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PubMed1995

Molecular mimicry by Mycoplasma pneumoniae to evade the induction of adherence inhibiting antibodies.

Specific regions of adherence binding sites and epitopes of the P1 adhesin of Mycoplasma pneumoniae were synthesised as octapeptides and used as targets in a modified enzyme-linked immunosorbent assay. Acute phase and convalescent sera from 10 patients with M. pneumoniae infection were tested for antibody reactivity to these octapeptides. In convalescent sera, antibody activities were directed against octapeptides of the epitope regions, whereas no antibody activity was found in acute or convalescent sera to octapeptides of adherence-mediating binding sites could be explained partially from the results of cross-reactivity experiments with adherence-inhibiting anti-P1 adhesin monoclonal antibodies (MAbs). Two of these MAbs showed cross-reactions with intracellular antigens of eukaryotic cell lines in immunofluorescence microscopy experiments. The cross-reacting antigens were isolated and characterised as glyceraldehyde-3-phosphate dehydrogenase and 2-phospho-D-glycerate hydrolyase. Antigenic mimicry of eukaryotic structures by functional sites of the P1 adhesin of M. pneumoniae may influence the pathogenesis of M. pneumoniae infection.

Adhesins, Bacterial↗
PubMed2004

Antibody response and viraemia during the course of severe acute respiratory syndrome (SARS)-associated coronavirus infection.

To understand the time-course of viraemia and antibody responses to severe acute respiratory syndrome-associated coronavirus (SARS-CoV), RT-PCR and ELISA were used to assay 376 blood samples from 135 SARS patients at various stages of the illness, including samples from patients who were in their early convalescent phase. The results showed that IgM antibodies decreased and became undetectable 11 weeks into the recovery phase. IgG antibodies, however, remained detectable for a period beyond 11 weeks and were found in 100 % of patients in the early convalescent phase. SARS-CoV viraemia mainly appeared 1 week after the onset of illness and then decreased over a period of 1 month, becoming undetectable in the blood samples of the convalescent patients. At the peak of viraemia, viral RNA was detectable in 75 % of blood samples from patients who were clinically diagnosed with SARS 1 or 2 weeks before the test.

Adolescent↗
PubMed1978

Complement-dependent cytotoxicity of sera obtained from subacute sclerosing panencephalitis patients.

Human lymphoid cells (NC-37) persistently infected with either measles virus (Schwarz and TYCSA strains) or subacute sclerosing panencephalitis (SSPE) virus (Halle and Mantooth strains) were destroyed in the presence of complement by anti-measles sera as well as by sera from SSPE patients. The cytotoxic activity was demonstrated in both IgG and IgM fractions of measles convalescent sera, but only in IgG fraction of SSPE sera. Measles convalescent sera completely lost the cytotoxic activity to all the cell lines, when absorbed with any one of the cell lines, indicating that the viral surface antigens of these cell lines infected with measles or SSPE virus are identical. On the other hand, the cytotoxic activity of SSPE sera could not be readily absorbed with these cells. Thus, the affinity of SSPE sera for the viral surface antigens might be lower than that of measles convalescent sera.

Cell Line↗
PubMed1981

Anticomplement immunofluorescence for the titration of antibody to varicella-zoster virus.

Anticomplement immunofluorescence (ACIF) was tested for its use for the titration of antibody against varicella-zoster virus (VZV). ACIF antibody responses of patients with VZV infection were specific for VZV antigen and heterotypic responses to herpes simplex virus type-1 and cytomegalovirus antigens were not observed. Comparative studies of ACIF, membrane immunofluorescence (MIF) and indirect immunofluorescence (IF), using acetone-fixed antigen, were carried out with nonimmune sera and convalescent sera of patients who had recovered from varicella, herpes zoster and Rumsey Hunt disease. Nonspecific staining occurred with some nonimmune sera at a 1:4 dilution in the MIF and IF tests, after freezing and thawing of the serum, but not in the ACIF test. The antibody titers in convalescent sera agreed well in these three methods and the highest titer was obtained by MIF. The titers in ACIF and IF were similar but the ACIF antibody decreased earlier than the IF antibody during convalescence. On the other hand there was a discrepancy between the titers of ACIF and those of MIF and IF antibody in the sera of healthy adults, all sera with titers higher than 10 in the MIF and IF tests had titers below 10 in the ACIF test. The average titer of ACIF antibody declined to less than 10 with increasing age (13 to more than 20 years), whereas the MIF antibody increased during the same period of life.

Adolescent↗
PubMed2006

Pneumococcal surface protein A (PspA) is effective at eliciting T cell-mediated responses during invasive pneumococcal disease in adults.

Humoral immune response is essential for protection against invasive pneumococcal disease and this property is the basis of the polysaccharide-based anti-pneumococcal vaccines. Pneumococcal surface protein A (PspA), a cell-wall-associated surface protein, is a promising component for the next generation of pneumococcal vaccines. This PspA antigen has been shown to stimulate an antibody-based immunity. In the present study, we evaluated the capacity of PspA to stimulate CD4+ T cells which are needed for the correct development of a B cell based immune response in humans. Cellular immunity to PspA was evaluated by whole-blood culture with different pneumococcal antigens, followed by flow cytometric detection of activated CD4+CD25+ T cells. T cell-mediated immune responses to recombinant PspA proteins were assessed in acute-phase and convalescent blood from adults with invasive pneumococcal disease and in blood from healthy subjects. All cases had detectable antibodies against PspA on admission. We found that invasive pneumococcal disease induced transient T cell depletion but adaptive immune responses strengthened markedly during convalescence. The increased production of both interleukin (IL)-10 and interferon (IFN)-gamma during convalescence suggests that these cytokines may be involved in modulating antibody-based immunity to pneumococcal disease. We demonstrated that PspA is efficient at eliciting T cell immune responses and antibodies to PspA. This study broadens the applicability of recombinant PspA as potent pneumococcal antigen for vaccination against S. pneumoniae.

Adult↗
PubMed1989

Glucose intolerance in acute infections.

To evaluate the influence of an infective agent, severity of infection and the age of the patient on infection-induced glucose intolerance, concentrations of fasting blood glucose, serum insulin (n = 31) and plasma glucagon (n = 22) were measured and the oral glucose tolerance test (OGTT) was carried out (n = 26) during acute and convalescence phases and after complete recovery in patients with viral (n = 17) or bacterial (n = 14) infections. Serum insulin was increased (P less than 0.001) but plasma glucagon was decreased (P less than 0.01) both during acute infection and the convalescence period. In the acute stage, 2-h values of blood glucose (P less than 0.01) and serum insulin concentrations (P less than 0.001) during OGTT were elevated. The index of insulin resistance (glucose x insulin) was increased by 33% during infection and by 28% during convalescence (P less than 0.001). The observed changes did not correlate with the severity of infection, were more pronounced in younger patients than in older ones and were not dependent on the infective agent. It is clinically important to recognize that the restoration of insulin sensitivity takes longer than the immediate recovery period from the infection.

Adolescent↗
PubMed1986

Antigenic diversity and size diversity of Plasmodium falciparum antigens in isolates from Gambian patients. I. S-antigens.

Ring-stage asexual parasites of P. falciparum were collected from six Gambian children and the S-antigens radiolabelled by 3H-glycine uptake during in vitro culture up to rupture of infected cells and merozoite release. Ouchterlony double diffusion of boiled culture supernatants against a panel of adult Gambian sera identified one S-antigen precipitin arc for five isolates and two precipitin arcs for one isolate. Five of the six isolates were serologically distinct. Analysis of S-antigens by comparison of SDS-polyacrylamide gel electrophoresis patterns of heat-treated soluble proteins revealed a more complex pattern of 3H-labelled S-antigens that was different for each isolate. There were between two and six different 3H-labelled bands for each isolate in the size range of molecular weight 137 000 to 285 000. This result confirms the large size range of S-antigens identified with culture adapted P. falciparum. Several bands were relatively weakly labelled with 3H-glycine, suggesting that natural isolates contain one or two predominant S-antigen phenotypes and several other S-antigen phenotypes expressed by minor parasite subpopulations. Immunoprecipitation was performed using a panel of sera from Gambian adults, or, acute and 3 week convalescent sera from the same patients used for S-antigen radiolabelling. Adult sera generally immunoprecipitated some of the S-antigens in each isolate, including antigens that must represent extremely minor parasite subpopulations since they could not be seen in the patterns of non-immunoprecipitated heat-stable proteins. Sera from convalescent children were generally negative on immunoprecipitation, even with the homologous isolate. In one case we observed the acquisition of specific immunoprecipitating antibody to one of the homologous S-antigens during the convalescent period. The antigenic and structural complexity of S-antigens in natural isolates that have not been submitted to the selection pressure of adaptation for in vitro culture is clearly greater than for culture adapted P. falciparum.

Adult↗
PubMed1991

Heat shock protein as a predisposing and immunopotentiating factor in Kawasaki disease.

Local inflammatory reactivation of a previous BCG inoculation site is a specific and early manifestations of Kawasaki disease (KD). It is postulated that the erythematous changes at the BCG site are part of a generalized activation of the immune system, and that molecules cross-reacting between the infectious agent involved and mycobacterial BCG antigens contribute to the inflammatory process. To detect the cross-reacting molecules, the antibody responses to mycobacterial antigens were examined in 21 patients by IgG-immuno-blotting using acute and convalescent phase sera. Markedly increased levels of antibody against mycobacterial 65kDa protein were detected in all convalescent but in none of the acute sera. Since the antibodies detected are also directed to recombinant 65kDa heat shock protein (HSP65), HSP65 may be related to the initial phase of KD. Moreover, peripheral lymphocytes from convalescent KD patients proliferated in culture in the presence of HSP65, suggesting the sensitization of lymphocytes to HSP65, and gamma delta TCR(+) T cells in the peripheral blood were increased in half the KD patients. Thus, HSP65 is a potent factor involved in activation of the immune system, and consequent endothelial damage in the initial phase of KD.

Antigens, Bacterial↗
PubMed1998

Surface marker patterns of T cells and expression of interleukin-2 receptor in measles infection.

The surface marker patterns of T cells of Ghanaian children during measles infection were studied and an attempt was made to demonstrate T cell activation and viability in vitro after activation in vivo by measles virus. The frequencies of CD4+ and CD8+ naive T cells in measles patients were high while their memory T cells were remarkably reduced with no sign of proliferation even at the acute phase of the illness. The reduction of memory T cells was prolonged during the convalescent phase (2 months after onset). The anti-CD3 monoclonal antibody-induced expression of interleukin-2 receptor alpha chain (IL-2R/CD25) was significantly suppressed; however, the addition of phorbol 12-myristate 13-acetate or ionomycin caused a remarkable recovery of CD25 expression. On simple culture, an appreciable proportion of T cells from measles patients died rapidly in contrast with only a few T cells from healthy controls doing so. The suppression of CD25 expression was still demonstrated during the convalescent phase of the disease. Taken together these results suggest unresponsiveness and activation-induced cell death of T cells during severe measles infection in Ghanaian children. Furthermore the prolonged abnormalities of T cells (i.e. decreased memory T cells and inhibition of CD25 expression during the convalescent phase) might be related to post-measles infection immunosuppressive status.

Antigens, Surface↗
PubMed1992

Increased protein absorption after acute gastroenteritis in children.

We investigated the intestinal absorption of the macromolecule human alpha-lactalbumin during and after an episode of acute gastroenteritis in children. Twenty children were studied in the acute phase and 17 excreted rotavirus. Eleven children were studied again 5-8 weeks later (convalescent phase). Human alpha-lactalbumin serum concentrations in the acute phase were similar but in the convalescent phase they were significantly (p less than 0.001) higher than those in the reference children. The serum concentrations were also higher in the convalescent than in the acute phase (p = 0.021). This study suggests that there is an increased absorption of proteins from the gut into the circulation 5-8 weeks after rotavirus gastroenteritis.

Acute Disease↗
PubMed1980

Group B meningococcal opsonins in serum measured by polymorphonuclear leukocyte chemiluminescence.

Serum opsonic activity against a strain of group B meningococci was measured by the initial increment in polymorphonuclear leukocyte chemiluminescence. Chemiluminescence was markedly enhanced by serum and was dependent upon both thermostable and thermolabile factors. It was higher in convalescent sera from patients with meningococcal disease than from other patients and normal controls. Absorption of sera with the homologous strain substantially decreased the opsonic activity. Convalescent serum from a patient with group C meningococcal pericarditis showed high opsonic activity towards a group B meningococcal strain. A convalescent serum showed significantly higher activity towards its homologous group B strain than towards another group B meningococcal strain. As measured by chemiluminescence, properties besides group specificity are of importance in opsonization of meningococci. The simplicity and precision of the chemiluminescence measurements seem to offer an improved tool for the study of meningococcal disease immunology.

Humans↗
PubMed1992

[Biochemical analysis of lipopolysaccharides from respiratory pathogenic Branhamella catarrhalis strains and the role of anti-LPS antibodies in Branhamella respiratory infections].

We characterized lipopolysaccharides (LPSs) from respiratory pathogenic Branhamella catarrhalis (BC) strains, and evaluated the protective property of anti-BC LPS antibody in BC respiratory infections. LPSs from four strains of BC were lipooligosaccharide having no O-side chain and a M(r) of 3 KDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). All of them produced different patterns, showing two to four bands on SDS-PAGE. We found high level of anti-BC IgG antibody in convalescent sera from a patient with BC respiratory tract infection by ELISA. This IgG antibody recognized BC LPSs on Western blots. Two respiratory pathogens of BC (strains; 87-122, 88-23) were tested in a bactericidal assay employing a convalescent sera. 87-122 strain was susceptible to antibody-dependent, complement-mediated killing, while 88-23 strain was resistant. The killing of 87-122 strain was inhibited by addition of the homologous BC LPS to the convalescent sera in a dose-dependent manner. These data support that anti-BC LPS antibody may mediate complement-lysis of some strains of BC, and play a protective role in BC respiratory infections.

Antibodies, Bacterial↗
PubMed2005

[Evaluation of ELISA kits for detection of Mycoplasma pneumoniae--specific IgG, IgA, IgM antibodies on the diagnosis of Mycoplasma pneumoniae infection in children].

A retrospective study was conducted to evaluate the utility of Mycoplasma pneumoniae IgG (quantitative), IgA (quantitative), IgM (qualitative) ELISA kits (Medac Diagnostika, Germany) for the diagnosis of M. pneumoniae pneumonia in children under 16 years of age. This study included a total of 159 serum samples from 113 patients with acute respiratory diseases such as bronchitis, pneumonia, which were classified into three groups according to the results of a particle agglutination (PA) test as a reference method, that is, Group I (Mycoplasma-definite cases): Group I-a (paired 52 samples from 26 cases); a four-fold or greater rise of antibody from an acute phase PA titer of < or = 1:80, Group I-b (paired 12 samples from 6 cases); a four-fold or greater rise of antibody from an acute phase PA titer of > or = 1:160, Group I-c (48 samples from 38 cases); a single high PA titer of > or = 1:640 either or both in acute or convalescent serum, Group II (Mycoplasma-probable cases, 18 samples from 17 cases): a PA titer of 1:160 or 320 was observed either or both in acute or convalescent serum, but the above serological criteria for Group I were not fulfilled, Group III (non-cases, 29 samples from 26 cases): a PA titer of any sample was < or = 1:80. The ELISA tests were performed according to the supplier's recommendations, and the results were classified according to the interpretation provided by the supplier: Early stage of infection (category 1,2), Acute- (3,4,5), Current- (6), Past- (7), and No-infection (8). The day of onset of fever (defined as a body temperature of > or = 37.5 degrees Celsius) was denoted as day 0. As a result from Group I, the category initially observed following the onset of fever was category 8 (triple negative), and the predominance of category 8 was replaced by category 1 (IgM solely positive) after day 4, followed by a shift of predominance to category 4 (IgM and IgG double positive) or 5 (triple positive) after day 10 or later. Specifically, category 1 was rather exclusively observed before day 21 following the onset of fever. These results suggest that category 1, when observed, is a useful marker of acute infection by Mycoplasma pneumoniae in children because it appears early in the acute phase and no longer observed beyond the convalescent phase. On the other hand, significance of detecting IgA antibody, which must be important for adults, was not remarkable in our study. Five samples in group II and 3 samples in group III fell into category 1. Whether or not such cases, in the absence of significant PA titers, can be taken actually as mycoplasmal infection remains to be clear. This study validated the utility of this ELISA methodology in terms of the acute phase diagnosis using a single point serum sample for Mycoplasma pneumoniae infection specifically in children.

Adolescent↗
PubMed2000

Antimalarial bioavailability and disposition of artesunate in acute falciparum malaria.

The pharmacokinetic properties of oral and intravenous artesunate (2 mg/kg of body weight) were studied in 19 adult patients with acute uncomplicated Plasmodium falciparum malaria by using a randomized crossover design. A sensitive bioassay was used to measure the antimalarial activity in plasma which results from artesunate and its principal metabolite, dihydroartemisinin. The oral study was repeated with 15 patients during convalescence. The mean absolute oral bioavailability of the antimalarial agent in patients with acute malaria was 61% (95% confidence interval [CI], 52 to 70%). The absorption and elimination of oral artesunate were rapid, with a mean elimination half-life of antimalarial activity of 43 min (95% CI, 33 to 53 min). Following oral administration to patients with acute falciparum malaria, peak antimalarial activity in plasma and the area under the plasma concentration-time curve were approximately double those during convalescence and the apparent volume of distribution and clearance were approximately half those during convalescence (P < or = 0.005). Acute malaria is associated with a significant reduction in the clearance of artesunate-associated antimalarial activity.

Administration, Oral↗
PubMed2005

Serum antibody responses in children with rotavirus diarrhea can serve as proxy for protection.

We examined sera from 42 patients 1 to 30 months of age for rotavirus immunoglobulin M (IgM), IgA, IgG, and IgG subclasses and sought to determine if serum antibody could serve as a reliable marker for prediction of disease severity. Infants in the first few months of life usually had high maternal IgG titers and, when they were infected with rotavirus, had low IgM titers or no IgM in acute-phase sera and poor seroconversions 3 weeks later, suggesting that maternal antibodies had inhibited viral replication and antibody responses. All patients > or =6 months of age had IgM in acute-phase sera, indicating that IgM is a good marker for acute rotavirus infection. IgG was the best overall predictor of an infection, as the convalescent-phase sera of 81% of the patients had a fourfold rise in the IgG titer. IgA titers in convalescent-phase sera and conversion rates were higher among patients > or =12 months of age than among children younger than 12 months. IgG1 was the predominant subclass detected in the acute-phase sera of some children and in all 28 convalescent-phase serum samples examined. Patients with preexisting acute-phase IgG titers of > or =100 or > or =200 had diarrhea that was less severe or of a shorter duration. These results indicate that serum IgG is the most reliable marker for seroconversion and is a consistent proxy for protection against severe disease.

Antibodies, Viral↗
PubMed2005

Immunoglobulin G subclass profile of antimeasles response in vaccinated children and in adults with measles history.

The aim of this study was to investigate measles-specific immunoglobulin G (IgG) subclass profile in vaccinated children and in adults with natural infection. Serum samples were collected before and 30 days after vaccination. The sera from 51 late convalescent adults and seven adults with natural measles infection at the 12th day after onset of rash have been also investigated. Measles IgG antibodies and specific IgG subclasses were tested by enzyme-linked immunosorbent techniques. In children younger than 3 years, the predominant subclass was IgG3, which contributed, on average, 63.3% of the total IgG response. The contributions of specific IgG1 and IgG4 to the total IgG antimeasles response were lower (19.9% and 16.8%, respectively), whereas IgG2 was not found. In contrast, in the group of children older than 4 years, just IgG2 was a predominant subclass; it contributed 42.6% of the total IgG response. Other subclasses were also present but the contribution was much lower. In adult volunteers with measles history, IgG2 was a predominant subclass of total IgG. Thus, in early convalescence IgG2 contributed 62% of the total IgG response, whereas in late convalescence the contribution was lower (41.4%). There were no visible differences in IgG subclass composition between subjects with natural infection and vaccinated children except those below 3 years of age. The humoral immune response of such subjects is immature and the IgG2 subclass of virus-specific antibodies has not been revealed in the sera.

Adult↗
PubMed1995

Antibody responses to the capsular polysaccharide of Neisseria meningitidis serogroup B in patients with meningococcal disease.

We measured antibody responses to meningococcal serogroup B (MenB) polysaccharide (PS) by enzyme-linked immunosorbent assay (ELISA) in sera from 94 patients from The Netherlands with disease caused by Neisseria meningitidis group B. The patients ranged in age from 3 to 73 years (mean age, 18.8 years). In initial studies we showed that the binding of a panel of MenB PS-reactive human immunoglobulin M (IgM) paraproteins to biotinylated MenB PS bound to avidin-coated microtiter wells was inhibited > 90% by the addition of soluble MenB PS or encapsulated group B meningococci. In contrast, inhibition of IgM anti-MenB PS antibody-binding activity in many of the patient sera was less than 50% (range, 20 to 94%). These data suggested a high frequency of nonspecific binding in the patient sera. Therefore, all serum samples were assayed in replicate in the presence or absence of soluble MenB PS, and only the inhibitable fraction of the binding signal was used to calculate the anti-MenB PS antibody concentrations. In 17 control patients with meningococcal disease caused by serogroup A or C strains, there was no significant difference in the respective IgM or IgG anti-MenB PS antibody concentrations in paired acute- and convalescent-phase sera. In contrast, in patients with MenB disease, the geometric mean IgM anti-MenB PS antibody concentration increased from 3.9 units/ml in acute-phase serum to 10.5 units/ml in convalescent-phase serum (P < 0.001). The corresponding geometric mean IgG anti-MenB PS antibody titers were 1:27 and 1:36 (P < 0.05). There was only a weak relationship between age and the magnitude of the logarithm of the antibody concentrations in convalescent-phase sera (for IgM, r2 = 0.06 and P < 0.05; for IgG, r2 = 0.08 and P < 0.01). Our data indicate that precautions are needed to avoid nonspecificity in measuring serum antibody responses to MenB PS by ELISA. Furthermore, although this PS is thought to be a poor immunogen, patients as young as 3 years of age recovering from MenB disease demonstrate both ImG and IgG antibody responses in serum.

Adolescent↗
PubMed2002

Widal test in diagnosis of typhoid fever in Turkey.

We studied the value of the Widal tube agglutination test for the diagnosis of typhoid fever. The subjects were all adults >18 years of age and were divided into four groups: (i) 317 healthy blood donor controls, (ii) 31 bacteriologically confirmed patients with Salmonella enterica serotype Typhi, (iii) 21 patients with a clinical diagnosis of typhoid fever, and (iv) 41 febrile nontyphoid patients. Blood donor controls were screened with a slide agglutination test for the Salmonella enterica serotype Typhi O and H antigens, and positives were then tested with the Widal test. Acute- and convalescent-phase sera from patients in groups 2, 3, and 4 were obtained 7 to 10 days apart and tested by the Widal test. Using a cutoff of >or = 1/200 for the O antigen test performed on acute-phase serum gave a sensitivity of 52% and a specificity of 88% with a positive predictive value (PPV) of 76% and a negative predictive value (NPV) of 71%. This increased to 90% sensitivity and specificity with a PPV of 88% and an NPV of 93% when the convalescent-phase serum was tested. We concluded that O and H agglutinin titers of > or = 1/200 are of diagnostic significance. The Widal test is easy, inexpensive, and relatively noninvasive. It can be of diagnostic value when blood cultures are not available or practical. The results must be interpreted cautiously because of the low sensitivity of the test. The Widal test done on convalescent-phase serum gave more-reliable results with higher specificity and sensitivity.

Adult↗