Extensive reinnervation of the hippocampus by embryonic basal forebrain cholinergic neurons grafted into the septum of neonatal rats with selective cholinergic lesions

The immunodominant surface antigen of Toxoplasma gondii, surface antigen 1 (SAG1), was expressed in Escherichia coli as a fusion protein containing a majority of the SAG1 protein supplied with six histidyl residues in the N-terminal end. The recombinant protein was purified on a Ni-chelate column and then on a fast-performance liquid chromatography column and was in a nonreduced condition. It was recognized by T. gondii-specific human immunoglobulin G (IgG) and IgM antibodies as well as by a mouse monoclonal antibody (S13) recognizing only nonreduced native SAG1. Antibodies induced in mice by the recombinant SAG1 recognized native SAG1 from the T. gondii RH isolate in culture. Recombinant SAG1 is suitable for use in diagnostic systems for detecting anti-SAG1-specific IgG and IgM antibodies.     

Serologic reactivity to purified recombinant and native 29-kilodalton peripheral membrane protein of pathogenic Entamoeba histolytica

The 29-kDa peripheral membrane protein of Entamoeba histolytica has recently been demonstrated to have epitopes on pathogenic clinical isolates which were not detected by monoclonal antibodies on nonpathogenic isolates. To analyze the serological response to this protein, we tested 93 serum specimens (from 33 patients with amebic liver abscess, 7 patients with colitis, 2 patients with ameboma, 18 individuals harboring a nonpathogenic zymodeme strain, 10 healthy Mexican migrant workers, and 23 healthy controls) by enzyme-linked immunosorbent assay (ELISA) using immunoaffinity-purified native or recombinant protein. When tested by ELISA with the native antigen, 79% (26 of 33) of the serum specimens from patients with amebic liver abscess, 4 of 9 serum specimens from symptomatic patients with colitis or ameboma, and serum from one migrant worker were positive. None of the 18 subjects harboring a nonpathogenic strain or 23 control individuals were seropositive to the native antigen (sensitivity, 71%; specificity, 98%). Of 30 serum specimens from patients with amebic liver abscess tested with recombinant antigen, 27 were seropositive (90%). In addition, six patients with colitis or ameboma and two individuals who harbored a nonpathogenic strain were seropositive to the recombinant antigen. One healthy Mexican migrant worker tested positive by both ELISAs (sensitivity, 87%; specificity, 94%). Immunoblotting of 51 serum specimens to sodium dodecyl sulfate-denatured native 29-kDa protein was less sensitive (65%) than ELISA in detecting serum antibodies to the antigen. These results suggest a similar antibody response to native and recombinant antigens (r = 0.86) and support the potential utility of a quantitative assay with defined recombinant antigen for the serodiagnosis of invasive amebiasis in nonendemic areas in conjunction with other diagnostic tools.     

Capillary haemangiomas in association with morning glory disc anomaly

The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.     

Chlamydia trachomatis antibody detection and diagnosis of reactive arthritis

OBJECTIVE: To investigate whether determining the presence of serum or synovial fluid (SF) IgG and IgA of anti-Chlamydia antibodies with two recent commercially available enzyme-linked immunosorbent assays (ELISA) using synthetic peptides or recombinant antigen could be helpful to detect possible Chlamydia trachomatis (CT)-involved disease in rheumatological patients without evidence of urogenital CT infection. METHODS: The prevalence of such antibodies was determined in samples from patients with well-defined disease, i.e. CT sexually acquired arthritis and from patients with other inflammatory arthropathies unrelated to CT. RESULTS: When considering IgG and/or IgA anti-MOMP or anti-LPS antibodies, a sensitivity of 100% was obtained for serum and SF samples, but with a low specificity. A sensitivity and a specificity equal or close to 80% were observed for the SF IgG anti-MOMP antibodies. CONCLUSION: Clinically, the most appropriate determination was the SF IgG anti-MOMP antibodies. This commercially available ELISA test could be useful for the diagnosis of probable CT reactive arthritis.     

Fine tuning of epitopic dominance induced by lung cancer on the IgG response to bovine betalactoglobulin: towards a paraneoplastic immune marker

BACKGROUND: Investigating the humoral immune response to mucosal antigens in patients with lung cancer, we have documented a preferential immunoglobulin G (IgG) binding to cryptic epitopes unmasked by the proteolysis of bovine beta-lactoglobulin (BLG). In contrast, IgG from healthy controls and patients with chronic bronchitis (COPD) bind preferentially to continuous epitopes presented on both native (n) and denaturated (d) forms of this antigen. The present study further characterized the differences in the epitope profiles recognized on BLG. METHODS: The capacity of individual sera from 65 lung cancer patients, tested before and after cancer removal for the patients with early stage lung carcinoma, 65 healthy controls, and 52 patients with COPD, to prevent the binding of pooled IgG fractions from each population as well as murine monoclonal antibodies (MoAb), specific for BLG, to solid phase bound antigen was evaluated in enzyme-linked immunoadsorbent assay using streptavidin-biotin technology. Some of these experiments were also performed with sera from 42 patients diagnosed with other cancers. RESULTS: Compared with control sera and sera from patients with other solid tumors, lung cancer patient sera showed distinct capacities to prevent the binding of murine MoAb as well as human pooled IgG fractions to n- and d-BLG. The inhibition capacities of lung cancer sera changed as soon as five weeks after cancer removal. CONCLUSIONS: The results indicate that the difference in epitope specificity exhibited by lung cancer sera is not restricted to cryptic epitopes, but also affects continuous and discontinuous epitopes, accessible only on the native antigen. A high level of binding discrimination between antibodies from the study populations is also observed at the level of the epitope. This deviation in the epitope specificity of antibodies changes soon after cancer removal, suggesting a tumor-dependent disturbance. Also documented in the Dermatophagoides pteronyssinus model, it opens the way to a new class of paraneoplastic immune markers for this malignancy, with, at first glance, a high specificity level.     

Immunological assessment of exposure to Echinococcus granulosus in a rural dog population in Uruguay

An ELISA was used to screen a dog population in Uruguay (Sarandi Del Yi, Durazno District) for the prevalence of specific serum antibodies (IgG, IgA and IgE) to Echinococcus granulosus. The sensitivity (61%) and specificity (97%) of the ELISA were determined using well-defined serum groups. A total of 408 dogs from Sarandi del Yi and environs were screened serologically, and 29.7% (8.6-13.8% for each antibody class) of dogs had positive levels of antibody to E. granulosus. This antibody prevalence (exposure) was significantly higher than the percentage of dogs found to be positive for E. granulosus worms by arecoline purgation (7.6%). This level of exposure to E. granulosus determined by ELISA is considered unacceptable from a public health perspective. Measures will now focus on obtaining data on the true prevalence of current infection in this dog population and on determining the transmission patterns of the disease in this endemic region.     

Mutagenicity and amount of chloroform after chlorination of bank filtered lake water

A one-step immunoassay for simultaneous detection of serum IgG and IgM antibodies to Borrelia burgdorferi has been developed. The assay is based on C1q, which binds to immune complexes containing IgG and/or IgM antibodies. Micro-beads pre-coated with antibodies to human C1q are mixed with human serum samples and fluorochrome-labelled B. burgdorferi flagellum antigen. In the presence of serum IgG and/or IgM antibodies to B. burgdorferi, fluorochrome-labelled antigen/antibody complexes are formed. These are then bound by serum C1q and are subsequently captured on the anti-C1q-coated beads. The sample is analysed on a flow cytometer and the presence of fluorescent beads is, thus, indicative of a positive test result. In the present study the sensitivity and specificity of the assay are compared to those of the indirect IDEIA B. burgdorferi IgG and the mu-chain capture IDEIA B. burgdorferi IgM ELISAs for separate determination of IgG and IgM. Detection using a flow cytometer can be performed without separation of the beads from the reaction mixture, which means that in practice, the method is carried out as a one-step assay and it is, thus, very suitable for automation. Other advantages of this kind of assay includes an antibody/antigen reaction which occurs in solution and the potential of using the method for the detection of antibodies against several antigens from the same or different infectious agents (multi-parameter screening).     

Radioimmunoassay for glutamic acid decarboxylase (GAD65) autoantibodies as a diagnostic aid for stiff-man syndrome and a correlate of susceptibility to type 1 diabetes mellitus

OBJECTIVE: To establish and validate a double-antibody radioimmunoassay (RIA) for detecting serum auto-antibodies against glutamic acid decarboxylase (GAD65). This enzyme catalyzes synthesis of the neurotransmitter gamma-aminobutyric acid in neurons and pancreatic islet cells. MATERIAL AND METHODS: We compared the frequency of GAD65 and other "thyrogastric" autoantibodies in adult patients with stiff-man (Moersch-Woltman) syndrome, type 1 diabetes, or polyendocrine disorders and in healthy subjects. The frequency of pancreatic islet cell antibody (ICA) detection was also assessed. The GAD65 RIA was validated by testing blinded samples, by confirming the specificity of low-titered positive results by "cold" antigen inhibition, and by comparing the RIA results with results of a kit assay incorporating staphylococcal protein A as immunoprecipitant. Recombinant GAD65 protein labeled with 125I was used as antigen, and a combination of anti-human IgG and IgM was used as immunoprecipitant. Seropositivity was determined for ICA and gastric parietal cell antibodies by indirect immunofluorescence assays and for thyroid peroxidase (microsome) and thyroglobulin antibodies by agglutination assays. RESULTS: We detected GAD65-specific antibodies in all but 1 of 46 local patients with stiff-man syndrome (98%); 16 had evidence of diabetes. Positive values exceeded 20 nmol/L in 96%, and 89% were ICA-positive; 76% had additional thyrogastric antibodies. Of 41 patients with type 1 diabetes (17 local and 24 workshop serum specimens), 33 were GAD65 antibody-positive (80%); 85% of these positive values were 20 nmol/L or lower. Only 18% of sera from patients with type 1 diabetes were ICA-positive, but 59% had other thyrogastric autoantibodies. Of 20 patients with autoimmune endocrinopathies without diabetes or stiff-man syndrome, 35% were GAD65 antibody-positive, 5% were ICA-positive, and 90% were thyrogastric antibody-positive. Of 117 healthy control subjects, 8% were GAD65 antibody-positive, and a third of those had other thyrogastric antibodies (14% overall); none was ICA-positive. CONCLUSION: Seropositivity in the double-antibody RIA for GAD65 autoantibody is a sensitive and specific marker of predisposition to type 1 diabetes and related organ-specific autoimmune disorders. As such, this RIA is complemented by assays for thyroid and gastric parietal cell autoantibodies.     

Detection of serum antibodies against Chlamydia pneumoniae by ELISA

Chlamydia pneumoniae causes pneumonia and other respiratory infections in children, adolescents and adults. We tried to evaluate the diagnostic value of detection of serum antibodies by ELISA for C. pneumoniae infections in Japanese children. Serum IgG, IgA and IgM antibodies to C. pneumoniae were determined by the microimmunofluorescence (MIF) test. Serum IgG and IgA antibodies were also determined by ELISA test kits. Results obtained by ELISA were compared with those obtained by MIF test. IgG antibody to C. pneumoniae was detected in 135 (39.5%) by ELISA and in 125 (36.5%) by MIF out of 342 sera from Japanese infants and children without respiratory infections (aged from 2 months old to 15 years old). IgA antibody to C. pneumoniae was detected in 129 (37.7%) by ELISA and in 117 (34.2%) by MIF out of 342 sera tested. Of 342 specimens 113 were IgG-positive by ELISA and MIF (sensitivity: 90.4%, specificity: 89.9%, r = 0.853). Of 342 sera 28 had IgG antibody titers of 1:256 and none had titers 1:512 or higher by MIF. Of 28 infants and children a total of nine were less than 4 years of age. On the other hand, of 342 specimens 99 were IgA-positive by ELISA and MIF (sensitivity: 84.6%, specificity: 86.7%, r = 0.769). Of 342 sera 16 had IgA antibody titers of 1:256 or higher by MIF. Of 16 infants and children, ten were less than 4 years of age. ELISA had excellent sensitivity and specificity relative to MIF test for detection of IgC and IgA antibodies to C. pneumoniae. It was suggested that C. pneumoniae infection in Japanese infants and children under 4 years of age was not infrequent.     

Prevalence of cryoglobulinemia in chronic hepatitis C virus infection and response to treatment with interferon-alpha

A recombinant Schistosoma mansoni protein has been identified as a useful antigen for the detection of S. mansoni and Schistosoma haematobium antibodies. The purified recombinant protein, Sm22.3, was assayed using an enzyme-linked immunosorbent assay format against a battery of 491 well defined sera, including S. mansoni, S. haematobium, and Schistosoma japonicum infection sera, normal human sera, sera from 9 other parasitic infections, and sera from 2 additional infections. The sensitivity for detecting S. mansoni and S. haematobium infections with this single recombinant protein is 80.1%. The specificity is 94.8%. However, 15 of the 16 cross-reactive sera are malaria infection sera, and we have data suggesting that these malaria sera are actually recognizing an epitope on the vector-derived 6Xhistidine tag of recombinant Sm22.3. If this is the case, then, the actual specificity of the assay is 99.6%.     

Structural characterization of recombinant human erythropoietins by fluorophore-assisted carbohydrate electrophoresis

Two enzyme immunoassay (EIA) systems were compared for their ability to detect Borrelia burgdorferi sensu lato specific IgG and IgM antibodies and to differentiate between symptomatic (83 patients with neuroborreliosis) and asymptomatic seropositive subjects (80 healthy controls). Antibody concentrations were determined by EIA; the antigens used were either a sonicate of B. burgdorferi or three recombinant borrelial proteins: the 14-kDa flagellin fragment, the outer surface protein C (22 kDa) and the high molecular mass protein p83 (83 kDa). In the sonicate, EIA, IgG or IgM antibodies to B. burgdorferi, or both, were detected in all patients with neuroborreliosis and in all controls. Pre-absorption of sera with Treponema phagedenis sonicate diminished the sensitivity of detection of borrelial specific IgG (IgG or IgM or both) antibodies in patients with neuroborreliosis from 80 to 57% (100 to 82%) and in the controls from 100 to 32% (100 to 37%). While being specific for B. burgdorferi, the recombinant EIAs proved to be significantly more sensitive than the sonicate EIA: IgG or IgM, or both antibodies against any of the recombinant antigens were detected in 92% of patients with neuroborreliosis and in 24% of controls. The increase in sensitivity in patients with neuroborreliosis was mostly due to the higher detection rate of IgM antibodies in the recombinant EIA (77% versus 48% in the sonicate EIA), while IgG antibodies were demonstrated with similar frequencies in both EIA systems (57% versus 60%). It was concluded that the recombinant EIAs are superior to the sonicate EIA with pre-absorption of cross-reactive antibodies in the confirmation of an acute borrelial infection and in the differentiation between symptomatic and asymptomatic infections.     

Reactivity of twenty-two cytotoxic human monoclonal HLA antibodies towards soluble HLA class I in an enzyme-linked immunosorbent assay (PRA-STAT)

An ELISA, PRA-STAT was recently introduced for the detection of HLA class I specific antibodies of IgG isotype in patients' sera. We studied the antigenicity of the soluble HLA (sHLA) preparations that are used in this ELISA as the detection matrix, with the aid of a panel of complement binding human HLA monoclonal antibodies (HuMAbs). A total of 22 HuMAbs, including both IgG and IgM were used. CDC and PRA-STAT ELISA were in complete agreement on 9 of the mAbs tested, with 16 HLA-A and 16 HLA-B locus antigens or their splits identified identically on CDC and PRA-STAT. In 7 of the remaining 13 HuMAbs, there was a difference of one antigen in the specificity pattern of the two techniques three times a specificity call not made by CDC, and four times a call not made by PRA-STAT. For the remaining 6 HuMAbs the differences involve 2 antigens (4 HuMAbs), and 3 or 4 antigens (1 HuMAb each). This study shows the validity of PRA-STAT for detection of HLA-class I antibodies, irrespective of isotype, in serum. The immunological integrity of the sHLA preparations used in PRA-STAT is also confirmed, albeit with some slight discrepancies in antibody specificity seen between PRA-STAT and CDC.     

Increased synthetic peptide specificity of tissue-CSF bound anti-MBP in multiple sclerosis

Free and bound hydrosoluble protein extracts were prepared from four anatomical areas of a multiple sclerosis (MS) cerebrum and from corresponding anatomical areas of a normal (non-MS) control. Increased levels of IgG and anti-myelin basic protein antibodies (anti-MBP) were detected in all MS samples and they were undetectable in the controls. IgG and anti-MBP from free (unbound) hydrosoluble protein extracts are defined as free IgG and free anti-MBP while IgG and anti-MBP from tissue bound protein extracts are defined as bound IgG and bound anti-MBP. IgG was purified from free protein extracts by protein G Sepharose affinity chromatography and anti-MBP was further isolated from purified IgG by antigen specific (MBP) Sepharose affinity chromatography. Free and bound anti-MBP were reacted with 20 synthetic peptides of human MBP prepared by the Fmoc method. Free anti-MBP, whether in the context of whole protein extracts, or as purified IgG or as purified antibody was completely neutralized by peptides #12, #15, #56 and #56* containing overall residues 75-106, partially neutralized by peptides #27, #16 and #21 containing overall residues 61-83 and did not react with the remaining 13 peptides. Tissue bound anti-MBP was completely neutralized only by peptides #12, #15, #56 and #56* (overall residues 75-106) and showed no reactivity towards the remaining 16 peptides including peptides #27, #16 and #21. Synthetic peptide specificity of free anti-MBP purified from MS cerebrum was identical to previously reported specificity of free anti-MBP from MS cerebrospinal fluid (CSF), while tissue bound anti-MBP, as well as bound anti-MBP from CSF had a more restricted synthetic peptide specificity than free anti-MBP. This suggests that the most likely epitope of anti-MBP is located between residues 84 and 95 of human MBP just proximal to the tri-proline sequence (99-101).     

Evaluation of two nonculture antigen tests and three serotests for detection of anti-chlamydial antibodies in the diagnosis of ocular chlamydial infections

BACKGROUND: Diagnosis of chlamydial conjuctivitis is difficult in chronic diseases because chlamydial elementary bodies are mostly undetectable in conjunctival scrapings by cell culture. We therefore compared two nonculture antigen tests and three different serotests for anti-chlamydial antibodies with McCoy cell culture, the "gold standard" of chlamydial diagnosis. Conjunctival scrapings and serum samples of 93 patients attending the outpatient eye clinic in Graz because of chronic follicular conjunctivitis were tested. METHODS: A total of 558 conjunctival scrapings and 93 serum samples were investigated. Chlamydial antigen detection was done by McCoy cell culture, polymerase chain reaction (PCR; Amplicor, Roche), and direct immunofluorescence assay (DFA; Microtrak, Syva). Antichlamydial IgA and IgG antibodies in the sera were detected by an immunoperoxidase assay (IPAzyme, Savyon) and two different enzyme-linked immunosorbent assays (SeroELISA, Savyon and rELISA, medac). RESULTS: Cell culture and PCR yielded identical results. The positivity rate for chlamydial conjunctivitis was 8.6% (8 of 93 patients). PCR proved most sensitive and most specific. IPAzyme was 75% sensitive for IgA and 100% for IgG; SeroELISA and rELISA were less sensitive. IPAzyme was 81% specific for IgA and 47.3% for IgG. SeroELISA and rELISA were less specific for IgA, but more specific for IgG. Post-test likelihood of disease was greatest in IPAzyme. CONCLUSIONS: PCR proved to be a good alternative to cell culture; DFA is useful for quick diagnosis. Genus-specific serotests cannot compete with chlamydial antigen detection. They differ in sensitivity and specificity because of the antigen type they present. They are still of only supportive value in cases where chlamydial antigen detection is not possible. Recently introduced species-specific antibody tests should be of greater value.     

General anesthesia in surgical interventions in children]

The glycoprotein (15-18 kDa) antigen of Mycobacterium tuberculosis H37Rv was affinity isolated on the immunosorbent with monoclonal antibodies S4C1G4 (specific to M. tuberculosis H37Rv; Avdiyenko V. G., Kondrashov S.Yu, Lyashenko S.M.@Probl. Tuberk. 1996, v. 1, p. 6-8 (in Russian). This antigen and PPD (Batch RT 45, Stattens Seruminstitute, Denmark) that was a standard antigen were used for enzyme-linked immunosorbent assay (ELISA), by detecting serum IgG antibodies in patients with pulmonary tuberculosis, and control groups of patients with lung diseases other than tuberculosis (bronchitis and/or asthma, pneumonia) as well as healthy volunteers. The diagnostic parameters of specificity and sensitivity for titers and the same parameters for optic density (OD) (in serum dilution with maximum differences for groups of patients and donors) were compared. The new monoantigen method provided 86.11% specificity and 87.87% sensitivity, which were higher those obtained for optic density (63.89 and 80%, respectively). With PPD, the specificity and sensitivity were 58.04 and 78.78 (for the new titer method) versus 50 and 78.78% (OD data). The method error for titer determination was 10% and for standard OD determination was 27%. The new approach offers additional possibilities of enhancing the quality of ELISA for diagnosis of tuberculosis.     

Evaluation of a stage-specific proteolytic enzyme of Schistosoma mansoni as a marker of exposure

Cercarial elastase (CE) is one of the first proteins released in the host by invading schistosome cercariae. An enzyme-linked immunosorbent assay (ELISA)-formatted immunoassay has been developed to detect antibodies to the stage-specific CE antigen of Schistosoma mansoni as marker of exposure. We have evaluated this test system as an epidemiologic tool, using well-characterized sera collected from S. mansoni- and S. haematobium-infected subjects residing in endemic areas and from control subjects living in nonendemic areas in Egypt. Urine, stool specimens, and blood samples were collected from a sample of 272 endemic subjects randomly selected to represent different age groups in the range of 2-20 years of age. Of 47 S. mansoni-infected subjects, 41 (87.2%) had anti-CE IgG antibodies. Of 52 S. haematobium-infected cases, 38 (73.0%) had IgM antibodies to CE and 43 (82.7%) had IgG antibodies to CE. Of 173 egg-negative people in the endemic area, 84 (48.6%) were IgM positive and 99 (57.2%) were IgG positive. The mean IgM and IgG antibody levels were similar in the infected groups but were significantly lower in the egg-negative group (P = 0.001). All sera from young children (2-3 years of age) were uniformly ELISA negative. The prevalence of IgM and IgG antibodies to CE in children less than six years of age were significantly lower than in other age groups. There was no significant difference in prevalence rates of IgM and IgG anti-CE antibodies between subjects having other parasites present in the endemic area (Ascaris lumbricoides, Entrobius vermicularis, Hymenolepis nana, H. diminuta, Trichostrongylus spp., and Entamoeba histolytica) and those without any parasitic infection. All nonendemic sera (58), including those with other helminth infections, were uniformly ELISA negative for antibodies to CE. These findings suggest that antibodies to elastase indicate exposure, but not necessarily active schistosome infection.     

Immune response and in vivo production of cytokines in patients with liver hydatidosis

Cytokines play an important role in the human immunological response, but the exact role of cytokines in the human immune response against parasites, especially against Echinococcus granulosus, remains unclear. IL-1, IL-2, IL-4 and tumour necrosis factor (TNF) levels in peripheral blood of 21 patients with liver hydatidosis were evaluated before surgical treatment, and the levels of IgA, IgM, IgG, IgE, specific IgE against E. granulosus, C3, C4 and DF complement fractions and CD20, CD3, CD4, CD8 and CD16 cell percentages were also determined, as was the relationship between these variables and cytokine levels. Data from hydatid patients were compared with data obtained from 21 healthy volunteers. Hydatid patients showed increases of IgG, IgE, IgEs and IL-2 (P < 0.01), and decreases of IL-1 and TNF levels (P < 0.001), but these variables (respectively) increased in patients showing cysts in the central area of the liver or with a wide opening of cysts in the biliary tract. The increase of IL-1, IL-2 and IL-4 showed a close relationship with the number, characteristics and above all the location of cysts within the liver itself. IgG and IL-4 levels and also IgG and IgE levels showed a significant correlation (P < 0.05).     

Evaluation of serological diagnosis of Borna disease virus infection using recombinant proteins in experimentally infected rats

We produced two recombinant Borna disease virus (BDV) proteins, p40 and p24, by using a baculovirus vector as a diagnostic antigen. Antigenicities of these recombinant proteins were evaluated by immune rabbit sera. Recombinant p40 was a more sensitive antigen than p24 for the detection of antibodies in infected rats. Rats inoculated with BDV within 24 hr after birth showed higher detection rates of viral RNA and viral proteins from the brain than rats inoculated at 4 weeks-old. Depending on the age of infection and the time postinfection, the detection of BDV RNA, protein, or anti-BDV antibody did not always correlate in individuals. We suggest both serological and molecular biological methods are needed in the diagnosis of BDV infection.     

Performance characteristics of an enzyme-linked immunosorbent assay for determining salivary immunoglobulin G response to Helicobacter pylori

We evaluated the salivary immunoglobulin G (IgG) immune response to Helicobacter pylori in 70 subjects by enzyme-linked immunosorbent assay (ELISA). Subjects with a positive H. pylori culture showed significantly higher titers of antibodies than subjects with no detectable H. pylori: the overall sensitivity and specificity of the test were 84 and 90%, respectively. The detection of salivary anti-H. pylori IgG antibodies may be considered as an alternative to serum IgG detection for ease of sample collection or when blood samples are not available in screening of patients with dyspepsia.     

Surgical management and seizure outcome in patients with tuberous sclerosis

The central nervous system is considered an early and common target for the human immunodeficiency virus type 1 (HIV-1). Serum and cerebrospinal fluid (CSF) from 20 HIV positive patients, including 14 with AIDS-dementia complex (CDC stage IV) and 6 asymptomatic individuals (CDC stage II) were analyzed by enzyme immunoassay for detection of antibodies to native myelin basic protein (MBP) and for the amino acid sequence 68-84 exposed after partial degradation of native MBP. Control groups included HIV-1 negative patients with degenerative and/or vascular dementia, chronic multiple sclerosis (MS) and individuals without any sign of neurological or cognitive disturbances. As opposed to control groups, serum and CSF samples from MS and HIV-1 infected patients showed several oligoclonal bands running in the gamma region. AIDS-dementia complex (ADC) patients had increasingly high intrathecal IgG specific antibody titers for the amino acid sequence 68-84 of MBP. Marked intrathecal antibody production for myelin components was also detected in the majority of HIV-1 infected asymptomatic individuals. Such alteration paralleled development of cognitive deficits, neurological abnormalities and appearance of CNS demyelinating plaques. Preferential immune recognition of this myelin epitope within the CSF during early stages of HIV-1 infection might point to an ongoing process of active demyelination and ultimately indicate subclinical CNS involvement.