Stage determination and therapeutic decision in human African trypanosomiasis: value of polymerase chain reaction and immunoglobulin M quantification on the cerebrospinal fluid of sleeping sickness patients in Côte d'Ivoire

In human African trypanosomiasis (HAT), two disease stages are defined: the first, or haemo‐lymphatic stage, and the second, or meningo‐encephalitic stage. Stage determination forms the basis of therapeutic decision and is of prime importance, as the drug used to cure second‐stage patients has considerable side‐effects. However, the tests currently used for stage determination have low sensitivity or specificity. Two new tests for stage determination in the cerebrospinal fluid (CSF) were evaluated on 73 patients diagnosed with HAT in Côte d'Ivoire. The polymerase chain reaction (PCR) detecting trypanosome DNA (PCR/CSF) is an indirect test for trypanosome detection whereas the latex agglutination test detecting immunoglobulin M (LATEX/IgM) is an indicator for neuro‐inflammation. Both tests were compared with classically used tests, double centrifugation and white blood cell count of the CSF. PCR/CSF appeared to be the most sensitive test (96%), and may be of use to improve stage determination. However, its value for therapeutic decision appears limited, as patients whose CSF was positive with PCR were successfully treated with pentamidine. This result confirms those of previous works that showed that some patients with trypanosomes in the CSF could be treated successfully with pentamidine. LATEX/IgM, which depending on the cut‐off, showed lower sensitivity of 76% and 88%, but higher specificity of 83% and 71% for LATEX/IgM 16 and LATEX/IgM 8 respectively, appears more appropriate for therapeutic decision making.     

Intrathecal immune response pattern for improved diagnosis of central nervous system involvement in trypanosomiasis

Diagnosis of central nervous system (CNS) involvement in human African trypanosomiasis is crucial in determination of therapy. Cerebrospinal fluid (CSF) and serum immunoglobulin concentrations, blood-CSF barrier dysfunction, pattern of intrathecal immunoglobulin synthesis, trypanosome-specific antibody synthesis, and CSF lactate concentrations were analyzed in 272 patients with Trypanosoma brucei gambiense infection. As part of the 2- or 3-class immune response, the predominant intrathecal IgM synthesis was the most sensitive (95%) marker for inflammation of the brain. We propose to replace the World Health Organization (WHO) criteria (white blood cell count >5 cells/microL and presence of trypanosomes in CSF) with a new approach for stage determination in trypanosomiasis: CNS involvement is diagnosed only in patients with >20 cells/microL or with intrathecal IgM synthesis, independent of the presence of trypanosomes in CSF. Compared with the use of these new criteria, the WHO criteria incorrectly classified 49 of 234 patients in the meningoencephalitic stage and 7 of 38 patients in the hemolymphatic disease stage. We also show that trypanosomiasis-related immunoglobulin patterns are of value in differential diagnosis.     

Increased CXCL-13 levels in human African trypanosomiasis meningo-encephalitis

Objectives  To determine the role of the B-cell attracting chemokine CXCL-13, which may initiate B-cell trafficking and IgM production in diagnosing HAT meningo-encephalitis.
Methods  We determined CXCL-13 levels by ELISA on paired sera and CSF of 26 patients from Angola and of 16 controls (six endemic and ten non-endemic). Results were compared to standard stage determination markers and IgM intrathecal synthesis.
Results  CXCL-13 levels in patients' sera had a median value of 386.6 pg/ml and increased levels were associated with presence of trypanosomes in the CSF but not with other stage markers. CXCL-13 levels in patients' CSF had a median value of 80.9 pg/ml and increased levels were associated with all standard stage determination markers and IgM intrathecal synthesis.
Conclusion  CXCL-13 levels in CSF increased significantly during the course of HAT. Hence the value of CXCL-13 for diagnosis, follow-up or as a marker of disease severity should be tested in a well-defined cohort study.     

Comparison of major immunoglobulins intrathecal synthesis patterns in Ecuadorian and Cuban patients with angiostrongyliasis

Angiostrongylus cantonensis meningitis was first reported in Cuba in 1981, and it was recently reported in South America. The aim of this paper is to evaluate the intrathecal immunoglobulin synthesis patterns from Cuba's and Ecuador's patients with angiostrongyliasis; 8 Ecuadorian patients from two different outbreaks and 28 Cuban patients were studied. Simultaneous blood and cerebrospinal fluid samples were taken. Immunoglobulin (Ig) A, IgM, IgG, and albumin were quantified by radial immunodiffusion. Corresponding Reibergrams were applied. A three-Ig pattern was the most frequent in the two groups, but IgM was presented in all Ecuadorian young mature patients; however, in the Cuban children, only 12 of 28 patients had intrathecal IgM, but about 90% had an IgA and IgG synthesis at time of later puncture. This indicates that, with a larger amount of parasites ingested, clinical symptoms are more severe, and a higher frequency of intrathecal IgM synthesis could be observed. This is discussed as a similarity with the intrathecal IgM synthesis in African trypanosomiasis.     

Cerebrospinal fluid IgM, IgA, and IgG indexes in systemic lupus erythematosus. Their use as estimates of central nervous system disease activity

Paired serum and cerebrospinal fluid (CSF) specimens from 13 patients with systemic lupus erythematosus (SLE) and central nervous system involvement (CNS-SLE) were studied for CSF IgM, IgA, and IgG indexes (indicators of intrathecal immunoglobulin synthesis) and CSF-serum albumin quotient (Q albumin) (an indicator of blood-brain-barrier function). We also studied 20 patients with noninflammatory neurologic diseases and seven patients with SLE without CNS involvement for comparison. In addition to an increase in the CSF IgG index, IgM and IgA indexes also were elevated in patients with CNS-SLE. All three indexes decreased significantly when CNS manifestations subsided by successful treatment. The Q albumin was normal in most patients. The elevation of CSF immunoglobulin indexes may be a result of polyclonal B-lymphocyte activation within the CNS, rather than the leak of immunoglobulins from the systemic circulation into the CNS. Since these indexes reflect CNS disease activity in SLE, they may be a successful tool for the management of SLE.     

Laboratory Findings in Tick-Borne Encephalitis – Correlation with Clinical Outcome

Summary Infection with the tick-borne encephalitis virus (TBEV) can result in various neurological complications. At present, there are little data available on laboratory findings that might help predict the clinical course and prognosis of tick-borne encephalitis (TBE). In the present study 100 patients with TBE were examined in respect to various laboratory parameters potentially characteristic for the disease and indicative for the prognosis in TBE. Pleocytosis, impairment of the blood-CSF barrier and intrathecal synthesis of immunoglobulins (IgM > IgG, IgA) were common findings in most patients. On admission to the hospital, 84% of the patients presented with an intrathecal synthesis of TBEV-specific IgM and/or IgG antibodies in the CSF. At follow-up, intrathecal synthesis of TBEV-specific antibodies was demonstrated in all patients studied within 15 days after the first examination, but changes of CSF parameters did not correlate with the clinical course of disease. In contrast to those with moderate courses of disease, patients with severe courses of TBE displayed higher cell counts in the CSF and lower concentrations of neutralizing antibodies in serum, and more frequently revealed an intrathecal synthesis of total IgG. TBE-specific oligoclonal IgG antibodies in the CSF were demonstrated only in three patients with prior, incomplete, vaccination against TBE. The severe course of disease in individual patients with TBE may result from a slow or low production of neutralizing antibodies. In these patients, the more intense damage of the CNS tissue is reflected by higher cell counts in the CSF. At onset of disease the presence of a low concentration of neutralizing antibodies in serum and a high cell count in the CSF might indicate an unfavorable course of TBE. Received: October 20, 1999 · Accepted: January 12, 2000     

Increased intrathecal immunoglobulin synthesis of both kappa and lambda types in patients with systemic lupus erythematosus and central nervous system involvement

The concentration of immunoglobulins in paired serum and cerebrospinal fluid (CSF) specimens from 15 patients with systemic lupus erythematosus (SLE) and central nervous system (CNS) involvement (CNS-SLE) were determined based on their light chain properties. Eight patients with SLE without CNS involvement and 20 patients with noninflammatory neurological diseases were also studied for comparison. We found that both of the CSF Ig-k (immunoglobulin kappa) and Ig-1 (immunoglobulin lambda) indices were significantly elevated, indicating the increased intrathecal immunoglobulin synthesis of both kappa and lambda types in patients with CNS-SLE. Moreover, the CSF Ig-k and Ig-1 indices correlated well with each other, and both indices were found to decrease remarkably when the CNS manifestations subsided after successful treatment. In 4 of 15 patients, intrathecal synthesis of free light chains in addition to Ig bound light chains seemed to be increased. These observations provide substantial evidence for polyclonal B lymphocyte activation within the CNS.     

Antiganglioside antibodies in patients with neuropsychiatric systemic lupus erythematosus.

Antiganglioside antibodies (AGA) were determined in sera and cerebrospinal fluids (CSF) from 50 systemic lupus erythematosus (SLE) patients, and age-matched normal controls. The SLE patients were subdivided according to the type of clinical manifestation into two groups: neuropsychiatric SLE and active SLE without neuropsychiatric manifestation. The presence of these antibodies showed a significant correlation between IgG AGA in the CSF and IgM AGA in the serum and neuropsychiatric SLE. Fifteen patients had this antibody in the CSF without detectable levels in the serum. No correlation was seen between anticardiolipin antibodies in the serum of CSF and neuropsychiatric SLE. The present work suggests that antibodies against gangliosides may be a marker for neuropsychiatric SLE and that intrathecal antibody production can result in the development of this manifestation.     

Evaluation of LATEX/T.b.gambiense for mass screening of Trypanosoma brucei gambiense sleeping sickness in Central Africa

We compared the Card Agglutination Test for Trypanosomiasis (CATT), which consists of lyophilized bloodstream form trypomastigotes of Trypanosoma brucei gambiense (T.b.g.) variable antigen type LiTat 1.3, with LATEX/T.b.g., which consists of a lyophilized suspension of latex particles coated with variable surface glycoproteins of T.b.g. variable antigen types LiTat 1.3, 1.5 and 1.6. This study was carried out during two mass screening surveys in 1998 in Campo, a sleeping sickness focus in Cameroon, with a low prevalence (0.3%) and in 1999 in Batangafo which belongs to the Central African focus of Ouham which has a higher prevalence (3%). In Campo, we compared the CATT performed on whole blood with the LATEX/T.b.g. on diluted blood. In Batangafo, both tests were performed on diluted blood. In all circumstances, the specificity of the LATEX/T.b.g. was higher than of CATT. The use of LATEX/T.b.g. on diluted blood instead of CATT results in an important decrease of workload and as a consequence, of costs related to parasitological examinations. In the case of Campo the workload was up to 12 times less than when using CATT 1.3 on whole blood and the cost divided by 3. In Batangafo the workload was decreased by nearly 20% with the LATEX/T.b.g. Finally, it should be noted that in Batangafo, one of the parasitologically confirmed sleeping sickness patients was negative in CATT and positive in LATEX/T.b.g. and that the reading of the test result in LATEX/T.b.g. is easier than in CATT.     

Cerebrospinal fluid in benign and malignant monoclonal dysglobulinemias

The authors compared 9 cases of myeloma and 2 cases of Waldenstr?m's disease with 10 cases of benign monoclonal gammapathies. None of the patients (except one) had neurological involvement; one patient hab diabetes. The study was focused on the immunoglobulins and proteins in the CSF. The following observations were made: - The CSF protein was raised in 8 out of the 11 malignant gammopathies and in 5 out of the 10 benign gammopathies; - An identical monoclonal protein was found in the CSF and serum in all the cases of malignant gammopathies. In the cases considered to be benign, the results were less concordant: one IgM lymphoma had normal CSF; three IgG dysglobulinemias had different immunoglobulins proteins in their CSF: - The mean CSF immunoglobulin level was much higher in the myeloma patients than in the benign gammopathies; - A number of findings suggest intrathecal secretion (at least in the malignant gammopathies). This is also true in a good number of the benign gammopathy cases. Direct tumoural invasion of the CSF may be more common than generally supposed. However, the study of CSF immunoglobulins alone does not establish the diagnosis of malignant dysglobulinemia. Nevertheless, the authors consider that the finding of intrathecal secretion of immunoglobulin, especially in patients with a discordance in FAB and FC fractions, should lead to a closer follow-up of these patients.     

Cerebrospinal fluid levels of lysozyme, IgM and C-reactive protein in the identification of bacterial meningitis.

Lysozyme (LZM), immunoglobulin M (IgM) and C-reactive protein (CRP) levels were determined in cerebrospinal fluid (CSF) from patients classified on the basis of clinical and laboratory findings into three groups: bacterial meningitis (n = 33), lymphocytic meningitis (n = 21) and controls (n = 54). IgM and CRP levels were determined by enzyme-linked immunosorbent assay (ELISA) and LZM by the lysoplate method. Discriminant analysis demonstrated that 93.94% (31/33) and 96.97% (32/33) of patients with bacterial meningitis were correctly classified on the basis of CSF determinations of IgM and LZM, respectively. However, the measurement of CRP levels in CSF correctly classified 100% of these patients (33/33), thus representing a useful additional marker for the screening of bacterial meningitis. Moreover, no more than 4% (3/75) of patients were incorrectly classified as belonging to the bacterial group on the basis of the CRP test. Thus, CRP titres less than or equal to 80 identify cases belonging to one of the non-bacterial groups, whereas titres greater than or equal to 640 classify the bacterial group, with a very low chance of misclassification. The authors recommend that CSF IgM or LZM levels be also measured for patients with CSF CRP titres of 160 and 320, for a more accurate diagnosis. The probability of these cases being of bacterial aetiology, as calculated from the combined results of these measurements, is presented.     

Involvement of central nervous system disclosing Waldenstrom's disease: demonstration of intrathecal secretion of immunoglobulin M]

We report the case of a 68-year old woman complaining of disorders of memory and persistent headaches in whom the diagnosis of Waldenstr?m's macroglobulinaemia (WM) was made. Computerized tomography of the brain showed a butterfly-shaped hyperdensity in the splenium of the corpus callosum, with ventricular dilatation. Magnetic resonance imaging displayed high-intensity signals on T2-weighted sequences. Protein immunoelectrophoresis elicited an IgM kappa peak. The CSF was found to contain proteins and lymphocytes in excess, and immunohistochemical staining confirmed the predominance of anti-kappa and the presence of intrathecal IgM secretion. Chemotherapy was temporarily effective on the memory disorders, but the patient died 26 months after the beginning of treatment. Central nervous system manifestations are seldom observed in WM, and they are now grouped under the name of Bing-Neel syndrome. Psychic disorders are rarely reported. It is suggested that IgM secretion plays a predominant role in the pathogenesis of leucoencephalitis, and this is supported by the finding of intrathecal IgM synthesis.     

Clinical significance of cerebrospinal fluid beta 2-microglobulin determination in central nervous system leukemia

Cerebrospinal fluid (CSF) beta 2-microglobulin (beta 2-MG) level was measured in 72 healthy subjects and the value (means +/- S means) was found to be 1.26 +/- 0.06 mg/L. The CSF beta 2-MG level in 33 patients who had simple acute leukemia without CNS involvement was 1.46 +/- 0.13 mg/L, which was not significantly different from that in the normal controls (P greater than 0.05). In 25 patients who had leukemia with involvement of CNS, the CSF beta 2-MG level (mean +/- S mean) was 3.94 +/- 0.30 mg/L, which was statistically different from that in the healthy controls (P less than 0.01). It is found that beta 2-MG level in CSF was one of the reliable criteria for diagnosis and indication of intrathecal chemotherapy in CNS leukemia.     

Neopterin concentrations in cerebrospinal fluid and serum of individuals infected with HIV-1

Neopterin, a biochemical marker for the activation of cell-mediated immune reactions, was determined in serum and cerebrospinal fluid (CSF) from patients infected with HIV-1. A significant correlation was found between serum and CSF neopterin concentrations, although concentrations of neopterin in serum were more closely correlated with the clinical severity of HIV-1 infection than those in CSF. However, higher CSF levels were observed in patients with neurologic/psychiatric symptoms than in unaffected patients. Also, quotients of CSF neopterin versus serum neopterin concentrations were increased, indicating intrathecal production of neopterin. Positive HIV-1 isolation from peripheral blood mononuclear cells (PBMC) was associated with higher neopterin concentrations in serum, when compared with negative HIV-1 isolation. Neopterin in CSF appears to be a suitable biochemical marker in patients with HIV-1 infection for detecting overt neurologic/psychiatric disturbances. The data suggest that in HIV-1 infected patients, cell-mediated immune reactions might be activated intrathecally and might contribute to neuropsychiatric disease.     

Laboratory diagnosis of acute measles infections in hospitalized children in Zambia

Summary Laboratory diagnosis of measles infection is rarely performed in developing countries and tends to depend on clinical symptoms alone. We evaluated detection of immunoglobulin M (IgM) antibodies for confirmation of acute measles infection in Zambia. In 149 hospitalized children with clinical diagnosis of measles, IgM antibodies were detected in 88.6% (132/149). The IgM-positive rate increased with time after onset of skin rash and all samples were positive after ₄ days. In addition to IgM antibody test, virus isolations from throat swabs using B₉₅a cells were also performed. These were positive in only 20.9% (14/67), and both IgM and virus isolation in combination increased the positive rate to 92.5% (62/67). Vaccinated children had higher neutralizing (Nt) antibody responses and, among IgM-negative patients, all ₄ vaccinated children had high Nt antibodies while all 10 unvaccinated children had negative or low Nt results. The IgM antibody test was proved to be a sensitive method for laboratory confirmation of measles virus infection in developing countries.     

Improved latex agglutination test for detection of antibodies in serum and cerebrospinal fluid of Trypanosoma brucei gambiense infected patients.

A rapid latex agglutination test (LATEX/T. b. gambiense) for detection of antibodies in patients infected with Trypanosoma brucei gambiense is presented. The reagent is coated with a mixture of three variable surface antigens of bloodstream form trypanosomes. Two hundred and forty sera and 79 CSF samples from patients with parasitologically confirmed trypanosome infection along with 173 sera and 38 CSF samples from non-trypanosomiasis patients have been tested. At 1:16 serum dilution, test specificity was 99%, while sensitivity ranged from 83.8 to 100% depending on the geographical origin of the samples. Undiluted CSF samples from non-trypanosomiasis and from first stage patients scored negative while 42 out of 66 CSF samples from second stage patients were positive. Stability and reproducibility of the lyophilized reagent were excellent.     

384例性病门诊就诊者荧光梅毒螺旋体抗体吸附试验结果分析

目的对384例荧光梅毒螺旋体抗体吸附试验（FTA_ABs）检测结果进行分析,更好地为临床提供梅毒诊断的依据。方法对2012年确诊梅毒并经FTA—ABS检测为阳性的384例患者的血清,同时用梅毒螺旋体明胶颗粒凝集法（TPPA）和酶联免疫吸附试验（ELISA）进行复核检测,并对其中的40例患者进行脑脊液FTAABS检测。结果384例FTAABS检测确诊的阳性病例中,梅毒抗体IgG、IgM同时阳性的59例（占15．4％）,抗体IgG阳性IgM阴性的325例（占84．6％）。40例脑脊液中抗体IgG阳性的12例,未见抗体IgM阳性。梅毒合并感染HIV的患者有29例（占7．6％）,其中RPR滴度〉1：8有21例（72．4％）。单纯梅毒感染的355例（92．4％）,其中RPR滴度〉1：8有97例（27．3%）。结论FTAABS试验敏感性和特异性高,值得在临床中推广。     

几种弓形虫IgM酶联免疫吸附试验的建立及评估

目的　建立和评估几种弓形虫ELISA -IgM的检测方法。 方法　应用酶联免疫吸附试验 (ELISA)的原理 ,建立四种检测方法 ,并进行比较。结果　检测 373例孕妇及正常献血员血清标本 ,方法Ⅰ、方法Ⅱ、方法Ⅲ、方法Ⅳ的阳性率分别为3 8%、3 2 %、2 9%、2 4 %。结论　利用抗人IgM、弓形虫抗原和抗体建立的捕获法具有快速、敏感、特异的特点 ,适合于育龄妇女的筛查和早期诊断。     

Evaluation of Cerebrospinal Fluid for the Presence of Anticardiolipin Antibodies (aCL) in NP-SLE Patients

Many neurological or psychiatric manifestations of SLE (NP-SLE) are related to the presence of anticardiolipin antibodies (aCL) in the patient’s sera. The aim of this study was to evaluate the presence of aCL in cerebrospinal fluid (CSF) in SLE patients with NP features. Fifteen SLE patients were studied, all with NP features. CSF was evaluated for intrathecal IgG synthesis, oligoclonal IgG, and blood–brain barrier impairment. Sera and CSF were tested by ELISA for the presence of aCL-IgG and aCL-IgM with and without β₂ glycoprotein (β₂ GPI) cofactor. CSF and sera of 50 low back pain patients served as controls. Six patients were aCL(+) and nine aCL(–). In all patients the general CSF examination was normal. In all patients the value of indices of intrathecal IgG synthesis were normal but oligoclonal protein was present in the CSF of three patients. In none of the patients was the blood–brain barrier impaired. Neither aCL-IgG nor aCL-IgM was detected in the CSF of any NP-SLE patient. Mean levels of aCL in patients without cofactor β₂ GPI and with cofactor were as follows: for IgG class 0.005 and 0.057 OD (negative); for IgM class 0.004 and 0.024 OD (negative). We could not detect aCL in the CSF of patients with NP-SLE, even if sera were positive for aCL. Received: 6 July 1999 / Accepted: 18 January 2000     

Consecutive evaluation of immunoglobulin M and G antibodies against hepatitis E virus

Hepatitis E virus (HEV) infection is sporadic in the Guangzhou city southern China. However, the evaluation of antibodies to HEV during consecutive time periods after infection has not been reported. We utilized enzyme immunosorbent assay (ELISA) to detect IgM and IgG anti-HEV in consecutive serum specimens from patients with acute hepatitis E and compared that data with detection rates of IgM and IgG anti-HAV in patients with acute hepatitis A. IgM anti-HEV can be detected as early as 4 days after onset of disease symptoms in some patients. The detection rate of IgM anti-HEV is significantly higher in specimens collected within 4 weeks (95%) of onset than in those specimens collected 4 to 18 weeks after onset (67.6%) (P<0.005). IgM anti-HEV had a similar pattern to IgM anti-HAV and can be used as a marker of acute HEV infection. In contrast with IgG anti-HAV, 56.8% of the specimens did not contain detectable levels of IgG anti-HEV (P<0.005). One should be cautioned against making a diagnosis of HEV infection solely by the currently available assays for IgG anti-HEV. In conclusion, IgM anti-HEV can be used as a reliable and sensitive marker for recent HEV infection, but serum specimens should be collected within 4 weeks after onset of symptoms to avoid false-negative results. In contrast, we should be aware of the failure to develop IgG anti-HEV in some patients. These patients carry the risk of reinfection.