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Clinical Chemistry 53: 955-962, 2007. First published March 15, 2007; 10.1373/clinchem.2006.077107
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(Clinical Chemistry. 2007;53:955-962.)
© 2007 American Association for Clinical Chemistry, Inc.

Clinical Immunology

Cytokine Profiling in Human Colostrum and Milk by Protein Array

Miloslav Kverka1,a, Jaroslava Burianova1, Raja Lodinova-Zadnikova2, Ingrid Kocourkova2, Jana Cinova1, Ludmila Tuckova1 and Helena Tlaskalova-Hogenova1

1 Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
2 Institute for the Care of Mother and Child, Prague, Czech Republic.

aAddress correspondence to this author at: Fax 420-24172-1143; e-mail: kverka{at}biomed.cas.cz, immuno{at}biomed.cas.cz.

Background: Human colostrum and milk contain components that influence development. Our aim was to use a protein array to determine the cytokine profile of human lacteal secretions and changes that occur during the early postpartum period.

Methods: We collected 17 samples of colostrum during the first 2 days postpartum and a 2nd group of 5 sets of 2 to 3 sequential colostrum or milk samples (at 20- to 30-h intervals). We analyzed the samples with array membranes consisting of 42 or 79 antibodies directed against cytokines.

Results: In most samples, we detected the previously described cytokines interleukin-8 (IL-8)/CXCL8, epidermal growth factor (EGF), growth-related oncoprotein (GRO)/CXCL1–3, angiogenin, transforming growth factor ß-2, and monocyte chemotactic protein 1 (MCP-1/CCL2). In addition, we found 32 cytokines that have not been described before in colostrum. Cytokine concentrations differed among mothers, and the spectrum of cytokines changed with time after delivery. A significant decrease occurred in IL-12 and macrophage inflammatory protein-1{delta}/CCL15 and a significant increase in MCP-1/CCL2. The production of angiogenin, vascular endothelial growth factor, GRO/CXCL1–3, EGF, and IL-8/CXCL8 remained high throughout. The concentrations of 2 selected cytokines measured with the array technique and ELISA showed moderate to strong correlation (r = 0.63 for EGF and r = 0.84 for IL-8/CXCL8).

Conclusion: Despite the lack of precise quantification, the protein array might be suitable for cytokine screening. It allows simultaneous detection of a broad spectrum of cytokines (including those not described before) in lacteal secretions.




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