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Biomedical subjects

M Leinonen

Publications and source records attributed to M Leinonen.

At least 127 records · Page 7Linked to original sources
PubMed1996

PCR assay for detecting Streptococcus pneumoniae in the middle ear of children with otitis media with effusion.

We compared a newly developed pneumococcal polymerase chain reaction (PCR) for Streptococcus pneumoniae (Pnc) to bacterial culture in 123 middle ear effusion (MEE) samples of 123 children with otitis media with effusion (OME). For the pneumococcal PCR assay, DNA of MEE samples was purified by a QIAamp blood kit. The outer primers used amplified a 348 basepair region of the pneumolysin gene, and the inner a 208. Pnc was cultured in 14 (11%) and pneumolysin PCR was positive in 57 (46%) of the 123 MEE samples. All the culture positive samples were also PCR-positive. Both the samples with culturable Pnc and with positive pneumolysin PCR increased with shorter duration of OME and a greater number of acute otitis media during the preceding 6 months. In conclusion, pneumolysin PCR suggests pneumococcal involvement in MEE even in OMEs with no evidence of Pnc in culture, and thus offers a good diagnostic tool when a more accurate and sensitive pneumococcal diagnosis is needed.

Acute Disease↗
PubMed1995

The antibody response to a prototype liposome vaccine containing Neisseria meningitidis outer membrane protein P1 produced in Bacillus subtilis.

Monoclonal antibodies to the class 1 outer membrane protein P1 of Neisseria meningitidis B:15:P1.7,16 have been shown to be bactericidal and protective in an infant rat meningitis model. We have produced the P1 protein in Bacillus subtilis as inclusion bodies. When the purified and denatured protein (BacP1) was reconstituted with phosphatidylcholine into liposomes, native antigenic epitopes were formed. Such liposomes were reproducibly immunogenic in mice and guinea pigs at a low dose (1-10 micrograms of BacP1 protein) and without any other adjuvant. The resulting antisera contained high titers (enzyme immunoassay) of antibodies directed to native P1 epitopes exposed on the surface of meningococcal cells. The sera were also active with live N. meningitidis in bactericidal assays and protective in the infant rat meningitis model; all these activities were specific to the serosubtype of the P1 protein.

Animals↗
PubMed1995

Experimental Chlamydia pneumoniae infection in mice: effect of reinfection and passive immunization.

NIH/S mice were infected intranasally with Chlamydia pneumoniae isolate Kajaani 6 and rechallenged after either 28 or 70 days. A partial resistance to reinfection, indicated by a reduced recovery of live organisms, was noted at both time points of rechallenge: positive isolations from lung homogenates and/or bronchoalveolar lavage fluids were observed in fewer mice and the yields of isolated chlamydiae remained smaller, as compared to primary infection. However, a previous infection did not confer any protection against inflammatory changes. A strong peribronchial and perivascular inflammation with infiltrating lymphocytes and plasma cells was noted in the lungs of primary infected, as well as reinfected, mice. The effect of passive immunization was also studied. When mice were given convalescent or hyperimmune sera intraperitoneally before inoculation, lower C. pneumoniae isolation yields were detected. As in the rechallenge experiment, marked inflammation could still be seen in the lungs, now with polymorphonuclear leukocyte infiltration. The results suggest that immunological reactions play a role in the pathogenesis of C. pneumoniae infection. Antibodies may be important in reducing the amount of infective elementary bodies, but complete clearing of C. pneumoniae could not be achieved in these experiments, even less a protection against inflammatory lung changes.

Animals↗
PubMed1995

High prevalence of antibodies to Chlamydia pneumoniae; determinants of IgG and IgA seropositivity among Jerusalem residents.

The prevalence of antibodies to Chlamydia pneumoniae was examined in a stratified random sample of 581 Jerusalem adult residents between August 1987 and March 1989. IgG and IgA titres were measured by microimmunofluorescence, and associations with smoking and socio-demographic variables were assessed. IgG antibodies were found in 84.5% (95% confidence interval (CI): 80.4-87.9) of men and 68.7% (95% CI: 61.6-75.0) of women (P < 0.0001 for sex difference), indicating a very high rate of exposure in this population. IgA antibodies, postulated to represent persistent infection, were present in 45.1% (95% CI: 40.1-50.2) of men and 23% (95% CI: 17.4-29.7) of women (P < 0.0001 for sex difference). Factors associated with IgG seropositivity included family size, education and social class. On the other hand age (in men) and smoking were associated with IgA seropositivity. These findings support the hypothesis that low socioeconomic status and household crowding may be predictive of exposure to or infection with this organism (IgG seropositivity), whereas they do not explain persistence of the infection putatively expressed as IgA seropositivity.

Adult↗
PubMed1995

Antimicrobial resistance of Streptococcus pneumoniae in Finland, 1987-1990.

A nationwide survey of the prevalence of antimicrobial resistance among Streptococcus pneumoniae isolates from the middle ear fluid of children with acute otitis media (639 strains) and from throat-swab samples of healthy children (149 strains) was conducted in Finland during 1987-1990. The MICs of penicillin, cephalothin, cefaclor, erythromycin, trimethoprim, and co-trimoxazole were determined by the agar dilution method. Low-level resistance to penicillin (MIC, 0.1-1 microgram/mL) was found in 1.7% of the otitis-related and 1.3% of the healthy-carrier strains. No highly penicillin-resistant strains (MIC, > or = 2 micrograms/mL) were found. Six multiresistant strains were detected, three of them possibly belonging to a previously identified clone present in Finland since 1985. Eighty-five percent of the resistant otitis-related strains, including 9 of the 11 moderately penicillin-resistant strains (4 of which were multiresistant), belonged to the three most common serogroups (6, 19, and 23).

Acute Disease↗
PubMed1995

Diagnosis of bacteremic pneumococcal pneumonia by amplification of pneumolysin gene fragment in serum.

A polymerase chain reaction (PCR) assay based on the amplification of pneumolysin gene fragments in patient sera was developed for the etiologic diagnosis of acute pneumococcal pneumonia. Two pairs of oligonucleotide primers were designed to amplify a 348-bp and a 208-bp fragment of the pneumolysin gene. Amplified products were analyzed by agarose gel electrophoresis and hybridization using a 24-mer probe internal to the amplified DNA. The nested PCR could detect 10 organisms as determined by serial dilutions of DNA from Streptococcus pneumoniae. All 20 serum samples from patients with acute pneumococcal pneumonia (confirmed by blood culture) were positive. Among 100 serum samples from healthy elderly controls, 94 were negative and 6 were positive by PCR. Thus, PCR may be a novel diagnostic method for pneumococcal pneumonia.

Adult↗
PubMed1995

Nasopharyngeal antibodies to pneumococcal capsular polysaccharides in children with acute otitis media.

Antibodies to pneumococcal capsular polysaccharides were measured by EIA in acute- and convalescent-phase nasopharyngeal aspirates from 120 children with acute otitis media. Nasopharyngeal IgM- and IgG-class antibodies were rare, whereas IgA was detected more often, occurred independently from serum IgA, and correlated with the presence of the secretory component in pneumococcal antibody, indicating local production of IgA. Thirty-four percent of the children with pneumococci in middle ear fluid developed a nasopharyngeal IgA response to the polysaccharide pool of serotypes 6B, 14, 19F, and 23F compared with 7% of the children with pneumococci only in the nasopharynx or not found at all (P = .004). The responses were observed in children of all ages, the youngest 6 months of age. This supports the hypothesis that mucosal immunity to bacterial polysaccharides matures earlier than systemic immunity.

Acute Disease↗
PubMed1995

The etiology of community-acquired pneumonia among hospitalized patients during a Chlamydia pneumoniae epidemic in Finland.

The etiology of community-acquired pneumonia during a Chlamydia pneumoniae epidemic was studied among 125 hospitalized patients. Etiologic investigations included blood and sputum cultures, pneumococcal antigen detection, and serologic investigations for common respiratory viruses and for Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Mycoplasma pneumococcal antigen detection, and serologic investigations for common and for Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Mycoplasma pneumoniae, and Chlamydia species. A specific microbial agent was identified in 110 patients (88%). Some 48% of the patients had mixed infections. S. pneumoniae was the most common pathogen (55%), followed by C. pneumoniae (43%). C. pneumoniae was found both as a single etiologic agent and as a mixed infection, most often with S. pneumoniae. In conclusion, S. pneumoniae is the most common cause of community-acquired pneumonia, even during a C. pneumoniae epidemic. The organism, alone or with other pathogens (especially S. pneumoniae), may cause community-acquired pneumonia that requires hospital treatment.

Adolescent↗
PubMed1995

Bacterial antibody assays in the diagnosis of acute lower respiratory tract infection in children.

Bacterial antibodies were studied in acute, intermediate and convalescent phase sera (mean duration from first to last sample 36 days) of 121 children hospitalized for acute lower respiratory tract infection. Antibody responses were observed in 45% of all cases and in 29% of the 21 children < 1 year old. A total of 15 responses to Streptococcus pneumoniae (pneumolysin), 20 to Haemophilus influenzae, 9 to Moraxella catarrhalis, 3 to chlamydiae and 8 to Mycoplasma pneumoniae were found. In 79 patients with 4 consecutive samples available, 52% of the 31 responses were measurable within 5 days from admission. Overall the responses were not associated with upper respiratory tract bacterial findings or acute otitis media. Significantly more responses were found in the 121 children with acute lower respiratory tract infection than in healthy controls (P < 0.007). We conclude that bacterial antibody assays provide a useful tool in the study of the etiology of acute lower respiratory tract infection in young children, even if the interval between paired serum samples is short.

Acute Disease↗
PubMed1995

Erythrocyte sedimentation rate, white blood cell count and serum C-reactive protein in assessing etiologic diagnosis of acute lower respiratory infections in children.

The clinical signs, symptoms and host responses (erythrocyte sedimentation rate, white blood cell count and C-reactive protein) were studied to distinguish bacterial from viral acute lower respiratory infection (ALRI) in 121 children hospitalized for ALRI. Etiologic diagnosis was based on blood culture, antibody assays and antigen detection. Children with bacterial involvement only were older than those with viral involvement alone (mean, 5.1 vs. 2.5 years), and their duration of respiratory symptoms had lasted longer (mean, 4.6 vs. 3.3 days). Children with unknown etiology had a shorter duration of fever before hospitalization than those with etiology identified with the methods used (mean, 1.6 vs. 2.9 days). The host response ranged widely within etiologic groups. The mean erythrocyte sedimentation rate did not differ significantly between the bacterial and viral ALRI (38 vs. 28 mm/hour); neither did white blood cell count (13.2 vs. 13.6 x 10(9)/liter) or C-reactive protein (68 vs. 49 mg/liter). No combination of clinical signs and host responses or any cutoff values could be shown to differentiate reliably bacterial from viral ALRI.

Acute Disease↗
PubMed1995

Human immune response against outer membrane proteins of Moraxella (Branhamella) catarrhalis determined by immunoblotting and enzyme immunoassay.

The role of Moraxella (Branhamella) catarrhalis as a respiratory tract pathogen is increasingly recognized. We looked at the human immune response against individual outer membrane proteins of M. catarrhalis and against the 81-kDa CopB protein, which has previously been shown to be a target for protective antibodies. Paired serum samples from six elderly patients with pneumonia were tested by Western blot (immunoblot) analysis by using outer membrane vesicles of M. catarrhalis 035E as antigen. All of the six convalescent-phase serum samples reacted with a protein which migrated at the position of the CopB protein and with a high-molecular-weight protein of M. catarrhalis; three serum samples also reacted with a 34-kDa outer membrane protein. Paired serum samples from 18 patients, 10 of which had M. catarrhalis infection on the basis of previous serology results, were tested by enzyme immunoassay (EIA) with the CopB protein and whole cells of M. catarrhalis 035E as antigens. Nine patients showed a significant rise in EIA titer between acute- and convalescent-phase sera when whole bacterial cells were used as antigens. Six (67%) patient samples that were positive by the EIA with the whole-cell antigen were also positive by the EIA with the CopB antigen, and six of nine patient samples negative by the EIA with the whole-cell antigen were also negative by the EIA with the CopB antigen. These results suggest that both the CopB and a high-molecular-weight protein are major targets of the immune response against M. catarrhalis, and further studies with greater amounts of patient materials are needed to elucidate the usefulness of CopB as an antigen in etiologic studies.

Aged↗
PubMed1995

Measurement of sputum antibodies in the diagnosis of acute and chronic respiratory infections associated with Chlamydia pneumoniae.

The aim of this study was to develop methods for the measurement of sputum antibodies in the laboratory diagnosis of acute and chronic lower respiratory tract infections caused by Chlamydia pneumoniae. Paired serum specimens, sputum specimens, and pharyngeal or nasopharyngeal swabs were obtained from 97 patients; 51 of them had community-acquired pneumonia, and 46 had chronic obstructive pulmonary disease (COPD). C. pneumoniae-specific serum immunoglobulin G (IgG), IgA, and IgM antibodies were measured by the microimmunofluorescence (micro-IF) test. For sputa, specific IgA and IgG antibodies were measured by the micro-IF test and secretory IgA (sIgA) was measured by enzyme immune assay (EIA) with C. pneumoniae elementary bodies as the antigen. Sputum IgA and sIgA antibodies to C. pneumoniae were found, respectively, in 52 and 51% of the COPD patients. Elevated levels of stable serum IgG and IgA antibodies (IgG titer of > or = 128 and IgA titer of > or = 40), suggesting chronic infection, were found in 54% of the COPD patients. The sensitivity for the sputum IgA micro-IF test compared with elevated serum antibody levels was 87.5%, and that for the sputum sIgA EIA was 88%; the respective specificities were 90 and 95%. Acute C. pneumoniae infection was diagnosed in seven pneumonia patients, and two (29%) of these patients were positive by sputum EIA antibody measurements. Two pneumonia patients without acute infection had stable elevated IgG and IgA levels in their sera, and both of them were sputum antibody positive. We conclude that the measurement of IgA antibodies to C. pneumonia in sputum is a useful additional diagnostic tool for chronic C. pneumoniae infections.

Acute Disease↗
PubMed1995

Nasopharyngeal antibodies to pneumococcal pneumolysin in children with acute otitis media.

Pneumolysin, an intracellular protein toxin of all clinically relevant pneumococcal serotypes, is released in vivo during the autolysis of pneumococci and is believed to pave the way for intact pneumococci to invade and cause disease. Therefore, antibodies to pneumolysin should prevent its destructive function. We measured antibodies to pneumococcal pneumolysin in acute- and convalescent-phase nasopharyngeal aspirate samples of 120 children (median age, 2.5 years) with acute otitis media by enzyme immunoassay. Nasopharyngeal immunoglobulin M (IgM) and IgG class antibodies to pneumolysin were rarely detectable, whereas IgA class antibody was detected often, occurred independently of serum IgA antibody in serum, and correlated with the presence of the secretory component in pneumococcal antibody, indicating local production of IgA antibodies. Nasopharyngeal IgA antibody to pneumolysin was detected in 93% of the children already in the acute phase of otitis. Twenty percent of the children developed at least a threefold rise in the pneumolysin-specific IgA antibody concentration by the convalescent phase of otitis, with the youngest at 6 months of age, regardless of the pneumococcal findings in the nasopharynx or middle ear fluid. We suggest that nasopharyngeal IgA antibody to pneumolysin can be produced early in life by pneumococcal colonization.

Acute Disease↗
PubMed1995

Evaluation of sampling sites for detection of upper respiratory tract carriage of Streptococcus pneumoniae and Haemophilus influenzae among healthy Filipino infants.

Two sampling techniques, nasal swabbing and oropharyngeal swabbing, for detection of the upper respiratory tract carriage of Streptococcus pneumoniae and Haemophilus influenzae were studied prospectively with 296 healthy Filipino infants at various ages: 6 to 8, 10 to 12, 14 to 17, 18 to 22, 32 to 39, and 46 to 65 weeks. In all age groups S. pneumoniae was isolated significantly more often (P < 0.0001) from the nasal site than from the oropharyngeal site. H. influenzae was found equally often at both sites.

Carrier State↗
PubMed1995

Antibodies to pneumolysin and pneumococcal capsular polysaccharides in middle ear fluid of children with acute otitis media.

Antibodies to pneumococcal pneumolysin and capsular polysaccharides were measured by enzyme immunoassay in 169 acute phase middle ear fluid samples of 116 children with acute otitis media. Antibodies to pneumococcal pneumolysin were detected in 84% and to capsular polysaccharides in 50% of the MEF samples. The Ig class detected most often was IgA to both types of pneumococcal antigens, and it was present in MEF even with non-detectable levels of serum IgA of the same specificity. 59% of the MEF samples positive for IgA to pneumolysin were also positive for secretory component of the same specificity, and 53% of IgA to capsular polysaccharide pool (containing serotypes 6B, 14, 19F, and 23F), respectively. This suggests both leakage of specific IgA from serum to the middle ear and local production of it. In contrast, specific IgG was detected in MEF only with concomitant IgG in serum. Antibodies to pneumolysin occurred in no relation to bacterial findings in MEF. On the contrary, IgG class antibodies to capsular polysaccharides, most likely serum-derived, were detected less often in MEF samples positive for pneumococcus than for other bacteria.

Antibody Formation↗
PubMed1994

Chlamydia pneumoniae multiplies in human endothelial cells in vitro.

The ability of three C. pneumoniae isolates, Kajaani 6, Helsinki 12 and TW-183, to grow in human umbilical vein endothelial cells (HUVEC) and in an immortalized endothelial cell line EA.hy 926 was studied. All C. pneumoniae isolates were capable of multiplying in endothelial cells. EA.hy926 cells could support the growth of C. pneumoniae better than HUVEC, yet less efficiently than HL and HEp-2 cells that are conventionally used in C. pneumoniae culturing. Although centrifugation of the inoculum greatly increased the inclusion yields, it was not necessary for infectivity. In addition, a persistent infection of C. pneumoniae in EA.hy 926 and HL cells ensued and it was followed up for two months. The fact that endothelial cells can serve as hosts to C. pneumoniae might be a significant contributing factor in the pathogenesis of atherosclerosis, a disease which recent studies show to be associated with chronic C. pneumoniae infection.

Cells, Cultured↗
PubMed1994

Use of serology to diagnose pneumonia caused by nonencapsulated Haemophilus influenzae and Moraxella catarrhalis.

Antibodies against nonencapsulated Haemophilus influenzae and Moraxella (Branhamella) catarrhalis were measured by ELISA in paired sera from 158 adult patients with pneumonia. A mixture of 10 clinical isolates of each species was used as antigen. Eleven patients (7%) showed significant increases in antibody to H. influenzae. In 3 of them, the organism was isolated from transtracheal aspirate and in another 7 from sputum, nasopharynx, or both. Six patients with nonencapsulated H. influenzae in transtracheal aspirate cultures did not show any antibody increase. Six patients had significant increases in antibody to M. catarrhalis. The organism was isolated in transtracheal aspirates from 1 of them and in sputum and nasopharynx (or both) from another 3. Two patients with M. catarrhalis in transtracheal aspirate cultures showed no antibody response. In conclusion, the serologic methods increased the possibility to diagnose infections caused by the two agents but had low sensitivity.

Adult↗