Optimization of the weck-Cel collection method for quantitation of cytokines in mucosal secretions.
Measurement of immune components in mucosal secretions is important for the evaluation of local immunity at the mucosal surfaces. The Weck-Cel ophthalmic sponge provides a method for the collection of these secretions. The sponge absorbs a relatively large volume of material, therefore allowing for quantitation of multiple immune components. Additionally, it provides a method in which the same device may be used to collect specimens from different mucosal sites, such as the genital tract and oral cavity. This sampling technique has successfully been applied for collection and measurement of antibody in oral and genital tract secretions. The purpose of this work was to optimize the extraction of protein from the sponge matrix. Of particular interest was the recovery of cytokines from the sponge. Satisfactory recovery of the cytokines interleukin 1beta (IL-1beta), IL-2, IL-5, IL-12, IL-6, IL-8, IL-10, and granulocyte-macrophage colony-stimulating factor was obtained. However, IL-4 and gamma interferon recovery rates remained low. Using an alteration of the published extraction method, cytokine concentrations were measured in cervical secretions from women using oral contraceptives. The data revealed detectable concentrations of IL-6, IL-10, IL-8, and IL-12 on cycle days 9 and 20. The proposed technique provides an easy, practical, and consistent method for collection of nonconventional body fluids, such as cervicovaginal fluids and saliva, for the assay of immunoglobulins and several cytokines.
Rohan
L C
LC
Department of Obstetrics, Gynecology and Reproductive Sciences, Magee-Womens Research Institute, University of Pittsburgh, Pittsburgh, Pennsylvania.
Edwards
R P
RP
Kelly
L A
LA
Colenello
K A
KA
Bowman
F P
FP
Crowley-Nowick
P A
PA
D000328:Adult; D002584:Cervix Uteri / Q000378:metabolism; D016207:Cytokines / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D005836:Genitalia, Female / Q000378:metabolism*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D007378:Interleukins / Q000032:analysis / Q000302:isolation & purification*; D009092:Mucous Membrane / Q000378:metabolism; D012463:Saliva / Q000737:chemistry*; D013048:Specimen Handling / Q000379:methods*; D015503:Surgical Sponges; D014621:Vagina / Q000378:metabolism
D016207:Cytokines; D007378:Interleukins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor