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Biomedical subjects

K W Morley

Publications and source records attributed to K W Morley.

6 recordsLinked to original sources

Interaction between rheumatoid factor and antibody/DNA complexes: enhancement of complement fixation.

The influence of rheumatoid factor (RF) on the complement mediated binding of antibody/double-stranded DNA immune complexes to red blood cells has been investigated. Our results indicate that RF enhances this binding reaction, apparently by fixing complement via its own Fc region. These findings suggest that under certain circumstances, RF may play an exacerbating role in the antibody/DNA induced glomerulonephritis of systemic lupus erythematosus.

Antigen-Antibody Complex↗

DNA/anti-DNA complexes: correlation of size and complement fixation.

The hydrodynamic sizes of antibody/3H-dsDNA immune complexes are correlated with their ability to fix complement in a red blood cell-complement linked binding assay. The results with PM2 dsDNA and a sonicated preparation (with a molecular weight of approximately 5.10(5) suggest that packing considerations are also important in the complement binding reaction. The relevance of these findings with respect to the role of antibody/dsDNA immune complexes in the pathogenesis of glomerulonephritis in systemic lupus erythematosus is discussed.

Antigen-Antibody Complex↗

Studies of artificial cryoprecipitates containing anti-DNA antibody activity.

Artificial cryoprecipitates have been prepared from SLE sera by incubation of anti-double stranded DNA with 3H-PM2 DNA. IgM rheumatoid factor, and complement to study further the relationships between natural cryoprecipitation and the DNA/anti-DNA system in this disorder. Addition of rheumatoid factor and/or complement to a mixture of SLE serum and 3H-DNA results in increased precipitation of the 3H-DNA following incubation at 4 degrees C. Precipitation formed with added complement or complement plus rheumatoid factor could be rapidly resolubilized or reprecipitated by warming to 37 degrees C or recooling to 4 degrees C, i.e., exhibited the behavior of natural cryoprecipitates. The resolubilized cryoprecipitates contained excess antibody, because they bound significant quantities of additional 3H-DNA in the Farr assay. Of special interest was the observation that DNA binding activity frequently could be recovered from solubilized cryoprecipitates even though 3H-DNA was not added during the original cryoprecipitate preparation. With certain cryoprecipitations prepared in this way, the DNA binding activity was further increased by deoxyribonuclease treatment, raising the possibility that endogenous DNA may be constituent of such cryoprecipitates. The implications of these findings with respect to the nature of antibody-DNA immune complexes in SLE are discussed.

Antibodies, Antinuclear↗

Quantitative studies of the interaction of 3H-dsDNA/anti-DNA immune complexes with complement: comparison and evaluation of the Raji cell, the solution phase C1Q, and the red blood cell linked complement fixation radioimmunoassays.

We have prepared antibody/3H-dsDNA immune complexes and have used three independent radioimmunoassays to quantitate their interaction with complement; the solution phase C1q assay, the Raji cell assay, and a complement-based red blood cell adherence assay (RBC-CF). Our results indicate that although there is reasonable qualitative agreement between the Raji cell assay and the RBC-CF assay, there are some differences in the quantitative range of sensitivities of the two assays. On the other hand, we find that most of the complement-fixing antibody/3H-dsDNA complexes are not detected in the solution phase C1q assay. The results suggest this is because the absolute concentrations of the immune complexes were too low to achieve significant precipitation under the standard conditions used in the C1q assay. The implication of these findings with respect to the potential detection and analysis of antibody/dsDNA immune complexes is discussed.

Antibodies↗

Stability of DNA/ANTI-DNA complexes. IV. Complement fixation.

We describe an in vitro assay to study complement fixation of antibody/dsDNA immune complexes formed from SLE sera and radiolabeled dsDNA. The method measures the amount of radiolabeled dsDNA which is part of an immune complex bound to red blood cells via the C3b complement component receptor. The assay is dependent upon active complement, red blood cells, and anti-dsDNA antibodies, but it is independent of the red blood cell donor (type O) and the age of the red blood cells (up to 10 days). The method has been compared in some detail with the Farr assay with respect to the antibody/dsDNA ratio in the immune complexes and the relative stability of the complexes as measured by their resistance to dissociation by excess unlabeled dsDNA. Our results indicate that multiple binding of antibodies to dsDNA is required for complement fixation, and that a significant percentage of those antibodies which fix complement are of high avidity. Finally, a double label assay using both [3H]- and [14C]-dsDNA indicates that the complement fixing potential of the anti-dsDNA antibodies in an SLE serum is strongly influenced by the order of mixing of the isotopes with the serum. The DNA isotope which is added to the serum first is considerably more effective at fixing complement than the isotope which is added 1 h later. The implications of these results with respect to the pathogenesis of SLE are discussed.

Antigen-Antibody Complex↗