The relation between the bactericidal activity of complement and the character of the bacterial surfaces

The bactericidal activity of sera not containing antibodies (sera from precolostral piglets and calves) was tested with strains of gram-negative bacteria with different surfaces. The accuracy of the method of bactericidal test was evaluated statistically the bactericidal unit of complement was defined for comparing the activity of sera of different animals and different species. Various methods used for estimating the character of bacterial surface were compared. It was found that the bactericidal activity of piglet sera is directly dependent on the content of complement in the sera tested and the character of the bacterial surface (in the R-form). In selected strains there is a correlation in all criteria characterizing the surfaces of bacteria, and their susceptibility to bactericidal activity of sera; in a group of 37 strains selected at random, correlation with only one of the surface characteristics (stability in solution after heating to 100°C for 1 hour) was found. In calf sera a component was found which increases the effect of complement to some strains (e.g.Shigella shigae). This component may by absorbed from the serum only in the presence of complement. The nature of this factor is discussed.     

Humoral immunity aspects of meningococcal infection. I. A method of performing and the characteristics of the immune bacteriolysis reaction]

Immune bacteriolysis test with meningococcus, group A, was used for the purpose of serum antibody study. Meningococcus cultures with a bright orange fluorescence of the colonies in oblique illumination (the I type) proved to possess the greatest lysability. Guinea pig serum sorbed with meningococcus suspension was found to be the best source of the complement. Sera obtained after 1 to 3 days of rabbit immunization, containing mostly IgM antibodies, had the greatest bactericidal capacity. Only those fractions which contained IgM possessed bactericidal activity in the hyperimmune rabbit sera with a high IgG antibody concentration. No lytic activity was displayed against meningococcus by unfractionated hyperimmune sera.     

Antibodies to meningococcal H.8 (Lip) antigen fail to show bactericidal activity

Purified H.8 (Lip) antigen was coupled to tresyl-activated Sepharose 4B and used in affinity columns to purify anti-Lip antibodies from convalescent patient sera and from immune rabbit sera. Affinity-purified anti-Lip antibodies isolated from two convalescent patient sera contained 1000 and 1280 ELISA units of antibody and included antibodies of IgG, IgA, and IgM isotypes. An anti-Lip mouse monoclonal ascites (2-1-CA2) had 28,400 ELISA units of antibody. Bactericidal assays were performed using three different case strains of Neisseria meningitidis group B, namely 44/76, 8532, and 8047. Neither preparation of purified human anti-Lip antibodies had detectable bactericidal activity against strains 44/76 and 8532, but one of the two had a titer of 1:4 against strain 8047. Anti-Lip antibodies that were purified from immune rabbit serum and contained 1600 ELISA units of anti-Lip antibodies also failed to show detectable bactericidal activity. The rabbits were immunized with purified Lip antigen and showed specific antibody levels of 2000-2200 units by ELISA, but even the unfractionated sera had little or no bactericidal activity against the test strains. The high titer mouse monoclonal ascites had no bactericidal activity against the test strains. The poor bactericidal activity associated with monoclonal and polyclonal antibodies to the Lip antigen suggest that in spite of other attractive properties it may not be useful as a meningococcal vaccine.     

Variability of the lytic susceptibility of Neisseria gonorrhoeae to human sera.

The bactericidal activity of human sera for Neisseria gonorhoeae was studied. Sera were obtained from a group of patients with gonococcal infections who had acute urethritis, acute pelvic inflammatory disease, disseminated gonococcal infection, or who were asymptomatic carriers. The homologous and heterologous strains were tested with these sera. The development of serum bactericidal antibodies was not a universal event. With few exceptions, the susceptibility of a particular strain to human antibody and complement appeared to be largely independent of the particular person from whom the serum was obtained and was due instead to antigenic properties intrinsic to each individual strain. Lipopolysaccharide appeared to be the predominant antigen against which bactericidal antibodies were directed. The principal bactericidal antibody class was IgM. Blocking antibodies were not found to account for the lack of lytic activity. A correlation of bactericidal antibodies with protection from developing gonococcal infection could not be demonstrated in three pateints.     

The enhancement of the immunogenicity of streptococcal group-A M protein by conjugation with a synthetic polyelectrolyte]

Immunization with the polypeptide fragment of group A streptococcal protein M conjugated with the copolymer of acrylic acid and N-vinylpyrrolidone in complete Freund's adjuvant has been found to lead to a sharp increase in the level of antibodies to the type-specific determinants of protein M, detected in the enzyme immunoassay (EIA). The possibility of the application of such sera to preliminary typing of streptococci in EIA with the use of whole microbial cells as antigens has been shown. The data on high activity of the sera thus obtained in the bactericidal test with streptococci of the homologous type are presented. Recommendations on the use of sera obtained by the above method for highly precise typing of the virulent cultures of group A streptococci in the bactericidal test are given.     

Comparison of the bactericidal activity of different vertebrate sera

Schwab, G. E. (University of Adelaide, Adelaide, South Australia), and P. R. Reeves. Comparison of the bactericidal activity of different vertebrate sera. J. Bacteriol. 91:106-112. 1966.-The bactericidal activity for gram-negative bacteria of normal sera from eight species of vertebrates was investigated to compare complement-mediated killing in sera from animals representing various classes of vertebrates. Although all the sera were bactericidal, there was considerable variation in the range of bacterial species killed and in the bactericidal titers. The temperature dependence of the bactericidal and hemolytic complement activities was also studied. The curves relating activity to temperature were similar in shape for sera from a homeotherm and poikilotherms, but those for the homeotherm reached their maximum at temperatures of 5 to 10 C higher than the poikilotherms. The role of lysozyme and complement in killing rough gram-negative bacteria was examined, and the results suggest that, as for smooth organisms, killing is due to antibody plus complement. These natural antibodies, like those for smooth strains, were shown to be specific.     

Impaired bactericidal activity of serum of a child suffering from focal proliferative glomerulonephritis.

The serum of a child with focal proliferative glomerulonephritis was found to exhibit a weaker bactericidal activity against Pseudomonas aeruginosa, Salmonella typhimurium, Salmonella enteritidis and Escherichia coli strains as compared with sera of the child's parents. The child's serum showed a low haemolytical activity of complement as well as a low C3 concentration. The authors believe that the abnormal complement concentration could cause the impaired bactericidal activity of the patient serum.     

Bactericidal antibody response against P6, protein D, and OMP26 of nontypeable Haemophilus influenzae after acute otitis media in otitis-prone children

The bactericidal antibody response to three nontypeable Haemophilus influenzae (NTHi) outer membrane proteins (D, P6, and OMP26) was studied in 24 otitis-prone children (aged 7-28?months) after an acute otitis media (AOM) caused by NTHi. The study was carried out to understand the contribution of antigen-specific bactericidal antibody responses in the class of children who are most vulnerable to recurrent otitis media infections. Levels of protein D (P?=?0.005) and P6 (P?=?0.026) but not OMP26 antibodies were higher in bactericidal sera compared with nonbactericidal sera. For five (24%) and 16 (76%) of 21 bactericidal sera tested, removal of anti-protein D and P6 antibody, respectively, resulted in a two- to fourfold drop in bactericidal antibody. Antibodies to OMP26 did not make any contribution to the overall bactericidal activity in any serum samples. Eleven of 21 sera (52%) had bactericidal activity against a heterologous NTHi (86-028 NP) strain but the titers were significantly lower (P?相似文献   

The bactericidal activity of sera of healthy neonates and of newborns with hyperbilirubinaemia toEscherichia coli

Cord and newborn sera were found to possess a significantly decreased in vitro bactericidal activity to Escherichia coli as compared to sera of adult individuals. Hyperbilirubinaemia in neonates suffering from both Rh and ABO incompatibility results in a further significant impairment of the bactericidal activity of their sera in comparison with healthy newborn infants. Exchange transfusion improved considerably the serum bactericidal activity. The possible mechanism of toxic action of unconjugated bilirubin on cells of the immune system is discussed.     

Rapid Assay for Bactericidal Antibodies

Microtiter plates were successfully substituted for tubes in the bactericidal test for opsonic antibodies. This method is efficient and should prove useful for screening large numbers of sera.     

Comparative evaluation of the immunological activity of new typhoid vaccines with established prophylactic effectiveness (data from a controlled experiment)]

The authors present the results of studying the immunological efficacy of a dry alcoholic typhoid vaccine enriched with S. typhi Vi-antigen in the assessment of this vaccine in controlled epidemiological trial during the immunization of children aged 7--8 years. O- and Vi-antibodies were tested in the reaction of hemagglutination, H-antibodies--in the agglutination reaction with the microbial diagnostic agent, the properties of antibodies--in a test with cystein, and bactericidal properties of the sera--against the virulent S. typhi strain. Examination of 355 coupled sera obtained before and 3 weeks after the immunization demonstrated a high level of Vi-(1:47) and of the O-(1:580) antibodies and high bactericidal properties of the sera in persons vaccinated with the alcoholic vaccine enriched with the S. typhi Vi-antigen. The results obtained and the data on the formation of prolonged immunity following a simgle immunization suggested that a high protective effect was caused by a combined action of the O- and Vi-antigens contained in the vaccine in optimal doses.     

Assessment of bactericidal and growth-inhibiting activity of blood serum using flow cytometry and photometry

Method of measurement of biological fluids bactericidal activity against Staphylococcus aureus using laser flow cytometry has been developed and proposed for clinical use. Overall bactericidal activity of sera of healthy donors has been assessed by this method. Strong positive correlation between bactericidal activity measured by flow cytometry and ability of the sera of healthy donors to inhibit bacterial growth assessed by photometric method was determined. High degree of positive correlation between results of cytometry and classical microbiological method of measurement of mentioned parameters has been shown.     

The sensitivity of<Emphasis Type="Italic">Escherichia coli</Emphasis> strains with K1 surface antigen and rods without this antigen to the bactericidal effect of serum

The susceptibility of Escherichia coli strains with K1 surface antigen (K1+) and rods without this antigen (K1-) to the bactericidal action of normal bovine serum and human normal cord serum was determined. Seventy E. coli strains (35 K1+ and 35 K1-) were isolated from urine obtained from children with urinary tract infections. The strains investigated showed variable sensitivity to the bactericidal action of the sera. E. coli K1+ strains were characterized by lower sensitivity to bactericidal effect of the sera in comparison with K1- rods. The role of the particular mechanisms of complement activation in the process of killing of the E. coli strains was also determined.     

Bactericidal activity of human sera with a physiologic or genetic defect of the complement system

The serum of patient suffering genetically conditioned C2 defect showed a weak bactericidal activity against Pseudomonas aeruginosa strains. Out of 23 tested strains one was susceptible comparing with 16 ones sensitive to normal human serum. The normal cord serum exerted a bactericidal effect on 8 strains. All sera were found to be active against Salmonella strains tested.     

Serum bactericidal activity in a secondary school population following an outbreak of meningococcal disease: effects of carriage and secretor status

Abstract Sera obtained from 106 children following an outbreak of Neisseria meningitidis (B:4:P1.15) were screened for bactericidal antibodies against isolates of meningococci and Neisseria lactamica . Most had high titres of antibodies to N. lactamica and N. meningitidis NG:4:- but not to capsulate isolates: B:4:P1.15; B:15:P1.16; B:4:-; C:4:-. Bactericidal activity was higher for both carriers and secretors but the differences were not significant. Bactericidal activity was not associated with total or specific IgA or IgM. Carriers had significantly higher levels of IgG to N. lactamica but not to NG:4:- in sera with bactericidal activity for each of the capsulate strains. Among non-carriers, higher levels of IgG to N. lactamica were associated with killing of B:4:P1.15 and B:4:-. Secretors' sera with bactericidal activity had significantly higher levels of IgG to N. lactamica compared with sera that were not bactericidal. This was not observed among non-secretors. Antibodies to the outbreak strain were adsorbed by all Neisseria isolates tested and absorption of sera with N. lactamica alone completely removed the bactericidal activity against the outbreak strain.     

The opsonic activity of complement to rough strains ofEscherichia coli in vivo in newborn precolostral pigs

The participation of complement of newborn precolostral piglets (in the absence of antibody) in the opsonization of roughEscherichia coli was investigated using blood clearance test. It was found that the complement inhibitor-yeast mannan inhibits thein vivo blood clearance when injected i. venously into newborn precolostral piglets. This inhibition takes place at the serum level and not by saturation of RES cells by mannan: sera from mannan-treated piglets are not bactericidal in anin vitro bactericidal assay against roughEscherichia coli; secondly, the presence of antibody removes the blockade of phagocytosis by mannan indicating that the RES cells are functionally capable to perform phagocytosis. On the basis of our findings we postulate that in newborn precolostral piglets in which antibody is not present in their sera, complement alone (or with cooperation with other unknown serum component) acts as an opsonin on roughEscherichia coli and is responsible for the blood clearance rate of these bacteria.     

Immunogenicity of a chaotropically extracted protective antigen(s) of Pasteurella multocida type A (bovine origin) against experimental pasteurellosis in mice.

Immunization of mice with a potassium thiocyanate extract of Pasteurella multocida type A (bovine strain) imparted complete protection against an experimental challenge infection with 1.6 x 10(3) colony-forming units of the homologous strain. Formalin-treated vaccine (one or two doses) was much less effective. Significantly higher bactericidal antibody titres were observed in the sera of mice immunized with the extract compared with those given the vaccine. The bactericidal activity was localized in the IgM and IgG fractions. The extract consisted of protein(s) and carbohydrate(s) and on immunoelectrophoresis displayed at least three antigenetically distinct components.     

State of the nonspecific humoral factors of the body's natural resistance in patients with suppurative and septic infections having various ABO system blood groups]

The results of surveying 140 patients with severe purulent and septic infections of staphylococcal etiology, when compared with the distribution of the blood groups (as classified according to the ABO system) in 180 healthy donors, revealed that generalized purulent infections occurred most frequently in patients with blood groups A (II) and AB (IV), and more seldom in patients with blood groups O (I) and B (III). The average content of lysozyme, complement and normal antibodies to E. coli, as well as the average level of general bactericidal activity in the blood sera of the patients were considerably lower than in the blood sera of healthy donors; at the same time content of lysozyme, complement and normal antibodies in the blood sera of patients having different groups of blood did not reflect the degree of their predisposition or resistance to staphylococcal infections. The general bactericidal activity of the blood serum was found to correlate with the degree of predisposition or resistance to purulent septic infections of staphylococcal etiology to a greater extent than other characteristics.     

A rapid microtiter plate serum bactericidal assay method for determining serum complement-mediated killing of Mannheimia haemolytica

In this study, we describe a rapid microtiter serum bactericidal assay (RMSBA) that can be used to measure the functionality of immune sera. It quantifies bactericidal activity of immune sera in the presence of complement against a homologous bacterium, M. haemolytica in this case. There is high correlation between data from RMSBA and standard complement-mediated bacterial killing assay (r=0.756; p<0.0001). The RMSBA activity of sera can be generated in less than 5 h instead of overnight incubation. RMSBA costs substantially less in terms of time, labor, and resources and is highly reproducible.