RESUMEN
BACKGROUND: Diagnosis of tuberculous meningitis (TBM) is difficult. Rapid confirmatory diagnosis is essential to initiate required therapy. There are very few published reports about the diagnostic significance of 65 kD heat shock protein (hsp) in TBM patients, which is present in a wide range of Mycobacterium tuberculosis species and elicits a cellular and humoral immune response. In the present study we have conducted a prospective evaluation for the demonstration of 65 kD hsp antigen in cerebrospinal fluid (CSF) of TBM patients, by indirect ELISA method using monoclonal antibodies (mAb) against the 65 kD hsp antigen, for the diagnosis of TBM. METHODS: A total of 160 CSF samples of different groups of patients (confirmed TBM {n = 18}, clinically suspected TBM {n = 62}, non TBM infectious meningitis {n = 35} and non-infectious neurological diseases {n = 45}) were analyzed by indirect ELISA method using mAb to 65 kD hsp antigen. The Kruskal Wallis test (Non-Parametric ANOVA) with the Dunnett post test was used for statistical analysis. RESULTS: The indirect ELISA method yielded 84% sensitivity and 90% specificity for the diagnosis of TBM using mAb to 65 kD hsp antigen. The mean absorbance value of 65 kD hsp antigen in TBM patients was [0.70 +/- 0.23 (0.23-1.29)], significantly higher than the non-TBM infectious meningitis group [0.32 +/- 0.14 (0.12-0.78), P < 0.001] and also higher than the non-infectious neurological disorders group [0.32 +/- 0.13 (0.20-0.78), P < 0.001]. A significant difference in the mean absorbance of 65 kD hsp antigen was noted in the CSF of culture-positive TBM patients [0.94 +/- 0.18 (0.54-1.29)] when compared with clinically suspected TBM patients [0.64 +/- 0.20 (0.23-0.98), P < 0.05]. CONCLUSION: The presence of 65 kD hsp antigen in the CSF of confirmed and suspected cases of TBM would indicate that the selected protein is specific to M. tuberculosis and could be considered as a diagnostic marker for TBM.
Asunto(s)
Proteínas Bacterianas/líquido cefalorraquídeo , Líquido Cefalorraquídeo/metabolismo , Líquido Cefalorraquídeo/microbiología , Chaperoninas/líquido cefalorraquídeo , Tuberculosis Meníngea/líquido cefalorraquídeo , Tuberculosis Meníngea/diagnóstico , Biomarcadores/líquido cefalorraquídeo , Líquido Cefalorraquídeo/química , Chaperonina 60 , Diagnóstico Diferencial , Ensayo de Inmunoadsorción Enzimática/economía , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/normas , Humanos , Meningitis Aséptica/líquido cefalorraquídeo , Meningitis Aséptica/diagnóstico , Meningitis Viral/líquido cefalorraquídeo , Meningitis Viral/diagnóstico , Mycobacterium tuberculosis/metabolismo , Valor Predictivo de las Pruebas , Estudios Prospectivos , Tuberculosis Meníngea/microbiologíaRESUMEN
Although systemic immune reactivity to 65-kD mycobacterial hsp65 (m-hsp65) has been shown previously in Behçet's disease (BD), local immune response was not investigated. We studied anti-m-hsp65 IgG, IgM and IgA antibodies in the serum and cerebrospinal fluid (CSF) of 25 BD patients with cerebral parenchymal involvement (p-NBD), seven BD patients with intracranial hypertension (ih-NBD), eight BD patients without central nervous system (CNS) involvement, 30 patients with multiple sclerosis (MS) and 24 patients with non-inflammatory CNS disorders (NIC). Significantly higher CSF IgG responses were detected in p-NBD patients (ELISA ratio 1.3 +/- 0.9) compared with NIC (0.7 +/- 0.4, P < 0.01). In p-NBD patients' IgG, IgM or IgA CSF anti-m-hsp65 positivity rate was 48% (12/25); this was significantly higher when compared with MS (3/30; P < 0.03) and NIC (3/24; P < 0.01). CSF anti-m-hsp65 IgG ratios correlated with the duration of BD (r = 0.4, P < 0.04) but not with the duration of neurological involvement. Serum IgM and IgA responses were elevated in ih-NBD, suggesting a different type of involvement than p-NBD. These results implicate an increased local humoral response to m-hsp65 in the CSF of p-NBD patients, which might be related to the pathogenesis of neurological involvement.
Asunto(s)
Antígenos Bacterianos/líquido cefalorraquídeo , Antígenos Bacterianos/inmunología , Proteínas Bacterianas , Síndrome de Behçet/líquido cefalorraquídeo , Síndrome de Behçet/inmunología , Chaperoninas/líquido cefalorraquídeo , Chaperoninas/inmunología , Adulto , Formación de Anticuerpos , Chaperonina 60 , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina A/análisis , Inmunoglobulina G/análisis , Masculino , Persona de Mediana EdadRESUMEN
Parkinson's disease (PD) is an age-related neurodegenerative movement disorder of unknown etiology. In PD immune abnormalities were reported, but the cause of such abnormalities has not been resolved. Recently, the increased proportion of gamma delta + T cells out of all T cells has been found in patients with PD. Heat shock proteins (hsps) could be targets for gamma delta + T lymphocytes. We examined serum and CSF of patients with PD, age-matched patients with other non-inflammatory neurological diseases (OND old), young patients with other non-inflammatory neurological diseases (OND young), and donors of blood (DB). Antibodies were detected using the enzyme-linked immunosorbent assay. The plates were coated with recombinant mycobacterial hsp 65 and hsp 70. The present study showed that the mean ELISA ratio of CSF from patients with PD was significantly greater than that of CSF from patients with OND old (tested against IgG anti-hsp 65 and IgG anti-hsp 70) and OND young (tested against IgG anti-hsp 70). There was no difference between the mean ELISA ratio of sera from patients with PD, OND old and OND young (tested against IgG anti-hsp 65 and IgG anti-hsp 70). The significance of hsps immunity is not completely clear. Increased hsps expression, which is induced by stress, provides cells with protection against the environmental insults. Alternatively, the antibodies may be present as a consequence of prior infections.