Collection Instructions
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MUST USE FUTHAN COLLECTION KIT
Container/Tube; Lavender Top Tube
Specimen: 1 mL Plasma, EDTA with Futhan preservative
Transport Temperature: Frozen
Collection
Collect EDTA whole blood sample into a chilled lavender-top tube. Immediately add Futhan preservative as directed in the Futhan collection kit (LabCorp N° 78946). Recap lavender tube and invert several times to mix well. Centrifuge the whole blood specimen to separate the plasma. Transfer the plasma into the plastic screw-cap tube that is included in the collection kit. The specimen must be submitted in this transfer tube, which is labeled with the words "Futhan Added." Freeze immediately and maintain frozen until tested.
Patient Preparation
No radioactive isotopes administered within 48 hours prior to venipuncture.
Samples that are not properly collected and stored will produce erroneously elevated results due to in vitro activation. Blood must be drawn directly into tubes containing EDTA at a final concentration of at least 10 mM with the addition of nafamostat mesilate (Futhan, FUT-1750) to further reduce in vitro complement activation.
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Result Interpretation
Males: 152.0–1559.4 ng/mL
Females: 215.7–2025.9 ng/mL
Results for this test are for research purposes only by the assay's
manufacturer. The performance characteristics of this product have
not been established. Results should not be used as a diagnostic
procedure without confirmation of the diagnosis by another medically
established diagnostic product or procedure.
Complement activation can occur through three separate mechanisms. The first mechanism to be discovered, referred to as the classical complement cascade, is activated by antigen-antibody complexes.4-6 This was the basis for the name of this system as they serve to "complement" humoral immunity. Alternatively, the complement cascade can be activated directly by contact with bacterial cell surface molecules, including lipopolysaccharide from gram-negative outer membranes, teichoic acid from gram-positive cell walls, zymosan from fungal and yeast cell walls, and some parasite surface molecules. Recently, a third activation mechanism has been characterized in which mannose-binding lectin synthesized by the liver in response to inflammatory macrophage cytokines stimulates the activation of complement.
Complement cascade activation results in the formation of complement split products C3a, C4a, and C5a.4 These proteins, referred to as anaphylatoxins, facilitate the phagocytosis of immune complexes, viral particles, toxic cell debris and apoptotic corpses. Anaphylatoxins promote an inflammatory response by binding to complement receptors on granulocytes and macrophages.2 Anaphylatoxins also bind to receptors on mast cells, which trigger the release of histamine, increasing blood vessel permeability and smooth muscle contraction.6 They control the local inflammatory response through activation of leukocytes and stimulating their chemotaxis to the site of infection.4
Complement C4a levels can become increased in any condition associated with inflammation. Normal human pregnancy is associated with evidence of complement activation, with an increase in concentrations of the anaphylatoxins C3a, C4a, and C5a in the maternal circulation.7 Levels of anaphylatoxins C3a and C4a have also been found to be elevated in patients with antiphospholipid syndrome relative to healthy controls.8 Ingram and associates have shown that levels of C4a are increased in patients with multiple sclerosis, especially during relapse.
Complement activation split products are present only in trace amounts in normal plasma in vivo.10 It is crucial that samples be collected and stored properly in order to avoid in vitro activation.10 Blood must be drawn directly into tubes containing EDTA at a final concentration of at least 10 mM.10 Citrate and heparin do not block complement activation efficiently and should not be used.10 The addition of nafamostat mesilate (Futhan, FUT-175) further reduces in vitro complement activation.
Limitations
Results for this test are for research purposes only by the assay's manufacturer. The performance characteristics of this product have not been established. Results should not be used as a diagnostic procedure without confirmation of the diagnosis by another medically established diagnostic product or procedure.
Elevation of C4a levels is not predictive of any specific disease. Complement C4a levels can become increased in any condition associated with inflammation.
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