Achim Kaufhold

Reactogenicity and immunogenicity of a new live attenuated combined measles, mumps and rubella vaccine in healthy children

OBJECTIVE To compare the reactogenicity and immunogenicity of a novel live attenuated measles-mumps-rubella vaccine, SB MMR (Priorix; SmithKline Beecham Biologicals), with a widely used MMR vaccine, Merck MMR (M-M-R II; Merck & Co. Inc). METHODS A total of 4702 healthy children, ages 9 to 24 months, were enrolled in 8 single blind, randomized, controlled trials. Reactogenicity (local and general solicited symptoms and all unsolicited symptoms) was assessed for up to 42 days postvaccination. Immunogenicity [seroconversion rates and geometric mean titers (GMT)] was assessed at 42 or 60 days postvaccination in 1912 subjects in 7 studies. In two studies the persistence of the antibodies at Month 12 postvaccination was assessed in 201 subjects. RESULTS Local symptoms (pain on or immediately after injection; pain, redness and swelling within 4 days of injection) were reported less frequently after SB MMR than Merck MMR (P < 0.0001). General symptoms and all other events were similar between the two groups. Fever >39.5 degrees C was reported after 9.5 and 11.9% of the SB MMR and Merck MMR doses, respectively. At Days 42 to 60 postvaccination seroconversion rates for antimeasles antibodies were higher with SB MMR than with Merck MMR (98.7% vs. 96.9%, P < 0.031) but similar in both groups for anti-mumps and anti-rubella antibodies, GMTs being approximately 10% higher (P < 0.05) with Merck MMR than with SB MMR. At the Month 12 assessment the seropositivity rates and GMTs were similar in both groups. CONCLUSION When administered as primary vaccination in children in the second year of life, the new SB MMR vaccine has been shown to be superior to a comparator vaccine in terms of local reactogenicity, with equivalent immunogenicity.

Ochrobactrum anthropi Bacteremia: Report of Four Cases and Short Review

SummaryOchrobactrum anthropi, formerly“Achromobacter” CDC group Vd, is a nonfermentative, nonfastidious gram-negative bacillus, that only recently has been given attention as a potential human pathogen. Over a 2-year period, we observed four patients with multiple blood cultures that were positive for the organism. The patients had acute leukemia as underlying disease, and presented with clinical and microbiologic features consistent with catheter-related bacteremia. In three of the patients the infection initially appeared to be unrelated to chemotherapy-associated profound neutropenia and occurred early after, or was the reason for, hospital admission. The antimicrobial susceptibility of the isolates varied: unlike previously reported cases, resistance in some of our isolates included aminoglycosides, newer fluoroquinolones, and trimethoprim-sulfamethoxazole. Despitein vitro susceptibility to imipenem in initial isolates, treatment of two patients with this agent obviously failed to eradicate the organism, and the patients either relapsed with bacteremia shortly after discontinuation of treatment or remained persistently febrile and bacteremic.O. anthropi appears to be increasingly recognized as a human opportunist pathogen associated with intravascular catheters and unpredictable multiple antibiotic resistance.ZusammenfassungOchrobactrum anthropi, früher als„Achromobacter“-Gruppe CDC Vd bezeichnet, ist ein nicht-fermentatives, anspruchslos wachsendes gram-negatives Stäbchen, das erst seit kurzer Zeit als potentiell humanpathogen gilt. Während einer 2-Jahresperiode beobachteten wir vier Patienten, bei denen dieses Bakterium aus mehreren Blutkulturen isoliert wurde. Die Patienten hatten als Grunderkrankung eine akute Leukämie. Bei drei Patienten trat die Infektion jedoch unabhängig von einer schweren Neutropenie auf. Vielmehr war sie hier entweder die Ursache der Krankenhausaufnahme, oder sie trat bereits sehr kurz nach Krankenhausaufnahme auf. Die Infektion erschien nach klinischen und mikrobiologischen Befunden in allen Fällen als Katheter-assoziierte Bakteriämie. Die antimikrobielle Empfindlichkeit der Isolate war sehr uneiheitlich: im Unterschied zu früheren Berichten waren einige unserer Isolate auch resistent gegenüber Aminoglykosiden, neueren Fluorochinolonen und gegenüber Trimethoprim-Sulfamethoxazol. Die Therapie mit Imipenem bei zwei Patienten war trotzIn-vitro-Empfindlichkeit nicht erfolgreich. Ein Patient hatte erneute Bakteriämien durch den Erreger jeweils kurz nach Ende der Therapie, der andere Patient blieb unter Therapie febril und bakteriämisch.Ochrobactrum anthropi muß als nicht selten multiresistenter, opportunistischer humanpathogener Erreger vor allem Katheter-assoziierter Infektionen angesehen werden.

Early responses to nonconjugated polyribosylribitol phosphate challenge as evidence of immune memory after combined diphtheria-tetanus-pertussis-polio-Haemophilus influenzae type b primary vaccination

OBJECTIVES A high risk of invasive Haemophilus influenzae type b (Hib) disease exists in the first few years of life. A reduction in anti-polyribosylribitol phosphate (PRP) antibody concentrations follows the administration of certain diphtheria-tetanus-acellular pertussis (DTPa)-based Hib conjugate combined vaccines. However, these combined vaccines prime the immune memory, which is an important factor in protection. As yet there is no direct evidence of the time scale involved in the development of the immune memory post-primary vaccination. In this report we investigated the presence of immune memory at 10 and 12 months of age, 4 and 6 months after primary vaccination of young infants with a pentavalent combination of DTPa, inactivated poliovirus vaccine (IPV) and Hib (DTPa-IPV/Hib) vaccine. METHODS In two trials (A and B) infants received DTPa-IPV combined with Hib-tetanus conjugate (PRP-T) vaccine at 2, 4 and 6 months of age. The presence of immune memory was assessed by measuring anti-PRP concentrations 7 to 10 days after a nonconjugated PRP challenge given at 10 months in Trial A and at 12 months in Trial B. RESULTS Administration of a nonconjugated PRP challenge 4 and 6 months after primary vaccination in Trials A and B, respectively, elicited an increase in anti-PRP geometric mean concentrations (4.5 and 5.8 microg/ml, respectively) within 7 to 10 days. These concentrations exceed those reported in the literature involving unprimed children who had received a single dose of nonconjugated PRP at the same age. CONCLUSION The results demonstrate the development of anti-PRP immune memory at an early age, 4 and 6 months after completion of a three dose primary vaccination course of combined DTPa-IPV/Hib vaccine. The ability of primed infants to mount a rapid response is an important observation given the high risk of Hib infection at this critical age.

Endocarditis caused by Gemella haemolysans.

SummaryGemella haemolysans, a coccus related to the“Streptococcaceae”, was isolated from the blood of a patient with endocarditis. The patient was successfully treated with a combination of penicillin G and tobramycin, followed by clindamycin. The taxonomy of this organism, especially its relationship to“Streptococcus morbillorum” is discussed and previously reported cases ofGemella infections are reviewed.ZusammenfassungAus Blutkulturen einer Patientin mit einer Endokarditis wurde einGemella haemolysans-Stamm angezüchtet, ein zu den„Streptococcaceae“ gehöriges Bakterium. Die Patientin konnte mit einer Kombination von Penicillin G und Tobramycin sowie anschließender Clindamycin-Therapie erfolgreich behandelt werden. Die Taxonomie des Erregers, insbesondere seine Verwandtschaft zu„Streptococcus morbillorum“, wird diskutiert und eine Übersicht über kürzlich beschriebeneGemella-Infektionen gegeben.

Measuring resistance to phagocytosis of group A and G streptococci: comparison of direct bactericidal assay and flow cytometry

M protein is thought to contribute to the ability of non-opsonized group A and group G streptococci (GAS and GGS, respectively) to resist phagocytosis by polymorphonuclear leukocytes. In previous studies, correlation between M protein expression and phagocytosis was determined by incubating these pathogens in human blood and comparing colony-forming bacterial counts prior to and after exposure to blood (direct bactericidal assay; DBA). Here, we report the application of flow cytometry to measure GAS and GGS resistance to phagocytosis. The results of the assays were in complete agreement with those from DBAs. Nevertheless, flow cytometry was regarded as superior to DBA because of its speed and potential uses for quantitative studies. In addition, the use of anti-CD1 1b monoclonal antibody for granulocyte staining guaranteed a non-compromized granulocyte function. The optimized protocol for flow cytometry presented here could be utilized to directly measure the involvement of individual protein types in bacterial resistance to phagocytosis

Few-Minutes Tests for the Identification of Group A Streptococci and Enterococci with Chromogenic Substrates

The usefulness of paper strip tests for rapid identification of Streptococcus pyogenes and enterococci cultured on blood agar plates was investigated. The paper strips used contain dried chromogenic substrates for pyrrolidonyl peptidase (PYRase) and beta-glucosidase (beta-Gluc). Material from only a few colonies needed to be applied to the test strips. The reactions could be read after two min (PYRase) and 5-7 min (beta-Gluc), respectively. Results from testing 503 streptococcal strains were evaluated. The reactions proved very useful for rapid differentiation of S. pyogenes and enterococci from human sources. Nearly all strains of these streptococcal species showed positive reactions in the PYRase test whereas only the enterococci (E. faecalis and E. faecium) were positive for both enzymes.

Different alleles of the fcrA/mrp gene of Streptococcus pyogenes encode M-related proteins exhibiting an identical immunoglobulin-binding pattern

Abstract  The majority of group A streptococci (GAS, Streptococcus pyogenes) express immunoglobulin (Ig)-binding proteins. The genes encoding these proteins belong either to the emm or the emm-related (fcrA/mrp and enn) gene family and are located in close proximity on the GAS genome, where they form part of the vir regulon. In the present study analysis of sequence data of the 5′ terminal portions of the fcrA/mrp genes from GAS isolates representing 37 different M serotypes led to a classification of six different types. Thus, although fcrA/mrp genes exhibit an allelic polymorphism, they do not display the high degree of N-terminal sequence diversity found among emm genes. The nucleotide sequences of the fcrA/mrp genes from 3 GAS isolates, belonging to serotypes M8, M9, and M13 and representing newly characterized fcrA/mrp gene types, are reported. Analysis of the Ig-binding properties of recombinant FcrA/Mrp8, 9, and 13 proteins, demonstrated a similar Ig-binding profile being reactive with human IgG subclasses 1, 2, and 4. This pattern is identical to that previously described for other recombinant fcrA/mrp4, 49, 64/14 and 76 gene products, indicating that this property is not affected by the N-terminal variability. Evidence for recombination between an fcrA/mrp and an mga gene was observed in an M-type 33 strain isolate providing further support for the concept of gene rearrangement contributing to the diversity of vir regulon gene products.

Penicillin-resistant pneumococcus in community-acquired bacteremic pneumonia in Germany

For many years clinical isolates of Streptococcus pneumoniae showed uniformly high sensitivity to antibiotics, including benz3,1penicillin, which had extremely low minimum inhibitory concentration (MIC) values against pneumococcal strains. Therefore, penicillin was generally recommended as the antibiotic of choice in the treatment of pneumococcal infections. However, recently a high incidence of pneumococci resistant to penicillin has been reported from several countries [1-4]. In contrast, in the Federal Republic of Germany the rate of relative penicillin resistance (M1C 0.125-1 rag/l) has been very low and, of note, pneumococci exhibiting high-level penicillin resistance (MIC -> 2 mg/1) have not been described to date [5-7].

Species identification and antibiotic susceptibility of enterococci isolated from clinical specimens of hospitalized patients

Over a 4-month period, a total of 315 enterococci were isolated from various clinical specimens of hospitalized patients. By applying an array of biochemical tests, all strains were accurately identified to the species level, and their susceptibilities to clinically relevant antibiotics were determined by a standardized agar dilution technique. E. faecalis and E. faecium accounted for 87.3% and 9.2% of isolates, respectively. E. avium (1%), E. gallinarum (1%), E. durans (0.6%), E. hirae (0.6%), and E. casseliflavus (0.3%) isolates were also identified. Eleven strains of E. faecium and 1 E. hirae isolate were resistant to ampicillin, but none of the isolates produced beta-lactamase. Twenty-three E. faecium and 3 E. faecalis strains as well as 1 E. hirae isolate revealed imipenem resistance. A total of 25.4% enterococci (60 E. faecalis and 19 E. faecium isolates, 1 E. hirae strain) were erythromycin-resistant. Twelve strains (11 E. faecium and 1 E. avium) exhibited ciprofloxacin resistance. High-level resistance to streptomycin was found in 58 (21.1%) E. faecalis, 9 (31%) E. faecium, and both E. hirae strains, whereas high-level gentamicin resistance (HLGR) was exclusively seen in the species E. faecalis (11.6% of isolates belonging to this species). A simple agar screening test containing 500 micrograms of gentamicin per ml proved to be highly reliable for detection of HLGR. The structural gene coding for HLGR was specifically amplified by the polymerase chain reaction in all isolates showing this resistance trait. Moreover, the gene was specifically detected by a nonradioactively labelled oligonucleotide probe in colony blot hybridization assays, indicating the potential application of these molecular approaches as a diagnostic tool.

The group A streptococcal M-type 3 protein gene exhibits a C terminus typical for class I M proteins.

The M protein gene (emm gene) from a reference group A streptococcal strain of serotype M3 was amplified by the polymerase chain reaction and partially sequenced. Hybridization assays using an oligonucleotide probe derived from the N-terminal sequence revealed that this gene segment is highly homologous among M-type 3 isolates. Of note, analysis of the nucleotide sequence data from the C terminus of the gene confirmed that the emm 3 gene exhibited all the features characteristic for group A streptococcal M-class I molecules. Recently published sequence data that were assigned to emm 3 resulted from a strain confusion and were shown to be the first one derived from an emm gene of an M-untypable isolate.