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. 1973 Aug;70(8):2299–2303. doi: 10.1073/pnas.70.8.2299

Enumeration of Activated Thymus-Derived Lymphocytes by the Virus Plaque Assay

Shogo Kano *, Barry R Bloom *, Michael L Howe †
PMCID: PMC433722  PMID: 4365371

Abstract

Lymphocytes activated by antigens or mitogens acquire the capacity to replicate viruses, and the number of activated lymphocytes can be estimated by the virus plaque assay. Concanavalin A and pokeweed mitogen produced 33-fold and 17-fold increases in virus plaqueforming cells (V-PFC), respectively, above background, while lipopolysaccharide produced only a 2- to 3-fold increase. T (thymus-derived lymphocyte)-depleted lymphocyte populations, derived from anti-θ-treated or nude (arthymic) mouse spleens, failed to produce V-PFC after culture with concanavalin A or pokeweed mitogen. The present studies thus demonstrate that the virus plaque assay measures activated T-lymphocytes.

A dissociation between the V-PFC response and cell proliferation was previously observed in antigen-stimulated cells cultured in the presence of mitotic inhibitors. In the present studies, while stimulation of CBA (H2k) lymphocytes by DBA/2 (H2d) cells produced high levels of thymidine incorporation, lymphocyte target-cell cytotoxicity, and V-PFC, stimulation of BALB/c (H2d) lymphocytes against DBA/2 (H2d) cells resulted in even higher levels of thymidine incorporation with a virtual absence of cytotoxic lymphocytes or V-PFC. These results indicate that proliferation is not a sufficient condition for permitting lymphocytes either to exert cytotoxicity on target cells or to replicate viruses, and suggest that there may be a correlation between the development of V-PFC and cytotoxic lymphocytes. They are consistent with the view that there are at least two functional subpopulations of T-lymphocytes.

Keywords: cell-mediated immunity, mitogen stimulation, mixed lymphocyte culture

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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