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1.
Eur J Neurol ; 10(6): 711-9, 2003 Nov.
Article in English | MEDLINE | ID: mdl-14641518

ABSTRACT

Quantitative and qualitative techniques for assessment of the intrathecal humoral immune response in human African trypanosomiasis were compared, and their diagnostic potential for detection of the meningo-encephalitic stage of the disease was evaluated. Total and trypanosome specific immunoglobulin G (IgG) and IgM intrathecal synthesis were studied in paired cerebrospinal fluid (CSF) and blood samples of 38 trypanosomiasis patients and in three controls using Reiber's formulae. The presence of CSF-specific oligoclonal IgG and of trypanosome-specific antibodies was determined using iso-electric focusing followed by immunoblotting and antigen-driven immunoblots. The intrathecal IgG fraction (16% positive) and oligoclonal IgG detection (24% positive) were insensitive for detection of an intrathecal humoral immune response. Trypanosome-specific IgG synthesis, reflected by the IgG antibody index (AI) (26% positive), was confirmed by the presence of oligoclonal specific IgG (47% positive), but the latter was more sensitive. Although the detection technique failed for oligoclonal IgM, the intrathecal IgM fraction (42% positive) and the IgM AI (32% positive) indicated that the meningo-encephalitic stage of the disease is characterized by a dominant intrathecal IgM response, which was higher than the IgG response. The highest combination of diagnostic sensitivity and specificity for the meningo-encephalitic stage of trypanosomiasis was observed for quantitative IgM determinations.


Subject(s)
Trypanosomiasis, African/cerebrospinal fluid , Trypanosomiasis, African/immunology , Albumins/cerebrospinal fluid , Animals , Antibody Formation/immunology , Antibody Specificity , Cote d'Ivoire , Disease Progression , Enzyme-Linked Immunosorbent Assay , Humans , Immunoblotting , Immunoglobulin G/cerebrospinal fluid , Immunoglobulin M/cerebrospinal fluid , Parasite Egg Count , Trypanosoma brucei gambiense/immunology , Trypanosomiasis, African/diagnosis
2.
Brain Res Bull ; 61(3): 299-308, 2003 Aug 15.
Article in English | MEDLINE | ID: mdl-12909300

ABSTRACT

Polymerase chain reaction assays are a powerful tool for detecting the presence of infectious genomes in the cerebrospinal fluid. Positive results always mean a current or pending infection of the central nervous system. Subacute (>7 days) or chronic infections induce an intrathecal humoral immune response and the appearance of oligoclonal IgG antibodies directed against the causal infectious agent. This local synthesis may be observed even in cases of severe systemic immunodeficiency. The use of polymerase chain reactions in combination with the detection of a specific intrathecal immune response should represent the most reliable strategy for the diagnosis of viral and chronic infections of the central nervous system. The authors describe their experience, using this approach, in herpetic encephalitis, acute and recurrent herpetic meningitis, varicella zoster-induced neurological diseases, cytomegalovirus encephalitis, progressive multifocal leukoencephalitis and tuberculous meningitis.


Subject(s)
Central Nervous System Infections/cerebrospinal fluid , Central Nervous System Infections/diagnosis , Immunoblotting , Polymerase Chain Reaction , Aged , Aged, 80 and over , Central Nervous System Infections/immunology , Cytomegalovirus/genetics , Cytomegalovirus/isolation & purification , Cytomegalovirus Infections/cerebrospinal fluid , Cytomegalovirus Infections/diagnosis , DNA Primers , Diagnosis, Differential , Encephalitis, Herpes Simplex/cerebrospinal fluid , Encephalitis, Herpes Simplex/diagnosis , Encephalitis, Varicella Zoster/cerebrospinal fluid , Encephalitis, Varicella Zoster/diagnosis , Female , Herpesvirus 3, Human/genetics , Herpesvirus 3, Human/isolation & purification , Humans , Immunoblotting/methods , Immunoglobulin G/cerebrospinal fluid , Infant, Newborn , JC Virus/genetics , JC Virus/isolation & purification , Leukoencephalopathy, Progressive Multifocal/cerebrospinal fluid , Leukoencephalopathy, Progressive Multifocal/diagnosis , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Polymerase Chain Reaction/methods , Sensitivity and Specificity , Simplexvirus/genetics , Simplexvirus/isolation & purification , Tuberculosis, Meningeal/cerebrospinal fluid , Tuberculosis, Meningeal/diagnosis
3.
Clin Chem Lab Med ; 39(4): 333-40, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11388658

ABSTRACT

In normal conditions, albumin and immunoglobulin (Ig)G in the cerebrospinal fluid (CSF) originate from the blood, and there is no antibody production within the central nervous system. Up to 20% of CSF proteins are intrathecally synthesized, but the major fraction is blood-derived. The CSF/serum albumin quotient (QAlb) is the best marker of the blood-CSF barrier function. The corresponding immunoglobulin quotients (QIGG, QIGA, QIGM) are not linearly related to QAlb and their correlations are defined by an hyperbolic equation. This equation is used to discriminate between a blood-derived and a locally produced fraction of immunoglobulins in case of an intrathecal humoral immune response. The detection of CSF-specific oligoclonal IgG is more sensitive than the quantitative comparison between QIGG and QAlb. A further step is the determination of antibody indices and the detection of specific oligoclonal antibodies by antigen-driven immunoblots. CSF analysis remains a cornerstone for the diagnosis of various neurological disorders, including multiple sclerosis and infectious diseases of the central nervous system.


Subject(s)
Cerebrospinal Fluid/chemistry , Cerebrospinal Fluid/metabolism , Chemistry, Clinical/methods , Immunoglobulins/blood , Immunoglobulins/cerebrospinal fluid , Central Nervous System/metabolism , Humans , Immunoblotting , Immunoglobulin G/metabolism , Models, Statistical , Polymerase Chain Reaction , Sensitivity and Specificity
4.
Acta Neurol Belg ; 99(1): 11-20, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10218088

ABSTRACT

We amplified the mRNA for cytokines in peripheral blood mononuclear cells (PBMC) and cerebrospinal fluid (CSF) cells from 18 multiple sclerosis (MS) patients and 21 other neurological patients, using the reverse transcription polymerase chain reaction (RT-PCR). Radioactive hybridization of the amplified DNA allowed quantitation of mRNA levels. Expression of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma and interleukin (IL)-10 mRNA was elevated in CSF cells from MS patients. IFN-gamma and IL-10 mRNA levels were higher in MS patients than in other inflammatory neurological diseases. mRNA coding for transforming growth factor (TGF)-beta was detectable in the majority of cases, with higher expression in CSF cells of MS and other inflammatory neurological diseases than in noninflammatory controls, and higher expression in PBMC of MS patients than in all other cases. In many MS patients both proinflammatory and immunoregulatory cytokine messages were detected in the CSF compartment without correlation with the clinical activity of the disease. In contrast, mRNA for the costimulatory molecule B7.1 was only detected in the CSF cells of some MS patients, who showed clinical signs of acute relapse at the time of the spinal tap.


Subject(s)
Cytokines/metabolism , Leukocytes, Mononuclear/metabolism , Multiple Sclerosis/metabolism , Cytokines/blood , Cytokines/cerebrospinal fluid , Humans , Interferon-gamma/metabolism , Interleukins/metabolism , Multiple Sclerosis/blood , Multiple Sclerosis/cerebrospinal fluid , Reverse Transcriptase Polymerase Chain Reaction , Stimulation, Chemical , Transforming Growth Factor beta/metabolism , Tumor Necrosis Factor-alpha/metabolism
5.
J Neuroimmunol ; 76(1-2): 100-4, 1997 Jun.
Article in English | MEDLINE | ID: mdl-9184638

ABSTRACT

The intrathecal synthesis of antibodies against recombinant VP1, the major structural protein of JC virus (JCV), was studied in 18 patients with progressive multifocal leukoencephalopathy (PML) and in 31 patients with various neurological disorders. Two methods were used, the calculation of an antibody specific index (ASI) on one hand and an antigen-driven immunoblotting for the detection of oligoclonal antibodies on the other. Most PML patients displayed an elevated (> 1.5) ASI (78%) and anti-VP1 oligoclonal antibodies restricted to the cerebrospinal fluid (55%). Only two other patients (one case each of multiple sclerosis and of neuroborreliosis) also showed an intrathecal synthesis of anti-VP1 oligoclonal antibodies, likely as a result of a 'polyspecific' reaction within the central nervous system.


Subject(s)
Antibodies, Viral/cerebrospinal fluid , JC Virus/immunology , Leukoencephalopathy, Progressive Multifocal/immunology , Viral Structural Proteins/immunology , Enzyme-Linked Immunosorbent Assay , Humans , Immunoblotting , Recombinant Proteins/immunology
6.
Clin Diagn Lab Immunol ; 1(3): 353-6, 1994 May.
Article in English | MEDLINE | ID: mdl-7496976

ABSTRACT

Tuberculous meningitis cases were analyzed by an immunoblotting test based on Mycobacterium bovis BCG antigen complex A60. Anti-A60 immunoglobulin G (IgG) in cerebrospinal fluid (CSF) allowed early diagnosis, and concentrations decreased after recovery. In primary meningitis forms, anti-A60 IgGs were intrathecally synthesized and specific oligoclonal IgGs were present in CSF. In meningeal complications of pulmonary tuberculosis, there were matching titers of anti-A60 IgG in blood and CSF (mirror pattern). Correlation between CSF-restricted patterns and CSF pleocytosis was shown.


Subject(s)
Antigens, Bacterial/blood , Antigens, Bacterial/cerebrospinal fluid , Mycobacterium bovis/immunology , Tuberculosis, Meningeal/cerebrospinal fluid , Tuberculosis, Meningeal/immunology , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Immunoblotting , Infant , Male , Middle Aged
7.
J Neurol Neurosurg Psychiatry ; 53(8): 662-6, 1990 Aug.
Article in English | MEDLINE | ID: mdl-2120390

ABSTRACT

Cerebrospinal fluid (CSF) and serum samples from eight patients with bacteriologically proven (6) or clinically suspected (2) tuberculous meningitis were tested for the presence of anti-mycobacterial IgG antibodies by an affinity-mediated immunoblot technique. This technique is based on agarose gel isoelectric focusing of paired CSF and serum samples diluted to the same IgG concentration, and transfer of the specific IgG antibodies onto mycobacterial antigen-loaded nitrocellulose sheets. An intrathecal synthesis of anti-mycobacterial oligoclonal IgG antibodies, often superimposed on diffuse polyclonal production was shown in all patients but not in patients with tension headache or other neurological disorders. Similar results were obtained when a purified mycobacterial antigen, A60, was used for coating the nitrocellulose sheets in place of a whole mycobacterial homogenate, indicating that A60 was a major immunogen. The number of anti-mycobacterial oligoclonal IgG bands increased with time, and persisted for years even in clinically cured patients. Some IgG bands had no detectable anti-mycobacterial activity, at least with the antigens preparations used in this study. The demonstration of such anti-mycobacterial IgG bands in the CSF could be a useful adjunct for the diagnosis of tuberculous meningitis, especially in the case of negative cultures.


Subject(s)
Antibodies, Bacterial/cerebrospinal fluid , Antigens, Bacterial , Heat-Shock Proteins , Mycobacterium tuberculosis/immunology , Tuberculosis, Meningeal/immunology , Adolescent , Adult , Bacterial Proteins/immunology , Child , Child, Preschool , Female , Follow-Up Studies , Humans , Immunoblotting , Immunoglobulins/cerebrospinal fluid , Infant , Male , Middle Aged , Oligoclonal Bands , Tuberculosis, Meningeal/diagnosis
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