Title

Multiparameter Kinetic Analysis Of Killer Cell Initiation Using Separated Lymphocyte Immune Rna

Date of Award

1981

Availability

Article

Degree Name

Doctor of Philosophy (Ph.D.)

Abstract

Initial studies were performed using whole spleen cell Immune RNA (I-RNA) from lymphocytes exposed to EL-4. By treating nonsensitized C57Bl/6J spleen derived lymphocytes with EL-4 tumor cell directed xenogeneic extracted RNA we were able to monitor early changes in cellular DNA content by flow cytometric (FCM) analysis and 3H-thymidine uptake. RNA content was monitored by 3H-uridine uptake. These kinetic parameters were correlated with cell mediated cytotoxicity (CMC) which appeared as early as 8 hours after activation as measured by release of chromium-51 from labeled EL-4 target cells. FCM analysis, 3H-thymidine and 3H-uridine uptake data showed peak S phase activity at 72 hr.Immune RNA derived from EL-4 sensitized glass adherent (GADH) and non-adherent (NGADH) Hartley Guinea Pig (HGP) cell fractions was used to monitor the transfer of CMC. Unsensitized C57Bl/6J mouse lymphocytes were treated with the resulting CADH and NGADH extracted I-RNA at 100 ug/ml for 1 hour at 37(DEGREES)C. GADH I-RNA treated lymphocytes showed the most elevated cytotoxic activity which was statistically significant at 24 hours, peaked at 72 hours and fell off by 96 hours. The NGADH I-RNA treated lymphocytes showed no significant cytotoxicity until 48 hours after initial exposure and dropped off by 72 hours. The NGADH kinetics resemble the cytotoxic kinetics of whole spleen cell I-RNA as reported previously (J.Hist.Cytochem., 27:491, 1979). Control RNA preparations did not show significant stimulation over normal C57Bl/6J RNA and RNASE control values.C57Bl/6J syngeneic tumor cell lines, a melanoma (B-6) and a mammary adenocarcinoma (M-3560), were used as targets for GADH and NGADH treated lymphocytes. The cellular fractions of the I-RNA treated lymphocytes showed selective cytotoxicity toward the EL-4 target. The cytotoxicity toward the M-3560 and B-16 targets fell within the control range, indicating a specificity of transfer of cytotoxic effector cell function by I-RNA. Supernatant fractions of both the GADH and NGADH cell cultures in the presence of normal C57Bl/6J lymphocytes also proved to be selectively cytotoxic toward the EL-4. This could indicate that some component may be released into the supernatant after I-RNA stimulation.

Keywords

Health Sciences, Immunology

Link to Full Text

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