Solid phase enzyme immunoassays for the quantification of serum amyloid P (SAP) and complement component 3 (C3) proteins in acute-phase mouse sera.

Simple, reliable and sensitive enzyme immunoassays have been developed for the quantification of the mouse acute-phase SAP and C3 proteins. The ELISA systems were validated using sera from mice injected with S. dysenteriae endotoxin, and detected 500 pg protein/ml. The assays use 96-well microtitre plates permitting rapid processing of a large number of samples.

Measurement of cytokine production by the monocytic cell line Mono Mac 6 using novel immunoradiometric assays for interleukin-1 beta and interleukin-6.

Immunoradiometric assays for interleukin-1 beta and interleukin-6 were developed using affinity-purified IgG fractions from antisera initially raised for radioimmunoassay. Capture and detector functions were carried out by the same antibody preparation within each assay. The assays were precise, rapid and 6-8-fold more sensitive than the RIA systems previously employed. IRMAs were used for the initial characterisation of a candidate cell line (Mono Mac 6), under investigation in a 'monocyte test' for pyrogen detection, and permitted rapid and specific determination of the cytokines under stimulatory and inhibitory conditions.

Assay of pyrogens by interleukin-6 release from monocytic cell lines.

A novel in-vitro system has been developed for the detection and quantification of pyrogen in pharmaceutical products. The measured variable was evoked secretion of the pyrogenic cytokine interleukin-6 from MONO MAC 6 monocytic cells incubated with the product. The interleukin-6 was detected using a specific and sensitive ELISA developed for this purpose. The test system detected pyrogenic contamination in 3 batches of therapeutic human serum albumin which had caused adverse reactions in recipients. The contamination was not detected in conventional tests: the rabbit pyrogen test and the limulus amoebocyte lysate test.

A solid phase enzyme immunoassay for serum amyloid A (SAA) protein. Clinical evaluation.

An enzyme-linked immunosorbent assay for serum amyloid A protein using commercially available antisera has been developed. Immulon II microtitre plates are coated with undenatured sera by overnight incubation at 4 degrees C. The assay combines the advantages of simplicity, rapidity, precision, sensitivity and safety and confirms the suitability of measuring serum amyloid A protein as a sensitive marker during the acute-phase response. This method permits the rapid processing of a large number of samples.

The use of the rat hepatoma cell line MH1C1 to investigate the stimulus-secretion response of hepatic acute phase proteins.

The rat hepatoma cell line MH1C1 has been characterized to show a stimulated secretion of C-reactive protein in response to both leukocyte supernatant and a purified human interleukin-1 preparation. The time-dependency and dose-response relationship of CRP secretion were comparable to and somewhat more sensitive than the effects of leukocyte supernatant and purified human interleukin-1 on the proliferative rate of murine thymocytes; the proliferative rate of the hepatoma cell line MH1C1 was unchanged under these conditions. Agents which affect the thymocyte bioassay response to interleukin-1 namely interleukin-2, lipopolysaccharide, concanavalin A and phytohemagglutinin showed no effect on the C-reactive protein release of the MH1C1 cell line. These data strongly support the suitability of this cell line for the in vitro study of the hepatic acute phase stimulus-secretion response.