IMMUNOCHEMICAL EVIDENCE FOR A GAMMA GLOBULIN PECULIAR TO CEREBROSPINAL FLUID

1961 ◽  
Vol 39 (10) ◽  
pp. 1567-1574 ◽  
Author(s):  
Catherine F. C. MacPherson ◽  
James B. R. Cosgrove
Keyword(s):  
Multiple Sclerosis ◽  
Cerebrospinal Fluid ◽  
Human Serum ◽  
Gamma Globulin ◽  
Two Fluids ◽  
Human Sera ◽  
Normal Human

Two immunologically different gamma globulins were revealed in "normal" cerebrospinal fluid (NCSF) and in multiple sclerosis cerebrospinal fluid (MSCSF) by immunoelectrophoretic analyses, which employed rabbit antisera prepared against NCSF and MSCSF. Since only one gamma-globulin arc was formed on immunoelectrophoretic analyses of normal human sera (NHS) with rabbit anti-CSF sera and repeated absorptions of anti-CSF sera with human serum failed to remove antibodies to the minor gamma globulin, it would appear that the minor gamma globulin is peculiar to CSF. The major gamma globulin of CSF was shown to be immunologically identical with the single gamma globulin of serum as judged by the fusion of their respective precipitin arcs. The major and minor gamma globulins of NCSF could not be distinguished from those of MSCSF by any of the rabbit antisera used. However, the ends of the precipitin arc formed by the major gamma globulin of NCSF with its homologous antibody became frayed and sometimes doubled indicating it to be immunologically less homogeneous than the gamma globulin of serum. The two gamma globulins of MSCSF and the minor gamma globulin of NCSF migrated more slowly than the major gamma globulin of NCSF. In comparison with NHS, the immunoelectrophoretic patterns of CSF developed with anti-CSF sera were characterized, in the alpha-2 zone, by a greater number of stronger arcs. Experiments on absorbed anti-CSF sera indicated that this was due to differences in the relative concentrations of the various components of this zone in the two fluids.

Use of polyethylene glycol in separating bound from unbound ligand in radioimmunoassay of thyroxine.

1976 ◽  
Vol 22 (3) ◽  
pp. 299-304 ◽  
Author(s):  
M C Cheung ◽  
W R Slaunwhite
Keyword(s):  
Polyethylene Glycol ◽  
Human Serum ◽  
Salt Concentration ◽  
Room Temperature ◽  
Gamma Globulin ◽  
Separation Process ◽  
Normal Human Serum ◽  
Normal Sheep Serum ◽  
Normal Human ◽  
Entire Procedure

Abstract In most rapid radioimmunoassay methods, absorbents are used that disturb the equilibrium of the ligand-antibody interaction; thus the separation process must be rigidly timed. We show here that polyethylene glycol (mol wt. 6000) abolishes this constant. We measured gamma-globulin precipitation by polyethylene glycol (150g/liter) in a thyroxine radioimmunoassay system involving use of a sheep anti-thyroxine antiserum, quantitatively and qualitatively, with either normal human serum or completely precipitated at pH 6-9 at 4 and 25 degrees C, but incompletely if salt concentration was high (1 mol/liter). Specificity of gamma-globulin precipitation increased with increasing pH and temperature. All gamma-globulin-bound thyroxine was precipitated, and also some not bound to gamma-globulin. This was taken into account by including "blank" tubes (no antibody, but with normal sheep serum) in all assays. Because this reaction is predominantly entropic and temperature independent, the entire procedure can be done at 37 degrees C and room temperature without rigid timing. The assay requires 10 mul of serum, its reproducibility is 4-8% (CV) in the euthyroid and hyperthyroid range, and its accuracy is close to 100%.

scholarly journals Human Serum Contains a Protease That Protects against Cytotoxic Activity of Bacillus anthracis Lethal Toxin In Vitro

2008 ◽  
Vol 15 (6) ◽  
pp. 970-973 ◽  
Author(s):  
David L. Goldman ◽  
WangYong Zeng ◽  
Johanna Rivera ◽  
Antonio Nakouzzi ◽  
Arturo Casadevall
Keyword(s):  
Heat Treatment ◽  
Bacillus Anthracis ◽  
Human Serum ◽  
Protective Antigen ◽  
Lethal Toxin ◽  
Protective Activity ◽  
Calcium Chelation ◽  
Human Sera ◽  
Normal Human

ABSTRACT The role of innate immunity in the host response to Bacillus anthracis is poorly understood. We found that normal human serum contains an antitoxin mechanism that is capable of protecting macrophages in vitro from B. anthracis lethal toxin-mediated killing. This protective activity was limited to defined amounts of toxin and was lost by heat treatment or serum dilution. Some person-to-person variation in the protective activity of serum was noted, especially with higher concentrations of lethal toxin. A similar protective activity was found in murine serum, though human serum consistently neutralized more toxin than did murine serum. The protective activities of both murine and human sera correlated with cleavage of the protective antigen into two fragments with approximate molecular sizes of 20 and 50 kDa that were recognized by the monoclonal antibodies 7.5G and 10F4, respectively. This pattern of fragmentation is consistent with cleavage at multiple sites, including the furin-susceptible site. Cleavage was abolished by heat treatment and calcium chelation. These findings highlight a potential role for serum proteases in protection against the lethal toxin of B. anthracis.

Proteomic analysis of multiple sclerosis cerebrospinal fluid

2004 ◽  
Vol 10 (3) ◽  
pp. 245-260 ◽  
Author(s):  
B N Hammack ◽  
K YC Fung ◽  
S W Hunsucker ◽  
M W Duncan ◽  
M P Burgoon ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Cerebrospinal Fluid ◽  
Calcium Binding ◽  
Cell Signalling ◽  
Protein A ◽  
Peptide Mass Fingerprinting ◽  
Two Dimensional Gel Electrophoresis ◽  
Ph Gradients ◽  
Peptide Mass ◽  
Normal Human

Two-dimensional gel electrophoresis and peptide mass fingerprinting were used to identify proteins in cerebrospinal fluid (C SF) pooled from three patients with multiple sclerosis (MS) and in C SF pooled from three patients with non-MS inflammatory central nervous system (C NS) disorders. Resolution of C SF proteins on three pH gradients (3-10, 4-7 and 6-11) enabled identification of a total of 430 spots in the MS C SF proteome that represented 61 distinct proteins. The gels containing MS C SF revealed 103 protein spots that were not seen on control gels. A ll but four of these 103 spots were proteins known to be present in normal human C SF. The four exceptio ns were: C RTAC -1B (cartilage acidic protein), tetranectin (a plasminogen-binding protein), SPARC -like protein (a calcium binding cell signalling glycoprotein), and autotaxin t (a phosphodiesterase). It remains unknown whether these four proteins are related to the cause and patho genesis of MS.

scholarly journals DECREASED PERCENTAGE OF POLYMORPHONUCLEAR NEUTROPHILS IN MOUSE PERIPHERAL BLOOD AFTER INOCULATION WITH MATERIAL FROM MULTIPLE SCLEROSIS PATIENTS

1972 ◽  
Vol 136 (3) ◽  
pp. 618-629 ◽  
Author(s):  
Richard I. Carp ◽  
Pamela C. Licursi ◽  
Patricia A. Merz ◽  
George S. Merz
Keyword(s):  
Multiple Sclerosis ◽  
Cerebrospinal Fluid ◽  
Human Brain ◽  
Brain Homogenate ◽  
Polymorphonuclear Neutrophils ◽  
Human Material ◽  
Normal Human ◽  
Multiple Sclerosis Patients ◽  
Absolute Decrease ◽  
Differential Counts

Mice inoculated with brain homogenates from multiple sclerosis (MS) cases showed marked changes in their leukocyte differential counts, with a decrease in per cent polymorphonuclear neutrophils (PMN) and an increase in the per cent lymphocytes. These changes were based upon an absolute decrease in the number of circulating PMN. The decrease in PMN percentages was apparent at 16 hr after infection and persisted for at least 11 months. The factor responsible for the decrease in PMN was (a) recoverable from 12 hr to 8½ months after inoculation, (b) present in human brain homogenate at a concentration of 3 x 1012, and (c) between 25 and 50 nm in diameter. Inoculation of 100 units of factor into mice and subsequent titration showed that the factor had undergone a net increase in the mouse of at least 109-fold. The factor causing the PMN decrease was found in all MS material thus far tested: three brains, one spleen, three sera, and two cerebrospinal fluid (CSF) from nine cases of MS. The factor was not found in normal human material that included two brains, one spleen, two sera, and two CSF.

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