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E D Kuhlmann

Publications and source records attributed to E D Kuhlmann.

30 records · Page 2Linked to original sources

Opsin for immunological studies.

Opsin for immunological studies can be prepared free from S-antigen by affinity chromatography on Con A-Sepharose. This preparation, however, contains 2-6% Con A originating from the affinity medium. Con A as impurity disturbs lymphocyte transformation tests carried out with opsin as test antigen. We describe a method for the removal of Con A by immuno-adsorption to Protein A/anti-Con A/IgG in detergent. In addition, we have selected some detergents and detergent concentrations in which opsin can be purified and added to the lymphocyte culture medium.

Adsorption↗

Opsin-induced experimental autoimmune retinitis in rats.

Experimental autoimmune retinitis has been induced in Lewis rats by injection of opsin in mycobacterial adjuvant and Hemophilus pertussis adjuvant. Clinical, histopathological and immunological parameters of the disease are reported. Two types of opsin were prepared from purified bovine retina outer segments, one type in Triton X-100 and the other in lithium dodecyl sulfate. Both preparations were free from S-antigen. Dodecyl sulfate-denaturated-opsin displayed lower antigenicity and pathogenicity than Triton-opsin. Triton-opsin (250 micrograms) induced moderate to severe non-granulomatous uveitis (predominantly retinitis) in 70% of the Lewis rats at the end of the second week after injection. The photoreceptor cell layer was destructed within a few days. This group displayed high responses to opsin in the lymphocyte transformation test. In view of observed histological features, the possible early involvement of vasoactive factors is discussed. Low opsin doses (50 or 100 micrograms) seldomly induced severe retinitis, while the incidence of mild pathology was low. Lewis rats appeared to be more susceptible for the development of experimental autoimmune retinitis than Wistar rats.

Animals↗

Gamma-crystallin-dodecyl sulfate complexes show discrete precipitation in non-ionic detergents.

Gamma-crystallin-sodium dodecyl sulfate (SDS) complexes are precipitated by Triton X-100 and other non-ionic detergents. This is accomplished as discrete lines by double diffusion, radial diffusion and other immunotechniques. Many other protein-SDS complexes including those of alpha- and beta-crystallins remain soluble. Immunoglobulin G tends to precipitate under similar conditions, a reason why the use of detergents in immunoprecipitation reactions has to be carefully controlled. The phenomenon can be used for the estimation of SDS or non-ionic detergents in the presence of gamma-crystallin.

Animals↗