Mucous peristalsis, mucus and immunity proteins, such as lysozyme and lactoferrin, are part of humoral innate immunity. The aim of this study was to develop a quantitative method, a time-resolved-immunofluorometric assay, to measure lysozyme and lactoferrin in sera, saliva, stools and cervico-vaginal secretions. This method was validated in 51 healthy subjects. Linearity for lysozyme was between 1.02 and 25 μg/l and for lactoferrin between 1.02 and 100 μg/. The detection limit was 0.5 μg/l for lysozyme and 1 μg/l for lactoferrin. Albumin and α1-antitrypsin were measured by immuno-nephelometry to calculate salivary, intestinal and cervico-vaginal coefficients of excretion. Lysozyme and lactoferrin were present in all types of mucosal surfaces. Very high concentrations of lysozyme and lactoferrin were found in cervico-vaginal fluid (166.2 and 72.7 mg/l, respectively), compared to the concentrations found in the other mucosal fluids. Lysozyme in stools was produced at the rate of 0.42 mg/d compared to 0.02 mg/d lactoferrin production. Lysozyme and lactoferrin greatly exceeded the values expected from the molecular weight-affected seepage from plasma, suggesting primarily local synthesis in healthy subjects. Quantitative measurement of lysozyme and lactoferrin can aid in the assessment of the activity of mucus-associated lymphoid tissues in innate immunity, and can help in further understanding of the role of these proteins in mucosal diseases.
The aim of this study was to explore anti-Candida albicans systemic and mucosal humoral responses against Candida virulence antigens such as somatic antigen and secreted aspartic proteases (Saps) in HIV-infected patients with oral candidiasis.
Twenty-eight subjects were included in the study: 11 HIV-positive patients without oral candidiasis (group A), 6 HIV-positive patients with oral candidiasis (group B) and 11 HIV-negative healthy controls (group C). Total IgA, IgG and IgM concentrations and antibodies to C. albicans (somatic antigen, Sap1, Sap6) were measured in serum and saliva. We developed a time-resolved immunofluorometric assay with biotin and europium-labeled streptavidin for this purpose.
Salivary total IgA, IgG and IgM concentrations were higher in group B. IgA, IgG and IgM anti-C. albicans antibodies (against somatic antigen, Sap1, Sap6) were higher in saliva and serum from patients from group B compared with patients from group A and controls.
Our results suggest that, in oral candidiasis, HIV-infected patients have a high mucosal response, specifically directed against C. albicans virulence antigens, such as somatic antigen, Sap1 and Sap6.
The aim of this study was to explore lysozyme and lactoferrin concentrations in human immunodeficiency virus (HIV)-infected patients with oropharyngeal candidiasis (OPC). These proteins were measured by time-resolved immunofluorometric assay, validated in Part I of this study, in paired serum and salivary secretions of 30 patients. Eleven HIV-positive patients without OPC, eight HIV-positive patients with OPC and eleven HIV-negative healthy subjects were included in the study. The relative coefficient of excretion of salivary albumin was used to establish protein origin. In serum, the low lactoferrin concentrations in HIV-infected patients with and without OPC (0.610 mg/l (p < 0.05) and 0.896 mg/l (p < 0.01) vs. 1.439 mg/l in healthy subjects) were probably due to a decrease in nonspecific immunity, particularly the polymorphonuclear cells. In HIV-infected patients with OPC, the high salivary lysozyme and lactoferrin concentrations (170.94 mg/l and 66.48 mg/l vs. 23.35 mg/l and 10.20 mg/l in healthy subjects, respectively) and their mean relative coefficient of excretion of above 1 indicated a high local production of lysozyme and lactoferrin in saliva. The development of OPC in HIV-infected patients could be a consequence of inefficient lysozyme and lactoferrin concentrations and of decreased cooperation between innate and adaptative immune systems.