ABSTRACT
Toll-like receptors (TLRs) are implicated in the intracellular killing of Mycobacterium tuberculosis, and their expression is modulated by interleukin-4 (IL-4) in vitro. Our aim was to examine the expression of TLRs at the site of pathology in tuberculous lung granulomas and to explore the effect of the immune response on TLR expression. Immunohistochemistry was performed on lung granulomas from nine patients with tuberculosis undergoing lobectomy for haemoptysis. All nine patients expressed all of the TLRs studied (TLRs 1-5 and 9), whereas only five out of the nine patients had any granulomas positive for IL-4. Statistical analysis of TLR and cytokine staining patterns in 183 individual granulomas from the nine patients revealed significant associations between pairs of receptors and IL-4. A positive association between TLR2 and TLR4 (P < 0.0001) and a negative association between TLR2 and IL-4 (P < 0.0001) was observed. The associations between TLRs 1, 5, and 9 were significantly different in IL-4-negative compared with IL-4-positive patients. In conclusion, TLRs are expressed by various cell types in the human tuberculous lung, and their expression patterns are reflected by differences in the immune response.
Subject(s)
Interleukin-4/metabolism , Lung/metabolism , Membrane Glycoproteins/metabolism , Receptors, Cell Surface/metabolism , Tuberculosis, Pulmonary/metabolism , Adolescent , Adult , Antibody Specificity , Biomarkers , DNA-Binding Proteins/immunology , DNA-Binding Proteins/metabolism , Female , Granuloma, Respiratory Tract/metabolism , Granuloma, Respiratory Tract/pathology , Humans , Immunohistochemistry/methods , Interleukin-4/immunology , Lung/pathology , Male , Membrane Glycoproteins/immunology , Middle Aged , Necrosis , Receptors, Cell Surface/immunology , Toll-Like Receptor 2 , Toll-Like Receptor 4 , Toll-Like Receptor 9 , Toll-Like Receptors , Tuberculosis, Pulmonary/immunology , Tuberculosis, Pulmonary/pathologyABSTRACT
The use of large-scale microarray expression profiling to identify predictors of disease class has become of major interest. Beyond their impact in the clinical setting (i.e. improving diagnosis and treatment), these markers are also likely to provide clues on the molecular mechanisms underlining the diseases. In this paper we describe a new method for the identification of multiple gene predictors of disease class. The method is applied to the classification of two forms of arthritis that have a similar clinical endpoint but different underlying molecular mechanisms: rheumatoid arthritis (RA) and osteoarthritis (OA). We aim at both the classification of samples and the location of genes characterizing the different classes. We achieve both goals simultaneously by combining a binary probit model for classification with Bayesian variable selection methods to identify important genes.We find very small sets of genes that lead to good classification results. Some of the selected genes are clearly correlated with known aspects of the biology of arthritis and, in some cases, reflect already known differences between RA and OA.
ABSTRACT
The resistance of HIV clinical isolates with or without M184V was analysed in relation to plasma HIV-1-RNA level and time on therapy. The number of thymidine analogue mutations (TAMs) was lower in isolates with M184V, this was independent of plasma HIV-1-RNA level and time on therapy for T215F/Y, D67N and L210W. This suggests a direct effect of M184V on the reduced selection of TAMs. Lamivudine use was significantly associated with lower median fold resistance to zidovudine and stavudine.
Subject(s)
Drug Resistance, Microbial/genetics , Mutation , Reverse Transcriptase Inhibitors/pharmacology , Stavudine/pharmacology , Thymidine/analogs & derivatives , Zidovudine/pharmacology , Genotype , HIV Infections/drug therapy , HIV Infections/virology , HIV-1/drug effects , Humans , Phenotype , Retrospective Studies , Reverse Transcriptase Inhibitors/therapeutic use , Stavudine/therapeutic use , Viral Load , Zidovudine/therapeutic useABSTRACT
In order to examine the immune response at the site of pathology in tuberculosis, we analysed cytokines present in lung granulomas, their associations with each other and with caseous necrosis as well as the phenotype of the cellular infiltrate. Paraffin-embedded tissue from the lungs of seven patients with pulmonary tuberculosis was analysed by immunohistochemistry and in situ hybridization to detect interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4) proteins and IL-12p40 mRNA. All seven patients had granulomas staining positive for IFN-gamma, TNF-alpha and IL-12p40, but only four stained positive for IL-4. Cells with the morphology of lymphocytes, macrophages and giant cells expressed TNF-alpha, IFN-gamma and IL-4 protein. Furthermore, CD68-positive myeloid cells expressed IL-12p40 mRNA, as expected, but a subset of CD3-positive lymphocytes also expressed this mRNA. These lymphocytes producing IL-12p40 also stained positive for CD8 but not CD4. A total of 141 granulomas were scored for the presence or absence of cytokine or necrosis and two major associations were identified. The first association was between IFN-gamma and IL-12, with 76% of granulomas staining positive for both cytokines. Unexpectedly, those granulomas positive for IL-4 were always positive for IFN-gamma. The second association was between TNF-alpha and caseous necrosis, where all necrotic granulomas were TNF-alpha positive. This association was modulated by IL-4. Therefore, heterogeneity of cellular infiltrate and cytokine expression is observed between adjacent granulomas in the same patient.
