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PubMed1984

Diagnosis of postnatally acquired rubella by use of three enzyme-linked immunosorbent assays for specific immunoglobulins G and M and single radial hemolysis for specific immunoglobulin G.

Three commercial indirect enzyme-linked immunosorbent assays (ELISAs) (RUBELISA, Enzygnost-Rubella, and Rubazyme) and a commercial single radial hemolysis (SRH) test (Rubazone) were evaluated for the diagnosis of acute rubella by testing 41 acute- (less than 7 days postonset) and convalescent-phase (8 to 82 days postonset) serum pairs from cases of rubella previously confirmed by significant change in the hemagglutination inhibition test titer. Specificity was tested by using 10 acute- and convalescent-phase serum samples from patients with rash not confirmed as rubella (control group). In testing for rubella-specific immunoglobulin G (IgG) antibody, Enzygnost-Rubella and Rubazyme confirmed infection in 40 and RUBELISA in 39 of the 41 proven rubella patients. For one patient all three ELISAs failed to show a significant titer rise. No false-positive diagnoses occurred in the control group, although a suspected infection was shown by Rubazyme in one patient. No specific IgM could be detected in this case. Single radial hemolysis confirmed infection in 39 of the 41 proven rubella patients, and one false-positive diagnosis occurred in the control group patients. Of the 43 convalescent-phase serum samples, rubella-specific IgM was detected in 42 by Enzygnost-Rubella, in 41 by RUBELISA M, and in 39 by Rubazyme-M. For a rapid diagnosis with acute-phase sera, specific IgM detection by ELISA was most reliable in hemagglutination inhibition test-positive sera; of 18 such serum samples IgM was shown in 15 by Enzygnost-Rubella, in 13 by RUBELISA M, and in 11 by Rubazyme-M. False-positive specific IgM results were shown by Rubazyme-M in two serum samples from one patient in the control group. These serum samples were negative with the other two ELISA methods.

Adolescent↗
PubMed1987

Passive hemagglutination test for enteric fever.

A passive hemagglutination (PHA) test for serodiagnosis of enteric fever was developed by sensitizing glutaraldehyde-preserved erythrocytes with lipopolysaccharide from Salmonella serogroups A, B, C, and D singly or simultaneously. The lipopolysaccharide-sensitized erythrocytes were tested with sera from 200 blood donors, 100 patients whose hemoculture was positive for Salmonella species, and 10 patients septicemic for other members of the family Enterobacteriaceae. The PHA test was positive in 90% of 28 acute-phase serum samples from patients with enteric fever from one hospital and in 93% of 72 acute-phase serum samples from another hospital. It was also positive in 100 and 60% of early- and late-convalescent-phase sera, respectively. The PHA test was negative in all patients septicemic for other members of the Enterobacteriaceae. Absorption of sera from patients with enteric fever with lipopolysaccharide from other members of the Enterobacteriaceae did not reduce PHA titers, indicating the specificity of the PHA test. Simultaneous sensitization with lipopolysaccharide from Salmonella serogroups A, B, C, and D was useful as a screening test in a limited trial with 28 acute-phase sera, 10 early-convalescent-phase sera, and 17 late-convalescent-phase sera. The PHA test is indeed a simple, sensitive, specific, and rapid test supplementing hemoculture in laboratory diagnosis of enteric fever.

Enterobacteriaceae↗
PubMed1988

Influence of strain characteristics and immunity on the epidemiology of Campylobacter infections in Thailand.

To determine how strain differences and immunity affect the clinical expression of Campylobacter infections, we conducted a study of acute diarrheal disease in Thailand in which specimens from children with Campylobacter infections were cultured weekly for up to 12 weeks to determine the serotype-specific length of time of convalescent-phase excretion and rate of reinfection. Levels of immunoglobulin G to cell-surface antigens of C. jejuni were determined in another population of healthy children who were closely related by age and location to the children in the diarrheal disease study. Campylobacter species were initially isolated from 18% of 586 children under 5 years old with diarrhea; most isolates in Thailand belonged to serotypes commonly found in developed countries. C. coli was significantly less often associated with symptomatic infections and with bloody diarrhea than C. jejuni (P less than 0.001 and P = 0.045, respectively). The peak age of isolation and the peak level of immunoglobulin G to Campylobacter species occurred before 2 years of age. The mean duration of convalescent-phase excretion was 14 +/- 2 (standard error of the mean) days for children less than 1 year old and 8 +/- 2 days for children 1 to 5 years old (P = 0.02, t test). Infection with another Campylobacter serotype was found in 34% of 105 children during the 12-week follow-up period. The rate of reinfection in these children was 15% (range, 8 to 22%) each week. Hyperendemic exposure to Campylobacter species in Thailand confers immunity to infection that is associated with an early peak in specific serum antibodies and an age-related decrease in the case-to-infection ratio and duration of convalescent-phase excretion but does not prevent asymptomatic infections.

Age Factors↗
PubMed1988

Detection of nephropathia epidemica (Puumala virus)-specific immunoglobulin M by enzyme-linked immunosorbent assay.

Nephropathia epidemica (NE), a less severe form of hemorrhagic fever with renal syndrome, is caused by Puumala virus (PUU). This communication reports the development of a mu-capture enzyme-linked immunosorbent assay (ELISA) for detection of specific immunoglobulin M (IgM) antibodies to PUU virus in human sera. Acute- and early-convalescent-phase sera (collected 1 to 41 days after disease onset) from 29 Swedish patients with clinical NE were tested for PUU virus-specific IgG and IgM antibodies by the indirect immunofluorescence test and ELISA, respectively. Late-convalescent-phase serum was also collected from 18 of these patients 3 to 24 months postinfection and assayed. The IgM ELISA values were strongly positive in sera collected during the first 2 months; at 3 to 9 months, they were negative or in the lower range of significance, and at 24 months all sera were negative. Paired sera from NE patients often fail to show seroconversion or a significant titer rise when tested by indirect immunofluorescence. Since all acute- and early-convalescent-phase sera were positive by IgM ELISA, this test could become an important tool for early diagnosis of acute human NE infections.

Acute Disease↗
PubMed1989

Immunoglobulin M, A, and G antibody response to lipopolysaccharide O antigen in symptomatic and asymptomatic Shigella infections.

The antilipopolysaccharide antibody response in sera obtained from subjects involved in 10 outbreaks of shigellosis occurring in Israeli military field units was determined by an enzyme-linked immunosorbent assay and a passive hemagglutination test. Both tests were found to be sensitive and specific for the diagnosis of shigellosis. A significant antibody response was detected in 73 to 82% of the symptomatic and 48 to 60% of the asymptomatic subjects during the Shigella sonnei and Shigella flexneri outbreaks. Fifty percent of the symptomatic and none of the asymptomatic subjects showed a significant antibody response in the Shigella boydii outbreaks. An examination of the kinetics of the antibody levels over a 10-week period after the onset of disease revealed that immunoglobulin A (IgA) levels were highest 2 weeks after infection and had declined to initial levels within 2.5 months. In contrast, IgG levels at the late convalescent stage were half those measured at early convalescence, still being about twice as high as the initial titers. Although the IgM levels showed a pattern similar to that of IgA, their elevation at the early convalescent stage was less pronounced. We conclude that the detection of an increase in the level of the IgA fraction appeared to be the best indicator for recent symptomatic, as well as symptomatic, infections due to Shigella organisms.

Adult↗
PubMed1990

Homotypic serum antibody responses to rotavirus proteins following primary infection of young children with serotype 1 rotavirus.

The class-specific antibody responses to serotype 1 rotavirus structural proteins were examined by immunoblotting with sera obtained from young children hospitalized with acute rotavirus diarrhea caused by serotype 1. All were believed to be primary infections. Three consecutive samples were obtained from 16 patients during the acute and convalescent phases of the disease and then approximately 4 months later. Immunoglobulin G (IgG)-class antibody responses to two inner capsid proteins (VP2 and VP6) and to the major homologous outer capsid protein (VP7) were detected in all patients. Antibody responses to VP6 were rapid, increased in intensity during 20 to 40 days after the onset of symptoms, and persisted for more than 4 months. Responses to VP2 and VP7 were more delayed, were maximal in convalescent-phase sera, and decreased markedly in intensity 4 months after the onset of symptoms in the majority of children. Two patients with evidence of mixed infection showed persisting high levels of antibody to VP7. Responses to the outer capsid protein VP4 were detected in 67% of patients, peaked at 20 to 40 days after the onset of symptoms, and were no longer detected at 4 months in the majority of patients. It is likely that the immunoblotting technique underestimated responses to VP4. Acute- and convalescent-phase sera (known to contain antirotavirus IgM or IgA measured by enzyme immunoassay) were also examined by immunoblotting. IgM- and IgA-class antibody responses to viral proteins VP2, VP4, and VP7 appeared to be qualitatively identical to those observed for IgG in the same serum samples.

Antibodies, Viral↗
PubMed1993

Immunoglobulin G subclass responses against the structural components of Puumala virus.

Sera from sequentially bled nephropathia epidemica patients (acute-phase, convalescent-phase, and 2-year sera) and sera from 10 to 20 years postinfection were examined by immunoglobulin G (IgG) subclass-specific ELISAs for reactivities against each of the Puumala virus structural proteins (N, G1, and G2). IgG1 was found to be the dominating antiviral subclass, and most of the patients had IgG1 directed to all three structural proteins at all times with a continuous increase over time. IgG2 to the three viral proteins was detected in very low amounts in only a few of the samples. All of the serially bled patients developed IgG3 against each of the proteins, but the responses against the different viral components varied; anti-N- and anti-G1-specific IgG3 showed a peak in the convalescent-phase samples, while the anti-G2 IgG3 response was highest in the acute-phase sera. Eight of ten serially bled patients developed specific IgG4. The IgG4 responses showed similar patterns against all three viral proteins with very low optical density values in the acute- and convalescent-phase samples, followed by higher optical densities in the sera drawn 2 years after infection. Seven of nine 10-year sera contained virus-specific IgG4. These results support the theory concerning the persistence of Puumala virus or viral antigens several years after infection.

Antibodies, Viral↗
PubMed1993

Correlation of serum antibody titers against hepatitis C virus core protein with clinical features by western blot (immunoblot) analysis using a recombinant vaccinia virus expression system.

In order to study the relationships among the clinical features of hepatitis C patients, the presence of hepatitis C virus (HCV) RNA in their blood, and their serum antibody titers against the core protein of virus and to study the antibody levels in asymptomatic HCV carriers, a recombinant vaccinia virus containing a core protein gene was constructed. The recombinant virus expressed a protein with a molecular mass of 22 kDa in RK-13 cells as determined by Western blot (immunoblot) analysis. By using the cell lysate of virus-infected cells and serially diluted serum samples, core antibody titers in the groups of patients in the chronic hepatitis phase and in the convalescent phase as well as in asymptomatic carriers were determined by enhanced chemiluminescence Western blot analysis. Almost all patients in the chronic phase were shown to have high antibody titers of more than 1:500,000 and with no exception had of HCV RNA in their sera. On the other hand, patients who had recovered naturally and were in the convalescent phase were shown to have significantly lower antibody titers, and the antibody was not detected in the lowest serum dilution of 1:500 in 43% of these patients (three of seven total patients). Antibody levels of patients who showed a good response to interferon treatment decreased to intermediate levels between those of patients in the chronic phase and those of patients in convalescent phase. The antibody titers in asymptomatic carriers varied considerably from 1:500,000 to 1:500, and 41% (11 of 27 total individuals) of these carriers showed a high titer equivalent to that of those in the chronic phase. Core antibody was detected consistently in the individuals in whom HCV RNA was detected. This system for core antibody might be useful for identifying the stage of an apparent HCV infection.

Base Sequence↗
PubMed2004

Rapid diagnosis of human herpesvirus 6 infection by a novel DNA amplification method, loop-mediated isothermal amplification.

A novel nucleic acid amplification method, termed loop-mediated isothermal amplification (LAMP), which amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions, may be a valuable tool for the rapid detection of infectious agents. LAMP was developed for human herpesvirus 6 (HHV-6), and its reliability was evaluated in this study. Although LAMP products were detected in HHV-6 B and HHV-6 A DNA, they were not detected in HHV-7 and human cytomegalovirus DNA. The sensitivity of the original HHV-6 LAMP protocol was 50 copies/tube. In order to increase the method's sensitivity, HHV-6 LAMP was modified by increasing the primer concentration. As a result of the modification, sensitivity increased to 25 copies/tube. After these initial validation studies, 13 patients with fever were tested for HHV-6 by viral isolation, serological analysis, and HHV-6 LAMP. In three of the eight patients with primary HHV-6 infection, HHV-6 DNA was detected in whole blood by the original HHV-6 LAMP protocol in not only the acute phase but also the convalescent phase. HHV-6 DNA was detected by modified HHV-6 LAMP in all eight plasma samples collected in the acute phase; however, no HHV-6 DNA was detected in plasma samples collected in the convalescent phase. Although HHV-6 DNA was detected in both the acute and convalescent phases of whole-blood samples in patients with past HHV-6 infection, it was not detected in plasma samples that did not contain latent viral DNA. Thus, detection of HHV-6 DNA in plasma by using this modified HHV-6 LAMP protocol is appropriate for diagnosis of active HHV-6 infection.

Base Sequence↗
PubMed2003

Immunologic memory in Haemophilus influenzae type b conjugate vaccine failure.

AIMS: To compare the convalescent antibody response to invasive Haemophilus influenzae type b (Hib) disease between conjugate vaccine immunised and unimmunised children, to look for evidence of priming for immunologic memory. METHODS: Unmatched case-control study in the UK and Eire 1992-2001 and Victoria, Australia 1988-1990. A total of 93 children were identified as having invasive Hib disease following three doses of conjugate vaccine in infancy through post licensure surveillance throughout the UK and Eire; 92 unvaccinated children admitted to an Australian paediatric hospital with invasive Hib disease were used as historical controls. Convalescent serum was taken for measurement of Hib antibody concentration, and clinical information relating to potential disease risk factors was collected. The geometric mean concentrations of convalescent Hib antibodies were compared between immunised and unimmunised children, using raw and adjusted data. RESULTS: Hib conjugate vaccine immunised children had higher serum Hib antibody responses to disease (geometric mean concentration (GMC) 10.81 microg/ml (95% CI 6.62 to 17.66) than unimmunised children (1.06 microg/ml (0.61 to 1.84)) (p < 0.0001). However, following adjustment for the significant confounding influences of age at presentation and timing of serum collection, a difference persisted only in children presenting with meningitis (vaccinated GMC 3.78 microg/ml (2.78 to 5.15); unvaccinated GMC 1.48 microg/ml (0.90 to 2.21); p = 0.003). CONCLUSIONS: Higher antibody responses to invasive Hib disease in vaccinated children with meningitis reflect priming for immunologic memory by the vaccine. Although a majority of children in the UK are protected from Hib disease by immunisation, the relative roles of immunologic memory and other immune mechanisms in conferring protection remain unclear.

Age Factors↗
PubMed1977

Factors in resistance to and recovery from M. pulmonis-induced arthritis in mice.

A previous infection of C3H mice with viable Mycoplasma pulmonis organisms protected them from arthritis induced by intravenous inoculation of these organisms, whether or not their initial arthritis had resolved at the time of challenge. Although spleen cells obtained from such animals conferred resistance to acute arthritis in syngeneic recipient mice, convalescent-phase serum was much more effective in this respect. In addition, the transfer of 'immune' spleen cells to M. pulmonis-infected mice treated with cyclophosphamide did not prevent them from dying, whereas the transfer of convalescent-phase serum to such animals reduced the incidence of mortality. Since M. pulmonis organisms are rapidly inactivated in mice treated with convalescent-phase serum it is suggested that the transferred serum may act by promoting phagocytosis of the mycoplasmas. Although the reason for some mice developing a prolonged arthritis is not clear, the results show that humoral immune mechanisms are more important than cell-mediated ones in preventing acute arthritis and that the humoral mechanisms are of greater importance than the cell-mediated in resistance to and recovery from a generalized mycoplasma infection.

Animals↗
PubMed2004

Acute dehydrating disease caused by Vibrio cholerae serogroups O1 and O139 induce increases in innate cells and inflammatory mediators at the mucosal surface of the gut.

BACKGROUND AND AIMS: The general concept is that as Vibrio cholerae is not invasive, it mediates a non-inflammatory type of infection. This is being re-evaluated based on available data that natural cholera infection or cholera toxin induces a Th2-type of immune profile and stimulates the humoral immune response, innate cells, and mediators in the host. METHODS: To perform a comprehensive analyses of the inflammatory components, we studied mucosal biopsies from patients, both adults and children with acute watery diarrhoea caused by V cholerae O1 and O139. Patients with cholera, adults (n = 30) and children (n = 18), as well as healthy controls (n = 24) were studied. Histochemical, immunohistochemical, and ultrastructural studies were carried out to elucidate the contribution of the different factors using paraffin and frozen duodenal and/or rectal sections as appropriate. Samples were collected during the acute stage and during early and/or late convalescence. RESULTS: Following natural cholera infection, patients responded with increases in neutrophil polymorphs during the acute stage (p<0.001) compared with healthy controls whereas mucosal mast cells (MMC) (p = 0.008) and eosinophils (p = 0.034) increased in the gut during convalescence. Electron microscopic analyses of duodenal biopsies from adult patients showed increased piecemeal degranulation in both MMC and eosinophils and accumulation of lipid bodies in MMC. Duodenal biopsies from V cholerae O1 infected patients showed upregulation of myeloperoxidase, lactoferrin, PGHS-1, SCF, tryptase, tumour necrosis factor alpha, alpha-defensin, and eotaxin during the acute stage and chymase, interleukin 3 and major basic protein during convalescence. CONCLUSION: We have shown that innate cells and their mediators are upregulated in acute watery diarrhoea. These cells and factors of the innate arm may be important in the host's defence against cholera. Such effects may need to be simulated in a vaccine to achieve long lasting protection from cholera.

Acute Disease↗
PubMed2002

Surgical and transcatheter (Amplatzer) closure of atrial septal defects: a prospective comparison of results and cost.

OBJECTIVE: To compare effectiveness, complications, and cost of Amplatzer with surgical atrial septal defect (ASD) closure. DESIGN: Prospective study. SETTING: Tertiary cardiac referral centre. PATIENTS: 43 consecutive patients (excluding non-UK residents) aged between 2.1 and 56.8 years (median 7) undergoing ASD closure. MAIN OUTCOME MEASURES: Procedural success, complications, regression of right ventricular dilatation (up to one year postprocedure), cost, inpatient stay, and home convalescent time. RESULTS: Amplatzer ASD closure was successful in 24 of 27 (89%) patients. Surgical closure was successful in all 19 cases. Cardiac complications affecting management occurred in three (11%) of the Amplatzer group (two procedural failures, one device embolisation) and 4 of 19 (21%) surgical patients (one pericardial pain, one global pericardial effusion requiring drainage, and one patient with anaemia requiring haematinics in addition to an incidental pericardial effusion and one further incidental pericardial effusion) (p = NS). There were complications that did not affect management in a further 5 of 19 surgical patients. There was no significant difference in regression of right ventricular dilatation by six months postprocedure (median right ventricular end diastolic diameter decrease: Amplatzer group 17.5%, surgical group 15.1%; median cardiothoracic ratio decrease: Amplatzer 7.9%, surgical 7.5%). Both hospital stay and home convalescent times were significantly shorter after Amplatzer closure (median hospital stay: Amplatzer one day, surgery six days; median convalescent time: Amplatzer two weeks, surgery 5.5 weeks). Median cost was similar for both groups (Amplatzer pound5375, surgical pound5412). CONCLUSIONS: Amplatzer ASD closure has a lower chance of success with a single procedure than surgery. Overall, there were more complications in the surgical group but the majority of these were minor and did not require any change in management. Resolution of right ventricular dilatation over the study period was similar for both techniques. Time spent in hospital and away from work or school was shorter for the Amplatzer group. The cost of both techniques was similar.

Adolescent↗
PubMed1986

Antigenic specificity of serological response in Chlamydia trachomatis urethritis detected by immunoblotting.

Sera from 19 patients with Chlamydia trachomatis culture positive non-gonococcal urethritis were studied for the presence of antibodies to chlamydial proteins by immunoblotting. Ten C trachomatis negative patients with non-gonococcal urethritis and 10 healthy controls were also studied. Acute phase sera from C trachomatis positive patients with non-gonococcal urethritis reacted only with the major outer membrane protein whereas all the convalescent phase serum samples reacted with the major outer membrane protein and with a 60,000 and a 62,000 molecular weight protein. Some sera also reacted with a 45,000 molecular weight protein. Five of 10 convalescent phase samples from patients with C trachomatis negative non-gonococcal urethritis showed a reaction pattern comparable with that observed in convalescent sera from C trachomatis from C trachomatis positive patients with non-gonococcal urethritis. Sera from healthy seronegative subjects were negative by blotting.

Antibodies, Viral↗
PubMed1997

Quantitative assessment of IgG and IgA subclass producing cells in rectal mucosa during shigellosis.

AIMS: To assess quantitatively both the morphological changes in the rectal mucosa and the changes in the relative frequency of IgA and IgG subclass producing cells found in the rectal mucosa during the acute phase of shigellosis and at convalescence. METHODS: Rectal biopsies from 25 Shigella dysenteriae 1 infected patients, 10 Shigella flexneri infected patients, and 40 uninfected controls were studied. Morphological changes in the mucosa were graded. The frequency of IgA and IgG subclass producing cells was assessed. In addition, immunostaining for secretory component in epithelial cells was analysed. RESULTS: Using morphological grading, 20% of the 35 patients studied had advanced inflammation (grade 3) in the acute phase of the disease. At convalescence, grade 1 inflammation was seen in 37% of the patients and in 10% of the controls. In the acute phase, as well as at convalescence, the number of IgA1, IgA2, and IgG2 positive cells was significantly higher than in the controls. The results were related to the histopathological degree of inflammation. CONCLUSIONS: In shigellosis, there is evidence for a prolonged humoral response residing in the mucosa long after the clinical symptoms have resolved, suggesting that shigellosis induces persisting mucosal humoral immune and inflammatory responses, remaining at least until 30 days after the infection.

Adult↗
PubMed1978

Economic aspects of day care after operations for hernia or varicose veins.

In a trial of 360 patients with hernia or varicose veins, day care surgery provided an economic alternative to the provision of surgical aftercare either in the surgical wards of a district hospital or in a convalescent hospital. There was only slightly more work for general practitioners. Most of the additional work for the community services was carried out by district nurses, with an average contact time in the postoperative period of 325 minutes for day care patients, compared with 186 minutes and 204 minutes respectively for patients admitted for 48 hours to the surgical or convalescent wards. Day care produced estimated savings of 30 pounds compared with the costs of a 48-hour stay in the surgical wards, and savings of 22 pounds compared with a 48-hour stay in the convalescent wards.

Aftercare↗
PubMed2003

Increased circulating 92 kDa matrix metalloproteinase (MMP-9) activity in exacerbations of asthma.

BACKGROUND: The 72 kDa matrix metalloproteinase 2 (MMP-2) and the 92 kDa matrix metalloproteinase 9 (MMP-9) are type IV collagenases implicated in various aspects of inflammation including accumulation of inflammatory cells, tissue injury, and development of remodelling. The role of these enzymes in the pathogenesis of asthma exacerbations is unknown. METHODS: Circulating levels of MMP-2 and MMP-9 proteins and the expression of their inhibitor, tissue inhibitor of metalloproteinase 1 (TIMP-1), were measured in 21 patients experiencing an asthma exacerbation and 21 age matched patients with stable asthma. Circulating gelatinolytic activity was compared during the asthma exacerbation and during subsequent convalescence by gelatin zymography in the same individuals. In addition, MMP-9 specific activity was quantified with a colorimetric assay which uses an artificial proenzyme containing a specific domain recognised by MMP-9 in the same paired samples. RESULTS: A significant increase in the circulating level of MMP-9 was seen in patients with an asthma exacerbation compared with patients with stable asthma (202.9 (22.0) v 107.7 (9.9) ng/ml, p=0.0003). There were no significant differences in the circulating levels of MMP-2 or TIMP-1. Gelatin zymography identified two major circulating gelatinolytic activities corresponding to MMP-2 and MMP-9, and showed that asthma exacerbations are characterised by markedly increased MMP-9 activity with no significant change in MMP-2 activity compared with the activities during convalescence in the same individuals. Direct measurement showed that MMP-9 specific activity is significantly increased during asthma exacerbations compared with subsequent convalescence (269.6 (31.7) v 170.4 (12.6) ng/ml, p=0.0099). CONCLUSIONS: Asthma exacerbations are characterised by increased circulating MMP-9 activity. This increased activity may be related to exaggerated airway inflammation and airway remodelling.

Adult↗
PubMed1988

Human antibody response to outer membrane proteins and fimbriae of Haemophilus influenzae type b.

We investigated the prevalence of antibodies in childrens' sera directed against outer membrane proteins (OMP) and fimbriae of Haemophilus influenzae type b. Invasive isolates of H. influenzae type b were enriched for fimbriae production; OMP and fimbriae were resolved by SDS-PAGE. After blotting to nitrocellulose, the proteins were incubated with homologous patient sera or with sera from healthy children. IgG antibodies bound to OMP were detected by immunoperoxidase staining. Immunoblotting was also performed using purified, nondenatured fimbriae as antigen. Nine of the 10 patients studied had antibodies in the acute serum directed against one or more of the OMP. Neither the acute nor the convalescent serum of the remaining patient contained antibodies against OMP. Antibodies against a greater number of OMP were present in the convalescent serum, in comparison to the acute serum, in 4 of the 10 patients. Five of 10 patients had antibodies against the purified fimbriae of an unrelated invasive isolate in either the acute or the convalescent serum. Acute sera from patients more frequently contained antibodies directed against OMP 60K (p less than or equal to 0.01) and OMP 51K (p less than or equal to 0.003) compared with the sera of healthy controls. In contrast, the sera of healthy children more frequently contained antibodies directed against OMP 40K (p less than or equal to 0.04). Sera from both patients and controls contained antibodies against commensal Haemophilus. We conclude that although antibodies against OMP are commonly present in healthy children, antibodies against certain OMP may be markers for susceptibility or protection.

Antibodies, Bacterial↗