The conglutination phenomenon IX. The production of immuno-conglutinin in rabbits

Abstract

1. An investigation has been carried out into the conditions necessary for the production of immuno-conglutinin in rabbits.2. The inoculation of complement adsorbed on sensitized cells stimulates the production of immuno-conglutinin. We have called this procedure hetero-stimulation.3. Immuno-conglutinin also results from the inoculation of untreated but killed bacteria. This procedure for its production we call auto-stimulation. Gram-negative bacteria appear to be more effective stimulants than Gram-positive bacteria. Soluble antigens did not appear to be as effective as bacterial suspensions.4. The necessity for clarifying the part played by immuno-conglutinin in in vivo immune processes has been stressed.The authors would like to thank the following for gifts of experimental materials: Mr H. I. Field, strains of S. pullorum; Dr J. Boissard, strain of C. hofmanni; Dr C. H. Lea and Dr R. S. Hannan, purified casein; Miss Dunkerley, Bence Jones protein. Cx polysaccharide for testing acute phase protein in rabbit serum was made available by the kindness of Dr McCarty of the Hospital of the Rockefeller Institute for Medical Research, New York; and much of the work on this aspect was done in consultation with Dr H. C. Anderson, to whom we are most grateful.

The conglutination phenomenon: X. Conglutinin and immuno-conglutinin in guinea-pigs, pigs, donkeys, rats and mice

“…(Coombs & Coombs, 1953) experiments were reported which confirmed the observations of earlier workers on the production of immunoconglutinin in rabbits (Streng, 1930;Wartiovaara, 1932). Conglutinin was found to be present in very small amounts in the sera of normal uninoculated rabbits and the production of immuno-conglutinin was induced by injecting them with complement from a different animal species which had been adsorbed either specifically on to sensitized bacteria or non-specifically on to kaolin; the term hetero-stimulation was suggested for this procedure.…”

“…They were injected intravenously into one of the tail veins and bled by cardiac puncture. Inocula The preparation of the stock bacterial suspensions of Salmonella pullorum, Proteus OX 19 and Staphylococcus aureus has been described in a previous paper (Coombs & Coombs, 1953). The preparation of a suspension of sensitized bacteria which had adsorbed complement was also described in that paper; the procedure was similar in the present experiment except that, when guinea-pigs were being injected, a guinea-pig anti-proteus serum was used to sensitize the proteus organisms.…”

“…Method of testing for conglutinating activity Method 1 as described by Coombs & Coombs, (1953) in which sensitized sheep cells and a dilution of fresh horse serum were added consecutively to the test serum, was found to give equivocal results with guinea-pig serum and to fail to detect any conglutinating activity in pig, rat or mouse serum. Because in this method the horse complement is exposed, before or during adsorption, to any anticomplementary action of the serum being tested, method 2b as described by Coombs & Coombs (1953) was used routinely since in this method the complement was adsorbed on to the cells in the absence of test serum.…”

“…Because in this method the horse complement is exposed, before or during adsorption, to any anticomplementary action of the serum being tested, method 2b as described by Coombs & Coombs (1953) was used routinely since in this method the complement was adsorbed on to the cells in the absence of test serum. The indicator system in method 2 b was sheep cells sensitized with the antibody from normal bovine serum and then exposed to a dilution of horse complement.…”

“…(2) AN ATTEMPT TO ELUCIDATE THE REASONS FOR THE FAILURE OF GUINEA-PIGS TO PRODUCE IMMUNO-CONGLUTININ (AUTO-STIMULATION) The hypothesis has been advanced in a previous paper (Coombs & Coombs, 1953) that the animal's own complement, fixed in vivo on to injected bacteria, might be the direct antigenic stimulant for the production ofimmuno-conglutinin. The failure ---I I -1 V-I I I I I I I I of immuno-conglutinin (auto-stimulation) production in guinea-pigs, when considered in the light of this hypothesis, could be due to any of at least three causes.…”