Abstract
The transfer of tumour-specific cytotoxicity against a murine fibrosar-coma has been demonstrated in vitro using xenogeneic RNA extracted from tumour-cell-immune animals. Poly(A)-tailed messenger RNA from immunogenic RNA was isolated by passage through an oligo(dT)-cellulose column, and evaluated to determine whether the same tumour-specific cytotoxicity could be transferred. Aliquots of normal C3H mouse lymphocytes were treated with poly(A)-containing immune RNA, whole-cell immune RNA lacking poly(A) and total cellular immune RNA. Treated cells were tested in vitro using an adaptation of the Takasugi and Klein microcytotoxicity assay. Percent cytotoxicity was calculted using cells treated with fractions of normal RNA as control. An increase in tumour cytotoxicity was found with poly(A)-containing immune RNA. The optimum dose of poly(A)-tailed immune RNA was estimated as 6.5 microgram of RNA per 4 x 10(6) lymphocytes. Populations of lymphocytes were separated using glass and nylon wool. T- and B-enriched populations were treated with various RNA components. The adherent cell population showed no significant cytotoxicity, whilst treatment of the nonadherent population with poly(A)-tailed immune RNA produced high levels of cytotoxicity.
Highlights
Summary.-The transfer of tumour-specific cytotoxicity against a murine fibrosarcoma has been demonstrated in vitro using xenogeneic RIBONUCLEIC ACID (RNA) extracted from tumourcell-immune animals
Wlhile it is apparent that immunogenic RNA (IRNA) is capable of transferring a variety of immune responses, the active component of IRNA seems to be confined to a narrow range of sedimentation coefficients
This study investigates the ability of this poly(A)+ IRNA to transfer specific cytotoxicity to nylon-wool-purified populations of lymphocytes
Summary
LM is a nontumorigenic cell of C3H mouse origin Another tumour cell line, S91, was used as a control of specificity. Immunization of mice.-Mice (C3H/HeJ) were given 3 i.p. injections of 4 x 106 4198V tumour cells every 5-6 days. Non-immune mice will produce palpable tumours in 10-14 days after i.m. injection of 5 x 105 4198V cells. C-control mean wells test wvells of control wx-ells x00 fractionator, the sucrose gradient was fractionated and absorbance (A) read at 254 nm through the flow cell of an ISCO Model 222 spectrophotometer. The percent 0+ cells was calculated using lymphocytes treated with normal rabbit serum as control. The percent rosettes was calculated on the basis of counting 100 cells after incubation of equal volumes of 106/ml lymphocytes Awith 100 EAC.
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