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Messenger RNA Expression of IL-8, FOXP3, and IL-12 Differentiates Latent Tuberculosis Infection from Disease¹

Bo Wu^*, Chunhong Huang^*, Midori Kato-Maeda, Philip C. Hopewell, Charles L. Daley^,, Alan M. Krensky^* and Carol Clayberger^2,*

^* Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305; Division of Pulmonary and Critical Care Medicine, University of California, San Francisco, CA 94110; and Division of Mycobacterial and Respiratory Infections, National Jewish Medical and Research Center, Denver, CO 80206

Differentiation of active from latent tuberculosis (TB) is a major challenge in the control of TB. In this study, PBMC from latent TB-infected subjects, TB patients, and tuberculin skin test-negative donors stimulated with the Mycobacterium tuberculosis (Mtb)-specific Ag, early secretory antigenic target 6, and mRNA for 45 immune-related genes was measured by quantitative real-time PCR. Univariate analysis showed significant differences in the expression of 10 genes (IFN-, FOXP3, IL-1, IL-1, IL-2, IL-6, IL-8, IL-12, IL-12, and IL-24) in PBMC from TB patients vs latent TB-infected subjects (p < 0.01). Multivariate logistic regression and classification and regression tree analyses revealed that expression of three genes, IL-8, FOXP3, and IL-12, is predictive for TB vs latent Mtb infection. Thus, measurement of Ag-specific expression of these three genes may offer a specific and noninvasive means of differentiating between latent Mtb infection and TB.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by grants from the National Institutes of Health (RO1 AI051356 to C.C. and RO1 AI034238 to P.C.H.).

² Address correspondence and reprint requests to Dr. Carol Clayberger, Department of Pediatrics, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305. E-mail address: cclay{at}stanford.edu

³ Abbreviations used in this paper: Mtb, Mycobacterium tuberculosis; TB, tuberculosis; LTBI, latent TB infected; Treg, T regulatory cell; TST, tuberculin skin test; PPD, purified protein derivative; BCG, bacille Calmette-Guérin; ESAT-6, early secretory antigenic target 6; CPF-10, culture filtrate protein 10; qPCR, quantitative real-time PCR; CART, classification and regression tree; ROC, receiver-operating-characteristic; AUC, area under the curve; Ct, cycle threshold; CI, confidence interval.