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A One-Tube Polymerase Chain Reaction Protocol Demonstrates CC Chemokine Overexpression in Graves’ Disease Glands

Immunology (Y.A., M.S., C.R.-M., A.L.-M., R.P.-B.) and Endocrinology (A.L.-M.) Divisions, University Hospital Germans Trias i Pujol, Badalona, 08916 Barcelona; and Almirall-Prodesfarma Research Center (Y.A., O.D.), 08980 Barcelona, Spain

Address all correspondence and requests for reprints to: Prof. R. Pujol-Borrell, Immunology Unit, University Hospital Germans Trias i Pujol, Carretera. del Canyet s/n, Badalona, 08916 Barcelona, Spain. E-mail: ricardo.pujol{at}uab.es

An adaptation of mixed oligonucleotide primed amplification of complementary DNA to detect the profile of CC chemokines in biological samples is presented. By introducing normalization, two correction coefficients, performing a single amplification reaction, and five parallel hybridizations, intrasample and intersample comparisons can be reliably made. This protocol of single tube PCR CC chemokine profiling was applied to tissue samples from an autoimmune thyroid condition, Graves’ disease, and from a nonautoimmune condition, multinodular goiter. Results demonstrate overexpression of CC chemokines in Graves’ disease, statistically significant for macrophage inflammatory protein-1 and -1ß, which correlated with the aberrant human leukocyte antigen class II expression by thyrocytes, as assessed by flow cytometry. Overexpression of CC chemokines probably plays a major role in determining the characteristics of the lymphocytes migrating to the thyroid gland and influences the course of the disease. The study of chemokine profile should be more informative than the study of isolated chemokines and cytokines, and as it can be applied to fine needle aspiration biopsies, it may be useful to clinical research.

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