Evaluation of the "Long Chain Reaction" as a Means for Detecting Type-Specific Antibody to Group a Streptococci in Human Sera

Stollerman, Gene H.; Siegel, Alan C.; Johnson, Eloise E.

doi:10.1084/jem.110.6.887

Cited by 22 publications

(21 citation statements)

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“…Most virulent strains exhibited the long chaining phenomenon. Suitable representatives of Types 1,3,4,5,6,12,14,19,30, and "Red Lake" were preserved by lyophilization in rabbit blood. Frozen stocks of some strains also were prepared by resuspending the cells sedimented from 40 ml.…”

Section: Methodsmentioning

confidence: 99%

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Factors Affecting the Chain Length of Group a Streptococci

Ekstedt

Stollerman

1960

The Journal of Experimental Medicine

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At the close of the nineteenth century much attention was given to the effect of immune serum upon the growth of microorganisms. In some of these early studies it was noted that certain bacteria not only clumped together but sometimes formed threads or chains when grown in homologous antiserum.Thus, in 1889 Charrin and Roget (1) commented that B. pyocyaneus grown in immune rabbit serum formed "chalnettes" of 6 to 10 segments. Two years later Metchnikoff (2) observed that Vibrio metchnikowii formed de spirilles allong~e, in addition to thick masses, when grown in immune serum and that le microbe de la tmeumonie also grew in long chains (des paquets de streptocoques trbs longs) in the serum of vaccinated rabbits.In 1897 Bordet (3) observed that streptococci formed long, tortuous chains in rabbit anfisera. The following year "long chaining" received specific attention in the studies of Pfaundler (4) who described the phenomenon as the "thread reaction" (Fadenbildung). He observed the formation of long threads when strains of E. coli and B. proteus were grown in sera obtained from the same patients from whom the organisms were isolated. This was interpreted as a special form of agglutination in which organisms failed to divide along their long axes but remained as rods adherent end-toend.During the early part of the twentieth century several investigators studying the effect of immune serum upon the pneumococcus referred to the "thread reaction." By 1928 the sensitivity and potential usefulness of this phenomenon were appreciated by Bailey (5) who wrote: "The presence of such antibodies ( . . . . agglutinins and precipitins . . . . ) in very small amounts, not demonstrable by ordinary serological methods, may be revealed if the organism be grown in the presence of serum or blood (the thread reaction of Pfaundler). For example, the sera of patients with pneumonia frequently show no agglutinins by the usual methods, but if pneumococci are sown in such sera they grow out in masses with chain formation." *

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Section: Methodsmentioning

confidence: 99%

“…It was shown that this phenomenon was dependent upon the interaction of M protein and its antibody. The long chain reaction was found useful for detecting human type specific antibody with almost the same sensitivity as the bactericidal test (12). Long chaining was also used recently by Wilson (13) to identify M protein in strains of streptococci.…”

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Factors Affecting the Chain Length of Group a Streptococci

Ekstedt

Stollerman

1960

The Journal of Experimental Medicine

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“…Long-Chain test. The test was performed according to a method previously described (25). Tests for Type-specific Antibody Responses by Enzyme-linked Immunosorbent Assays (ELISA ).…”

Section: Preparation Ofpep M Antigens For Injection For Skin Testingmentioning

confidence: 99%

Human immune response to immunization with a structurally defined polypeptide fragment of streptococcal M protein.

Beachey¹,

Stollerman²,

Johnson³

et al. 1979

The Journal of Experimental Medicine

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The mechanism of the pathogenesis of rheumatic fever and rheumatic carditis remains unknown (1). Efforts to vaccinate against group A streptococci have been hampered by toxicity of most preparations of M protein and the fear that streptococcal vaccines may contain some of the antigens shown to be cross-reactive with heart tissues (2-4) and, therefore, theoretically may trigger, rather than prevent, rheumatic fever and carditis.It has been recognized for a long time that the resistance of group A streptococci to phagocytosis is largely attributable to the presence on the surface of virulent organisms of a type-specific antigen, the M protein. Hosts who are immune to a particular M serotype of streptococci possess antibodies that render organisms bearing the homologous type of M protein on their surfaces susceptible to ingestion and killing by host phagocytes (5). Efforts to purify the M antigen in the past, however, have failed to separate heterologous protein antigens from the type-specific determinant of M protein (6-11). Moreover, after extensive purification, the M antigen often became less immunogenic and was of questionable value in affording protective immunity (12).In our search for more effective methods for the extraction and purification of M protein for use as vaccines, we found that the antigen could be extracted in satisfactory yields by subjecting intact streptococci to limited digestion with pepsin (13). Upon purification of the peptic extracts of types 6 and 24 M protein (pep M) 1 by ammoniumsulfate precipitation, ion-exchange chromatography, and isoelectric focusing, the heterologous, heart-reactive, and toxic materials were readily separated from the type-* Supported by program-directed research funds from the United States Veterans Administration and by research grants AI-10085 and AI-13550 from the United States Public Health Service.Presented in part at the annual meeting of the Association of American Physicians, Washington, D. C., May 1979.Medical Investigator of the Veterans Administration. § Research Associate of the Veterans Administration. i Abbreviations used in th,s paper." CF, complement fixation; CFU, colony-forming units; DCH, delayed cutaneous hypersensitivity; ELISA, enzyme-linked immunosorbent assay; NTSM, non-type-specific M; PBS, phosphate-buffered saline; PBS-Tween, PBS supplemented with 0.05% Tween and 0.02% NAN3; pep M, purified peptic extracts of types 6 and 24 M protein; pep M6, type 6 M protein pepsin extracts; pep M24, type 24 M protein pepsin extracts; saline, 0.85% solution of NaCI; saline-Tween, I ml of 0.15 M NaC1 containing 0.05% Tween; SDS, sodium dodecyl sulfate. 862J. ExP. MED.

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“…Because virulent encapsulated streptococci grow in long chains in the presence of homologous type antiserum, and in very short chains in serum which does not contain type specific antibody, this index is a semiquantitative expression for the amount of anti-M antibody in the system. A threefold increase in mean chain length (index = 3), or greater, was considered a statistically significant test for type specific antibody (26,27).…”

Section: Methodsmentioning

confidence: 99%

“…At critical levels, therefore, the bactericidal test could be positive and the long chain test negative. Occasionally, the reverse situation could occur due to nonspecific factors affecting phagocytosis (26,27). The degree of virulence of the test strain of streptococci also had an effect on the resulting indices when titers of serum antibody were low (Table I).…”

Section: Methodsmentioning

confidence: 99%

Recall of Type Specific Antibodies in Man by Injections of Streptococcal Cell Walls*

Potter¹,

Stollerman²,

Siegel³

1962

J. Clin. Invest.

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In the early part of this century several attempts were made to immunize human beings against beta hemolytic streptococci (1-4). These attempts generally were considered to be unsuccessful. The results were difficult to evaluate, however, because of the multiplicity and toxicity of streptococcal antigens and the limited understanding of their relationship to virulence. Subsequently, immunity was correlated with antibody against type specific M protein contained in streptococcal cell walls (5-9), and sensitive methods were developed for the assay of these type specific antibodies (6, 7).Partially purified preparations of M protein were employed thereafter as vaccines (10-15). Large doses proved antigenic in rabbits (10-11). Toxic reactions, however, limited vaccination in human beings to the administration of very small amounts of M protein. These quantities rarely provoked an immune response (12-15). Investigators, therefore, have been discouraged by the possibility that the amount of M protein which can be tolerated in single doses may be below the threshold of antigenicity.In the course of a controlled study of the immune response to streptococcal pharyngitis (16,17), patients with infections of known serologic type have been observed for the development of type specific antibody. During the 4 years that this study has been in progress, type specific antibody has disappeared from the blood of approxi-*

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Evaluation of the "Long Chain Reaction" as a Means for Detecting Type-Specific Antibody to Group a Streptococci in Human Sera

Cited by 22 publications

References 9 publications

Factors Affecting the Chain Length of Group a Streptococci

Factors Affecting the Chain Length of Group a Streptococci

Human immune response to immunization with a structurally defined polypeptide fragment of streptococcal M protein.

Recall of Type Specific Antibodies in Man by Injections of Streptococcal Cell Walls*

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