Abstract
From the earliest work on the lytic action of serum, the cells of the tissues, particularly the phagocytes, have been considered a source of complement. However, convincing proof is still lacking that these cells either contain or liberate active complement. The possibility that they may contribute substances inactive alone but nevertheless essential to complete complement action has received little consideration.
It seemed of interest to determine whether or not a suspension of leucocytes would contribute complementing substances, either heat-labile or heat-stable, which, like heated serum, 1 might restore the hemolytic activity of complement made inactive by ammonia.
Fresh guinea pig serum was inactivated with half its volume of N/5 ammonium hydroxide at 37°C. for 1 1/2 hours and then neutralized to pH 7.6 with N/5 hydrochloric acid. The final dilution was 1:2.
Leucocytes from guinea pig blood were collected in strata and washed in physiological salt solution by centrifugalization at low speed, according to the technic of Wadsworth and Hoppe. 2 They were washed, however, not once but 5 times. Sufficient salt solution was used in washing so that the dilution of plasma in the final suspension of leucocytes was at least 1:200,000. Also, in order to compensate for the unavoidable mutilation of the cells during the additional washings, the leucocytes from 10 cc. of blood were suspended in 0.5 cc., rather than in 1 cc., of salt solution.
In order to control the reactivating effect of traces of plasma which might be retained in the preparation of the leucocyte suspension, a portion of the supernatant fluid from which these cells had been separated was diluted to the same degree as were the leucocytes in the process of washing, and tested in parallel with them for its reactivating effect. For comparison, the reactivating properties of very small amounts of heated serum were also determined.
Get full access to this article
View all access options for this article.