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. 2000 Mar;7(2):307-11.
doi: 10.1128/CDLI.7.2.307-311.2000.

Measurement of T-lymphocyte responses in whole-blood cultures using newly synthesized DNA and ATP

P R Sottong  1 , J A Rosebrock, J A Britz, T R Kramer
Affiliations

Measurement of T-lymphocyte responses in whole-blood cultures using newly synthesized DNA and ATP

P R Sottong et al. Clin Diagn Lab Immunol. 2000 Mar.

Abstract

The proliferative response is most frequently determined by estimating the amount of [(3)H]thymidine incorporated into newly synthesized DNA. The [(3)H]thymidine procedure requires the use of radioisotopes as well as lengthy periods of incubation (>72 h). An alternative method of assessing T-lymphocyte activation in whole-blood cultures involves the measurement of the nucleotide ATP instead of [(3)H]thymidine incorporation. In addition, the Luminetics assay of T-cell activation measures specific T-lymphocyte subset responses through the use of paramagnetic particles coated with monoclonal antibodies against CD antigens. This assay permits rapid (24 h) analysis of lymphocyte subset activation responses to mitogens and recall antigens in small amounts of blood.

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Figures

FIG. 1
FIG. 1
Lymphocyte responses to PHA-L at two time periods (P1 and P2) at an interval of 6 weeks. (A) Proliferative responses of lymphocytes to PHA-L. (B) Measurement of CD4+ T-lymphocyte responses to PHA-L via determination of ATP. SE, standard error.
FIG. 2
FIG. 2
Lymphocyte responses to ConA at two time periods (P1 and P2) at an interval of 6 weeks. (A) Proliferative responses of lymphocytes to ConA. (B) Measurement of CD4+ T-lymphocyte responses to ConA via determination of ATP. SE, standard error.
FIG. 3
FIG. 3
(A) Proliferative responses of lymphocytes to TT before and after vaccine boost, compared by paired-sample t test. (B) Proliferative response of lymphocytes from preboosted and unboosted controls to TT antigen.
FIG. 4
FIG. 4
(A) Measurement of CD4+ T-lymphocyte responses to TT before and after vaccine boost (6-week interval) via determination of ATP (comparisons by paired-sample t test). (B) Measurement of CD4+ T-lymphocyte responses of preboosted and unboosted controls to TT antigen via determination of ATP.
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