The Agglutination of Tumour Cells In Vitro by Sera from Tumour Patients and Pregnant Women

Abstract

IN the course of study on the possibility of autoimmune mechanisms against tumour, the level of circulating antibodies to the autologous tumour cells was estimated by cell agglutination titres according to a technique previously developed in this laboratory (Pikovski, Tal, Schlesinger and Margoliash, 1957). It was soon found that cross agglutination occurred. It appeared that the sera from tumour patients contained a factor (AF) which could cause agglutination of, and be adsorbed by, tumour cells from a variety of sources. This paper represents an analysis of this phenomenon. MATERIALS AND METHODS Cell preparationCell suspensions were prepared from some human tumours, several normal human tissues and some mouse tumours by a previously described technique (Pikovski, Tal, Schlesinger and Margoliash, 1957). Suspensions of tissue cultures of HeLa cervical carcinoma, KB buccal carcinoma and Chang normal liver cells were also used. The preparation of the tissue culture suspensions is given in the following detail using HeLa cultures as the example.HeLa cultures were grown in Eagle's lact-yeast medium plus 15 per cent of normal human serum and horse serum, respectively, in milk bottles. After 7-8 days of growth, the medium was pipetted off, and the cell layer washed with 10 ml. of warm (370 C.) phosphate-buffered saline (PBS). The PBS was replaced by 10 ml. of 0-02 per cent Ethylenediamine Tetracetic acid (EDTA) in PBS devoid of Ca and Mg, and the culture kept at 370 C. for [15][16][17][18][19][20] minutes. This procedure loosened the cells from the wall of the culture bottle and the EDTA cell suspension was transferred to a test tube. Clumps of cells were dispersed by repeated pipetting with a fine Pasteur pipette, taking care to avoid foaming. The suspension was then centrifuged at 500 r.p.m. for 5 minutes. The supernate was carefully removed so as not to disturb the cell pellet. The cells were then resuspended in 8 ml. of warm RBS and centrifuged as above. The washed cells were counted and suspended in sufficient normal saline to give a concentration of 4 x 106 cells per ml. Agglutination procedureSera were diluted fourfold with 0-2 M phosphate buffer (pH 7.0). The pH of the system was critical. 2 x 105 Cells were added to 0 4 ml. of serum dilutions.

The toxic effects of human serum on Ehrlich ascites tumour cells inin vitro culture

“…Our findings are all consistent with these points 1--6 which stem from the following research groups, in Israel: Tal (1965), Ginsburg et al (1961), Tal et al (1964), Laskov et al (1968a, b, c), in Canada: Fedoroff (1958, 1963, Fedoroff and Cook (1961), Fedoroff and Doerr (1962), Fedoroff and Webb (1962); other important contributions: Lumsden and Kohn-Speyer (1929), Landschiitz (1954Landschiitz ( , 1956), Penttinen and Sax6n (1956a, b), Saxdn andPenttinen (1962, 1965), Bolande (1960), Bolande and McClain (1960), Landy et al (1960), Takada et al (1961), Stambuk and Burk (1961), Hartveit (1964, 1966), and Hotham-Iglewski and Ludwig (1967a.…”

A Tumour Cell Aggregation Promoting Substance from Rat Ascites Hepatoma Cells

“…The potency of 11l,tg of the aggregation promoting factor seemed to correspond to that of 250,ug ofJack bean concanavalin A. The power to induce tumour cell aggregation has been seen in the serum (Tal, Dishon and Gross, 1964) and ascitic fluid of cancer patients (Mori, Akedo and Tanigaki, 1970) and of tumour bearing mice (Oppenheimer and Humphreys, 1971). However, the problem of whether such potency may be related to an aggregation promoting factor of cancer cell origin has not yet been established.…”

Agglutination of Murine Tumor Cells by Sera from Patients with Autoimmune Diseases

“…It was not possible, however, to assign a diagnostic value to the agglutination test since the sera from patients with other diseases such as cancer and those from pregnant women also exhibited this activity [8,9,13]. Also, the agglutination activity could not be correlat ed to clinical stages of autoimmune diseases.…”