhydrase isoenzyme I1 (CA 11) (Komoly et al, 1987) and anti-human glutamine synthetase from Dr N. Gregson, and anti-rat myelin associated glycoprotein (MAG) from Dr R. Quarles (Newcombe et al, 1982).…”