Abstract

BackgroundPatients with uncharacteristic inflammatory symptoms such as long-standing fatigue or pain, or a prolonged fever, constitute a diagnostic and therapeutic challenge. The aim of the present study was to determine if an extended immunophenotyping of lymphocytes and monocytes including activation markers can define disease-specific patterns, and thus provide valuable diagnostic information for these patients.MethodsWhole blood from patients with gram-negative bacteraemia, neuroborreliosis, tuberculosis, acute mononucleosis, influenza or a mixed connective tissue disorders, as diagnosed by routine culture and serology techniques was analysed for lymphocyte and monocyte cell surface markers using a no-wash, no-lyse protocol for multi-colour flow cytometry method. The immunophenotyping included the activation markers HLA-DR and CD40. Plasma levels of soluble TNF alpha receptors were analysed by ELISA.ResultsAn informative pattern was obtained by combining two of the analysed parameters: (i), the fractions of HLA-DR-expressing CD4+ T cells and CD8+ T cells, respectively, and (ii), the level of CD40 on CD14+ CD16- monocytes. Patients infected with gram-negative bacteria or EBV showed a marked increase in monocyte CD40, while this effect was less pronounced for tuberculosis, borrelia and influenza. The bacterial agents could be distinguished from the viral agents by the T cell result; CD4+ T cells reacting in bacterial infection, and the CD8+ T cells dominating for the viruses. Patients with mixed connective tissue disorders also showed increased activation, but with similar engagement of CD4+ and CD8+ T cells. Analysis of soluble TNF alpha receptors was less informative due to a large inter-individual variation.ConclusionImmunophenotyping including the combination of the fractions of HLA-DR expressing T cell subpopulations with the level of CD40 on monocytes produces an informative pattern, differentiating between infections of bacterial and viral origin. Furthermore, a quantitative analysis of these parameters revealed the novel finding of characteristic patterns indicating a subacute bacterial infection, such as borreliosis or tuberculosis, or a mixed connective tissue disorder. The employed flow cytometric method is suitable for clinical diagnostic laboratories, and may help in the assessment of patients with uncharacteristic inflammatory symptoms.

Highlights

  • Patients with uncharacteristic inflammatory symptoms such as long-standing fatigue or pain, or a prolonged fever, constitute a diagnostic and therapeutic challenge

  • The aim of the present study is to determine if an extended immunophenotyping of lymphocytes and monocytes, including cellular activation markers, can define disease-specific patterns, and provide valuable diagnostic information for patients with uncharacteristic inflammatory symptoms

  • Several lymphocyte and monocyte subtypes were analysed with regard to their relative frequencies among peripheral blood leucocytes, and for activation status, in order to evaluate if any of these data can be of clinical value

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Summary

Introduction

Patients with uncharacteristic inflammatory symptoms such as long-standing fatigue or pain, or a prolonged fever, constitute a diagnostic and therapeutic challenge. The aim of the present study was to determine if an extended immunophenotyping of lymphocytes and monocytes including activation markers can define diseasespecific patterns, and provide valuable diagnostic information for these patients. The aim of the present study is to determine if an extended immunophenotyping of lymphocytes and monocytes, including cellular activation markers, can define disease-specific patterns, and provide valuable diagnostic information for patients with uncharacteristic inflammatory symptoms. Gram-negative enterobacteriacae strongly stimulate neutrophil phagocytosis and cytokine production by monocytes, in addition to effects on B and CD4+ T lymphocytes [5] Another strong immunostimulator is Epstein-Barr virus (EBV), with as much as 50% of all peripheral blood T cells being specific for this virus during the acute phase of the infection [6]. The relative frequencies of CD4+ T helper cells and CD8+ T cytotoxic cells is known to become altered by many microbes, for example CD4+ T lymphocytopenia has been documented in some cases of tuberculosis [9]

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