Rectal reaction to injected ulcerative colitis leucocytes and plasma.

Abstract

EDITORIAL COMMENT In patients with ulcerative colitis viable leucocytes from their peripheral blood injected into the rectal mucosa caused a local reaction which was not seen after injection with plasma, a phenomenon which needs further study.Leucocytes from the peripheral blood of patients with chronic ulcerative colitis are cytotoxic for foetal colon cells in tissue culture (Perlmann and Broberger, 1963). This report describes the effect of autologous ulcerative colitis leucocytes and cell-free plasma upon intact rectal mucosa. MATERIALS AND METHODSPATIENTS Seventeen ambulatory adult males with wellestablished diagnoses of chronic ulcerative colitis were studied. Rectal biopsies were compatible in each case. Patients were chosen who were in clinical remission, and without friable or ulcerating mucosa at the time of the investigation. Fifteen patients had been taking salicylazosulphapyridine (Azulfidine) for six to 24 months. None had been taking adrenal steroids during the preceding three months (Table I). Four patients with Laennec's cirrhosis and three with duodenal ulcer were also tested.PREPARATION OF MATERIAL Autologous leucocytes were obtained from venous blood by the method of Friedman, Bardawil, Merril, and Hanau (1960). A 30 ml. syringe containing 100 mg. of heparin (Liquaemin sodium, Organon) and 400 mg. of dextran in 20% aqueous solution was used to collect 27 ml. of blood. The needle was capped and the syringe placed in a vertical position at 4°C. for one hour. The buffy coat and plasma layers were then expressed into a centrifuge tube and centrifuged at 1,000 r.p.m. for 10 minutes. The supernatant clear plasma was pipetted off and approximately 1 ml. of cellular sediment-containing plasma was left. This 1 ml. contained 2.5 x 107 white blood cells, platelets, and a small number of red blood cells. In seven experiments, 0-5 ml. of this fluid was transferred to a sonic oscillator1 and oscillated at 9 kc. per second for 10 minutes. These 'Raytheon magnetostriction sonic oscillator model S-102A. preparations were examined by direct phase microscopy and repeatedly oscillated until all the cells seen were pyknotic and showed marked morphological change. These suspensions were used for injection when the morphological changes fulfilled the criteria of Schrek (1958) for non-viability, and are henceforth referred to as 'non-viable leucocytes' in this report. An aliquot of the supernatant plasma was placed in a Neubauer counting chamber and scanned for white blood cells. This plasma was re-centrifuged if necessary and used for injection when the microscopic examination failed to demonstrate leucocytes. This plasma is hereafter referred to as leucocyte-free.PROCEDURE Two hours after a 135 ml. sodium biphosphate enema (Fleet)2, 33 separate injections of 0-1 ml. of leucocyte-free plasma and of 0.1 ml. of plasma containing 2-5 x 106f intact leucocytes were made into two areas of rectal mucosa approximately 9 cm. from the anus. The lowest valve of Houston was taken as a reference point, and the injections were gi...

Aetiology of ulcerative colitis.

“…This finding too has been recently confirmed by Watson, Quigley, and Bolt (1966) and in vivo by Fink, Donnelly and Jablokow (1967).…”

Ulcerative colitis: definition, historical background, aetiology, diagnosis, naturel history and local complications

“…But this argument was in part refuted by their further studies (Perlmann & Broberger, 1963) showing that the leucocytes from patients with ulcerative colitis had a cytotoxic effect upon the foetal colon cells in tissue culture, an effect which was inhibited by pre-treatment with colon antigen. Broberger & Perlmann's pioneer work has recently been confirmed and extended (Fink, Donnelly & Jablokow, 1967;Watson, Quigley & Bolt, 1966;de Dombal, 1967).…”