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15 July 1994 | Volume 121 Issue 2 | Pages 149-150
Casey and colleagues [1] note the potential importance of measuring cytokine levels in patients with systemic sepsis, whereas Dinarello and Cannon [2] detail many of the pitfalls associated with assaying cytokine levels. We report some other difficulties inherent in measuring "immunoreactive" tumor necrosis factor (TNF)- LETTER
Pitfalls in Measuring Cytokines
TO THE EDITOR: 
by enzyme-linked immunosorbent assay (ELISA). As noted by Casey and colleagues [1], the assessment of TNF- immunoreactivity reflects the level of the biologically active and inactive forms of TNF-. However, the level of immunoreactive TNF- is greatly influenced by the type of monoclonal antibody used in the ELISA kit, as well as by the presence of type I and type II TNF-binding proteins (soluble receptors) in the serum during septic shock [3]. This point is illustrated in Figure 1. In the ELISA kit used by Casey and colleagues [1] (T Cell Science, Cambridge, Massachusetts), the level of TNF- immunoreactivity was masked by 75% and 60%, respectively, in the presence of type 1 TNF-binding proteins and type 2 TNF-binding proteins. In contrast, the level of immunoreactive TNF- measured by an ELISA kit from a second vendor (Genzyme Corp., Cambridge, Massachusetts) was not affected by the type 1 proteins, whereas TNF- immunoreactivity was masked by 80% in the presence of the type 2 proteins. Finally, levels of immunoreactive TNF- measured by an ELISA kit from a third vendor (R & D Systems, Minneapolis, Minnesota) were not masked by either type of protein.
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Although Casey and colleagues [1] have made an important contribution regarding the clinical significance of cytokine profiles in septic shock, it is critically important for the scientific community to standardize the methods used to measure cytokine levels in future trials.
References
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 Top References |
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1. Casey LC, Balk RA, Bone RC. Plasma cytokine and endotoxin levels correlate with survival in patients with the sepsis syndrome. Ann Intern Med. 1993; 119:771-8.
2. Dinarello CA, Cannon JG. Cytokine measurements in septic shock. Ann Intern Med. 1993; 119:853-4.
3. Van Zee KJ, Kohno T, Fischer E, Rock CS, Moldawer LL, Lowry SF. Tumor necrosis factor soluble receptors circulate during experimental and clinical inflammation and can protect against excessive tumor necrosis factor in vitro and in vivo. Proc Natl Acad Sci U S A. 1992; 89:4845-9.
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