Military Strategies for Sustainment of Nutrition and Immune Function in the Field (1999) / Chapter Skim
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10 Application of Whole-Blood Cultures to Field Study Measurements
10 Application of Whole-Blood Cultures to Field Study Measurements
Pages 249-262
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From page 249... ...
This chapter describes the use of whole-blood cultures in evaluating the functional activity of blood lymphoid cells In vitro, with emphasis on the feasibility of their use in field studies. Results are presented from experiments to establish optimal mitogenic lymphocyte proliferation, interleukin production, and release of interleukin receptors in whole-blood cultures.
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From page 250... ...
Using whole-blood cultures has helped to establish associations between suppressed mitogenic proliferative responsiveness of T-lymphocytes in vitro and each of the following: marathon running (Eskola et al., 1978) , intense treadmill exercise (Nieman et al., 1994)
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From page 251... ...
. Table 10-2 lists results of a representative experiment to determine the optimal blood volume needed for maximum mitogenic proliferative responsiveness of blood lymphocytes to PHA in whole-blood cultures.
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From page 252... ...
9 Tabulation of lymphocyte proliferative responsiveness.
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From page 253... ...
plasma. Because whole-blood cultures contain both of these blood components, a series of experiments was conducted to determine the optimal duration of incubation for maximum mitogenic proliferative responsiveness of TABLE 10-3 Duration of Incubation for Mitogenic Proliferative Responsiveness of Lymphocytes to PHA in Whole-Blood Cultures Duration (hours)
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From page 254... ...
for maximum mitogenic proliferative responsiveness of blood lymphocytes cultured for a total of 120 hours ex vivo. PHA-stimulated whole-blood cultures labeled with 3H-thymidine during the final 18 hours of the 120-h incubation showed a higher mean 3H-thymidine incorporation than did those labeled for 3, 6, 12, and 24 hours.
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From page 255... ...
Lymphocyte proliferation in whole-blood cultures is most frequently presented as becquerels of 3H-thymidine incorporated per culture, thus, per volume of blood. However, when the number of absolute lymphocytes is known by automated cell count, the proliferative responsiveness also can be presented as activity per TABLE 10-5 Optimized Duration of Cell Culture Incubation and 3H-Thymidine Labeling for Maximum Proliferative Responsiveness of Lymphocytes to PLEA in Whole-Blood Cultures Hours of 3H-thymidine Incubationt Meant CV§ 18 96 5,961 13 24 96 5,050 16 18 120 4,815 19 24 120 4,672 23 1.0 psi 3H-thymidine.
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From page 256... ...
Elects of Holding and Shipping Blood on Whole-Blood Lymphocyte Proliferation The traditional practice of setting up cell cultures for lymphocyte proliferation within a few hours after blood collection has limited the use of this in vitro test in many studies. In efforts to expand the use of in vitro lymphocyte proliferation tests in field studies, this laboratory has studied the effects of 24-h holding and shipping of blood on PHA-induced lymphocyte proliferation in whole-blood cultures.
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From page 257... ...
maximum proliferative responsiveness of blood lymphocytes to PHA in whole-blood cultures. There was, however, suppressed activity in cultures Dom the blood that had been held for 24 hours and stimulated with suboptimal concentrations of PHA for maximum lymphocyte proliferation in vitro (data not presented)
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From page 258... ...
The mean concentrations of INF-y, IL-10, and sIL-2R were higher in whole-blood cultures stimulated with 16 ,ug than in those stimulated with 32 fig of PHA per culture. It is encouraging TABLE 10-9 Dose-Response Cytokine Production and Release of Soluble Interleukin Receptor Cytokine/Receptor Hours ~gPHA*
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From page 259... ...
Studies are currently under way to determine whether the variation can be reduced, even as cytok~ne production is maintained. AUTHOR'S CONCLUSION As an alternative to the traditional use of PBMCs to determine the functional activity of blood lymphocytes in vitro, the use of whole-blood cultures presents several advantages: they require less blood, less laboratory work time, and less worker expertise, and thus they are more cost effective than are cultures of PBMCs.
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From page 260... ...
1997. Modulated mitogenic proliferative responsiveness of lymphocytes in whole-blood cultures by low carotene diet and mixed-carotenoid supplementation of women.
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From page 261... ...
SYDNE CARLSON-NEWBERRY: What is the number of peripheral blood lymphocytes needed to reliably measure the proliferative responsiveness of lymphocytes in whole-blood cultures? TIM KRAMER: We have not done a definitive study to determine the minimal absolute lymphocyte number needed for reliable lymphocyte proliferation results in whole-blood cultures.
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From page 262... ...
Except for probably the skin test response, the whole- blood culture system is probably the closest that we can get to what is going on in the body. This is one of the reasons that we have been interested in it, because it keeps the milieu constant.
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