Synthesis and immunologic properties in mice of vaccines composed of Staphylococcus aureus type 5 and type 8 capsular polysaccharides conjugated to Pseudomonas aeruginosa exotoxin A

Fattom, Ali; Schneerson, Rachel; Szu, Shousun C.; Vann, Willie F.; Shiloach, Joseph; Karakawa, Walter W.; Robbins, John B.

doi:10.1128/iai.58.7.2367-2374.1990

Cited by 124 publications

(65 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Cultures which did not show agglutination at a titre of Ͻ 1:4 of the original sera were considered fully capsulated and could be tested for capsular typing. Two different rabbit sera were used in the direct agglutination assay to identify the capsular type of each isolate: sera from animals vaccinated with a whole cell vaccine (Karakawa et al 1985), and sera prepared in rabbits against either type 5 or type 8 conjugated vaccine (Fattom et al 1990. Monoclonal antibodies to capsular types 5 and 8 produced and standardized by Nabi (Rockville, Maryland) were used to confirm the capsular serotype of the isolates.…”

Section: Capsular Polysaccharide Typingmentioning

confidence: 99%

Molecular, antibiogram and serological typing of Staphylococcus aureus isolates recovered from Al‐Makased hospital in East Jerusalem

Essawi¹,

Nawas²,

Hawwari³

et al. 1998

Tropical Med Int Health

View full text Add to dashboard Cite

SummaryStaphylococcus aureus is a major cause of nosocomial infections and a risk in patients who have either undergone surgery or are on haemodialysis. Pulsed-field gel electrophoresis of all isolates with the restrictionendonuclease enzymes Sma I revealed 34 patterns demonstrating that no single methicillin-sensitive S. aureus strain was endemic in the hospital. However, all MRSA isolates with a type 8 capsule showed identical PFGE patterns using the 2 restriction-endonuclease enzymes Sma I and SST II. Moreover, type 5 isolates showed identical patterns (one isolate differed from the rest with one band only). These data suggest and confirm the clonality of type 5 and type 8 MRSA isolates. Analysing the results of the capsular and antibiogram typing schemes in conjunction proved useful and suggested that such an analysis can be employed as a helpful epidemiological tool in hospitals with limited resources.keywords Staphylococcus aureus, nosocomial infection, epidemiology, pulsed-field-gel electrophoresis correspondence Dr Ali Fattom, Nabi Rockville,

show abstract

Section: Capsular Polysaccharide Typingmentioning

confidence: 99%

Molecular, antibiogram and serological typing of Staphylococcus aureus isolates recovered from Al‐Makased hospital in East Jerusalem

Essawi¹,

Nawas²,

Hawwari³

et al. 1998

Tropical Med Int Health

View full text Add to dashboard Cite

show abstract

“…Conjugation was carried out by thiolation of T5/ 8 PS followed by coupling to rEPA via an N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) spacer. 6 Samples of bulk concentrate and final fill conjugate vaccines in 200 mM NaCl, 10 mM sodium phosphate buffer, pH 7.4 with 0.01% polysorbate 80 (Tween 80) were stored at -70˚C and 4˚C, respectively. The corresponding purified T5/ 8 PS and rEPA lots were stored without detergent at -70˚C until used.…”

Section: Methodsmentioning

confidence: 99%

“…4,5 Like many other bacterial capsular polysaccharides, S. aureus types 5 and 8 polysaccharides (T5 and 8 PS) alone did not elicit an antibody response in animal models. 6 A bivalent conjugate vaccine using S. aureus capsular polysaccharides T5 and 8 conjugated to a non-toxic genetic mutant of Pseudomonas aeuriginosa exotoxin A (ETA) was first reported by Fattom et al in 1990 6 as the first prototype vaccine for S. aureus. A genetically detoxified recombinant P. Aeuriginosa exoprotein A (rEPA) expressed in Escherichia coli was later used for this conjugate to eliminate all residual toxicity due to ADP-ribosylating activity of ETA.…”

Section: Introductionmentioning

confidence: 99%

Preclinical Laboratory Evaluation of a BivalentStaphylococcus aureusSaccharide: Exotoxin A Protein Conjugate Vaccine

Bolgiano²,

Martino³

et al. 2006

Human Vaccines

View full text Add to dashboard Cite

A bivalent, unadjuvanted conjugate vaccine composed of Staphylococcus aureus capsular polysaccharides type 5 and 8 (T5 and T8 PS) conjugated to a novel carrier protein, the mutant nontoxic recombinant Pseudomonas aeruginosa exotoxin A (rEPA), has been the subject of recent clinical trials. A program of preclinical laboratory evaluation was carried out in support of the clinical trials conducted by the National Vaccine Evaluation Consortium. This involved physical chemical characterization and limited assessment of toxicity and immunogenicity. The carrier protein showed good stability and its conformation was essentially maintained when conjugated. The T5- and T8-rEPA conjugates were of a size range (1-3 x 10(6) g/ mol) consistent with polysaccharide conjugates. Fluorescence spectroscopy and molecular sizing showed good batch-to-batch consistency. Although all batches of final fill preparations elicited positive immune responses in the mouse model with three schedule doses of 0.25 microg of each T5/8 conjugate per dose, the mouse serum IgG response to T8 PS varied from batch to batch. Storage temperature at 37 degrees C or below or with repetitive temperature fluctuations did not significantly affect the IgG responses to T5 or T8 PS. Storage at 56 degrees C, however, diminished the mouse serum IgG response to T5 PS. The conformation of the conjugated protein and size of the conjugates correlated well with mouse immunogenicity in the thermal stability samples; significant unfolding of the protein and downshifts in molecular size of the conjugate were only observed when stored at 56 degrees C. The relatively high stability of the novel carrier protein when conjugated to large polysaccharides makes this an attractive candidate carrier protein for other conjugate vaccines. When assayed for serum IgG concentration, the bivalent T5/ 8 conjugate was found to evoke an IgG response well over the threshold value of 10 microg/ ml anti-T5 and -T8 IgG established for the ELISA immunogenicity assay.

show abstract

“…Capsule-conjugate technology has been expanded to generate vaccines against other pathogens, including Neisseria meningitidis (Frasch, 2005), Streptococcus pneumoniae (Black et al, 2000), and Salmonella typhi (Lin et al, 2001). The discovery of CP in S. aureus prompted the development of capsular-conjugate vaccines (Fattom et al, 1990). All clinical S. aureus isolates can be categorized as two CP types (CP5 and CP8) or capsule negative (CP − ; Fig.…”

Section: Ghost Of Vaccines Pastmentioning

confidence: 99%

Staphylococcus aureus vaccines: Deviating from the carol

Missiakas

Schneewind

2016

Journal of Experimental Medicine

View full text Add to dashboard Cite

Staphylococcus aureus, a commensal of the human nasopharynx and skin, also causes invasive disease, most frequently skin and soft tissue infections. Invasive disease caused by drug-resistant strains, designated MRSA (methicillin-resistant S. aureus), is associated with failure of antibiotic therapy and elevated mortality. Here we review polysaccharide-conjugate and subunit vaccines that were designed to prevent S. aureus infection in patients at risk of bacteremia or surgical wound infection but failed to reach their clinical endpoints. We also discuss vaccines with ongoing trials for combinations of polysaccharide-conjugates and subunits. S. aureus colonization and invasive disease are not associated with the development of protective immune responses, which is attributable to a large spectrum of immune evasion factors. Two evasive strategies, assembly of protective fibrin shields via coagulases and protein A–mediated B cell superantigen activity, are discussed as possible vaccine targets. Although correlates for protective immunity are not yet known, opsonophagocytic killing of staphylococci by phagocytic cells offers opportunities to establish such criteria.

show abstract

Synthesis and immunologic properties in mice of vaccines composed of Staphylococcus aureus type 5 and type 8 capsular polysaccharides conjugated to Pseudomonas aeruginosa exotoxin A

Cited by 124 publications

References 49 publications

Molecular, antibiogram and serological typing of Staphylococcus aureus isolates recovered from Al‐Makased hospital in East Jerusalem

Molecular, antibiogram and serological typing of Staphylococcus aureus isolates recovered from Al‐Makased hospital in East Jerusalem

Preclinical Laboratory Evaluation of a BivalentStaphylococcus aureusSaccharide: Exotoxin A Protein Conjugate Vaccine

Staphylococcus aureus vaccines: Deviating from the carol

Contact Info

Product

Resources

About