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Journal Article•DOI•

New Classification of Neisseria meningitidis by Means of Bactericidal Reactions

01 May 1970-Infection and Immunity (American Society for Microbiology (ASM))-Vol. 1, Iss: 5, pp 479-484
TL;DR: A bactericidal assay is described which allows identification of distinct serotypes within a serogroup of Neisseria meningitidis, and six different serotypes, containing one or two factors, were identified among 16 group C strains examined.
Abstract: A bactericidal assay is described which allows identification of distinct serotypes within a serogroup of Neisseria meningitidis. Antisera produced in rabbits against seven group C strains by two intravenous inoculations of live organisms were found to contain two types of bactericidal antibodies. One, directed against the group-specific polysaccharide, caused various degrees of killing of all strains. Absorption of this antibody by purified group C polysaccharide revealed the presence of the second bactericidal antibody. This antibody was directed against antigenically distinct factors associated with serotype specificity. Extensive cross-absorption yielded antisera with activity directed against four separate factors. The presence of a factor in a strain was indicated by its susceptibility to killing by antisera containing antibody to that factor. A serotype was defined by the particular combination of factors. Six different serotypes, containing one or two factors, were identified among 16 group C strains examined.
Citations
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Journal Article•DOI•
TL;DR: A new serotyping nomenclature and scheme based on these two proteins and the lipopolysaccharide are proposed, which are predominant proteins, show a useful degree of antigenic variation, and are epidemiologically relevant.
Abstract: Neisseria meningitidis serogroups B and C were originally subdivided into serotypes by the use of two different classification systems based upon type-specific bactericidal antibodies and immunoprecipitation in agar gels, respectively. The serotype specificities were later found to be associated with different major outer membrane proteins and with the lipopolysaccharide. Physiochemical characterization of the four to five major outer membrane proteins has resulted in designation of classes 1-5 on the basis of peptide mapping. The class 2 (41,000-dalton) or class 3 (38,000-dalton) protein is present in all meningococcal strains. They both are predominant proteins, show a useful degree of antigenic variation, and are epidemiologically relevant. A new serotyping nomenclature and scheme based on these two proteins and the lipopolysaccharide is therefore proposed. With use of this scheme, a meningococcal strain could be identified by serogroup to protein-serotype to lipopolysaccharide serotype, as in B:2a:L3. The class 1 (46,000-dalton) protein can be used to further define a meningococcal strain, as in B:2a:P1.2:L3.

499 citations


Cites background or methods or result from "New Classification of Neisseria men..."

  • ...The previous serotyping schemes of Gold and Wyle [1], Kasper et al....

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  • ...[3] and Gold and Wyle [1], respectively....

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  • ...These initial observations were extended by Gold and Wyle [1], who in 1970 reported that group C meningococci could be subdivided into...

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  • ...A number of serotyping schemes have been used to identify antigenically related strains of Neisseria meningitidis serogroups B and C [1-4] and have been the subject of earlier reviews [5, 6]....

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Journal Article•DOI•
TL;DR: The standardized SBA was reliable in all laboratories regardless of experience in performing SBAs, and will facilitate interlaboratory comparisons of the functional antibody produced in response to current or developing serogroup A and C meningococcal vaccines.
Abstract: A standardized serum bactericidal assay (SBA) is required to evaluate the functional activity of antibody produced in response to Neisseria meningitidis serogroup A and C vaccines. We evaluated assay parameters (assay buffer, target strains, growth of target cells, target cell number, complement source and concentration, and methods for growth of surviving bacteria) which may affect the reproducibility of SBA titers. The various assay parameters and specificity of anticapsular antibody to five serogroup A strains (A1, ATCC 13077, F8238, F9205, and F7485) and four serogroup C strains (C11, G7880, G8050, and 1002-90) were evaluated with Centers for Disease Control and Prevention meningococcal quality control sera. The critical assay parameters for the reproducible measurement of SBA titers were found to include the target strain, assay incubation time, and complement. The resulting standardized SBA was used by 10 laboratories to measure functional anticapsular antibody against serogroup A strains F8238 and serogroup C strain C11. In the multilaboratory study, SBA titers were measured in duplicate for 14 pairs of sera (seven adults and seven children) before and after immunization with a quadrivalent polysaccharide (A, C, Y, and W-135) vaccine. The standardized SBA was reliable in all laboratories regardless of experience in performing SBAs. For most sera, intralaboratory reproducibility was +/- 1 dilution; interlaboratory reproducibility was +/- 2 dilutions. The correlation between median titers (interlaboratory) and enzyme-linked immunosorbent assay total antibody concentrations was high for both serogroup A (r = 0.86; P < 0.001; slope = 0.5) and serogroup C (n = 0.86; P < 0.001; slope = 0.7). The specified assay, which includes the critical parameters of target strain, incubation time, and complement source, will facilitate interlaboratory comparisons of the functional antibody produced in response to current or developing serogroup A and C meningococcal vaccines.

410 citations


Cites background or methods from "New Classification of Neisseria men..."

  • ...N. meningitidis serogroup A and C polysaccharide-specific antibodies were measured by a standardized ELISA (3, 8)....

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  • ...In addition, there was a significant (P , 0.001) correlation between SBA titers and ELISA antibody concentrations for both serogroup A and C....

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  • ...These meningococcal QC sera (CDC QC sera) and the ELISA reference serum are maintained as a lyophilized collection at CDC and were rehydrated with sterile endotoxin-free water just before use in the assay....

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  • ...This standardized serogroup A and C meningococcal SBA, modified from the method recommended by WHO, gave serogroupspecific, reproducible bactericidal titers that correlated with ELISA antibody concentrations....

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  • ...In addition, the serum titers were higher with GBSS-gelatin....

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Journal Article•DOI•
TL;DR: The application of 13-C nuclear magnetic resonance to the analysis of some sialic acid-containing meningococcal polysaccharide antigens is described and complete assignments of the spectra of both the native serogroup B and the de-O-acetylated serogroups C polySaccharides have been made.

331 citations

Journal Article•DOI•
TL;DR: Carriage of N. lactamica may assist in the development of natural immunity to N. meningitidis by induction of cross-reactive antibodies.
Abstract: Asymptomatic carriage of Neisseria meningitidis and Neisseria lactamica was studied in a total of 2,969 healthy infants and children in Danbury, Conn., between October 1971 and June 1975. The prevalence of N. meningitidis averaged 0.71% during the first four years of life and increased to 5.4% by 14--17 years. Rates of carriage of N. lactamica increased from 3.8% in three-month-old infants to a peak of 21.0% at 18 months and then declined to 1.8% by 14--17 years of age. Of the children who acquired N. lactamica, 66% developed fourfold or greater rises in titers of IgG antibody to groups A, B, and/or C meningococci as determined by immunofluorescence compared with only 5% of control children. Of new carriers of N. lactamica, 40% developed increased titers of bactericidal antibody to groups A, B, and/or C meningococci as compared with 7% of noncarriers. Carriage of N. lactamica may assist in the development of natural immunity to N. meningitidis by induction of cross-reactive antibodies.

315 citations


Cites background from "New Classification of Neisseria men..."

  • ...The noncapsular serotypespecific antigens, which are present in the outer membrane protein of meningococci [28], can be differentiated by bactericidal [29] and a variety of other methods [28]....

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01 Jan 1997
TL;DR: The specified assay, which includes the critical parameters of target strain, incubation time, and complement source, will facilitate interlaboratory comparisons of the functional antibody produced in response to current or developing serogroup A and C meningococcal vaccines.
Abstract: A standardized serum bactericidal assay (SBA) is required to evaluate the functional activity of antibody produced in response to Neisseria meningitidis serogroup A and C vaccines. We evaluated assay parameters (assay buffer, target strains, growth of target cells, target cell number, complement source and concentration, and methods for growth of surviving bacteria) which may affect the reproducibility of SBA titers. The various assay parameters and specificity of anticapsular antibody tofive serogroup A strains (A1, ATCC 13077, F8238, F9205, and F7485) and four serogroup C strains (C11, G7880, G8050, and 1002-90) were evaluated with Centers for Disease Control and Prevention meningococcal quality control sera. The critical assay parameters for the reproducible measurement of SBA titers were found to include the target strain, assay incubation time, and complement. The resulting standardized SBA was used by 10 laboratories to measure functional anticapsular antibody against serogroup A strain F8238 and serogroup C strain C11. In the multilaboratory study, SBA titers were measured in duplicate for 14 pairs of sera (seven adults and seven children) before and after immunization with a quadrivalent polysaccharide (A, C, Y, and W-135) vaccine. The standardized SBA was reliable in all laboratories regardless of experience in performing SBAs. For most sera, intralaboratory reproducibility was 61 dilution; interlaboratory reproducibility was 62 dilutions. The correlation between median titers (interlaboratory) and enzyme-linked immunosorbent assay total antibody concentrations was high for both serogroup A (r 50.86;P< 0.001; slope 50.5) and serogroup C (n 50.86;P< 0.001; slope 5 0.7). The specified assay, which includes the critical parameters of target strain, incubation time, and complement source, will facilitate interlaboratory comparisons of the functional antibody produced in response to current or developing serogroup A and C meningococcal vaccines.

315 citations

References
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Journal Article•DOI•
TL;DR: Results of the present study suggest that natural immunity to meningococcal disease is initiated, reinforced, and broadened by intermittent carriage of different strains ofMeningococci throughout life.
Abstract: Results of the present study suggest that natural immunity to meningococcal disease is initiated, reinforced, and broadened by intermittent carriage of different strains of meningococci throughout life. In young adults, carriage of meningococci in the nasopharynx is an efficient process of immune sensitization. 92% of carriers of serogroup B, C, or Bo meningococci were found to develop increased titers of serum bactericidal activity to their own meningococcal isolate, and 87% developed bactericidal activity to heterologous strains of pathogenic meningococci. The rise in bactericidal titer occurred within 2 wk of onset of the carrier state, and was accompanied by an increase in titer of specific IgG, IgM, and IgA antibodies to meningococci. In early childhood, when few children have antibodies to pathogenic meningococci, active immunization seems to occur as a result of carriage of atypical, nonpathogenic strains. Immunity to systemic meningococcal infection among infants in the neonatal period is associated with the passive transfer of IgG antibodies from mother to fetus. The antigenic determinants which initiate the immune response to meningococci include the group-specific C polysaccharide, cross-reactive antigens, and type-specific antigens.

1,028 citations


"New Classification of Neisseria men..." refers background in this paper

  • ...(7), also using a bactericidal assay, subsequently reported that there is complex antigenic diversity within group C strains....

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  • ...The essential difficulty in recognizing strain differences within a serogroup lies in the fact that there are two distinct kinds of bactericidal antibodies present in rabbit anti-meningococcal serum (7)....

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  • ...(7) that distinct antigenic differences exist between strains within a serogroup have been confirmed....

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  • ...(7), seven group C strains were initially chosen for study: 60E, 1381, 1714, 913, 126E, 1185, and 321....

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Journal Article•DOI•
TL;DR: A passive hemagglutination test developed to measure antibodies to the polysaccharides demonstrated the specificity of these antigens, and this test could be used for serogrouping meningococcal isolates.
Abstract: The group-specific polysaccharides of group A and group C meningococci have been isolated by a new procedure which employs the cationic detergent Cetavlon to precipitate these polysaccharides from the whole culture. The A and C polysaccharide prepared by this method are noteworthy because they are of high molecular weight. The main constituent of the A polysaccharide is N-acetyl, O-acetyl mannosamine phosphate; of the C polysaccharide N-acetyl, O-acetyl neuraminic acid. This purification procedure, when applied to cultures of group B organisms, yields a polysaccharide consisting primarily of N-acetyl neuraminic acid. A passive hemagglutination test developed to measure antibodies to the polysaccharides demonstrated the specificity of these antigens. Using a hemagglutination inhibition test, these antigens were again found to be group-specific, and this test could be used for serogrouping meningococcal isolates.

391 citations


"New Classification of Neisseria men..." refers background or methods in this paper

  • ...With fixed sheep red blood cells sensitized with C polysaccharide (9), transitory low titers (1:32 or less) of hemagglutinating antibody were detected within 1 week after immunization....

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  • ...(9) and referred to hereafter as C polysaccharide] was prepared in New Zealand rabbits by the method of Plescia et al....

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Journal Article•DOI•
TL;DR: This work highlights the importance of knowing the carrier and removal status of canine coronavirus, as a source of infection for other animals, not necessarily belonging to the same breeds.
Abstract: * This work was supported by grant 23571 of the National Science Foundation, and grant AI04865-02 from the National Institutes of Health. Mortenson, L. E., Ann. Rev. Microbiol., 17, 115 (1963). 2 Mortenson, L. E., Biochim. Biophys. Acta, 81, 473 (1964). 3 Wilson, P. W., in Handbuch der P;flanzenphysiologie, ed. W. Ruhland (Berlin: Springer, 1958), vol. 8, p. 9. 4 McNary, J. E., and R. H. Burris, J. Bacteriol., 84, 598 (1962). 5 Arnon, D. I., M. Losada, M. Nozaki, and K. Tagawa, Nature, 190, 601 (1961). 6 Wilson, P. W., and S. G. Knight, Experiments in Bacterial Physiology (1952), 3rd ed. 'Carnahan, J. E., L. E. Mortenson, H. F. Mower, and J. E. Castle, Biochim. Biophys. Acta, 44, 520 (1960). 8 Umbreit, W. W., R. H. Burris, and J. F. Stauffer, Manometric Techniques (Minneapolis: Burgess, 1949). 9 Lux, H., in Anorganish-Chemishe Experimentier Kunst (Leipzig: J. A. Barth, 1959), 2d ed., p. 66. 10 Gornall, A. G., C. J. Bardawill, and M. M. David, J. Biol. Chem., 177, 751 (1949). 11 Stadtman, E. R., in Methods in Enzymology, ed. S. P. Colowick and N. 0. Kaplan (New York: Academic Press, 1957), vol. 3, p. 228. 12 Mortenson, L. E., Biochim. Biophys. Acta, 81, 71 (1964). 13 Mortenson, L. E., Anal. Biochem., 2, 216 (1961).

226 citations

Journal Article•DOI•

121 citations

Journal Article•
TL;DR: A rapid photometric assay method is presented for the titration of the bactericidal activity of antisera or of complement, which requires an initial reaction period during which the organisms, antibody, and complement interact.
Abstract: Summary A rapid photometric assay method is presented for the titration of the bactericidal activity of antisera or of complement. Like the conventional plate count assay method it requires an initial reaction period, during which the organisms, antibody, and complement interact. In our method, the relative numbers of organisms surviving are then determined by their growth rate properties on subculture, the quantitation of the relative total growth being made with a photoelectric colorimeter. Linear plots are obtained (on normal-deviate or probit graph paper) between the percentage kill and the dosage (in logarithmic units). From this the 50% end point and the response slope may be determined. If the culture density is standardized, the standard deviation of the absolute titer is about 10%; without such control, only the titers relative to a reference serum may be determined with this precision. Several factors which influence the test are discussed.

110 citations